Compositions and methods for treating halitosis

Abstract

The present invention relates to improved phosphopeptide stabilized amorphous calcium phosphate and / or amorphous calcium fluoride phosphate complexes and compositions containing these complexes. Methods of making the complexes of the invention and treating or preventing oral conditions are also provided. In one aspect, the invention provides a method for treating or preventing halitosis in an individual in need thereof, the method comprising administering to the oral cavity of the individual a stabilized amorphous calcium phosphate (ACP) and / or a stabilized amorphous calcium fluoride phosphate (ACFP), thereby treating or preventing halitosis. In another aspect, the invention provides a zinc-associated stabilized amorphous calcium phosphate (ACP) and / or a zinc-associated stabilized amorphous calcium fluoride phosphate (ACFP) complex.

Description

[0001] Related applications

[0002] This application claims priority to Australian Provisional Application No. 2023903602, filed on 9 November 2023, the entire disclosure of which is incorporated herein by reference. Technical Field

[0003] This invention relates to improved stable amorphous calcium phosphate and / or amorphous calcium fluoride phosphate complexes and compositions containing these complexes. Methods for preparing the complexes of this invention and for treating or preventing oral diseases are also provided. Background Technology

[0004] Halitosis, or bad breath, is a prevalent problem affecting quality of life. Various epidemiological studies report that approximately 25-37% of the adult population suffers from halitosis (Miyazaki et al. 1997, Ueno et al. 2007, Liu et al. 2006). About 90% of halitosis cases originate within the oral cavity, while only 10% originate outside the oral cavity (Ayers and Colquhoun 1998). Tongue coating, various forms of periodontal disease and other dental problems, as well as reduced saliva flow, can all contribute to halitosis originating within the oral cavity (bad breath) (Tangerman 2002, Papaioannou and Dereka 2009). It is now widely recognized that microorganisms play a crucial role in the pathogenesis of halitosis. The main sources of microorganisms associated with halitosis are the dorsum of the tongue and dental plaque (Quirynen et al. 2009). In fact, there is a correlation between the amount of microorganisms on the dorsum of the tongue and the intensity of halitosis (Hartley et al., 1996). The crypts of the tongue and dental plaque may be sites of increased numbers of Gram-negative anaerobic microorganisms with proteolytic activity, involved in the release of sulfur-containing amino acids to produce volatile sulfur compounds (VSCs). VSCs are the main odor substances associated with halitosis (Tonzetich 1977, Persson et al., 1990, Miyazaki et al., 1995). It has been shown that inhibiting enzymes that produce VSCs and reducing VSC levels can directly reduce halitosis (Takeuchi et al., 2010, Tsai et al., 2008).

[0005] Halitosis is a significant social problem, and new or improved methods are needed to reduce its levels. Zinc ions are widely used in mouthwash, chewing gum, and toothpaste formulations to reduce halitosis, but they are easily isolated by phosphates and other molecules in saliva and dental plaque, and therefore have limited effectiveness in inhibiting enzymes / bacteria that produce vesicular odor (VSCs).

[0006] New and / or improved treatments for halitosis are needed.

[0007] References to any prior art in this specification do not imply an acknowledgment or implication that such prior art constitutes part of common common knowledge in any jurisdiction, nor do they imply that such prior art can be reasonably understood, regarded as related to, and / or combined with other prior art by a person skilled in the art. Summary of the Invention

[0008] On one hand, the present invention provides a method for treating or preventing halitosis in an individual in need, the method comprising applying stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) to the oral cavity of the individual, thereby treating or preventing halitosis.

[0009] In any aspect or embodiment, the method further includes the step of identifying an individual suffering from halitosis. The individual may suffer from moderate or severe halitosis. For example, the individual's oral volatile sulfur compound (VSC) level may be approximately 150 ppb (parts per billion) or higher. The individual may be identified as 1, 2, 3, 4, or 5 according to the Rosenberg scale for halitosis, preferably 3, 4, or 5 according to the Rosenberg scale.

[0010] In no aspect or embodiment has the individual been diagnosed with periodontal disease.

[0011] In no respect of any aspect or embodiment does the individual suffer from periodontal disease.

[0012] In no aspect or embodiment has the individual been diagnosed with periodontitis or severe gingivitis.

[0013] In no aspect or embodiment does the individual suffer from periodontitis or severe gingivitis.

[0014] In no aspect or embodiment has the individual been diagnosed with gingivitis.

[0015] In no respect of any aspect or embodiment is the individual suffering from gingivitis.

[0016] On the other hand, the present invention provides a method for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of an individual in need, the method comprising applying stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) to the oral cavity of the individual, thereby reducing the generation of VSCs in the oral cavity of the individual.

[0017] In any aspect or embodiment, the method further includes the step of determining the level of volatile sulfur compounds (VSCs) in the oral cavity of the individual.

[0018] In any aspect or embodiment, the oral volatile sulfur compound (VSC) level of the individual may be at least 150 ppb (parts per billion). Preferably, the oral VSC level of the individual is at least 50 ppb, 60 ppb, 70 ppb, 80 ppb, 90 ppb, 100 ppb, 110 ppb, 120 ppb, 130 ppb, 140 ppb, 150 ppb, 160 ppb, 170 ppb, 180 ppb, 190 ppb, 200 ppb, 300 ppb, 400 ppb, 500 ppb, 600 ppb, 700 ppb, 800 ppb, 900 ppb, or 1000 ppb.

[0019] On the other hand, the present invention provides a method for inhibiting the activity of an enzyme (preferably a protease) produced by oral bacteria, the method comprising applying stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) to the oral cavity of an individual, thereby inhibiting the activity of the enzyme (preferably a protease) produced by the oral bacteria. Preferably, the oral bacteria are oral bacteria involved in the production of vegetative-oral stenosis (VSC). Preferably, the oral bacteria are Gram-negative oral bacteria associated with halitosis.

[0020] On the other hand, the present invention provides the use of stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) for the preparation of medicaments for treating or preventing halitosis in individuals in need.

[0021] On the other hand, the present invention provides a stable amorphous calcium phosphate (ACP) and / or a stable amorphous calcium fluoride phosphate (ACFP) for the treatment or prevention of halitosis in individuals in need.

[0022] On the other hand, the present invention provides the use of stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) for the preparation of a medicament for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of individuals in need.

[0023] On the other hand, the present invention provides a stable amorphous calcium phosphate (ACP) and / or a stable amorphous calcium fluoride phosphate (ACFP) for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of individuals in need.

[0024] On the other hand, the present invention provides the use of stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) for the preparation of a medicament for inhibiting the activity of enzymes (preferably proteases) produced by oral bacteria. Preferably, the medicament is formulated for or adapted for oral administration. Preferably, the oral bacteria are Gram-negative oral bacteria associated with halitosis.

[0025] On the other hand, the present invention provides a stable amorphous calcium phosphate (ACP) and / or a stable amorphous calcium fluoride phosphate (ACFP) for inhibiting the activity of enzymes (preferably proteases) produced by oral bacteria. Preferably, the stable amorphous calcium phosphate (ACP) and / or the stable amorphous calcium fluoride phosphate (ACFP) are applied to the oral cavity. Preferably, the oral bacteria are Gram-negative oral bacteria associated with halitosis.

[0026] On the other hand, the present invention provides a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex.

[0027] In any aspect or embodiment, the zinc ion content of the zinc-associated stable ACP or ACFP complex is, or at least 0.5 mol of zinc per mole of phosphopeptide. Preferably, the zinc ion content of the zinc-associated stable ACP or ACFP complex is, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mol of zinc per mole of phosphopeptide. Even more preferably, the zinc ion content ranges from about 0.5 mol to about 100 mol, about 0.5 mol to about 50 mol, about 0.5 mol to about 20 mol, or about 0.5 mol to about 10 mol of zinc per mole of water-soluble complex. The zinc ion content can range from 0.5 mol to 100 mol, 0.5 mol to 50 mol, 0.5 mol to 20 mol, or 0.5 mol to 10 mol of zinc per mole of phosphopeptide.

[0028] In any embodiment, the zinc ion content described above can be the zinc ion content that is tightly bound to a water-soluble complex (as described herein). When assessing the zinc ion content, the tightly bound zinc ion content is measured using the methods described herein, specifically the method described in Example 4.

[0029] In any embodiment, the zinc-associated stable ACP and / or ACFP complex contains zinc ions that remain associated with the complex after centrifugation at approximately 3000 g for 1 hour in a 1000 molecular weight cutoff filter at room temperature.

[0030] In any embodiment, the zinc-associated stable ACP or ACFP complex has at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% tightly bound zinc associated with the complex, as determined by the method in Example 4.

[0031] In any embodiment, the zinc-associated stable ACP or ACFP complex has at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% zinc incorporated into the complex for the preparation of the complex. The complex can be prepared as outlined in Example 1.

[0032] On the other hand, the present invention also provides a composition comprising, substantially comprising, or consisting of a zinc-associated stable ACP and / or ACFP complex as described herein. Preferably, the composition further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

[0033] On one hand, the present invention provides a method for treating or preventing halitosis in an individual in need, the method comprising applying a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex to the oral cavity of the individual, thereby treating or preventing halitosis.

[0034] On the other hand, the present invention provides a method for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of an individual, the method comprising applying a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex to the oral cavity of the individual, thereby reducing the generation of the volatile sulfur compounds in the oral cavity of the individual.

[0035] On the other hand, the present invention provides a method for inhibiting the activity of an enzyme (preferably a protease) produced by oral bacteria, the method comprising applying a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex to the oral cavity of an individual, thereby inhibiting the activity of the enzyme (preferably a protease) produced by the oral bacteria. Preferably, the oral bacteria are oral bacteria involved in the production of vegetative-smelling bacteria (VSCs). Preferably, the oral bacteria are Gram-negative oral bacteria associated with halitosis.

