Oral care product comprising stannous fluoride

Abstract

The invention provides a new oral care product comprising stannous fluoride and at least 0.01 wt% of a self-assembling peptide, wherein the self-assembling peptide is present in the oral care product essentially in an assembled form. The assembled peptide forms a three-dimensional matrix, which is able to stabilize the stannous fluoride by protecting it from oxidation. The oral care product can further comprise a polyphosphate, such as sodium tripolyphosphate, and a tin chelator, such as sodium gluconate. They can be provided in the form of a dental gel, a toothpaste, a tooth foam, an oral rinse or a preventive paste. Also disclosed is a method of stabilizing stannous fluoride in an oral care product. Finally, the invention relates to a method of treatment for protecting teeth from demineralization, remineralizing the enamel of non-cariogenic dental lesions, disrupting biofilm formation on teeth and treating dentinal hypersensitivity.

Description

[0001] This invention provides a novel oral care product comprising stannous fluoride and at least 0.01 wt% of a self-assembled peptide, wherein the self-assembled peptide is primarily present in an assembled form within the oral care product. The assembled peptide forms a three-dimensional matrix capable of stannous fluoride stabilizing by protecting it from oxidation. The oral care product may also include polyphosphates, such as sodium tripolyphosphate, and tin chelating agents, such as sodium gluconate. These may be provided in the form of dental gels, toothpastes, preventative pastes, dental foams, or oral rinses. This invention also discloses a method for stabilizing stannous fluoride in an oral care product. Finally, this invention relates to methods for protecting one or more teeth from demineralization, remineralizing enamel, disrupting biofilms formed on one or more teeth, and treating dentin hypersensitivity.

[0002] The caries-preventing effect of fluoride is scientifically proven and therefore widely recognized. Fluoride reacts with calcium in dental plaque and saliva to form calcium fluoride on the tooth surface, thereby inhibiting tooth demineralization and promoting tooth remineralization. Calcium fluoride eventually dissolves, allowing calcium and fluoride ions to interact with the tooth and form fluorapatite within the tooth structure. By converting the calcium mineral apatite in the tooth into fluorapatite, fluoride also makes tooth enamel more resistant to acid erosion by bacteria. In addition, fluoride can directly inhibit dental plaque bacteria by interfering with bacterial metabolism through various mechanisms (https: / / en.wikipedia.org / wiki / Tin(II)_fluoride; Marquis, 1995; Nasser et al., 2023).

[0003] Given these known beneficial effects of fluoride, oral care products such as toothpaste (teeth cleaning agents) containing fluoride are now commonly used.

[0004] Historically, the first fluoride toothpastes provided fluoride in the form of tin(II) fluoride, commonly also known as stannous fluoride (SnF2). These tin(II) fluoride toothpastes showed considerable efficacy in reducing crown caries; however, their use in oral care compositions was limited due to their instability in water. Tin(II) fluoride ions (SnF2) 2+ It has high reactivity and can be rapidly oxidized to inactive tin ions (Sn). 4+ Sn 2+ Oxidation of stannous fluoride manifests as a yellowish-brown staining on the tooth surface in the oral cavity. Furthermore, tin in stannous fluoride is readily hydrolyzed at pH values ​​above 4, causing it to precipitate from the solution and thus lose its therapeutic properties. Importantly, the chemical reactions leading to the deactivation of stannous fluoride occur not only during storage but also directly during processing (White, 1995).

[0005] Therefore, manufacturers of fluoride toothpaste products have turned to alternative fluoride sources, such as monofluorophosphate or sodium fluoride, whose anti-caries activity is comparable to that of fluoride ions in stannous fluoride. However, unlike sodium monofluorophosphate or sodium fluoride, stannous fluoride is also known for its inherent antibacterial effects and its clinically proven efficacy in controlling gingivitis. Furthermore, it exhibits significant reactivity with the root surface and has been shown to reduce dentin hypersensitivity. Consequently, there has been renewed interest in manufacturing dental products containing stannous fluoride in recent years, as it is considered a "multi-functional" active ingredient with therapeutic benefits (White, 1995).

[0006] Therefore, scientists face the challenge of stannous fluoride stabilizing in dental care products and during administration. Stable stannous fluoride formulations significantly improve the bioavailability of stannous and fluoride ions, thereby enhancing their benefits to oral health. Stabilizing stannous fluoride requires preventing the formation of Sn. 2+ Oxidized to Sn 4+ It also protects SnF2 from hydrolysis. Early stannous fluoride dental cleanings proved to be generally stable enough to provide some caries prevention, but the bioavailable stannous fluoride provided was insufficient to exert the antibacterial activity required to control gingivitis. In other words, the stability requirements for stannous fluoride as a gingivitis activator appear to be more stringent than its caries prevention activity (White et al., 1995).

[0007] Over time, many strategies have been developed to obtain more stable stannous fluoride formulations:

[0008] For example, by significantly reducing or completely omitting the addition of water to SnF2-containing toothpaste formulations, Sn can be prevented from... 2+ It oxidizes to tin ions. However, a low water content of less than 10% will affect the taste and texture of the resulting toothpaste.

[0009] Stannous fluoride can also be dissolved in anhydrous materials such as glycerol to obtain non-hydrogel formulations. For example, US4418057A and US3433544A describe methods for formulating stannous fluoride into non-hydrogel mixtures containing anhydrous glycerol and hydroxyethyl cellulose as gelling agents.

[0010] Further attempts to stabilize stannous fluoride in oral care products include adding stannous salts that can react with oxygen to protect the stannous fluoride from oxidation. Stannous salts, such as stannous phosphate or stannous chloride, can also provide a reservoir source to replenish Sn lost from active SnF2. 2+ US5004597A discloses an exemplary composition containing stannous fluoride, which relies on stannous chloride and stannous gluconate as substitutes for unstable Sn. 2+A reservoir of tin ions. However, using some of these tin salts can negatively affect the taste of toothpaste, and more importantly, they have proven to be rather ineffective in preventing the formation of large amounts of tin ions.

