ENZYMATIC COMPOSITION FOR ORAL HEALTH AND NATURAL ENAMEL WHITENING

Abstract

ENZYMATIC COMPOSITION FOR ORAL HEALTH AND NATURAL ENAMEL WHITENING One aspect of the invention relates to an enzymatic complex for oral health, the prevention and removal of bacterial biofilm in dental plaque, and natural enamel whitening, comprising a combination of the following compounds: an enzyme hydrolyzing the β-2,6 and / or β-2,1 bonds of sucrose; a mixture of glucanohydrolase enzymes hydrolyzing (1-3)-α-D-glucans and (1-6)-α-D-glucans; a glycoside hydrolase enzyme hydrolyzing the (1-6)-α-D glycosidic bonds of sugar dimers and trimers; an oxidoreductase enzyme forming hydrogen peroxide from glucose; and a peroxidase enzyme transforming thiocyanate into hypothiocyanate. in the presence of hydrogen peroxide.

Description

Title of the invention: ENZYMATIC COMPOSITION FOR ORAL HEALTH AND NATURAL ENAMEL WHITENING TECHNICAL FIELD OF THE INVENTION

[0001] The technical field of the invention is that of oral hygiene and cosmetic products.

[0002] The present invention relates to a novel enzyme complex capable of preventing the formation of dental plaque, cavities and tartar, while achieving natural whitening of the enamel. The present invention also relates to a novel form of prevention and treatment of dental bacterial biofilm by said enzyme complex. TECHNOLOGICAL BACKGROUND OF THE INVENTION

[0003] The sugar provided by food, consisting of more than 90% sucrose, is degraded into glucans by cariogenic bacteria in the oral cavity, which form a bacterial biofilm on the tooth enamel. A bacterial biofilm is an organized and structured community of bacterial cells entangled in an extracellular matrix of variable density and composition. In the presence of sucrose, this bacterial biofilm grows in thickness on the teeth, first forming soft dental plaque, composed of soluble glucans (known as (l-6)-aD-glucan or dextrans), and then hard dental plaque formed by the transformation of said dextrans into water-insoluble glucans (known as (l-3)-aD-glucan or mutans).

[0004] In the case of a predominantly sugary diet, the pH of the dental plaque becomes acidic and promotes the formation of cavities, while in the case of a predominantly meat-based diet, the pH becomes basic and the hard plaque calcifies to form tartar which becomes very difficult to eliminate.

[0005] According to a 2021 WHO report, in 2020, 3.5 billion people worldwide suffered from dental problems related to the formation of cavities and / or tartar. Similarly, according to data from the American National Health and Nutrition Examination Survey in 2018, 59% of adolescents aged 12 to 19 and 92% of adults aged 20 to 64 suffered from dental caries in their permanent teeth. Therefore, there is still a need to reduce these phenomena.

[0006] Current approaches to reducing dental plaque include mechanical cleaning, chemicals, fluoride applications, and broad-spectrum antiseptic antibacterial agents such as chlorhexidine. Although effective in the short term, these therapies all come with side effects. Mechanical cleaning is known to cause tooth sensitivity and bleeding gums, chemicals are known to pose a health risk, fluoride use can cause major adverse human health problems while having only a modest effect on preventing dental caries, and chlorhexidine can cause tooth staining and fillings in cavities. Furthermore, the use of antiseptics and chemicals has been shown to disrupt the commensal microbiota, increasing the pathogenic characteristics and cytotoxicity of host cells and causing other adverse effects on oral health.

[0007] These solutions are therefore not satisfactory.

[0008] It is increasingly preferred to use more preventive treatments that are more respectful of the overall health of the consumer, with minimal side effects, such as the use of probiotics, prebiotics or natural substances, but their therapeutic usefulness is notoriously limited.

[0009] Among the pathogenic bacteria of the oral cavity, Streptococcus mutans is the main responsible for the formation of bacterial biofilm, the main cause of caries, because it uses dietary sucrose to synthesize extracellular polysaccharides, in particular (1-3) and (1-6)-aD-glucans, which promote the formation of biofilms and increase the resistance of microorganisms in the biofilm. The acidic biofilm formed by Streptococcus mutans is highly resistant to the hostile environment, host immunity and antimicrobial treatments, in particular lysozyme.