[0036] On the other hand, the present invention provides the use of a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for the preparation of a medicament for the treatment or prevention of halitosis in individuals in need.

[0037] On the other hand, the present invention provides a zinc-associated stable amorphous calcium phosphate (ACP) complex and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for the treatment or prevention of halitosis in individuals in need.

[0038] On the other hand, the present invention provides the use of a zinc-associated stable amorphous calcium phosphate (ACP) complex and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for the preparation of a medicament for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of individuals in need.

[0039] On the other hand, the present invention provides a zinc-associated stable amorphous calcium phosphate (ACP) complex and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of individuals in need.

[0040] On the other hand, the present invention provides the use of a zinc-associated stable amorphous calcium phosphate (ACP) complex and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for the preparation of a medicament for inhibiting the activity of an enzyme (preferably a protease) produced by oral bacteria. Preferably, the medicament is formulated for or adapted for oral administration. Preferably, the oral bacteria are Gram-negative oral bacteria associated with halitosis.

[0041] On the other hand, the present invention provides a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for inhibiting the activity of enzymes (preferably proteases) produced by oral bacteria. Preferably, the zinc-associated stable amorphous calcium phosphate (ACP) and / or the zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex is applied to the oral cavity. Preferably, the oral bacteria are Gram-negative oral bacteria associated with halitosis.

[0042] In any embodiment, the method or use of the present invention causes a reduction in intraoral VSC levels of at least 50 ppb, 60 ppb, 70 ppb, 80 ppb, 90 ppb, 100 ppb, 110 ppb, 120 ppb, 130 ppb, 140 ppb, or 150 ppb. Alternatively or additionally, the method or use of the present invention causes a reduction in the individual's Rosenberg Scale measurement by at least 1. VSC can be measured using any of the methods described herein, including those described in the examples, and can be measured at least 30 minutes, 1 hour, 2 hours, 3 hours, or 4 hours after application of: stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP); zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complexes; or compositions comprising stable ACP and / or ACP or zinc-associated stable ACP and / or zinc-associated stable ACFP. In one embodiment, the reduction was observed 14 days after treatment.

[0043] In any embodiment, the stable amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) are phosphopeptides stabilized. Preferably, the phosphopeptide (as defined below) is a casein phosphopeptide.

[0044] In any embodiment, the zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated amorphous calcium fluoride phosphate (ACFP) complex is phosphopeptide-stabilized. Preferably, the phosphopeptide (as defined below) is a casein phosphopeptide.

[0045] In any method or use of this invention, a stable ACP or ACFP complex may be applied to an individual for 5 to 60 minutes, 10 to 45 minutes, 10 to 30 minutes, or 20 minutes. Alternatively, the stable ACP or ACFP complex may be applied 4, 5, or 6 times daily or every 24 hours. Preferably, the stable ACP or ACFP complex is applied for a period of 1 to 2 weeks.

[0046] In any method or use of this invention, the zinc-associated stable ACP or ACFP complex can be applied to an individual for 5 to 60 minutes, 10 to 45 minutes, 10 to 30 minutes, or 20 minutes. Alternatively, the zinc-associated stable ACP or ACFP complex can be applied 4, 5, or 6 times daily or every 24 hours. Preferably, the application of the zinc-associated stable ACP or ACFP complex lasts for 1 to 2 weeks.

[0047] In any method or use of the present invention, stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) can be applied to the oral cavity in the form of compositions, such as those described herein.

[0048] In any method or use of the present invention, zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complexes can be applied to the oral cavity in the form of compositions, such as those described herein.

[0049] In any respect, the composition can be prepared and used in various forms suitable for oral use, such as dental cleaning agents, including toothpaste, tooth powder and liquid dental cleaning agents, mouthwash, rinses, oral sprays, varnishes, dental cements, lozenges, chewing gum, dental pastes, gum massage creams, mouthwash tablets, dairy products and other foods, including yogurt. Preferably, the composition is a mouthwash. Preferably, the composition is a mint or chewing gum, preferably a sugar-free mint or sugar-free chewing gum.

[0050] In any aspect or embodiment, the calcium ion content of the stable ACP or ACFP complex is greater than about 30 mol per mole of PP. Preferably, the calcium ion content is in the range of about 30 mol to 100 mol of calcium per mole of PP. More preferably, the calcium ion content is in the range of about 30 mol to about 50 mol of calcium per mole of PP.

[0051] In any aspect or embodiment, the calcium ion content of the zinc-associated stable ACP or ACFP complex is greater than about 30 mol per mole of PP. Preferably, the calcium ion content is in the range of about 30 mol to 100 mol of calcium per mole of PP. More preferably, the calcium ion content is in the range of about 30 mol to about 50 mol of calcium per mole of PP.

[0052] In any respect, the stable ACP complex is a stannous phosphopeptide (PP)-stabilized amorphous calcium phosphate (ACP) complex, and the stable ACFP complex is a stannous phosphopeptide (PP)-stabilized amorphous calcium fluoride phosphate (ACFP) complex.

[0053] In every respect, a stable ACP complex is a non-tin-associated stable ACP complex, and a stable ACFP complex is a non-tin-associated stable ACFP complex.

[0054] In any aspect or embodiment, the phase of ACP is predominantly (i.e., > 50%) a basic phase, wherein ACP primarily comprises the species Ca. 2+ PO4 3- and OH - The basic phase of ACP can have the general formula [Ca3(PO4)2]. x [Ca2(PO4)(OH)], where x 1. Preferably, x = 1-5. More preferably, x = 1, that is, the two components in the formula exist in equal proportions. Thus, in one embodiment, the basic phase of ACP has the formula Ca3(PO4)2Ca2(PO4)(OH).

[0055] In any aspect or embodiment, the phase of ACFP is predominantly (i.e., > 50%) a basic phase, wherein ACFP mainly comprises the species Ca. 2+ PO4 3- and F - The basic phase of ACFP can have the general formula [Ca3(PO4)2]. x [Ca2(PO4)F] y Where when y = 1, x 1, or where when x = 1, y 1. Preferably, y = 1 and x = 1-3. More preferably, y = 1 and x = 1, that is, the two components in the formula exist in equal proportions. Thus, in one embodiment, the basic phase of ACFP has the formula Ca3(PO4)2Ca2(PO4)F.

[0056] In one embodiment, the ACP complex is substantially composed of phosphopeptides, calcium, phosphate and hydroxide ions, and water. Preferably, the complex further includes zinc ions.

[0057] In one embodiment, the ACFP complex generally comprises: phosphopeptide, calcium, phosphate, fluorine and hydroxide ions, and water. Preferably, the complex further comprises zinc stannous ions.

[0058] On the other hand, the present invention also relates to a kit for use in the method or application of the present invention, the kit comprising: (a) the composition as described herein; or (b) Stable ACP or ACFP complexes as described herein.

[0059] On the other hand, the present invention also relates to a kit for use in the method or application of the present invention, the kit comprising: (a) the composition as described herein; or (b) Zinc-associated stable ACP or stable ACFP complexes as described herein.

[0060] Preferably, the kit further includes instructions for using the kit in the method or application of the present invention.

[0061] On the other hand, the present invention provides a method or process for forming zinc-associated phosphopeptide-stabilized ACP or ACFP complexes, the method or process comprising or consisting of the following: (i) Obtaining a solution containing at least one phosphopeptide; and (ii) Mixing a solution containing phosphopeptides, calcium ions, phosphate ions, hydroxide ions, and optionally fluoride ions while maintaining the pH at about 7 or higher, preferably about 9, to form a solution containing phosphopeptide-stabilized ACP or ACFP; and (iii) Mix the water-soluble zinc salt with the solution containing phosphopeptide-stabilized ACP or ACFP at a pH of 5.5 to 7.5, adding slowly, mixing and maintaining the pH of the solution. This results in the formation of zinc-associated phosphopeptide-stabilized ACP or ACFP complexes.

[0062] In one embodiment, the solution is added slowly at a rate of less than about 1% by volume per minute and thoroughly mixed.

[0063] On the other hand, the present invention provides a method or process for forming zinc-associated phosphopeptide-stabilized ACP or ACFP complexes, the method or process comprising or consisting of the following: (i) Provide a solution of phosphopeptide-stabilized ACP or ACFP; and (ii) Mix the water-soluble zinc salt with the solution. This results in the formation of zinc-associated phosphopeptide-stabilized ACP or ACFP complexes.

[0064] In any aspect or embodiment, the water-soluble zinc salt is in the form of zinc acetate, zinc lactate, zinc sulfate, zinc nitrate, zinc gluconate, zinc chloride, zinc citrate, or any other water-soluble zinc salt.

[0065] As used herein, unless the context otherwise requires, the term “comprise” and variations thereof, such as “comprising,” “comprises,” and “comprised,” are not intended to exclude additional additives, components, wholes, or steps.

[0066] Further aspects of the invention and further embodiments of the aspects described in the foregoing paragraphs will become apparent from the following description, which is given by way of example and with reference to the accompanying drawings. Detailed Implementation

[0067] It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more features mentioned or apparent in the text or drawings. All these different combinations constitute various alternative aspects of the invention.

[0068] Further aspects of the invention and further embodiments of the aspects described in the foregoing paragraphs will become apparent from the following description, which is given by way of example and with reference to the accompanying drawings.

[0069] Reference will now be made to certain embodiments of the invention. Although the invention will be described in conjunction with embodiments, it should be understood that it is not intended to limit the invention to those embodiments. Rather, the invention is intended to cover all alternatives, modifications, and equivalents that may be included within the scope of the invention as defined in the claims.