[0011] Chelating agents are also considered to protect stannous fluoride chelates from hydrolysis and oxidation, thereby reducing or even eliminating the precipitation of insoluble tin compounds. Suitable stannous chelating agents include sodium gluconate, stannous gluconate, citrates such as zinc citrate, and hydroxylated aliphatic dicarboxylic acids and tricarboxylic acids such as citric acid or malic acid. Formulations containing these chelating agents are disclosed, for example, in US5716600A, US5004597A, US9968803B2, or US3282792A. Alternatively, copolymers of maleic anhydride or maleic acid with polymerizable vinyl unsaturated monomers (preferably lower alkyl vinyl ethers, such as methoxyethylene) have been found to be effective with Sn. 2+ Formation of chelates, thereby protecting Sn 2+ Ions are protected from oxidation. For example, the use of such polymeric chelating agents in tin fluoride formulations is disclosed in US5017363A, US4960586A, or US4961924A. However, the addition of some of the aforementioned chelating agents may result in a coarse toothpaste texture and a bitter, metallic, astringent, or salty taste (Li et al., 2019).

[0012] Finally, Colgate Total SF, produced by Clogate-Palmolive, relies on a formulation in which SnF2 is stabilized with zinc phosphate, which chelates SnF2 to prevent its oxidation.

[0013] Despite the different methods and strategies for stabilizing stannous fluoride in oral care formulations, there is still a need for improved stannous fluoride oral care products that are characterized by both a stable composition and enhanced efficacy. Summary of the Invention

[0014] The present invention, and particularly the subject matter of the present invention, solves this problem.

[0015] This invention provides an oral care product comprising:

[0016] i. stannous fluoride (II), and

[0017] ii. At least 0.01 wt% of a self-assembled peptide comprising the sequence X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid, aspartic acid, glutamine and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan and glutamine, wherein the self-assembled peptide is present in the oral care product in a substantially assembled form.

[0018] The oral care products of the present invention include tin(II) fluoride, which, in the context of the present invention, is also referred to as stannous fluoride or SnF2. Tin(II) fluoride may be present in the oral care products at a concentration of 0.1 wt% to 1.0 wt%.

[0019] Preferably, stannous fluoride is present in the oral care product at a concentration of 0.2 wt% to 0.7 wt%. Previous studies have shown that concentrations of 0.4 wt% to 0.7 wt% (equivalent to about 1000 ppm to 1500 ppm of fluoride), particularly 0.454 wt% stannous fluoride, provide optimal therapeutic protection against caries, gingivitis, plaque, and dentin hypersensitivity (West et al., 2018; Parkinson et al., 2020). For children, a concentration of 0.227 wt% (about 500 ppm of fluoride) is recommended. Therefore, in a preferred embodiment, the oral care product of the present invention comprises stannous fluoride at a concentration of 0.1 wt% to 0.7 wt%, with a most preferred concentration of 0.454 wt%.

[0020] The inventors have surprisingly discovered that the dispersion of stannous fluoride in a three-dimensional matrix formed by self-assembled peptides effectively protects stannous ions (Sn). 2+ It is not oxidized to tin ions (Sn). 4+ This prevents the formation of extrinsic staining on the tooth surface due to oxidation.

[0021] In the context of this invention, self-assembling peptides (SAPs) are peptides capable of self-assembling in a pH-dependent manner to form three-dimensional scaffolds or matrices, thereby promoting tissue regeneration. As used herein, “self-assembly” of a peptide refers to the spontaneous and reversible organization of a peptide into a multimeric assembly with other peptides of the same class (or peptides with similar structures) through non-covalent interactions. The non-covalent interactions responsible for the formation of multimeric assemblies include van der Waals forces, π-stacking, hydrogen bonds, polarity, and ionic interactions between the amino acid backbone and / or amino acid side chains of the peptide.

[0022] For example, suitable self-assembling peptides are taught in WO 2004 / 007532 A1, US10 / 521,628, US12 / 729,046, US13 / 551,878, US14 / 062,768, WO 2010 / 041636 A1, or WO2014 / 027012 A1, all of which are incorporated herein by reference in their entirety. Self-assembling peptides can assemble into β-sheets in one dimension, as well as into higher-order assemblies such as filaments or ribbons. They can form three-dimensional supramolecular structures of self-assembling proteins with an affinity for calcium phosphate.

[0023] The size of the self-assembly peptides used in the oral care products of this invention is not particularly limited. They can be of any length, allowing for self-assembly in a pH-dependent manner. The size of the peptides can be about 5-200 amino acids, for example 9-100 amino acids, 10-50 amino acids, 10-30 amino acids, or 11-20 amino acids. Preferably, the length of the self-assembly peptides is about 27 amino acids, 24 amino acids, 21 amino acids, 15 amino acids, or 11 amino acids. In a particularly preferred embodiment, the length of the self-assembly peptide is 11 amino acids, i.e., it consists of 11 amino acids.

[0024] In order to adhere to the tooth surface, the matrix formed by the self-assembled peptides present in the oral care product of the present invention must be able to bind mineral particles present on the tooth surface. Therefore, the matrix contains binding sites for mineral particles, enabling it to bind particles (preferably containing calcium) to the tooth surface. For example, charged amino acid residues such as Glu on the surface of the self-assembled peptides bind to hydroxyapatite particles and the tooth surface, which is also substantially composed of hydroxyapatite. Three-dimensional self-organizing ability is important for binding. Generally, highly charged surfaces promote the adhesion of mineral particles. The protein matrix is ​​particularly effective when its surface displays glutamate residues that can attach to calcium phosphate or other mineral particles. Preferably, the protein contains 5% or more, 10% or more, 20% or more, or 30% or more of charged amino acid residues, such as glutamate residues.