[0010] There is therefore a need to prevent the proliferation of Streptococcus mutans.

[0011] Lactoperoxidase is an enzyme that is known for its bactericidal effect in the presence of hydrogen peroxide and thiocyanate, particularly against Streptococcus mutans. It is known to provide these three substances by providing exogenous lactoperoxidase, glucose oxidase and potassium thiocyanate. In this type of composition, however, the formation of hydrogen peroxide is insufficient for the bactericidal effect of lactoperoxidase to significantly affect the Streptococcus mutans biofilm from the point of view of reducing the formation of caries and / or tartar. Furthermore, the provision of exogenous glucose in this type of composition is not recommended in the field of oral hygiene. This solution is therefore not satisfactory for preventing the proliferation of Streptococcus mutans.

[0012] Generally speaking, the major problem in the fight against cavities and tartar, and to preserve healthy and beautiful teeth, is therefore to have dental plaque at a neutral pH every day and as physiological as possible. Recent research on dental pathologies shows that the best way to avoid them is to be able to eradicate bacterial biofilms. To this end, it is particularly important to prevent the formation of Streptococcus mutans biofilm and to disassemble existing biofilms in order to replace them with physiological plaque, i.e. the natural protective biofilm that surrounds a healthy tooth and which is made up of saliva and microbiota from the oral cavity.

[0013] In addition to having healthy teeth, it is also desirable to have the most beautiful teeth possible. There is therefore a constant need to whiten tooth enamel in order to combat yellowing, caused in particular by aging and the formation of insoluble dental plaque. Summary of the invention

[0014] The invention offers a solution to the problems mentioned above, by providing an enzyme complex whose composition, which is based in particular on a synergistic action of its various constituents, results from numerous studies carried out by the applicant. Of natural origin, this enzyme complex is respectful of the overall health of the consumer and does not present any side effects.

[0015] One aspect of the invention relates to an enzyme complex for oral health and enamel whitening comprising the following compounds: • an enzyme hydrolyzing the [3-2,6 and / or [3-2,1] bonds of sucrose, • a mixture of glucanohydrolase enzymes hydrolyzing (l-3)-aD-glucans and (l-6)-aD-glucans, • a glycoside hydrolase enzyme hydrolyzing the (l-6)-aD osidic bonds of ose dimers and trimers, • an oxidoreductase enzyme forming hydrogen peroxide from ose monomers. • a peroxidase enzyme transforming thiocyanate into hypothiocyanate in the presence of hydrogen peroxide.

[0016] According to one aspect of the invention, the enzyme hydrolyzing the [3-2,6 and / or [3-2,1] bonds of sucrose is an invertase.

[0017] According to another aspect of the invention, the mixture of glucanohydrolase enzymes hydrolyzing (l-3)-aD-glucans and (l-6)-aD-glucans comprises a mutanase and a dextranase, or a mixture of chimeric glucanases comprising a dextranase and a mutanase linked by genetic engineering.

[0018] According to a further aspect of the invention, the mutanase is a mutanase having exolytic activity, preferably a mutanase from a fungus Trichoderma Harzianum, or a mutanase having endolytic activity, preferably a mutanase from a bacterium Paracoccus Mutanolyticus, Paenibacillus or Streptomyces, or a mixture comprising a mutanase having exolytic activity, preferably a mutanase from a fungus Trichoderma Harzianum, and a mutanase having endolytic activity. dolytic, preferably a mutanase from a Paracoccus Mutanolyticus, Pae-nibacillus or Streptomyces bacterium.

[0019] According to one aspect of the invention, the glycoside hydrolase enzyme hydrolyzing the (l-6)-aD osidic bonds of ose dimers and trimers is an amyloglucosidase enzyme.

[0020] According to another aspect of the invention, the oxidoreductase enzyme forming hydrogen peroxide from ose monomers is a glucose oxidase.

[0021] According to a further aspect of the invention, the peroxidase enzyme transforming thiocyanate into hypothiocyanate in the presence of hydrogen peroxide is a lactoperoxidase.