[0070] Those skilled in the art will recognize that many methods and materials can be similar to or equivalent to those described herein, and can be used to practice this invention. This invention is by no means limited to the methods and materials described.

[0071] All patents and publications mentioned in this article are incorporated in their entirety by reference.

[0072] For the purposes of interpreting this specification, terms used in the singular will also include the plural form, and vice versa.

[0073] As used herein, unless the context otherwise requires, the term "comprise" and variations thereof, such as "comprising," "comprises," and "comprised," are not intended to exclude additional additives, components, wholes, or steps. As used herein, unless the context otherwise requires, "comprise" and "including" may be used interchangeably.

[0074] Bad breath

[0075] The terms “halitosis,” “bad breath,” “foul odor,” “breathmalodour,” “feter oris,” or “fege bosta” are used interchangeably and refer to a condition in which there is a noticeable unpleasant odor in the exhaled breath.

[0076] The vast majority of oral malodor originates from the degradation of sulfur-containing amino acids by anaerobic bacteria in the oral cavity, resulting in the emission of hydrogen sulfide (H2S), methyl mercaptan (CH3SH), and dimethyl sulfide (CH3SCH3) (collectively referred to as volatile sulfur compounds (VSC)). In any aspect or embodiment of the invention, VSC may be one or more of hydrogen sulfide (H2S), methyl mercaptan (CH3SH), and dimethyl sulfide (CH3SCH3).

[0077] Halitosis (bad breath) can be measured in a variety of ways, including studies using sensory intensity and sensory pleasure indices, as well as instruments that quantify the amount of volatile sulfur compounds or bacterial enzymes that contribute to the production of odorous compounds.

[0078] Sensory measurement, or smelling and ranking odor intensities, is considered the standard for measuring halitosis. A 5-point or 10-point scale can be used, and the degree of halitosis is typically assessed by two independent judges. The intensity of halitosis is based on the Rosenberg Scale for Halitosis (Rosenberg et al., *Journal of Periodontology*). J Periodontol (Reference: 《《》, 1991; 62:487-489》, this scale assesses odor intensity as follows: 0 - No odor detected 1- A suspicious, foul odor, almost undetectable. 2- Slightly foul odor, exceeding the odor detection threshold 3- A foul odor was clearly detected. 4- Strong stench 5 - Extremely strong stench Intraoral volatile sulfur compounds (VSCs) can be measured using electrochemical sensors (e.g., Halimeter). ® ) and portable gas chromatographs (e.g., OralChroma) ® TwinBreather ® These measurements are well known in the art. The concentration of VSC is typically measured in parts per billion (ppb).

[0079] If an individual is found to have an increased number of Gram-negative proteolytic bacteria on the dorsum of the tongue or in dental plaque, they may be identified as having halitosis or at risk of developing halitosis. The increased number can be compared to the number of the same bacterial species in individuals known not to have halitosis (i.e., healthy controls).

[0080] Zinc-associated stable amorphous calcium phosphate (ACP) or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complexes

[0081] As described herein, the present invention provides a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex.

[0082] In any aspect or embodiment, the zinc ion content of the zinc-associated stable ACP or ACFP complex is, or at least 0.5 mol of zinc per mole of phosphopeptide. Preferably, the zinc ion content of the zinc-associated stable ACP or ACFP complex is, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mol of zinc per mole of phosphopeptide. Even more preferably, the zinc ion content ranges from about 0.5 mol to about 100 mol, about 0.5 mol to about 50 mol, about 0.5 mol to about 20 mol, or about 0.5 mol to about 10 mol of zinc per mole of phosphopeptide. The range of zinc ion content can be from 0.5 mol to 100 mol, 0.5 mol to 50 mol, 0.5 mol to 20 mol, or 0.5 mol to 10 mol of zinc per mole of phosphopeptide.

[0083] In any embodiment, the zinc ion content described above can be the zinc ion content that is tightly bound to the complex (as described herein). When assessing the zinc ion content, the tightly bound zinc ion content is measured using the methods described herein, specifically the method described in Example 4.

[0084] In any embodiment, the zinc-associated stable ACP and / or ACFP complex contains zinc ions that remain associated with the complex after centrifugation at approximately 3000 g for 1 hour in a 1000 molecular weight cutoff filter at room temperature.

[0085] In any embodiment, the zinc-associated stable ACP or ACFP complex has at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% tightly bound zinc associated with the complex, as determined, for example, by the method in Example 4.

[0086] In any embodiment, the zinc-associated stable ACP or ACFP complex has at least 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% zinc incorporated into the complex for the preparation of the complex. The complex can be prepared as outlined in Example 1.

[0087] On the other hand, the present invention also provides a composition comprising, substantially comprising, or consisting of a zinc-associated stable ACP and / or ACFP complex as described herein.

[0088] In any aspect or embodiment, zinc ions are Zn 2+ Typically, during the formation of stable zinc-associated ACP and / or ACFP complexes, zinc ions are provided by Zn(NO3)2 or any other water-soluble zinc salt, including the zinc salts described herein.

[0089] Stable ACP or ACFP

[0090] Stable amorphous calcium phosphate (ACP) and stable amorphous calcium fluoride phosphate (ACFP) can also be referred to as stable ACP complex and stable ACFP complex, respectively, in this paper.

[0091] As mentioned herein, stable ACP or ACFP complexes include those described in PCT / AU2005 / 001781, the contents of which are incorporated herein by reference.

[0092] In a preferred embodiment, the phosphopeptide-stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex has both 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 pH 7.0. Optionally, the ACP or ACFP is primarily in an alkaline form.

[0093] As mentioned herein, stable ACP or ACFP complexes include stable ACP or ACFP complexes formed at pH levels below 7.0. Preferably, the complex is formed at a pH range of about 5.0 to below 7.0. More preferably, the complex is formed at a pH range of about 5.0 to about 6.0. In a preferred embodiment, the complex is formed at a pH of about 5.5. Preferably, the ACP or ACFP in the complex is primarily in a basic form.

[0094] A stable ACP can be generated by including the following steps: (i) Obtaining a solution containing at least one phosphopeptide; and (ii) Mix solutions containing calcium ions, phosphate ions and hydroxide ions while maintaining the pH at about 7.0 or below.

[0095] A stable ACFP can be generated by a method that includes the following steps: (i) Obtaining a solution containing at least one phosphopeptide; and (ii) Mix solutions containing calcium ions, phosphate ions, hydroxide ions and fluoride ions while maintaining the pH at about 7.0 or below.

[0096] The phosphopeptide-stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex may further include ACP in the complex having both tightly bound 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 pH 7.0, and the ACP or ACFP is primarily in a basic form, and may be obtained by methods comprising the following or by methods comprising the following: A first solution containing calcium ions, a second solution containing phosphate ions, and a third solution optionally containing fluoride ions are mixed into a solution containing phosphopeptides and a solvent with a pH of about 5 to below; and during the mixing, the pH of the solution is maintained at about 5.0 to below 7.0 by adding hydroxide ions.

[0097] The “tightly” and “loosely” bound calcium and phosphate in ACP or ACFP can be determined using analytical ultrafiltration. In short, a solution of phosphopeptides, calcium, phosphate, and optionally desfluoride, mixed while maintaining the pH at approximately 7.0 or below, can first be filtered through a 0.1-micron filter to remove free calcium and phosphate not associated with the complex. This free calcium and phosphate is present in the filtrate and is discarded. Any free calcium or phosphate not associated with the complex in any way is not bioavailable, i.e., delivered to the teeth via phosphopeptides. The retentate from the 0.1-micron filter can be further analyzed by centrifugation at 1,000 g for 15 minutes using a 3000 mw cutoff filter. The resulting filtrate contains calcium and phosphate loosely bound to or associated with the complex. Under this centrifugal force, calcium and phosphate 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. Then, by subtracting the amount of Ca and Pi in the filtrate from the total amount of Ca and Pi in the 0.1-micron filtrate, the amount of tightly bound Ca and Pi in the filtrate can be determined.

[0098] As mentioned herein, stable ACP or ACFP complexes include those described in PCT / AU2006 / 000885, the contents of which are incorporated herein by reference.

[0099] The complex is a “superloaded” phosphopeptide or phosphoprotein (PP)-stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complex. The complex can form at any pH (e.g., 3-10). Preferably, the phosphopeptide comprises the sequence -ABC-, where A is a phosphate amino acid, preferably phosphoserine, B is any amino acid including a phosphate amino acid, and C is glutamic acid, aspartic acid, or a phosphate amino acid. The phosphate amino acid may be phosphoserine. The PP is superloaded with calcium and phosphate ions. The calcium ion concentration can be in the range of 30-1000 mol Ca per mole of PP, or in the range of 30-100 or 30-50 mol Ca per mole of PP. In another embodiment, the mol Ca per mole of PP is at least 25, 30, 35, 40, 45, or 50.

[0100] The calcium ion content of amorphous calcium phosphate or amorphous calcium fluoride phosphate complexes stabilized by phosphopeptides or phosphoproteins (PP) can be 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.

[0101] Phosphopeptide or phosphoprotein (PP) stabilized amorphous calcium phosphate (ACP) or amorphous calcium fluoride phosphate (ACFP) complexes can be produced by a method comprising the following steps: (i) Obtain a solution containing calcium, inorganic phosphate, and fluorine (optionally); and (ii) Mix (i) with a solution containing PP-ACP.

[0102] In a preferred embodiment, PP is casein phosphopeptide (CPP).