[0025] In the context of this invention, self-assembling peptides may be able to self-assemble, such as peptides P11-4, P11-8, P11-2, and P11-5 mentioned below, but they may also choose to self-assemble in combination of two self-assembling peptides, such as polypeptides P11-13 / P11-14 and P11-28 / P11-29, P11-30 / P11-31 mentioned below.

[0026] The self-assembled peptide in the oral care product of this invention comprises a common sequence X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid, aspartic acid, glutamine, and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, and glutamine. Independent selection means, for example, that X1 at positions 1, 3, or 5 of the above sequence can be different from each other. Of course, they can also be the same.

[0027] In one embodiment, the self-assembled peptide comprises the sequence X1-X2-X1-X2-X1, wherein X1 is an amino acid having an acidic side chain and X2 is an amino acid having a hydrophobic or polar side chain.

[0028] Preferably, the self-assembled peptide further includes X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid and ornithine, and X2 is independently selected from tryptophan and phenylalanine.

[0029] The self-assembled peptide may also contain X3-F-X1-W-X1-X1-F-X1, wherein X1 is independently selected from glutamic acid and ornithine, and X3 is selected from arginine, glutamic acid and ornithine, wherein X3 is preferably arginine.

[0030] The self-assembled peptides used in the oral care products of this invention may comprise, or preferably comprise, X4-X4-X3-F-X1-W-X1-F-X1-X4-X4, wherein X1 is independently selected from glutamic acid and ornithine, wherein X3 is selected from arginine, glutamic acid, and ornithine, and wherein X4 is independently selected from glutamine, glutamic acid, serine, threonine, and ornithine. X3 is preferably arginine. Independently, X4 is preferably glutamine.

[0031] The self-assembled peptide may contain, or preferably consist of, SEQ ID NO: 5: QQRF-X1-W-X1-F-X1-QQ, wherein X1 is independently selected from glutamic acid and ornithine.

[0032] In the context of this invention, self-assembling peptides taught in WO 2004 / 007532 A1, US10 / 521,628, US12 / 729,046, US13 / 551,878, US 14 / 062,768, or WO2014 / 027012 A1 are preferred, all of which are fully incorporated herein by reference. Most preferably, the peptides comprise or consist of the specific peptides listed in Table 2. Of course, self-assembling peptides assembled in combination with another self-assembling peptide, for example as described above, can be formulated into compositions.

[0033] Preferably, the self-assembly peptide comprises or is composed of the sequence of SEQ ID NO:6. Peptides composed of the sequence of SEQ ID NO:6 are also designated as P11-4 and are preferred in this invention. In another preferred embodiment, the self-assembly peptide comprises or is composed of the sequence of SEQ ID NO:9 (P11-8). In another preferred embodiment, the self-assembly peptide comprises or is composed of the sequence of SEQ ID NO:15 (P11-20).

[0034] Peptides of SEQ ID NO: 6, 9 or 15 are particularly advantageous, for example, because they can be used at relatively low concentrations, they are highly compatible with cells and have a beneficial charge distribution.

[0035] Table 1: Common sequences of preferred self-assembled peptides

[0036] Table 2: Preferred self-assembled peptides. Position X1 is indicated by an underline, and "O" represents the amino acid ornithine.

[0037] The compositions of the present invention may further comprise at least one self-assembled peptide having at least 45% sequence identity with the peptide composed of SEQ ID NO:6. Preferably, the peptide has at least 54%, at least 63%, at least 72%, at least 81%, or at least 90% sequence identity with the peptide composed of SEQ ID NO:6, or is the peptide itself.

[0038] The self-assembled peptides used in the oral care products of the present invention can also be structurally modified at one or more amino acid positions, for example by introducing one or more modified amino acids. According to the present invention, these modified amino acids can be amino acids altered by, for example, biotinylation, phosphorylation, glycosylation, acetylation, branching, and / or cyclization. Furthermore, the self-assembled peptides present in the oral care products of the present invention may additionally or alternatively contain other modifications, such as terminal blocking groups, formyl groups, γ-carboxyglutamic acid, hydroxyl groups, methyl groups, phosphoryl groups, pyrrolidone carboxylic acid groups, and / or sulfate groups.

[0039] In a preferred embodiment, all self-assembled peptides present in the oral care products of the present invention are acetylated at their N-terminus and / or amidated at their C-terminus, for example with an NH2 group, and most preferably both are amidated. Since unblocked forms tend to undergo deamination, the ends of the self-assembled peptides described herein are preferably blocked to improve stability. Therefore, in a preferred embodiment, the self-assembled peptides present in the oral care products of the present invention, such as the peptides of SEQ ID NO:6, SEQ ID NO:9, and / or SEQ ID NO:15, comprise an Ac-N-terminus and an NH2-C-terminus. A particularly preferred embodiment is peptide P11-4 (SEQ ID NO:6), which has an N-terminus acetylated and a C-terminus amidated with an NH2 group.

[0040] The oral care product may contain a single type of self-assembly peptide or two or more types, such as three, four or five types, of different types of self-assembly peptides. For example, it may contain a mixture of self-assembly peptides composed of SEQ ID NO:6 and self-assembly peptides composed of SEQ ID ID:15.

[0041] In the oral care products of the present invention, the self-assembling peptides are present essentially in an assembled form. As used herein, this means that at least about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99%, or even about 100% of the peptides found in the oral care products are assembled, i.e., in polymer or multimer form.

[0042] Most self-assembling peptides in the oral care products of the present invention, such as P11-4 (SEQ ID NO:6) and / or P11-20 (SEQ ID ID:15) and their terminal-modified variants, undergo self-assembly once the pH of their environment drops below pH 7.5. This means that when the pH drops below 7.5, the self-assembling peptides present in the oral care products of the present invention begin to self-assemble to a large extent.