[0022] According to one aspect of the invention, the enzyme complex comprises the following compounds: • an invertase, • a mixture of mutanase and dextranase, • an amyloglucosidase, preferably an amylo-a-1,6 glucosidase, • a glucose oxidase, and • a lactoperoxidase.

[0023] According to another aspect of the invention, the enzymatic complex comprises the following compounds, the percentages indicated being percentages by volume: • from 10 to 30%, preferably from 15 to 25%, of an invertase, • from 10 to 40%, preferably from 15 to 30%, of mutanase, • from 1 to 10%, preferably from 4 to 7%, of dextranase • from 20 to 60%, preferably from 30 to 50%, of an amyloglucosidase, of preferably an amylo-a-1,6 glucosidase, • from 5 to 20%, preferably from 7 to 15%, of glucose oxidase, and • from 1 to 10%, preferably from 4 to 7%, of lactoperoxidase.

[0024] According to a further aspect of the invention, the enzyme complex further comprises a sucrase-iso-maltase or lysozyme or a mixture of sucrase-iso-maltase and lysozyme.

[0025] Another aspect of the invention relates to an oral composition for oral health and enamel whitening, which comprises an enzyme complex as described above.

[0026] According to one aspect of the invention, the oral composition comprises from 1.4 to 3%, preferably from 2.0 to 2.5%, of enzyme complex.

[0027] According to another aspect of the invention, the oral composition further comprises lactoferrin.

[0028] According to a further aspect of the invention, the oral composition further comprises: from 20 to 60%, preferably from 30 to 50%, of sorbitol, • from 1 to 20%, preferably from 5 to 10%, of xylitol, • from 0 to 3%, preferably from 1 to 2%, of polysorbate 80, • from 0 to 3%, preferably from 1 to 2%, of polysorbate 20, • from 0 to 0.5%, preferably from 0.1 to 0.2%, of sodium saccharin, • from 0 to 5%, preferably from 1 to 2%, of colloidal silica, • from 0 to 3%, preferably from 1 to 2%, of dimethicone, • from 0 to 5%, preferably from 1 to 2%, of mint aroma and / or mint essential oils, • from 0.1 to 0.5%, preferably from 0.1 to 0.3%, of zinc citrate, • from 0 to 0.5%, preferably from 0.1 to 0.2%, of potassium thiocyanate, • from 0 to 0.5%, preferably from 0.1 to 0.2%, of preservative, and • water, in a quantity sufficient to reach 100%.

[0029] According to one aspect of the invention, the oral composition is in the form of a paste, liquid, gel, mouthwash, nebula, spray, capsule, tablet or chewing gum.

[0030] Thanks to the enzyme hydrolyzing the [3-2,6 and / or [3-2,1] bonds of sucrose, the enzymatic complex according to the invention advantageously reduces the quantity of sucrose available in the oral cavity by transforming it into monomers of oses (glucose) and into fructose, which deprives the cariogenic bacteria of metabolic substrate, greatly reduces the production of soluble and insoluble glucans, and thus contributes to the reduction of the bacterial biofilm.

[0031] Thanks to the mixture of glucanohydrolase enzymes hydrolyzing the (1-3) and (1-6)-aD-glucan bonds, which are key structural and functional constituents of the Streptococcus mutans biofilm matrix, the enzyme complex according to the invention advantageously prevents the formation of bacterial biofilm and disassembles existing biofilms. The destruction of the bacterial biofilm further reduces the formation of dental plaque, which is malodorous and yellows teeth under the influence of tobacco or dyes such as coffee or tea. Soluble glucans are transformed into sugar dimers or trimers by the mixture of glucanohydrolase enzymes, these sugar derivatives being then transformed into sugar monomers by the action of amylo-a-1,6 glucosidase.

[0032] The preferential association of a dextranase with a mutanase, more preferably from different fungi or bacteria, or a chimeric glucanase (dextranase-mutanase), shows very satisfactory effects aimed at preventing the formation of Streptococcus mutans biofilm and at disassembling existing biofilms in a few minutes at nano molar concentrations. This association clearly improves the sensitivity of the biofilm to antimicrobial and antibacterial agents, such as lysozyme, highlighting its great potential in the fight against dental caries. related to acid biofilm.