[0103] The PP-stabilized ACP and / or ACFP complex may further comprise at least an equal amount of calcium phosphate by weight. Preferably, the calcium phosphate is CaHPO4. Preferably, the calcium phosphate (e.g., CaHPO4) is dry-mixed with the PP-stabilized ACP and / or ACFP complex. In a preferred embodiment, the PP-ACP and / or PP-ACFP complex:calcium phosphate ratio is about 1:1-50, more preferably about 1:1-25, and even more preferably about 1:5-15. In one embodiment, the PP-ACP and / or PP-ACFP complex:calcium phosphate ratio is about 1:10.

[0104] When used in the oral cavity, oral care formulations comprising a phosphopeptide or phosphoprotein (PP) stabilized amorphous calcium phosphate (ACP) and / or amorphous calcium fluoride phosphate (ACFP) complexes containing a calcium ion content greater than approximately 30 moles of calcium per mole of PP can be produced by a method including the following steps: (i) Obtaining a powder comprising PP-ACP and / or PP-ACFP composites; (ii) Dry mix with an effective amount of calcium phosphate; and (iii) Formulate the dry-mixed PP-ACP and / or PP-ACFP and calcium phosphate mixture into an oral care formulation.

[0105] Preferably, the calcium phosphate used for dry mixing is in the form of any soluble calcium phosphate, including but not limited to CaHPO4, Ca2HPO4 and calcium lactate.

[0106] The compositions described herein may further include free fluoride ions. Fluoride ions can be from any suitable source. Sources of fluoride ions may include free fluoride ions or fluoride salts. Examples of sources of fluoride ions include, but are not limited to, sodium fluoride, sodium monofluorophosphate, stannous fluoride, sodium fluorosilicate, and amine fluoride. These may be provided in the form of solutions (typically aqueous solutions) or suspensions.

[0107] Fluoride ions are preferably present in the composition in an amount greater than 1 ppm. More preferably, the amount is greater than 3 ppm. In another embodiment, the amount is preferably greater than 10 ppm. In the typical embodiments described below, the amount may be several hundred or several thousand ppm. Fluoride content is typically measured in ppm in oral compositions in a manner commonly used in the art. Where the fluoride is derived from a source with stable ACP, ppm refers to the concentration of fluoride in this source (typically a solution or suspension of bioavailable fluoride).

[0108] As mentioned herein, tin-associated ACP or ACFP complexes include any of those described in PCT / AU2014 / 050447, the entire contents of which are incorporated herein by reference.

[0109] The compositions used in the methods and applications of the present invention as described herein may include stannous-associated ACP or ACFP complexes. The compositions may include 2% CPP-ACP and 290 ppm fluorine, wherein 220 ppm fluorine is stannous fluoride and 70 ppm is sodium fluoride.

[0110] In the context of this invention, "phosphopeptide" means an amino acid sequence in which at least one amino acid is phosphorylated. Preferably, a phosphopeptide comprises one or more of the amino acid sequences -ABC-, where A is a phosphate amino residue, B is any amino acyl residue including a phosphate amino residue, and C is selected from glutamine, aspartic acid, or a phosphate amino residue. Any of the phosphate amino residues may independently be a phosphate serine residue. Ideally, B is a residue whose side chain is neither relatively large nor hydrophobic. It may be Gly, Ala, Val, Met, Leu, Ile, Ser, Thr, Cys, Asp, Glu, Asn, Gln, or Lys.

[0111] In another embodiment, at least two of the phosphate amino acids in the sequence are preferably consecutive. Preferably, the phosphopeptide comprises the sequence ABCDE, wherein A, B, C, D, and E are independently phosphoserine, phosphothreonine, phosphotyrosine, phosphohistidine, glutamic acid, or aspartic acid, and at least two, preferably three, of A, B, C, D, and E are phosphate amino acids. In a preferred embodiment, the phosphate amino 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.

[0112] In one embodiment, ACP or ACFP is stabilized by casein phosphopeptide (CPP), which is in the form of whole casein or casein fragments, and the resulting complex preferably has the formula [CPP(ACP)8]n or [(CPP)(ACFP)8]n, where n is equal to or greater than 1, for example, 6. Therefore, the resulting complex can be a colloidal complex in which the core particles aggregate to form large (e.g., 100 nm) colloidal particles suspended in water. Thus, PP can be casein or phosphopeptide.

[0113] PP can be from any source; it can exist in the context of a larger polypeptide (including full-length casein polypeptide), or it can be isolated by trypsin or other enzymatic or chemical digestion of casein or other phosphate-rich proteins (such as phosphatidylcholine, osteopontin) or by chemical or recombinant synthesis, provided that it contains the sequence -ABC- or ABCDE as described above. Sequences flanking this core sequence can be any sequence. However, α is preferred. s1 (59-79), β(1-25), α s2 (46-70) and α s2The flanking sequences in (1-21). The flanking sequences may optionally be modified by deletion, addition, or conservative substitution of one or more residues. The amino acid composition and sequence of the flanking region are not important. Preferably, the PP is obtained from a food source, such as phosphoprotein from egg yolk, osteopontin from milk, or any chemically or recombinantly synthesized PP containing the sequence -ABC-.

[0114] In one embodiment, the ACP or ACFP is stabilized by casein phosphopeptide (CPP), osteopontin phosphopeptide (OPN-PP), egg yolk high phosphoprotein phosphopeptide (PPP), or a chemically or recombinantly synthesized PP containing the sequence -ABC-. ​​Preferably, the ACP or ACFP is stabilized by casein phosphopeptide (CPP).

[0115] Table A below shows examples of conservative substitution.

[0116] Table A

[0117] Flanking sequences can also include amino acid residues that are not naturally occurring. Common amino acids not encoded by the genetic code include: 2-Amino-4-phosphonobutyric acid for phosphoserine [Ser(P)]; 2-Aminohexanoic acid (Aad) for Glu and Asp. 2-Aminopimelic acid (Apm) used in Glu and Asp. 2-Aminobutyric acid (Abu) is used for Met, Leu and other aliphatic amino acids. 2-Aminoheptanic acid (Ahe) is used for Met, Leu and other aliphatic amino acids. 2-Aminoisobutyric acid (Aib) used in Gly. Cyclohexylalanine (Cha) is used in Val, Leu, and Ile. High arginine (Har) for Arg and Lys; 2,3-Diaminopropionic acid (Dpr) for Lys, Arg and His. N-Ethylglycine (EtGly) used in Gly, Pro and Ala. N-Ethyl Aspartic Acid (EtAsn) for use in Asn and Gln; Hydroxylysine (Hyl) for Lys; Allohydroxylysine (AHyl) for Lys; 3-(and 4)hydroxyproline (3Hyp, 4Hyp) for Pro, Ser and Thr; Alloleucine (Alle) for Ile, Leu and Val; p-Aminophenylalanine used in Ala; N-methylglycine (MeGly, sarcosine) used in Gly, Pro, and Ala; N-methyl isoleucine (MeIle) for use in Ile; Used for valine (Nva) in Met and other aliphatic amino acids. Ortholeucine (Nle) is used for Met and other aliphatic amino acids. Ornithine (Orn) for Lys, Arg and His; Citrulline (Cit) and methionine sulfoxide (MSO) are used for Thr, Asn and Gln. N-methylphenylalanine (MePhe), trimethylphenylalanine, halogen (F, Cl, Br and I) phenylalanine, and trifluoromethylphenylalanine are used for Phe.

[0118] In one embodiment, PP is one or more phosphopeptides selected from the group consisting of: α s1 (59-79)[1], β(1-25) [2], α s2 (46-70) [3] and α s2 (1-21) [4]:

[0119] [1] Gln 59 -Met-Glu-Ala-Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-Ile-Val- Pro-Asn-Ser(P)-Val-Glu-Gln-Lys 79 α s1 (59-79) (SEQ ID NO: 1)

[0120] [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 β(1-25) (SEQ ID NO: 2)

[0121] [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 α s2 (46-70) (SEQ ID NO: 3)

[0122] [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 α s2 (1-21) (SEQ ID NO: 4).

[0123] Oral composition

[0124] In another embodiment of the invention, a stable ACP and / or a stable ACFP complex is incorporated into an oral composition, such as toothpaste, mouthwash, or oral formulation, to help prevent and / or treat gingivitis or periodontitis. The oral composition contains a sufficient amount of stable ACP and / or ACFP to form a layer on the tooth surface, preferably with a calcium:phosphate ratio equal to that of normal apatite, for example, a ratio of about 2:1. The layer may contain about 20 wt% calcium. The stable ACP and / or ACFP complex may comprise 0.01% to 50% by weight of the composition, preferably 1.0% to 50%, preferably 1.0% to 30%, preferably 1.0% to 20%, preferably 1.0% to 10%, preferably 2% to 10%. In a particularly preferred embodiment, the oral composition of the invention contains about 2% of a stable ACP or ACFP complex or a mixture of both. The oral compositions of the present invention containing the above-described agents can be prepared and used in various forms suitable for oral use, such as dental cleaning agents, including toothpaste, tooth powder and liquid dental cleaning agents, mouthwash, rinses, oral sprays, coatings, dental adhesives, lozenges, chewing gum, mints, dental pastes, gum massage creams, mouthwash tablets, dairy products and other foods, including yogurt and confectionery. Depending on the type and form of the particular oral composition, the oral compositions according to the present invention may further include other well-known ingredients. Some compositions of the present invention (such as toothpaste, tooth powder and liquid dental cleaning agents, mouthwash, rinses and oral sprays) have relatively low viscosity and are effective for treatment or prevention without significant residence time in the oral cavity.

[0125] In certain preferred forms of the invention, the oral composition may be substantially liquid in nature, such as a mouthwash, rinse, or spray. In such formulations, the mediator is typically a water-alcohol mixture, ideally including the humectants described below. Typically, the weight ratio of water to alcohol is in the range of about 1:1 to about 20:1. The total amount of the water-alcohol mixture in this type of formulation 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.