[0043] Assembly is primarily controlled by pH. The pH at which self-assembling peptides assemble depends on their sequence. For example, P11-4 assembles below pH 7.5. The pH of P11-4 and similar peptides in the oral care products of the present invention is preferably below 7.5, especially if the self-assembling peptide is P11-4. The pH is preferably between 4 and 7.5, for example 5-7.5 or 6-7.

[0044] Other self-assembling peptides, such as P11-8, self-assemble at pH 7.5 or higher. If such self-assembling peptides are used, the pH of the oral care product needs to be adjusted accordingly, i.e., the pH should be higher than the pH at which the self-assembling peptide assembles. For example, a pH higher than 7.5.

[0045] The assembly state of peptides is also affected by ionic strength. The ionic strength of a solution is a function of the concentration of all ions in that solution. Therefore, even at pH values ​​above where peptide self-assembly begins—that is, when the peptide is essentially a monomer in solution—particularly high ionic strengths can trigger peptide assembly.

[0046] Those skilled in the art will know how to determine and measure the ionic strength of a solution. Ionic strength I is typically calculated using the formula I = ½∑zi²bi, where z is the valence factor and bi is the molar concentration of the ions [mol / kg{H₂O}]. The sum ∑ is the sum of all ions in the solution. For example, the ionic strength of a 150 mM NaCl solution is approximately 0.15 mol / L. This is also the ionic strength of blood. The ionic strength of saliva in the oral cavity is typically much lower, for example, about 0.04 mol / L. In the context of this invention, the physiological range of ionic strength is considered to correspond to an ionic strength of 0.15 mol / L.

[0047] Those skilled in the art also know that peptide concentration can affect peptide assembly; that is, particularly high peptide concentrations may trigger assembly. Conversely, extremely low peptide concentrations may prevent the assembly of the peptides of the present invention, i.e., even under low pH conditions present in dental lesions and the oral cavity.

[0048] Those skilled in the art will be able to determine, through routine experiments, whether substantially all self-assembled peptides are in an assembled form. For example, the assembly state of peptides in solution can be determined by nuclear magnetic resonance (NMR) methods such as 1H-NMR, circular dichroism spectroscopy, dynamic light scattering (DLS) analysis, diffuse wave spectroscopy, native electrophoretic methods, viscosity measurements (rheology), and quartz crystal microbalances with dissipation monitoring (QCMD), with native electrophoresis being preferred. TEM can detect the presence of self-assembled peptide fibers.

[0049] Suitable buffers and pH adjusters for obtaining the desired pH are known in the art.

[0050] As discussed in this article, the stability of stannous fluoride involves preventing the formation of stannous ions (Sn). 2+ Premature oxidation to tin ions (Sn) 4+The inventors have surprisingly discovered that the dispersion of stannous fluoride in a three-dimensional matrix formed by the self-assembled peptides described herein protects stannous ions from oxidation within oral care products and when these products are applied to tooth surfaces. Unbound by theory, fibers formed from the self-assembled peptides with a net charge of -2 act as stabilizing factors, thereby preventing the oxidation of stannous ions. The three-dimensional matrix of the self-assembled peptides, through its affinity for hydroxyapatite, further contributes to extending the duration of stannous fluoride on the enamel surface. The increased stability of stannous ions due to the presence of the self-assembled peptides prevents the formation of tooth staining typically observed due to SnF2 oxidation.

[0051] It is noteworthy that the inventors have discovered that the remarkable ability of the self-assembled peptides described herein to stabilize and protect stannous fluoride from oxidative changes is largely dependent on the concentration of the self-assembled peptides in the oral care product: this beneficial effect is only visible in formulations containing 100 PPM (i.e. 0.01 wt%) or more of the self-assembled peptides, while this effect is not apparent in formulations with self-assembled peptide concentrations below 100 PPM.

[0052] Therefore, the self-assembled peptide is present in the oral care product of the present invention at a concentration of at least 100 PPM or at least 0.01 wt%, for example at a concentration of 0.01 wt%-5 wt%, such as 0.01 wt%-0.1 wt%, 0.1 wt%-1 wt%, 1 wt%-2 wt%, 2 wt%-3 wt%, 3 wt%-4 wt%, or 4 wt%-5 wt%. Preferably, the self-assembled peptide is present in the oral care product at a concentration of 0.01 wt%-0.1 wt%, for example about 0.01 wt%, 0.02 wt%, 0.03 wt%, 0.04 wt%, 0.05 wt%, 0.06 wt%, 0.07 wt%, 0.08 wt%, 0.09 wt%, or 0.1 wt%. Most preferably, the self-assembled peptide is present in the oral care product at a concentration of about 0.03 wt% to 0.07 wt%, for example, about 0.05 wt%.

[0053] While not essential, in a preferred embodiment, the oral care product of the present invention further includes polyphosphates. Polyphosphates are inorganic polymers composed of two or more phosphate molecules arranged primarily in a linear configuration, although cyclic derivatives may be used in some cases. As chelating agents, polyphosphates are able to prevent plaque formation by binding to the tooth surface. Therefore, polyphosphates reduce the adhesion of absorbed salivary glycoproteins and facilitate the desorption of salivary proteins from tooth enamel (Mason et al., 2019). Thus, polyphosphates help remove stains and control stain buildup during brushing. Therefore, polyphosphate compounds are frequently used in whitening toothpastes. Furthermore, due to their chelating ability, polyphosphates can bind to calcium ions in saliva and plaque, thereby inhibiting the formation of calcified deposits (called tartar) on the tooth surface.

[0054] Surprisingly, the inventors discovered that polyphosphates do not prevent self-assembling peptides from adhering to tooth enamel, so they can be incorporated into oral care products without compromising the dental benefits of the self-assembling peptides.