[0033] The residual sugar derivatives resulting from the degradation of glucans in dental plaque are then transformed into sugar monomers by a glycoside hydrolase enzyme hydrolyzing the (l-6)-aD sugars, while the sugar monomers are transformed by an oxidoreductase into D-glucono-delta-lactone, but also especially into hydrogen peroxide. The formation of hydrogen peroxide is advantageous because it has an antiseptic effect actively participating in eliminating the biofilm of Streptococcus mutans, and a natural whitening effect on the enamel, which contributes to a more beautiful appearance of the teeth.

[0034] These beneficial effects are advantageously reinforced by the fact that the enzymatic complex according to the invention provides several sources of hydrogen peroxide in the oral cavity, by the in situ transformation of sucrose, saccharide dimers and trimers, soluble glucans and insoluble glucans into ose monomers, i.e. into glucose. The glucose thus regularly available, allows, by its transformation by the gluco-oxidase enzyme, to produce hydrogen peroxide directly in contact with the teeth in a regular, gentle and lasting manner, for an optimal effect with regard to oral hygiene and enamel whitening. Since the glucose is transformed in real time, the quantity of glucose present in the oral cavity at any given moment advantageously remains much lower and therefore much less harmful than when exogenous glucose is supplied.

[0035] The hydrogen peroxide generated in situ thanks to the enzymatic complex according to the invention is produced in a much smaller quantity than that usually used for tooth whitening which is supplied exogenously, but it is also present in the oral cavity for a longer period, which makes it possible to provide a satisfactory progressive whitening effect, without being accompanied by the usual effects encountered with a massive and exogenous supply of hydrogen peroxide, namely: demineralization of the hard tissues of the tooth, irritation of the mucous membranes, reaction with the filling materials, premature wear of the enamel with hypersensitivity and weakening of the teeth.

[0036] Furthermore, in the presence of hydrogen peroxide, the peroxidase enzyme advantageously transforms the thiocyanate naturally present in saliva into hypo-thiocyanate, which is a powerful bactericide and further enhances the action of the enzyme complex of the invention with respect to oral hygiene. To enhance this effect, it is also possible to add exogenous thiocyanate.

[0037] The addition of a sucrase-iso-maltase to the enzyme complex according to the invention is advantageous in that this enzyme breaks down the starch and glycogen possibly present in the oral cavity into ose dimers (maltose) which are then transformed into ose monomers by an alpha amylase present naturally in saliva.

[0038] The addition of lysozyme to the enzyme complex according to the invention is advantageous in that it hydrolyzes the polysaccharide cell wall of bacteria and, in the absence of the bacterial biofilm generated by Streptococcus mutans, exhibits strong bactericidal activity actively participating in oral hygiene. Indeed, the bactericidal activity of the enzyme complex according to the invention is reinforced by the lysozyme which acts more effectively "after" the disappearance of the bacterial biofilm.

[0039] In summary, through the synergistic action of its components, the enzymatic complex according to the invention ensures the following beneficial effects while respecting the overall health of the consumer and without side effects: • an anti-cariogenic sugar action by inhibiting the acid transformation of dietary sugars and therefore a preventive effect against cavities, • an action of destructuring the bacterial biofilm of dental plaque and therefore a preventive effect against periodontal diseases and tartar, • a cleansing action of the microbiota of the oral cavity by reducing pathogenic bacteria under the effect of stimulation of the salivary lactoperoxidase system, • a gentle and prolonged action of whitening tooth enamel through natural production of hydrogen peroxide.

[0040] The addition of lactoferrin to a composition for oral use comprising an enzyme complex, by ensuring the chelation of iron necessary for the binding of the polysaccharides of the microbial envelope, necessary for the survival of cariogenic streptococci, advantageously leads to better eradication of the Streptococcus mutans biofilm and participates in the synergy mentioned above. Indeed, lactoferrin lyses the bacterial membrane of many organisms, including acidogenic streptococci, and thus greatly increases the body's immune defenses.