[0126] In other desirable forms of the invention, the composition may be substantially solid or paste-like in nature, such as tooth powder, dental tablets, or toothpaste (dental cream) or gel dental cleaning agents. The carrier of such solid or paste-like oral preparations typically contains dentally acceptable polishing materials. Examples of polishing materials are water-insoluble sodium metaphosphate, potassium metaphosphate, tricalcium phosphate, calcium phosphate dihydrate, anhydrous dicalcium phosphate, calcium pyrophosphate, magnesium orthophosphate, trimagnesium phosphate, calcium carbonate, hydrated alumina, calcined alumina, aluminum silicate, zirconium silicate, silica, bentonite, and mixtures thereof. Other suitable polishing materials include particulate thermosetting resins such as melamine-formaldehyde resin, phenolic resin, and urea-formaldehyde resin, as well as cross-linked polyepoxides and polyesters. Preferred polishing materials include crystalline silica, silica gel, or colloidal silica with a particle size of up to about 5 micrometers, an average particle size of up to about 1.1 micrometers, and a surface area of ​​up to about 50,000 cm² / g, and composite amorphous alkali metal aluminum silicates.

[0127] When using visually transparent gels, polishing colloidal silica (such as those sold under the trademarks SYLOID as Syloid 72 and Syloid 74 or under the trademark SANTOCEL as Santocel 100) and alkali metal aluminosilicate complexes are particularly useful because their refractive indices are close to those of gelling liquid (including water and / or humectants) systems commonly used in dental cleaning agents.

[0128] Many so-called “water-insoluble” polishing materials are anionic in nature and also contain small amounts of soluble material. Therefore, insoluble sodium metaphosphate can be formed in any suitable manner, for example, as described in Thorpe's Dictionary of Applied Chemistry, Volume 9, 4th Edition, pp. 510-511. Other examples of suitable materials are insoluble sodium metaphosphate in the form of Madrell's salt and Kurrol's salt. These metaphosphates exhibit very low solubility in water and are therefore commonly referred to as insoluble metaphosphates (IMPs). Small amounts of soluble phosphate material are present as impurities, typically a few percent, such as at most 4% by weight. If necessary, the amount of soluble phosphate material can be reduced or eliminated by washing with water; in the case of insoluble metaphosphates, soluble phosphate material is considered to include soluble sodium trimetaphosphate. Insoluble alkali metal metaphosphates are typically used in powder form, with particle sizes such that no more than 1% of the material is larger than 37 micrometers.

[0129] Polishing materials are typically present in solid or paste compositions at a concentration of about 10% to about 99% by weight. Preferably, they are present in toothpaste at an amount of about 10% to about 75%, and in tooth powder at an amount of about 70% to about 99%. In toothpaste, when the polishing material is essentially silica, it is typically present at an amount of about 10-30% by weight. Other polishing materials are typically present at an amount of about 30-75% by weight.

[0130] In toothpaste, the liquid carrier may contain water and a humectant, typically in amounts ranging from about 10% to about 80% by weight of the formulation. Glycerin, propylene glycol, sorbitol, and polypropylene glycol exemplify suitable humectants / carriers. Liquid mixtures of water, glycerin, and sorbitol are also advantageous. In transparent gels where refractive index is an important consideration, about 2.5-30% w / w water, 0% w / w to about 70% w / w glycerin, and about 20-80% w / w sorbitol are preferably used.

[0131] Toothpaste, creams, and gels typically contain natural or synthetic thickeners or gelling agents in proportions of about 0.1% w / w to about 10% w / w, preferably about 0.5% w / w to about 5% w / w. Suitable thickeners are synthetic lithium soapstone, a synthetic colloidal magnesium-alkali metal silicate composite clay, available as, for example, Laponite (e.g., CP, SP 2002, D) sold by Laporte Industries Limited. Laponite D is approximately 58.00 wt% SiO2, 25.40 wt% MgO, 3.05 wt% Na2O, 0.98 wt% Li2O, and some water and trace metals. Its true specific gravity is 2.53, and its apparent bulk density at 8% moisture is 1.0 g / ml.

[0132] Other suitable thickeners include Irish moss, carrageenan, tragacanth gum, starch, polyvinylpyrrolidone, hydroxyethylpropyl cellulose, hydroxybutylmethylcellulose, hydroxypropylmethylcellulose, hydroxyethyl cellulose (e.g., available as Natrosol), sodium carboxymethyl cellulose, and colloidal silica, such as finely ground Syloid (e.g., 244). Solubilizers may also be included, such as humectant polyols (e.g., propylene glycol, dipropylene glycol, and hexanediol), cellosols (e.g., methyl cellosol and ethyl cellosol), vegetable oils and waxes (containing at least about 12 carbons in the straight chain) (e.g., olive oil, castor oil, and petrolatum), and esters (e.g., amyl acetate, ethyl acetate, and benzyl benzoate).

[0133] Understandably, as is customary, oral preparations are typically sold or otherwise distributed in appropriately labeled packaging. Therefore, a mouthwash will have a label describing it as a mouthwash or rinse of its own nature, along with instructions for use; and toothpaste, creams, or gels are usually packaged in collapsible tubes (typically aluminum, lead-lined, or plastic) or other squeeze, pump, or pressurized dispensers for metering the contents, which are also labeled as toothpaste, gel, or toothpaste of its own nature.

[0134] Organic surfactants can be used in the compositions of the present invention to achieve enhanced preventative effects, help achieve thorough and complete dispersion of the surfactant throughout the oral cavity, and make the compositions of the present invention more cosmetically acceptable. The organic surfactant material is preferably anionic, nonionic, or amphoteric in nature, and preferably does not interact with the surfactant. Preferably, a cleaning material that imparts cleaning and foaming properties to the composition is used as the surfactant. Suitable examples of anionic surfactants are water-soluble salts of monoglycerides of higher fatty acids, 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 sulfonate; higher alkyl sulfonate salts; higher fatty acid esters of 1,2-dihydroxypropane sulfonate; and substantially saturated higher aliphatic acylamides of lower aliphatic aminocarboxylic acid compounds, such as those compounds having 12 to 16 carbons in the fatty acid, alkyl, or acyl group, etc. The last example of an amide mentioned is N-lauroyl sarcosine, and sodium, potassium, and ethanolamine salts of N-lauroyl, N-myristoyl, or N-palmitoyl sarcosine, which should be substantially free of soap or similar higher fatty acid materials. The use of these zinc soapstone compounds in the oral compositions of the present invention is particularly advantageous because these materials, in addition to reducing the solubility of tooth enamel in acidic solutions to a certain extent, exhibit a long-term significant inhibitory effect on acid formation in the oral cavity due to carbohydrate decomposition. Examples of suitable water-soluble nonionic surfactants are condensation products of ethylene oxide with various hydrogen-containing compounds that react therewith, said hydrogen-containing compounds having long hydrophobic chains (e.g., aliphatic chains of about 12 to 20 carbon atoms), said condensation products (“ethoxamers”) containing a hydrophilic polyoxyethylene moiety, such as 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).

[0135] Surfactants are typically present in an amount of about 0.1-5% by weight. It is worth noting that surfactants can help dissolve the active agents of this invention, thereby reducing the amount of solubilizing and moisturizing agents required.

[0136] Various other materials may be incorporated into the oral formulations of the present invention, such as whitening agents, preservatives, silicones, chlorophyll compounds, and / or ammoniating materials, such as urea, diammonium phosphate, and mixtures thereof. These adjuvants are incorporated into the formulation in amounts that do not substantially adversely affect the desired properties and characteristics.

[0137] Any suitable flavoring or sweetener material may also be used. Examples of suitable flavoring ingredients are flavoring oils such as those of spearmint, peppermint, wintergreen, sassafras, clove, sage, eucalyptus, marjoram, cinnamon, lemon, and orange, as well as methyl salicylate. Suitable sweeteners include sucrose, lactose, maltose, sorbitol, xylitol, sodium saccharin, perilla syrup, AMP (methyl aspartate), saccharin, etc. Appropriately, flavorings and sweeteners may each or together constitute approximately 0.1% to more than 5% of the preparation.

[0138] The compositions of the present invention can also be incorporated into lozenges, chewing gum, or other products, such as breath-freshening mints, for example, by stirring into a warm gum base or coating the outer surface of the gum base, such as jelutong, rubber latex, vinyl resin, etc., which ideally contains conventional plasticizers or softeners, sugars or other sweeteners, or such as glucose, sorbitol, etc. The compositions of the present invention can be biphasic compositions, wherein each phase allows for the release of components at different times.

[0139] An alternative composition may be a composition providing stable ACP or ACFP and stannous or zinc salts, which then form a stannous-associated stable ACP or ACFP complex or a zinc-associated stable ACP or ACFP complex in situ (e.g., in the oral cavity). An exemplary composition may be chewing gum containing stable ACP or ACFP in its granules and a water-soluble stannous or zinc salt in its central chewing material.

[0140] In another aspect, the present invention provides compositions comprising pharmaceutical compositions comprising a stable ACP or ACFP complex as described above, such as a zinc-associated stable ACP or ACFP complex, and a compound capable of increasing or maintaining the pH of the solution and a pharmaceutically acceptable carrier. Such compositions may be selected from the group consisting of dental anti-caries compositions and therapeutic compositions. Dental compositions or therapeutic compositions may be in the form of gels, liquids, solids, powders, creams, or lozenges. Therapeutic compositions may also be in the form of tablets or capsules. In one embodiment, the stable ACP or ACFP complex is essentially the only active component of such compositions. For example, a cream formulation containing the following may be used: water; glycerin; CPP-ACP / SnF2; D-sorbitol; silica; sodium carboxymethyl cellulose (CMC-Na); propylene glycol; titanium dioxide; xylitol; phosphoric acid; guar gum; sodium saccharin; ethylparaben; magnesium oxide; butylparaben; and propylparaben.