[0055] The U.S. Food and Drug Administration (FDA) has listed sodium pyrophosphate, sodium trimetaphosphate, and sodium hexametaphosphate as Generally Recognized As Safe (GRAS) food additives (Moon et al., 2019). Any one of these or a combination thereof can be used in the oral care products of this invention.

[0056] For example, oral care formulations containing polyphosphates (such as sodium hexametaphosphate) are disclosed in US6667027B2, US6350436B1, US6821507B2, US5578293A, US5145666A, US5281411A or US5281410A for reducing staining caused by oxidation of stannous fluoride.

[0057] In some embodiments, the polyphosphate present in the oral care product of the present invention can be a pyrophosphate, specifically a diphosphate characterized by two phosphorus atoms bonded to one oxygen atom. The pyrophosphate can be provided in the oral care product as a disodium or tetrasodium salt.

[0058] However, in a preferred embodiment, the polyphosphate present in the oral care product of the present invention is sodium tripolyphosphate (STP). STP may be present in the oral care product at a concentration of 1 wt%-10 wt%, for example, 2 wt%-5 wt%. Preferably, the oral care product of the present invention contains about 3 wt% STP.

[0059] Oral care products may optionally further contain additional ingredients known to have the ability to stabilize stannous fluoride.

[0060] For example, in some embodiments, oral care products may also contain one or more agents capable of chelating stannous ions. Chelation refers to a chemical reaction in which an ion or molecule binds to a metal ion. Suitable stannous chelating agents capable of forming stable chelates with stannous fluoride and thus protecting it from hydrolysis and oxidation are known in the art, including, for example, sodium gluconate, stannous gluconate, or zinc citrate.

[0061] The reagent capable of chelating stannous ions can be present in oral care products at a concentration of 0 wt%-5 wt%, such as 0.5 wt%-1 wt%, 1 wt%-2 wt%, 2 wt%-3 wt%, 3 wt%-4 wt%, or 4 wt%-5 wt%. More preferably, the reagent capable of chelating stannous ions is present in oral care products at a concentration of 2 wt%-4 wt%, such as 2 wt%, 3 wt%, or 4 wt%.

[0062] Preferably, the reagent capable of chelating stannous ions is sodium gluconate. Sodium gluconate is non-corrosive, non-toxic, and readily biodegradable. It forms stable chelates with calcium, iron, copper, aluminum, and other heavy metals (including tin). Therefore, sodium gluconate is preferably used in the oral care products of the present invention to further stabilize stannous fluoride.

[0063] Therefore, in a preferred embodiment, the present invention provides an oral care product comprising:

[0064] i. 0.1 wt% - 1.0 wt% stannous fluoride;

[0065] ii. At least 0.01 wt% of a self-assembled peptide consisting of SEQ ID NO:6 (P11-4);

[0066] iii. 1 wt%-10 wt% sodium tripolyphosphate; and

[0067] iv. Optionally, 0.5 wt%–5 wt% sodium gluconate,

[0068] The pH of the oral care products mentioned therein is below 7.5.

[0069] The oral care product according to the invention can be a dental gel, toothpaste (teeth cleaning agent), preventative paste, dental foam, or oral rinse, preferably a dental gel. The oral care product is intended for use by professionals in dental clinics, preferably using suitable oral instruments such as rubber polishers. It can also be applied using a syringe. However, the oral care product can also be used at home, preferably applied to the tooth surface using a finger or a toothbrush (e.g., an interdental toothbrush).

[0070] Oral care products may also include one or more common ingredients found in the product. These common ingredients may include:

[0071] - Abrasives, such as carbonates, phosphates, silicates, acrylates, and alumina.

[0072] - Suspension agents, such as glycerin, polyethylene glycol (PEG), sorbitol, xylitol and / or erythritol,

[0073] - Adhesives, such as cellulose and its derivatives, such as carboxymethyl cellulose (cellulose gum), carrageenan, paraffin, xylose,

[0074] - Detergents, such as hydrogenated castor oil, sodium lauryl sulfate,

[0075] -Flavors, such as caramel, vanillin, menthol,

[0076] - Preservatives, such as ethanol and sodium benzoate,

[0077] - Colorants, such as Solvent Red and Acid Blue 3,

[0078] - Additional fluoride sources, preferably in the form of tertiary amines, such as amine fluorides or organofluorine compounds, such as sodium monofluorophosphate.

[0079] Oral care products typically contain water, optionally more than 10%.

[0080] For example, a particularly preferred oral care product of the present invention, comprising stannous fluoride and the self-assembling peptides described herein, is a dental gel and further includes...

[0081] a. Sodium tripolyphosphate,

[0082] b. Sodium gluconate

[0083] c. Sorbitol,

[0084] d. Cellulose gum,

[0085] e. Xylitol,

[0086] f. Erythritol,

[0087] g. Hydrated silica,

[0088] h. Phosphoric acid,

[0089] i. Optionally, NaOH, and

[0090] j. Water.

[0091] This invention provides an oral care product in which stannous fluoride is stabilized using a matrix formed from the self-assembled peptides described herein. Therefore, this invention also relates to a novel method for stabilizing stannous fluoride in an oral care product, comprising mixing...

[0092] i) Tin fluoride, and

[0093] ii) at least 0.01 wt% of a self-assembled peptide comprising the sequence X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid, aspartic acid, glutamine, and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, and glutamine, wherein the self-assembled peptide is substantially present in an assembled form in the oral care product.

[0094] Preferably, the oral care product is the oral care product of the present invention.

[0095] The oral care product of the present invention is stable at room temperature (20°C) for at least 6 months, at least 12 months, at least 18 months, at least 24 months, or at least 36 months. In this case, stability means that no visible discoloration occurs due to oxidation of stannous fluoride.

[0096] Using the oral care products described in this article has a range of beneficial effects.

[0097] For example, the oral care products of the present invention can be used to prevent tooth demineralization or to remineralize tooth enamel, such as remineralization of non-carious dental lesions.