[0041] The invention and its various applications will be better understood upon reading the following description. DETAILED DESCRIPTION

[0042] By enzyme complex is meant here a stable association and according to a defined structure of a number of different proteins whose various enzymatic activities will contribute in an orderly manner to the development of a synergistic activity on a target substrate.

[0043] The enzyme complex according to the invention is intended for oral use in order to improve oral health and whiten tooth enamel.

[0044] It comprises a combination of the following constituents: • an enzyme capable of hydrolyzing the [3-2,6 and / or [3-2,1] bonds of sucrose, • a mixture of glucanohydrolase enzymes capable of hydrolyzing (l-3)-aD-glucans and (l-6)-aD-glucans, • a glycoside hydrolase enzyme capable of hydrolyzing the (l-6)-aD osidic bonds of ose dimers and trimers, • an oxidoreductase enzyme capable of breaking down ose monomers to form hydrogen peroxide, • a peroxidase enzyme capable of transforming thiocyanate into hypothiocyanate in the presence of hydrogen peroxide.

[0045] The enzyme capable of hydrolyzing the [3-2,6 and / or [3-2,1] bonds of sucrose, is an invertase of the family of saccharases degrading sucrose into glucose and fructose.

[0046] The mixture of glucanohydrolase enzymes degrades dental plaque into ose dimers or trimers. It is preferably a mixture of mutanase and dextranase. Dextranase degrades soft dental plaque by transforming soluble dextrans into ose-dimer or trimer 1,6-type osidic polymers, while mutanase degrades insoluble mutans of hard plaque by transforming them into ose-dimer or trimer 1,3-type osidic polymers.

[0047] In the glucanohydrolase enzyme mixture, a mutanase having exolytic activity and a mutanase having endolytic activity are preferred. As the mutanase having exolytic activity, a mutanase obtained from a fungus, especially the fungus Trichoderma Harzianum, can be used. As the mutanase having endolytic activity, a mutanase obtained from a bacterium, especially the bacterium Paracoccus Mutanolyticus, the bacterium Paenibacillus or the bacterium Streptomyces, can be used.

[0048] In the mixture of glucanohydrolase enzymes, instead of a mixture of mutanase and dextranase, a mixture of chimeric glucanases comprising a dextranase and a mutanase linked by genetic engineering can also be used.

[0049] The enzyme glycoside hydrolase is an enzyme that reduces ose dimers or trimers into ose monomers, i.e. glucose. It is preferably an amyloglucosidase, more preferably an amylo-a-1,6 glu-cosidase enzyme. Amyloglucosidase transforms ose derivatives into glucose.

[0050] The enzyme oxidoreductase is an enzyme that converts glucose into hydrogen peroxide. It is preferably a glucose oxidase, also referred to as gluco-oxidase. The enzyme gluco-oxidase converts glucose into D-glucono-delta-lactone and hydrogen peroxide. D-glucono-delta-lactone is an acidity regulator. In the oral cavity, it is partially converted into gluconic acid.

[0051] In the presence of hydrogen peroxide, the enzyme peroxidase transforms the thiocyanate to hypothiocyanate according to the following reaction:

[0052] SCN + H2O2 => OSCN + H2O

[0053] The peroxidase enzyme is preferably a lactoperoxidase.

[0054] Hypothiocyanate, a strong bactericide, limits the growth of bacteria in the oral cavity.

[0055] The enzymatic complex according to the invention preferably has the following composition (the percentages indicated are percentages by volume): • Invertase: from 10 to 30%, preferably from 15 to 25%, • Mutanase: from 10 to 40%, preferably from 15 to 30%, • Dextranase: from 1 to 10%, preferably from 4 to 7%, • Amyloglucosidase: from 20 to 60%, preferably from 30 to 50%, • Glucose oxidase: from 5 to 20%, preferably from 7 to 15% and • Lactoperoxidase: from 1 to 10%, preferably from 4 to 7%.

[0056] The table below gives an example of a control formulation CET of enzyme complex with a high quantity of amyloglucosidase and without mutanase, and three examples of formulations CE 1-3 for the enzyme complex according to the invention. The percentages indicated are percentages by volume.