[0141] The present invention further includes the formulations described above and instructions for use for the treatment or prevention of any or more of dental caries or tooth decay, tooth erosion and fluoride poisoning, dentin hypersensitivity, dental plaque, gingivitis or periodontitis.

[0142] In another embodiment, the compositions of the present invention as described herein do not include phosphate buffers and / or calcium chelating agents. For example, any dental cleaning agents described herein may not include phosphate buffers and / or calcium chelating agents.

[0143] In one embodiment of the present invention, a composition is provided, wherein the composition does not include a phosphate buffer and / or a calcium chelating agent.

[0144] In another embodiment, the composition of the present invention as described herein does not include a viscosity modifier, nor does it include 0.5% to 50% of a viscosity modifier.

[0145] In another embodiment, the compositions of the present invention as described herein do not include sodium carboxymethyl cellulose, or 0.01% to 10% of sodium carboxymethyl cellulose with a degree of esterification of 0.7 to 1.0.

[0146] In one embodiment, the active component of the composition is substantially composed of a stable ACP or ACFP complex.

[0147] In one embodiment, the active component of the composition is substantially composed of a zinc-associated stable ACP or ACFP complex.

[0148] It will be clearly understood that although this specification specifically relates to applications in humans, the invention can also be used for veterinary purposes. Therefore, the invention is applicable in all respects to livestock animals such as cattle, sheep, horses, and poultry; companion animals such as cats and dogs; and animals in zoos.

[0149] The present invention also provides a kit comprising stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP), said kit being adapted for use with the methods described above.

[0150] The kit may include: - A container that contains a composition comprising stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP); - Includes a label or packaging insert with an instruction manual.

[0151] The present invention also provides a kit comprising zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complexes, said kit being adapted for use with the methods described above.

[0152] The kit may include: - A container that contains a composition comprising a zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex; - Includes a label or packaging insert with an instruction manual.

[0153] In some embodiments, the kit may contain one or more additional active ingredients for treating diseases or symptoms.

[0154] The kit may include a container and a label or packaging insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, blister packs, etc. The container may be formed from various materials, such as glass or plastic. The container contains a therapeutic composition for effectively treating a condition and may have a sterile access port (e.g., the container may be an intravenous solution bag or vial with a stopper for hypodermal needle puncture). The label or packaging insert indicates that the therapeutic composition is intended to treat the selected condition. In one embodiment, the label or packaging insert includes instructions for use and indicates that the therapeutic composition can be used to treat a given disease or condition.

[0155] The kit may comprise (a) a therapeutic composition; and (b) a second container containing a second active ingredient. In this embodiment of the invention, the kit may further include a packaging insert indicating that the composition and other active ingredients can be used to treat symptoms as described herein or to prevent complications arising from dysbiosis. Alternatively or additionally, the kit may further comprise a second (or third) container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextran solution. The kit may further include other materials desired from a commercial and user perspective, including other buffers, diluents, filters, needles, and syringes.

[0156] The invention will now be further described with reference to the following non-limiting examples.

[0157] It should be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more features mentioned or apparent in the text or drawings. All these different combinations constitute various alternative aspects of the invention.

[0158] Example

[0159] Example 1 - Preparation of CPP-ACP and Zn-CPP-ACP complex

[0160] Add stock solutions of 3.25 M CaCl2 and 1.25 M NaH2PO4 (pH 5.5) in approximately thirty aliquots to a 10–15% w / v trypsin digest of casein until precipitation or gelation occurs (typically yielding approximately 78 mM to 124 mM CaCl2). 2+ (And a final concentration of 48 mM to 76 mM inorganic phosphate). Add the solution slowly (i.e., less than approximately 1% volume per minute) and mix thoroughly. First add an aliquot of phosphate solution, followed by an aliquot of calcium solution. Maintain the pH of the bulk solution at 9.0 using 1 M to 10 M NaOH and mix thoroughly. Add sodium hydroxide solution automatically via a pH adjuster, typically adding hydroxide ions after each addition of calcium ions. After the addition of calcium, phosphate, and hydroxide ions is complete, filter the solution through a 0.1 micron filter to concentrate it 1–2 times. Then, wash the retentate 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. This dried powder is then added to water after the addition of zinc to form a final concentration of 10% w / v CPP-ACP solution with a pH ranging from 5.5 to 7.5. Zinc(II) in the form of a water-soluble zinc salt (e.g., zinc acetate, zinc lactate, zinc sulfate, zinc nitrate, zinc gluconate, zinc chloride, zinc citrate, or any other water-soluble zinc salt) is added to the CPP-ACP solution such that the ratio of Zn mol to CPP mol is 0.5 to 10. The Zn(II) salt is added slowly and mixed under constant pH adjustment to allow the formation of a specific Zn-CPP-ACP complex or Zn-CPP-ACFP complex, wherein sodium fluoride (NaF) or sodium monofluorophosphate (NaMFP) has been incorporated into the complex. For example, a solution prepared with zinc nitrate hexahydrate containing 10% CPP-ACP and 15 mM CPP and 2.23% zinc nitrate hexahydrate containing 75 mM Zn has a Zn / CPP molar ratio of 5. For a mouthwash with a zinc level of 0.1% (15 mM Zn), it is formulated with 2% CPP-ACP and 0.45% zinc nitrate hexahydrate to achieve a Zn / CPP molar ratio of 5. For a toothpaste with 10% CPP-ACP and a zinc level of 1.0%, this is equivalent to 4.5% zinc nitrate hexahydrate and a Zn / CPP molar ratio of 10.

[0161] Example 2 - CPP-ACP and Zn-CPP-ACP complex against Porphyromonas gingivalis ( P. gingivalis Inhibition of VSC-related enzymes (proteases)

[0162] Whole cells of *Porphyromonas gingivalis* (W50 and ATCC33277) were grown and collected, and then... 650 The assay was performed at 0.006. Purified *Porphyromonas gingivalis* proteases (rKgp and rRgpB) were also used at 5.0 µg / mL in a 100 µL reaction volume. The buffer used was 50 mM Tris, 150 mM NaCl, 5 mM CaCl2, pH 6.5 or 7.0, containing 10 mM cysteine, 20 mM Gly-Gly, 1 mM substrate [N-(p-toluenesulfonyl)-Gly-Pro-Lys 4-nitroaniline acetate (GPK-NA) or α-benzoyl-L-arginine 4-nitroaniline hydrochloride (BApNA)], and inhibitor Zn. 2+ (e.g., Zn(NO3)2) 0-0.5 mM or Zn-CPP-ACP complex 0-0.5 mM total Zn(II). The substrate was added after mixing all other reagents and incubating at 37°C for 1 hour. Immediately after substrate addition, the absorbance change at 405 nm (A405) was monitored at 37°C.

[0163] 0.5 mM Zn(II) as zinc nitrate exhibited some inhibitory activity against whole *Porphyromonas gingivalis* cells and purified proteases (Table 1). However, at the same total zinc concentration of 0.5 mM, the Zn-CPP-ACP complex showed superior inhibition of both whole cells and purified proteases (Table 2). These results clearly demonstrate the advantage of the Zn-CPP-ACP complex in inhibiting the enzymatic processes involved in the formation of volatile sulfur compounds (VSCs) by common Gram-negative oral bacteria associated with halitosis.

[0164] Table 1. Zn as Zn(NO3)2 2+ Inhibition of VSC-related enzymes. Relative enzyme activities of whole cells (W50 and ATCC 33277) and purified rKgp and rRgpB in the presence and absence of different concentrations of Zn(II). Relative activity: ratio at pH 7.0 and pH 6.5 to the absence of Zn(II).

[0165]

[0166] Table 2. Inhibition of VSC-related enzymes by the Zn(II)-CPP-ACP complex prepared using CPP-ACP and Zn(NO3)2. Relative enzyme activities of whole cells of *Porphyromonas gingivalis* and purified rKgp and rRgpB proteases in the presence and absence of different concentrations of the Zn(II)-CPP-ACP complex. Relative activity: ratio at pH 7.0 and pH 6.5 to the absence of Zn(II)-CPP-ACP.

[0167]

[0168] Example 3 - Clinical trial of halitosis using CPP-ACP and Zn-CPP-ACP complex mouthwash

[0169] Clinical protocol

[0170] ● Three treatments, double-blind, randomized, crossover design, 20 participants.

[0171] ● Randomized mouthwash treatments were assigned to: (i) Zn 15 mM; (ii) 2% CPP-ACP; and (iii) 2% Zn-CPP-ACP complex (Zn 15 mM).

[0172] ● Use mouthwash (10 ml) three times a day for 60 seconds after your normal oral hygiene routine, for 14 consecutive days.

[0173] ● Use Halimeter to measure volatile sulfur compounds (VSC) at baseline and at the end of each treatment.

[0174] ● Selection criteria include a consistent baseline intraoral VSC level of 150 ppb or higher.

[0175] ● This study screened more than 80 participants.

[0176] ● Determine the VSC change for each participant from baseline to the end of treatment and average it for each group.

[0177] Oral hygiene

[0178] During the treatment period, each participant continued their oral hygiene practices, consisting of brushing twice daily with fluoride toothpaste (provided) and a toothbrush (provided). All participants were instructed not to use antibiotics, antibacterial agents (e.g., mouthwash or antibacterial toothpaste), or consume antibacterial mints, lozenges, or chewing gum during the treatment period. At the start of each leg, participants received a package containing a bottle of mouthwash, a tube of fluoride toothpaste, and a toothbrush assigned to them for that leg of treatment.