[0098] Dental caries is one of the most common bacterial infections in the world. It is caused by the breakdown of tooth structure by bacterial metabolic products, primarily acids produced when bacteria break down food debris or sugars on the tooth surface or in the biofilm. This leads to an imbalance between demineralization and remineralization processes. The hard tooth structure—enamel, dentin, and cementum—is damaged by continuous demineralization, resulting in caries lesions and eventually tooth decay. The earliest sign of new caries lesions is the appearance of chalky white spots on the tooth surface, known as leukoplakia (also called early caries), or subsurface caries. As demineralization progresses, the mineralized surface of the lesion (partially) collapses and fractures, creating microcavities or cavities—holes in the tooth. This is called (partial) caries lesions or cavitary lesions.

[0099] Stannous fluoride is renowned for its significant efficacy in reducing crown caries by preventing enamel demineralization and promoting enamel remineralization (Fiorillo et al., 2020). On the other hand, as disclosed in International Patent Application WO 2021 / 110923 A1, a matrix formed from the self-assembled peptides described herein can constitute a protective layer or film on the tooth to which it is applied. The protective layer formed from the assembled self-assembled peptides can effectively prevent further demineralization and protect the enamel from acid erosion.

[0100] Furthermore, matrices formed from self-assembled peptides such as P11-4 disclosed in this paper can lead to increased mineralization (Soares et al., 2017). It was further found that self-assembled peptide matrices can prevent artificial caries lesions and lead to remineralization of enamel around orthodontic brackets (Jablonsky-Momeni et al., 2019).

[0101] While both stannous fluoride and the self-assembled peptides protect against enamel demineralization and remineralization, they may act synergistically. Furthermore, by suspending itself in a three-dimensional matrix formed by the self-assembled peptides disclosed herein, stannous fluoride remains and stabilizes on the enamel surface for an extended period, resulting in sustained protection against tooth demineralization and enamel remineralization.

[0102] Therefore, the present invention provides methods for preventing tooth demineralization and remineralizing tooth enamel (e.g., enamel in non-carious dental lesions) in subjects in need, wherein both methods involve applying an effective amount of the oral care product of the present invention to the teeth. Of course, treating one tooth can also be treating multiple teeth, preferably all teeth of the subject.

[0103] In another embodiment, the oral care product of the present invention can be used to disrupt bacterial biofilms formed on teeth.

[0104] Stannous fluoride is known to exhibit antibacterial activity through a bactericidal mechanism and by inhibiting bacterial metabolic enzymes. Consistent with this, the efficacy of stannous fluoride in controlling gingivitis has been clinically demonstrated (Paraskevas et al., 2006; Parkinson et al., 2020; Acherkouk et al., 2021). The inventors have discovered that the presence of self-assembled peptides further enhances the antibacterial effect of stannous fluoride. These self-assembled peptides form a negatively charged three-dimensional matrix that repels net negatively charged bacteria. Therefore, the biofilm formed on the tooth surface has a lower density and is more easily disrupted by stannous fluoride. Thus, the oral care product according to the invention allows for more effective removal of tooth biofilm.

[0105] Therefore, the invention disclosed herein also provides a method for reducing or destroying biofilms formed on teeth, comprising applying an effective amount of the oral care product of the present invention to the teeth.

[0106] Enhancing the disruption of the biofilm on the tooth surface helps prevent gingivitis, periodontitis, and / or peri-implantitis.

[0107] Gingivitis is a non-destructive periodontal disease, most commonly plaque-induced gingivitis caused by a bacterial biofilm (also known as dental plaque) that adheres to the tooth surface. Good oral hygiene can reverse gingivitis.

[0108] However, if left untreated or uncontrolled, gingivitis can develop into periodontitis. Periodontitis, or periodontal disease, is a group of inflammatory diseases affecting the periodontal tissues—the tissues that surround and support the teeth. Periodontitis is caused by microorganisms that attach to and grow on the surface of the teeth, and an overactive immune response to these microorganisms. As the gingival fibers break down, the gingival tissue separates from the teeth, leading to a deepening of the gingival sulcus, called a periodontal pocket. Subgingival microorganisms, those that exist at the base of the gingival line, colonize the periodontal pockets, causing further inflammation of the gingival tissue and progressive bone loss. If left undisturbed, microbial plaque can calcify to form tartar, commonly known as dental plaque. Tissue destruction (such as periodontal ligament damage) and alveolar bone resorption can ultimately lead to tooth loosening and subsequent loss of the affected teeth.

[0109] Peri-implantitis is a destructive inflammatory process that affects the soft and hard tissues surrounding the implant, caused by the formation of plaques in the peri-implant tissue.

[0110] Therefore, in another aspect, the present invention provides a method for preventing gingivitis, periodontitis and / or peri-implantitis in subjects with such need, comprising applying an effective amount of the oral care product of the present invention to the enamel surface.

[0111] Clinical studies have further demonstrated that stannous fluoride can significantly reduce dentin hypersensitivity (Schiff et al., 2005; Hines et al., 2019). Unbound by theory, the stannous ions in stannous fluoride form insoluble tin compounds in the presence of saliva, namely, tin-containing, low-solubility hydroxyapatite, which precipitates in and seals open dentinal tubules.

[0112] On the other hand, when the self-assembling peptides described in this article self-assemble into a three-dimensional matrix structure, the matrix forms a stable adhesion barrier on the exposed dentinal tubules, physically blocking the pathway of external stimuli.

[0113] Therefore, the stannous fluoride and self-assembled peptides present in the oral care product of the present invention promote parallel desensitization. They may work synergistically to reduce dentin hypersensitivity. Furthermore, since the self-assembled peptides in the oral care product also help maintain stannous ions on the dentin surface, they ensure that the desensitizing effect of stannous fluoride can be prolonged.