[0057] [Tables 1] Compound Formulation CET Formulation CEI Formulation CE2 Formulation CE3 Invertase 10% 10% 30% 20% Glucose oxidase 10% 10% 20% 15% Amyloglucosidase 70% 60% 20% 35% Lactoperoxidase 5% 5% 10% 5% Dextranase 5% 5% 10% 5% Mutanase 0% 10% 10% 20%

[0058] During clinical trials carried out on patients for six months by the applicant with the participation of a research laboratory under the responsibility of a medical doctor, the CET formulation gave unsatisfactory results. Indeed, compared to the control subjects, in the subjects participating in the study for 6 months and having used this formulation, the presence of dental plaque as well as a very slight whitening of the tooth enamel (<1 tone) was noted.

[0059] The CEI formulation gave more satisfactory results than the CET formulation. Indeed, compared to the control subjects, in the subjects participating in the study for six months and having used this formulation, a lower quantity of dental plaque in the oral cavity, as well as a marked whitening of tooth enamel (+1 shade of white)

[0060] The CE2 formulation gave more satisfactory results than the CET formulation. In fact, compared to the control subjects, in the subjects participating in the study for six months and using this formulation, a lower quantity of dental plaque in the oral cavity was noted, as well as a clear whitening of the tooth enamel (+1 shade of white).

[0061] The CE3 formulation gave more satisfactory results than the CEI and CE2 formulations. Indeed, compared to the control subjects, in the subjects participating in the study for six months and using this formulation, a virtual absence of dental plaque in the oral cavity was noted, as well as a significant whitening of the tooth enamel (+2 shades of white).

[0062] The enzymatic complex according to the invention may also comprise a sucrase-iso-maltase, lysozyme or a mixture of these two enzymes.

[0063] The enzyme complex preferably comprises from 0.1 to 0.4% by weight of lysozyme, more preferably 0.2%, and / or preferably from 0 to 0.4% by weight of sucrase isomaltase, more preferably 0.2% by weight.

[0064] The enzyme complex according to the invention can be used in an oral composition for oral health and enamel whitening.

[0065] This oral composition may, for example, be in the form of a paste, liquid, gel, mouthwash, nebulosity, spray, capsule, tablet or chewing gum.

[0066] The oral composition for oral-dental use according to the invention preferably has the following composition (the enzymatic complex is that described previously and the percentages indicated are percentages by weight) • Enzyme complex: from 1.4 to 3%, preferably from 2.0 to 2.5% • Sorbitol: 20 to 60%, preferably 30 to 50% • Xylitol: from 1 to 20%, preferably from 5 to 10%, • Polysorbate 80: from 0 to 3%, preferably from 1 to 2%, • Polysorbate 20: from 0 to 3%, preferably from 1 to 2%, • Sodium saccharin: from 0 to 0.5%, preferably from 0.1 to 0.2%, • Colloidal silica: from 0 to 5%, preferably from 1 to 2%, • Dimethicone: from 0 to 3%, preferably from 1 to 2%, • Mint flavoring and / or mint essential oils: 0 to 5%, preferably 1 to 2%, • Zinc citrate: from 0.1 to 0.5%, preferably from 0.1 to 0.3%, • Potassium thiocyanate: from 0 to 0.5%, preferably from 0.1 to 0.2%, • Preservative: from 0 to 0.5%, preferably from 0.1 to 0.2%, • Water: sufficient quantity to reach 100%.

[0067] The preservative is preferably citric acid.

[0068] The oral composition according to the invention may also comprise lactoferrin, preferably from 0 to 0.4% by weight, more preferably 0.1%.

[0069] It may also include hydrogen peroxide, for example 0 to 0.1% and violet dye, for a more immediate whitening effect.

[0070] The table below gives an example of a control formulation COT of an oral composition for oral use with the absence of an enzyme complex, and three examples of formulations CO 1-3 for the oral composition according to the invention with the enzyme complex CE3. The percentages indicated are percentages by weight.