[0179] Participant selection

[0180] Twenty participants aged 18 to 65 years were recruited from 83 volunteers based on their oral VSC levels of 150 ppb or higher at pre-screening and baseline. Individuals were excluded if they were allergic to milk protein or zinc, had received antibiotic treatment within one month prior to study start, or had a history of requiring antibiotic treatment. Statistical power calculations using a 10% dropout rate indicated that 20 participants would allow for 90% statistical power.

[0181] Randomization and Blinding

[0182] A randomization plan was developed to ensure that all six treatment combinations (ABC, ACB, BAC, BCA, CAB, CBA) had equal probability. This study was blinded for both examiners and participants. Throughout the study, clinical examiners / recorders were unaware of which treatment sequence a participant was assigned to. Participants were also unaware of which mouthwash they were assigned, as it was provided in the same coded packaging (Mouthwash A, Mouthwash B, and Mouthwash C). Personnel distributing test materials or supervising their use were not involved in measuring participants' VSCs to minimize potential bias. Participants were assigned participant numbers. These participant numbers were recorded on their case report forms. Participants were randomly assigned to one of the three treatments. Randomization was determined according to a standard randomization table for the number of treatments in crossover studies. Each of the three treatment periods was a consecutive 14-day period. Each treatment period was separated by a 14-day washout period. After screening, eligibility for all participants was assessed according to inclusion / exclusion criteria. The primary analysis set consisted of all participants who completed the trial and had no significant protocol differences (according to protocol analysis). During the blinded review of the data, no participants were excluded due to significant protocol differences. The blinded review was conducted before the treatment code was cracked and before any analysis, so the Intention-To-Treat (ITT) and Per-Protocol (PP) groups used for the analysis were equivalent.

[0183] VSC Measurement Solution

[0184] At baseline and at the end of each treatment, volatile sulfur compounds (VSC) were measured using a Halimeter (Interscan, CA, USA) by an experienced clinician (inspector / recorder). The meter was calibrated to zero immediately in ambient air before each measurement. Each participant kept their mouth closed for one minute before measurement, then a disposable pipette connected to the meter was placed on the back of the tongue, slightly towards the midpoint, and held in place until the maximum VSC reading (in ppb) was obtained. Three readings were taken consecutively and then averaged.

[0185] Statistical analysis

[0186] Descriptive statistics (mean, standard deviation, and range) for all continuous variables and frequencies for all ordinal variables were calculated. All analyses were performed using Stata (StataCorp LP, College Station, TX, USA) or R statistical software, with a pre-specified significance level of P < 0.05. The mean VSC level for each participant was determined at baseline and at the 14-day visit. Differences between baseline and 14-day VSC readings were calculated, and these differences were then analyzed between treatment groups using the Mann-Whitney test. The Cochran-Mantel-Haenszel test was used for comparisons between treatment groups where necessary.

[0187] result

[0188] Clinical trial results showed that the use of three mouthwash formulations reduced VSC levels in participants' breaths, with the Zn-CPP-ACP complex mouthwash being significantly more effective than CPP-ACP or Zn mouthwash formulations alone in reducing VSC levels (Table 3).

[0189] Table 3. Relative changes in volatile sulfur compounds achieved via CPP-ACP and Zn-CPP-ACP complexes

[0190] a. Significantly different from the Zn control (p < 0.01).

[0191] b. Significantly different from other values ​​in the column (p < 0.01).

[0192] Example 4 - Ion Analysis in Complexes

[0193] The following is the protocol for ion analysis of PP-ACP or Zn-PP-ACP complex solutions. Total samples (tightly bound and loosely bound) and loosely bound samples are prepared as follows: Total (tightly bound and loosely bound): Take 1 ml of any solution containing PP-ACP (e.g., CPP-ACP) or Zn-PP-ACP (e.g., Zn-CPP-ACP complex) and place it in 19 ml of 1 M HNO3. Incubate at room temperature with continuous, slow, end-to-end mixing for 24 hours (20 rpm). Centrifuge the mixture at 1000 g for 15 minutes at room temperature. Analyze the supernatant for calcium, zinc, phosphate, and fluorine.

[0194] Loosely bound ions: Take a sample of the same solution as the 'total' analysis described above and place it in a centricon with a 1000 MWCO filter. Centrifuge at 3000 g for 1 hour at room temperature to produce sufficient filtrate (<10% of the total sample to avoid affecting equilibrium) for analysis by atomic absorption spectrophotometry (AAS) and ion chromatography (IC). The filtrate is then measured to obtain loosely bound ions.

[0195] Total and loosely bound calcium, zinc, phosphate, and fluorine in solution were determined by ion chromatography (for fluoride and phosphate) and atomic absorption spectrometry (for calcium and zinc).

[0196] The tightly bound (colloid-retained) ions of PP are calculated based on the difference between the total and loosely bound ions (as explained above).

[0197] Example 5

[0198] Oral spray

[0199] 2% Zn-CPP-ACP complex containing 15 mM zinc

[0200] Flavoring agent (1.5%)

[0201] Preservatives (e.g., sodium benzoate)

[0202] water

[0203] Example 6

[0204] Sugar-free mints

[0205] Sorbitol / Xylitol (95%)

[0206] Zn-CPP-ACP complex (2.0%)

[0207] Natural flavoring (1.98%)

[0208] Steviosides (0.48%)

[0209] Anti-caking agent (0.59%)

[0210] Example 7

[0211] mouthwash

[0212] Zn-CPP-ACP complex containing 15 mM zinc (2%)

[0213] Flavoring agent (1.5%)

[0214] Preservatives (e.g., sodium benzoate)

[0215] water

[0216] Example 8

[0217] Sugar-free chewing gum

[0218] Gel base (33.4%)

[0219] Sorbitol (41%)

[0220] Mannitol (12%)

[0221] Glycerin (10%)

[0222] Zn-CPP-ACP complex (2.0%)

[0223] Flavoring agent (1.5%)

[0224] Aspartame (0.1%)

[0225] Example 9

[0226] toothpaste

[0227] water

[0228] Sorbitol

[0229] Hydrated silica

[0230] Zn-CPP-ACP complex (10%)

[0231] Sodium dodecyl sulfate

[0232] Carrageenan

[0233] Flavorings

[0234] Xanthan Gum

[0235] Cocamidopropyl Betaine

[0236] Tin fluoride

[0237] Titanium dioxide

[0238] Example 10

[0239] toothpaste

[0240] water

[0241] Sorbitol

[0242] Hydrated silica

[0243] Zn-CPP-ACFP complex (10%)

[0244] Sodium dodecyl sulfate

[0245] Carrageenan

[0246] Flavorings

[0247] Xanthan Gum

[0248] Cocamidopropyl Betaine

[0249] Sodium fluoride

[0250] Titanium dioxide

[0251] Total fluoride level 1450 ppm

[0252] References

[0253] Miyazaki, H., Sakao, S., Katoh, Y. and Takehara, T. (1997) Oral Malodor in the general population of Japan. M. Rosenberg (ed.). Bad breath: research perspectives, 119-137. Ramat Aviv: Ramot Publishing-Tel Aviv University Press.

[0254] Ueno, M., Yanagisawa, T., Shinada, K., Ohara, S. and Kawaguchi, Y. (2007) Prevalence of oral malodor and related factors among adults in Akita Prefecture. Journal of Medical and Dental Sciences, 54, 159-165.

[0255] Liu, XN, Shinada, K., Chen, XC, Zhang, BX, Yaegaki, K. and Kawaguchi, Y. (2006) Oral malodor-related parameters in the Chinese general population. Journal of Clinical Periodontology 33, 31-36.

[0256] Ayers, KMS and Colquhoun, AUK (1998) Halitosis: causes, diagnosis, and treatment. New Zealand Dental Journal 94, 156-160.

[0257] Tangerman, A. (2002) Halitosis in medicine: a review. International Dental Journal, 52 (Supplement 3), 201-206.

[0258] Papaioannou, W. and Dereka, X. (2009) Halitosis: A demanding problem in clinical dentistry. Odontostomatological Progress 63, 82-93.

[0259] Quirynen, M., Dadamio, J., Van den Velde, S., De Smit, M., Dekeyser, C., Van Tornout, M. and Vandekerckhove, B. (2009) Characteristics of 2000 patients who visited a halitosis clinic. Journal of Clinical Periodontology 36, 970-975.

[0260] Hartley, MG, El-Maaytah, MA, McKenzie, C. and Greenman, J. (1996) The tongue microbiota of low odor and malodorous individuals. Microbial Ecology in Health and Disease 9, 215-223.

[0261] Tonzetich, J. (1977) Production and origin of oral malodor: a review of mechanisms and methods of analysis. Journal of Periodontology 48, 13-20.

[0262] Persson, S., Edlund, MB, Claesson, R. and Carlsson, J. (1990) The formation of hydrogen sulfide and methyl mercaptan by oral bacteria. Oral Microbiology and Immunology 5, 195-201.

[0263] Miyazaki, H., Sakao, S., Katoh, Y. and Takehara, T. (1995) Correlation between volatile sulfur compounds and certain oral health measurements in the general population. Journal of Periodontology 66, 679-684.

[0264] Takeuchi, H., Machigashira, M., Yamashita, D., Kozono, S., Nakajima, Y., Miyamoto, M., Takeuchi, N., Setoguchi, T. and Noguchi, K. (2010) The association of periodontal disease with oralmalodour in a Japanese population. Oral Diseases 16, 702-706.