[0114] Therefore, by combining the desensitizing effects of self-assembled peptides and stannous fluoride, the oral care product according to the invention provides more lasting and faster sensitivity relief because the self-assembled peptide matrix forms an immediate barrier on the exposed dentin. The stannous ions remain persistently present on the root surface through the self-organized peptide matrix, prolonging the relief time.

[0115] Therefore, in another aspect, the present invention also provides an improved method for treating dentin hypersensitivity in a subject with this need, comprising applying an effective amount of the oral care product of the present invention to the teeth.

[0116] For all methods described herein, the preferred subjects are humans. However, they may also be non-human mammals, such as domestic pets like cats or dogs, or other livestock such as cattle, pigs, horses, sheep, goats, or camels. Subjects may also be another domesticated animal, such as a non-human primate, or a lion, tiger, or leopard.

[0117] In the context of this invention, "treatment" of any of the aforementioned conditions should be understood to include curative medical treatment aimed at curing, improving, or stabilizing the condition in a subject. On the other hand, "prevention" of a condition refers to the prior exclusion, avoidance, elimination, prevention, cessation, or obstruction of the occurrence of the condition. Therefore, prevention of a condition also explicitly includes preventative treatment of the subject.

[0118] Oral care products can be applied, for example, by using a rubber polisher, an applicator (such as a syringe), a finger, or a toothbrush, as described elsewhere in this document.

[0119] In the various methods disclosed herein, the oral care product is preferably applied once, twice, or three times daily for 1, 2, 3, 4, 5, 6, 7, or more days; in one embodiment, it is applied daily. It can also be applied less frequently, such as once weekly or once monthly.

[0120] In summary, the inventors have unexpectedly discovered a novel method for stannous fluoride in oral care products. By dispersing stannous fluoride in a three-dimensional matrix formed from self-assembled peptides, the bioavailability of bioactive stannous fluoride and fluoride ions can be improved. The resulting oral care products can be advantageously used for improved methods of treating and / or preventing various dental diseases, including caries, gingivitis, periodontitis, peri-implantitis, and dentin hypersensitivity.

[0121] In this invention, the term "about" is intended to be understood as "+ / -10%". If "about" relates to a range, it indicates the lower and upper limits of that range. "A (A)" means "one or more" unless otherwise explicitly stated. The term "comprising" as used herein also includes the meaning of "consisting of".

[0122] The following examples are intended to illustrate, not limit, the invention. All references cited in this application are incorporated herein in their entirety. Detailed Implementation

[0123] Example:

[0124] Self-assembled peptides have been shown to remineralize tooth enamel, inhibit its demineralization, and treat dentin hypersensitivity. Stannous fluoride at concentrations between 0.4% and 0.5% has also shown clinical efficacy in treating dentin hypersensitivity and preventing tooth decay.

[0125] The inventors were surprised to observe that the combination with the self-assembled peptides significantly increased the stability of the contained stannous fluoride, thereby increasing its bioavailability, possibly due to its interaction with the self-assembled peptide fibers.

[0126] Historically, the use of stannous fluoride in oral care compositions has been influenced by stannous ions (Sn). 2+ Due to its instability, it will rapidly oxidize into inactive tin ions (Sn). 4+ In the oral cavity, this also manifests as yellowish-brown spots on the tooth surface.

[0127] Surprisingly, no such oxidation change was observed in the stannous ions dispersed in the self-assembled peptide matrix. Oxidation was hindered when exposed to atmospheric oxygen and during redox experiments in the laboratory.

[0128] A gel formulation with a pH value below 7.5 containing the following ingredients was prepared to evaluate this effect:

[0129] 1. 0.45% stannous fluoride

[0130] 2. 0.45% stannous fluoride + 50 PPM P11-4 self-assembled peptide matrix

[0131] 3. 0.45% stannous fluoride + 100 PPM P11-4 self-assembled peptide matrix.

[0132] 4. 0.45% stannous fluoride + 250 PPM self-assembled peptide matrix

[0133] 5. 0.45% stannous fluoride + 500 PPM self-assembled peptide matrix

[0134] No oxidative changes were observed in any formulation containing 100 PPM or more of self-assembled peptide matrix.