[0071] [Tables2] Compound COT Formulation COI Formulation CO2 Formulation CO3 Formulation Enzyme Complex 0% 1.0% 2.0% 2.5% Sorbitol 40% 40% 40% 40% Xylitol 10% 10% 10% 10% Polysorbate 80 1.5% 1.4% 1.4% 1.4% Polysorbate 20 1.5% 1.4% 1.4% 1.4% Sodium Saccharin 0.15% 0.15% 0.15% 0.15% Colloidal Silica 1.5% 1.5% 1.5% 1.5% Dimethicone 1.5% 1.5% 1.5% 1.5% Mint Flavor and / or Mint Essential Oils 1.5% 1.5% 1.5% 1.5% Citrate Zinc 0.2% 0.2% 0.2% 0.2% Potassium thiocyanate 0.15% 0.15% 0.15% 0.15% Preservative 0.2% 0.15% 0.15% 0.15% Water qsp 100% qsp 100% qsp 100%

[0072] The effectiveness of these different formulations was tested on Streptococcus salivarius, which is from the same Viridans family as Streptococcus mutons, and a Gram-positive cocci bacterium like the latter, also forming a biofilm in the presence of sucrose, and known to have the same sensitivity to antiseptics and antibacterials. bacterial than Streptococcus mutans. The efficacy of said formulations was compared to a placebo not comprising an enzyme complex according to the invention and to a mouthwash solution comprising 0.12% chlorhexidine

[0073] In in vitro clinical trials carried out by the applicant with the participation of another analysis laboratory under the responsibility of a doctor of pharmacy, carried out with the CE3 enzyme complex, the COT formulation gave unsatisfactory results. Indeed, compared to the placebo vs. COT formulation vs. chlorhexidine, a high quantity of Streptococcus salivarius was noted in the placebo, a still high quantity of Streptococcus salivarius in the COT formulation and a zero quantity in the chlorhexidine-based formulation.

[0074] The COI formulation gave more satisfactory results than the COT formulation. Indeed, compared to the placebo vs. COI formulation vs. chlorhexidine, a high quantity of Streptococcus salivarius was noted in the placebo, a low quantity of Streptococcus salivarius in the COI formulation and a zero quantity in the chlorhexidine-based formulation.

[0075] The CO2 formulation gave more satisfactory results than the COT formulation and the COL formulation. Indeed, compared to the placebo vs. CO2 formulation vs. chlorhexidine, a high quantity of Streptococcus salivarius was noted in the placebo, a zero quantity of Streptococcus salivarius in the CO2 formulation and a zero quantity in the chlorhexidine-based formulation.

[0076] The CO3 formulation gave results as satisfactory as the CO2 formulation. Indeed, compared to the placebo vs. CO3 formulation vs. chlorhexidine, a high quantity of Streptococcus salivarius was noted in the placebo, a zero quantity of Streptococcus salivarius in the CO3 formulation and a zero quantity in the chlorhexidine-based formulation.

[0077] Thus, for an oral composition for oral-dental use comprising an enzyme complex according to the enzymatic formula CE3, although clinical effects could be observed with an amount of enzyme complex less than 2%, the bactericidal effect against Streptococcus is completely effective with an amount of enzyme complex greater than or equal to 2%.

Claims

Claims

1. An enzyme complex for oral health, for use in the treatment of dental plaque and bacterial biofilm of dental plaque and the natural whitening of enamel, characterized in that it comprises the following compounds: - an invertase enzyme hydrolyzing the [3-2,6 and / or [3-2,1] bonds of sucrose, - a mixture of glucanohydrolase enzymes hydrolyzing (l-3)-aD-glucans and (l-6)-aD-glucans, this mixture comprising a mutanase having exolytic activity and obtained from a fungus and a mutanase having endolytic activity and obtained from a bacterium, - a glycoside hydrolase enzyme hydrolyzing the (l-6)-aD-saccharide bonds of ose dimers and trimers, - an oxidoreductase enzyme forming hydrogen peroxide from monomers of oses, - a peroxidase enzyme transforming thiocyanate into hypo-thiocyanate in the presence of hydrogen peroxide.

2. Enzyme complex according to claim 1, characterized in that the mixture of glucanohydrolase enzymes hydrolyzing (l-3)-aD-glucans and (l-6)-aD-glucans further comprises a mutanase and a dextranase, or a mixture of chimeric glucanases comprising a dextranase and a mutanase linked by genetic engineering.