[0265] Tsai, CC, Chou, HH, Wu, TL, Yang, YH, Ho, KY, Wu, YM and Ho, YP (2008). The levels of volatile sulfur compounds in mouth air from patients with chronic periodontitis. Journal of Periodontal Research 43, 186-193.

Claims

1. A method for treating or preventing halitosis in an individual in need, the method comprising applying stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) to the oral cavity of the individual, thereby treating or preventing halitosis.

2. The method of claim 1, further comprising the step of identifying an individual suffering from halitosis.

3. The method according to claim 1 or 2, wherein the individual suffers from moderate halitosis.

4. The method according to claim 1 or 2, wherein the individual suffers from severe halitosis.

5. The method according to any one of claims 1 to 4, wherein the level of volatile sulfur compounds (VSC) in the oral cavity of the individual is or is about 150 ppb (parts per billion) or higher.

6. The method according to any one of claims 1 to 5, wherein the individual is identified as 3 according to the Rosenberg scale.

7. The method according to any one of claims 1 to 5, wherein the individual is identified as 4 according to the Rosenberg scale.

8. The method according to any one of claims 1 to 5, wherein the individual is identified as 5 according to the Rosenberg scale.

9. The method according to any one of claims 1 to 8, wherein the individual has not been diagnosed with periodontal disease.

10. The method according to any one of claims 1 to 9, wherein the individual does not suffer from periodontal disease.

11. The method according to any one of claims 1 to 10, wherein the individual has not been diagnosed with periodontitis or severe gingivitis.

12. The method according to any one of claims 1 to 11, wherein the individual does not suffer from periodontitis or severe gingivitis.

13. The method according to any one of claims 1 to 12, wherein the individual has not been diagnosed with gingivitis.

14. The method according to any one of claims 1 to 13, wherein the individual does not suffer from gingivitis.

15. A method for reducing the generation of volatile sulfur compounds (VSCs) in the oral cavity of an individual, the method comprising applying stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) to the oral cavity of the individual, thereby reducing the generation of the volatile sulfur compounds in the oral cavity of the individual.

16. A method for inhibiting the activity of an enzyme produced by oral bacteria, the method comprising applying stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) to the oral cavity of an individual, thereby inhibiting the activity of the enzyme produced by the oral bacteria.

17. The method of claim 16, wherein the enzyme is a protease.

18. The method of claim 16 or 17, wherein the oral bacteria are oral bacteria involved in the generation of VSC.

19. The method according to any one of claims 16 to 18, wherein the oral bacteria are Gram-negative oral bacteria associated with halitosis.

20. The method according to any one of claims 15 to 19, further comprising the step of determining the level of volatile sulfur compounds (VSCs) in the oral cavity of the individual.

21. The method according to any one of claims 15 to 20, wherein the individual has not been diagnosed with or does not have gingivitis or periodontal disease.

22. The method according to any one of claims 1 to 21, wherein the stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) are phosphopeptide stable.

23. The method of claim 22, wherein the phosphopeptide is selected from casein phosphopeptide, egg yolk phosphopeptide, osteopontin phosphopeptide, and chemically or recombinantly synthesized phosphopeptides containing the sequence -ABC-.

24. The method of claim 23, wherein the phosphopeptide is a casein phosphopeptide.

25. The method according to any one of claims 1 to 24, wherein the stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) is a zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex.

26. The method of claim 25, wherein the zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex is phosphopeptide stable.

27. The method of claim 26, wherein the zinc ion content of the zinc-associated stable ACP or ACFP complex is about or at least 0.5 moles of zinc per mole of phosphopeptide.

28. The method of claim 27, wherein the zinc ion content is in the range of about 0.5 moles to about 100 moles per mole of phosphopeptide.

29. The method of claim 27, wherein the zinc ion content is in the range of about 0.5 moles to about 50 moles per mole of phosphopeptide.

30. The method of claim 27, wherein the zinc ion content is in the range of about 0.5 moles to about 20 moles per mole of phosphopeptide.

31. The method of claim 27, wherein the zinc ion content is in the range of about 0.5 moles to about 10 moles per mole of phosphopeptide.

32. The method according to any one of claims 27 to 31, wherein the zinc ion content of the zinc-associated stable ACP or ACFP complex is about or at least 1 mole, 2 mole, 3 mole, 4 mole, 5 mole, 6 mole, 7 mole, 8 mole, 9 mole or 10 mole of zinc per mole of phosphopeptide.

33. The method according to any one of claims 1 to 32, wherein the stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP) are applied to the oral cavity in the form of a composition comprising the stable amorphous calcium phosphate (ACP) and / or stable amorphous calcium fluoride phosphate (ACFP).

34. The method according to any one of claims 33, wherein the composition is a dental cleaning agent, the dental cleaning agent including toothpaste, tooth powder and liquid dental cleaning agents, mouthwash, mouthwash, oral spray, varnish, dental cement, lozenges, chewing gum, dental paste, gum massage cream, mouthwash tablets, dairy products and other foods, including yogurt.

35. Use of a stable amorphous calcium phosphate (ACP) and / or a stable amorphous calcium fluoride phosphate (ACFP) in the preparation of a medicament for treating or preventing halitosis.

36. Use of a zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex for the preparation of a medicament for the treatment or prevention of halitosis.

37. The method or use according to any one of claims 1 to 36, wherein the method or use causes a reduction in intraoral VSC levels of at least 50 ppb, 60 ppb, 70 ppb, 80 ppb, 90 ppb, 100 ppb, 110 ppb, 120 ppb, 130 ppb, 140 ppb, or 150 ppb.

38. The method or use according to any one of claims 1 to 36, wherein the method or use causes a decrease of at least 1 in the individual's Rosenberg scale measurement result.

39. A zinc-associated stable amorphous calcium phosphate (ACP) and / or a zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex.

40. The complex of claim 39, wherein the zinc-associated stable amorphous calcium phosphate (ACP) and / or zinc-associated stable amorphous calcium fluoride phosphate (ACFP) complex is phosphopeptide stable.

41. The complex of claim 40, wherein the phosphopeptide is a casein phosphopeptide.

42. The complex of claim 41, wherein the zinc ion content of the zinc-associated stable ACP or ACFP is about or at least 0.5 moles of zinc per mole of phosphopeptide.

43. The complex according to claim 42, wherein the zinc ion content is in the range of about 0.5 moles to about 100 moles per mole of phosphopeptide.

44. The complex according to claim 42, wherein the zinc ion content is in the range of about 0.5 moles to about 50 moles per mole of phosphopeptide.

45. The complex according to claim 42, wherein the zinc ion content is in the range of about 0.5 moles to about 20 moles per mole of phosphopeptide.

46. ​​The complex according to claim 42, wherein the zinc ion content is in the range of about 0.5 moles to about 10 moles per mole of phosphopeptide.

47. The complex according to any one of claims 39 to 46, wherein the zinc ion content of the zinc-associated stable ACP or ACFP is about or at least 1 mole, 2 mole, 3 mole, 4 mole, 5 mole, 6 mole, 7 mole, 8 mole, 9 mole or 10 mole of zinc per mole of phosphopeptide.

48. The complex according to any one of claims 39 to 46, wherein the zinc-associated stable ACP and / or ACFP complex comprises zinc ions that remain associated with the complex after centrifugation at about 3000 g for 1 hour in a 1000 molecular weight cutoff filter at room temperature.

49. A composition comprising, substantially consisting of or consisting of a zinc-associated stable ACP and / or ACFP complex according to any one of claims 39 to 48.

50. The composition of claim 49, wherein the composition is a dental cleaning agent, the dental cleaning agent comprising toothpaste, tooth powder and liquid dental cleaning agents, mouthwash, rinses, oral sprays, coatings, dental cements, lozenges, chewing gum, mints, dental pastes, gum massage creams, mouthwash tablets, dairy products and other food products, including yogurt.

51. The composition of claim 50, wherein the composition is a mint or chewing gum, preferably a sugar-free mint or sugar-free chewing gum.

52. A reagent kit comprising: (a) The composition according to claim 49 or 50, or (b) A zinc-associated stable ACP or ACFP complex according to any one of claims 39 to 48.

53. The kit of claim 49, wherein the kit further includes instructions for using the kit in the method of any one of claims 1 to 38.

54. A method or process for forming zinc-associated phosphopeptide-stabilized ACP or ACFP complexes, said method or process comprising or consisting of the following: (i) Obtain a solution containing at least one phosphopeptide; as well as (ii) Mixing a solution containing phosphopeptides, calcium ions, phosphate ions, hydroxide ions, and optionally fluoride ions while maintaining the pH at about 7 or higher, preferably about 9, to form a solution containing phosphopeptide-stabilized ACP or ACFP; and (iii) Mix the water-soluble zinc salt with the solution containing the phosphopeptide-stabilized ACP or ACFP. This results in the formation of zinc-associated phosphopeptide-stabilized ACP or ACFP complexes.

55. The method or process of claim 54, wherein the solution is added slowly at a rate of less than about 1% by volume per minute and is thoroughly mixed.

56. A method or process for forming zinc-associated phosphopeptide-stabilized ACP or ACFP complexes, said method or process comprising or consisting of the following: (i) Provide a solution of phosphopeptide-stabilized ACP or ACFP; as well as (ii) Mix the water-soluble zinc salt with the solution. This results in the formation of zinc-associated phosphopeptide-stabilized ACP or ACFP complexes.

57. The method or process according to any one of claims 54 to 56, wherein the water-soluble zinc salt is in the form of zinc acetate, zinc lactate, zinc sulfate, zinc nitrate, zinc gluconate, zinc chloride, zinc citrate, or any other water-soluble zinc salt.

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