[0135] References: White, 1995, A “Return” to stannous fluoride dentifrices. The Journalof Clinical Dentristry. Marquis RE. Antimicrobial actions of fluoride for oral bacteria. CanJ Microbiol. 1995 Nov;41(11):955-64. doi: 10.1139 / m95-133. PMID: 7497353. Nassar Y, Brizuela M. The Role of Fluoride on Caries Prevention.[Updated 2023 Mar 19]. In: StatPearls [Internet]. Treasure Island (FL):StatPearls Publishing; 2023 Jan-. Available from: https: / / www.ncbi.nlm.nih.gov / books / NBK587342 / . https: / / en.wikipedia.org / wiki / Tin(II)_fluoride Levine RS. Pyrophosphates in toothpaste: a retrospective andreappraisal. Br Dent J. 2020 Nov;229(10):687-689. doi: 10.1038 / s41415-020-2346-4. Epub 2020 Nov 27. PMID: 33247264. Stephen Mason, Sarah Young, Jimmy Qaqish, Guillaume Frappin, ChhajuGoyal, Stain control with two modified stannous fluoride / sodiumtripolyphosphate toothpastes: A randomised controlled proof of concept study,Journal of Dentistry, Volume 91, Supplement, 2019. West N, et al. Erosion protection efficacy of a 0.454% stannousfluoride dentifrice versus an arginine-containing dentifrice. AmericanJournal of Dentistry. 2018 Apr;31(2):63-66. PMID: 29630787. Fiorillo L, et al. Stannous Fluoride Effects on Enamel: A SystematicReview. Biomimetics (Basel). 2020 Aug 31;5(3):41. doi: 10.3390 / biomimetics5030041. PMID: 32878006; PMCID: PMC7559150. Parkinson CR, et al. Gingivitis efficacy of a 0.454% w / w stannousfluoride dentifrice: a 24-week randomized controlled trial. BMC Oral Health.2020 Mar 26;20(1):89. doi: 10.1186 / s12903-020-01079-6. PMID: 32216778; PMCID:PMC7098169. Moon JH, et al. Effects of Sodium Tripolyphosphate on Oral Commensaland Pathogenic Bacteria. Pol J Microbiol. 2019;68(2):263-268. doi: 10.33073 / pjm-2019-029. PMID: 31257792; PMCID: PMC7256694. Soares R, De Ataide IN, Fernandes M, Lambor R. Assessment of EnamelRemineralisation After Treatment with Four Different Remineralising Agents: AScanning Electron Microscopy (SEM) Study. J Clin Diagn Res. 2017 Apr;11(4):ZC136-ZC141. doi: 10.7860 / JCDR / 2017 / 23594.9758. Epub 2017 Apr 1. PMID:28571281; PMCID: PMC5449906. Jablonsky-Momeni et al., 2019. Randomised in situ clinical trialinvestigating self-assembling peptide matrix P11-4 in the prevention ofartificial caries lesions. Scientific Reports 9:269. Acherkouk Aet al. A randomised clinical study investigating efficacyof a stannous fluoride toothpaste in improving gingival health after 3 weeks'use. BMC Oral Health. 2021 Sep 12;21(1):441. doi: 10.1186 / s12903-021-01727-5.PMID: 34511098; PMCID: PMC8436562. Paraskevas S, van der Weijden GA. A review of the effects of stannousfluoride on gingivitis. J Clin Periodontol. 2006 Jan;33(1):1-13. doi:10.1111 / j.1600-051X.2005.00860.x. PMID: 16367849. Schiff T, et al. Desensitizing effect of a stabilized stannousfluoride / Sodium hexametaphosphate dentifrice. Compend Contin Educ Dent. 2005Sep;26(9 Suppl 1):35-40. PMID: 16999008. Hines D, et al. Effect of a stannous fluoride toothpaste on dentinalhypersensitivity: In vitro and clinical evaluation. J Am Dent Assoc. 2019Apr;150(4S):S47-S59. doi: 10.1016 / j.adaj.2019.01.006. PMID: 30797259. US4418057A US3433544A US5004597A US5716600A US5004597A US9968803B2 US3282792A US5017363A US4960586A US4961924A WO 2004 / 007532 A1 US10 / 521,628 US12 / 729,046 US13 / 551,878 US14 / 062,768 WO 2010 / 041636 A1 WO2014 / 027012 A1 US6667027B2 US6350436B1 US6821507B2 US5578293A US5145666A US5281411A US5281410A WO 2021 / 110923 A1

Claims

1. An oral care product, comprising: i. stannous fluoride, and ii. At least 0.01 wt% of a self-assembled peptide comprising the sequence X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid, aspartic acid, glutamine and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan and glutamine, wherein the self-assembled peptide is present in an assembled form in the oral care product.

2. The oral care product according to claim 1, wherein the stannous fluoride is present in the oral care product at a concentration of 0.1-1.0 wt%.

3. The oral care product according to claim 1, wherein the self-assembled peptide comprises or is composed of the amino acid sequence of SEQ ID NO:6 (P11-4), SEQ ID NO:9 (P11-8) or SEQ ID NO:15 (P11-20).

4. The oral care product according to claim 1, wherein the self-assembling peptide is composed of SEQ ID NO:6 (P11-4).

5. The oral care product according to claim 1, further comprising polyphosphate.

6. The oral care product according to claim 5, wherein the polyphosphate is sodium tripolyphosphate.

7. The oral care product according to claim 6, wherein the sodium tripolyphosphate is present in the oral care product at a concentration of 1 wt%-10 wt%.

8. The oral care product according to claim 1, further comprising sodium gluconate.

9. The oral care product according to claim 8, wherein the sodium gluconate is present in the oral care product at a concentration of 0.5 wt% to 5 wt%.

10. The oral care product of claim 1, wherein the oral care product is a dental gel, toothpaste, preventative paste, dental foam, or oral rinse.

11. The oral care product according to claim 1, further comprising: a. Sodium tripolyphosphate, b. Sodium gluconate c. Sorbitol, d. Cellulose gum, e. Xylitol, f. Erythritol, g. Hydrated silica, h. Phosphoric acid, i. Optionally, NaOH, and j. Water.

12. The oral care product according to claim 1, wherein the pH value of the oral care product is less than 7.

5.

13. An oral care product comprising: i. 0.1 wt%-1.0 wt% stannous fluoride; ii. At least 0.01 wt% of a self-assembled peptide consisting of SEQ ID NO:6 (P11-4); iii. 1 wt%–10 wt% sodium tripolyphosphate; and iv. 0.5 wt% - 5 wt% sodium gluconate The pH of the oral care products mentioned therein is below 7.

5.

14. A method for preventing tooth demineralization, comprising applying an effective amount of the oral care product as claimed in claim 1 to the teeth.

15. A method for remineralizing the enamel of a subject in need, comprising applying an effective amount of the oral care product as claimed in claim 1 to the tooth.

16. A method for disrupting a biofilm formed on a tooth, comprising applying an effective amount of the oral care product as claimed in claim 1 to the tooth.

17. A method of treating dentin hypersensitivity in a subject in need, comprising applying an effective amount of the oral care product as claimed in claim 1 to the teeth.

18. A method for stabilizing stannous fluoride in an oral care product, comprising mixing i) Tin fluoride, and ii) at least 0.01 wt% of a self-assembled peptide comprising the sequence X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid, aspartic acid, glutamine and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan and glutamine, wherein the self-assembled peptide is present in an assembled form in the oral care product.

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