3. Enzyme complex according to claim 2, characterized in that the mutanase having exolytic activity is obtained from a fungus Trichoderma Harzianum.

4. Enzyme complex according to claim 2 or 3, characterized in that the mutanase having endolytic activity is obtained from a Paracoccus Mutanolyticus, Paenibacillus or Streptomyces bacterium.

5. Enzyme complex according to claim 1 or 2, characterized in that the glycoside hydrolase enzyme hydrolyzing the (1-6)-aD osidic bonds of the ose dimers and trimers is an amyloglu-cosidase enzyme, preferably an amylo-a-1,6 glucosidase.

6. Enzyme complex according to any one of the preceding claims, characterized in that the oxidoreductase enzyme forming Hydrogen peroxide from ose monomers is a glucose oxidase.

7. Enzyme complex according to any one of the preceding claims, characterized in that the peroxidase enzyme transforming thiocyanate into hypothiocyanate in the presence of hydrogen peroxide is a lactoperoxidase.

8. Enzyme complex according to any one of the preceding claims, characterized in that it comprises the following compounds: - an invertase, - a mixture of mutanase having exolytic activity and obtained from a fungus, of mutanase having endolytic activity and obtained from a bacterium, and of dextranase, - an amyloglucosidase, preferably an amylo-a-1,6 glucosidase, - a glucose oxidase, and - a lactoperoxidase.

9. Enzyme complex according to claim 8, characterized in that it comprises the following compounds, the percentages indicated being percentages by volume: - from 10 to 30%, preferably from 15 to 25%, of an invertase, - from 10 to 40%, preferably from 15 to 30%, of a mixture comprising a mutanase having an exolytic activity and obtained from a fungus and a mutanase having an endolytic activity and obtained from a bacterium, - from 1 to 10%, preferably from 4 to 7%, of dextranase - from 20 to 60%, preferably from 30 to 50%, of an amylo glucosidase, preferably an amylo-a-1,6 glucosidase, - from 5 to 20%, preferably from 7 to 15%, of glucose oxidase, and - from 1 to 10%, preferably from 4 to 7%, lactope roxidase.

10. An enzyme complex according to any one of the preceding claims preceding, characterized in that it further comprises a sucrase-iso-maltase or lysozyme or a mixture of sucrase-iso-maltase and lysozyme.

11. An oral health composition for use in the treatment of dental plaque and bacterial biofilm of dental plaque and the natural whitening of enamel, characterized in that it comprises an enzyme complex according to any one of the preceding claims.

12. Oral composition according to claim 11, characterized in that it comprises from 1.4 to 3%, preferably from 2.0 to 2.5%, of enzymatic complex.

13. Oral composition according to claim 11 or 12, characterized in that it further comprises lactoferrin.

14. Oral composition according to any one of claims 11 to 13, characterized in that it further comprises: - from 20 to 60%, preferably from 30 to 50%, of sorbitol, - from 1 to 20%, preferably from 5 to 10%, of xylitol, - from 0 to 3%, preferably from 1 to 2%, of polysorbate 80, - from 0 to 3%, preferably from 1 to 2%, of polysorbate 20, - from 0 to 0.5%, preferably from 0.1 to 0.2%, of sodium saccharin, - from 0 to 5%, preferably from 1 to 2%, of colloidal silica, - from 0 to 3%, preferably from 1 to 2%, of dimethicone, - from 0 to 5%, preferably from 1 to 2%, of mint flavoring and / or oils mint essential oils, - from 0.1 to 0.5%, preferably from 0.1 to 0.3%, of Zinc Citrate, - from 0 to 0.5%, preferably from 0.1 to 0.2%, of potassium thiocyanate, - from 0 to 0.5%, preferably from 0.1 to 0.2%, of preservative, - water,in an amount sufficient to reach 100%.,

15. An oral composition according to any one of claims 11 to 14, characterized in that it is in the form of a paste, liquid, gel, mouthwash, nebulosity, spray, capsule, tablet or chewing gum.