ES2364759T3 - COMPOSITIONS FOR THE ORAL TREATMENT CONTAINING AN ANTI-ADHESION AGENT, ANTIBACTERIAL AGENT AND INCOMPATIBLE COMPOUND. - Google Patents

Abstract

A composition for oral treatment comprising: an anti-adhesive agent comprising a non-stick protease enzyme wherein the protease enzyme comprises at least one cysteine protease, a cationic antibacterial agent; and an anionic surfactant that is incompatible with the anti-adhesive agent and the cationic antibacterial agent, and is secreted from the anti-adhesive agent and the cationic antibacterial agent.

Description

The present invention relates to compositions for oral treatment.

BACKGROUND OF THE INVENTION

The dental plaque or biofilm of the plaque is a soft deposit that forms on the surfaces of the oral cavity, such as tissue and teeth, and is formed by an accumulation of bacteria and salivary substances, as well as food by-products. The plaque adheres firmly to the points of irregularities or discontinuities (eg on the surface of rough stones, on the gum line, or on the surface of the tongue, and within cracks, and the like). In addition to being unsightly, plaque is involved in the appearance of gingivitis and other forms of periodontal diseases.

Bacteria are present in the tongue. Bacteria are part of a protective biofilm that essentially makes them resistant to most treatments. Few people clean their tongues after brushing their teeth, although it has been shown that an amount as high as 50 percent of mouth bacteria can be found here. In addition, for many people, brushing or scraping their tongues is difficult because of the pharyngeal reflex (arches). Therefore, tongue cleaning by non-mechanical procedures is very desirable for those who cannot do it with a mechanical device.

Despite the abundant prior art related to antibacterial dentifrices and compositions for oral treatment, there is still a need in the art to formulate a product capable of obtaining an enhanced effect in delaying the accumulation of bacterial plaque.

US-A-2004/0042977 describes two component desensitizing dentifrice compositions, in which a first component of the dentifrice contains an enzyme such as papain and a second component of the dentifrice contains an anionic surfactant such as sodium lauryl sulfate. Non-cationic antibacterial agents may be present in the first dentifrice component.

WO-A-2004/019898 describes a double component composition containing anti-plaque enzymes, comprising a first and second dentifrice components containing an enzyme and an antibacterial metal salt, in a buccally acceptable vehicle.

BRIEF SUMMARY OF THE INVENTION.

The present invention provides a composition for oral treatment according to claim 1. Preferred features are defined in the appended claims. The present invention includes a composition for oral treatment containing a non-stick protease enzyme, the non-stick protease enzyme comprising at least one cysteine protease, most preferably ficin. The protease enzyme is in combination with one or more ingredients, in particular a cationic antibacterial agent and an anionic surfactant, but the anionic surfactant is secreted from the protease enzyme and the cationic antibacterial agent. The non-stick agent mitigates the interaction between a subject's oral cavity and plaque forming materials.

In another aspect, the anionic surfactant, which is relatively incompatible with the antiadhesive agent and the cationic antibacterial agent, is included in a limited amount and is controlled to minimize the decrease.

or degradation of the activity of the antiadhesive agent and the cationic antibacterial agent. In yet another aspect, the anionic surfactant is included and segregation is implemented to avoid or inhibit contact with the non-stick protective enzyme and the cationic antibacterial agent.

DETAILED DESCRIPTION OF THE INVENTION.

In one aspect of the present invention, the oral treatment composition contains an effective amount of a non-stick protease enzyme. In another aspect, the oral treatment composition is formulated using a vehicle that contains an effective amount of the non-stick protective enzyme or a mixture of said non-stick protest enzymes.

The enzymes of the present invention inhibit the formation of the bacterial layer that would cause plaque formation. Such enzymes of the invention bind and attach to buccal surfaces, including tissue, and thus inhibit plaque growth or subsequent development. The terms "non-stick" and "anti-fixation" are used interchangeably in this text. The enzymes of the invention chosen are non-stick protective enzymes, desirably cysteine protease and most desirably are chosen from the ficin, papain and crilase group. Preferably, the antiadhesive agent comprises ficin and one or more other enzymes, such as other anti-adhesive enzymes or another type of enzyme, such as bromelain, chymotrypsin, alklase, amylases, glucose oxidase, cellulases, lipases and / or other proteases in addition of the non-stick cysteine protease.

The ficin for use in the present invention can be obtained by drying and filtering the latex of the genus Ficus, for example Ficus glabrata. Papain for use in the present invention can be obtained from the fruit and leaves of Carica papaya. The crilase to be used in the present invention can be extracted from the Antarctic krill (Euphausia superba).

In another embodiment, the composition comprises a buccally acceptable antiplate agent, including plate breakers and non-stick agents, and combinations thereof.

The amount of non-stick agent present in the composition depends on the application to which it is intended. It is present in less than 100 parts by weight in a composition or in a suitable vehicle. The non-stick agent is desirably present in an amount by weight of at least 0.01 parts per 100 parts of the composition. In a broad aspect, the non-stick enzyme agent is present in an amount by weight of about 0.01 to about 10 parts by weight per 100 parts by weight of the total composition. More desirably, it is at least 0.05 parts, and preferably 0.03 to 0.30 parts suitable for a paste composition.

One or more other antiplate agents may be present in an amount effective as antiplate. Suitable agents include, but are not limited to, tin, copper, magnesium and strontium salts, dimethicone copolyols such as cetyl dimethicone copolyol, glucoamylase, glucose oxidase, urea, calcium lactate, calcium glycerophosphate, strontium polyacrylates and chelating agents such as citric and tartaric acids, and alkali metal salts thereof.

An enzyme chosen that can be formulated in combination with a protease enzyme is glucoamylase. Other enzymes that can be used in the implementation of the present invention include other carbohydrases such as alpha-amylase, beta-amylase, dextranase and mutanase, and lipases such as vegetable lipase, gastric lipase, pancreatic lipase, pectinase, lysozyme tanase and proteases. Serine

Other suitable enzymes that may comprise the present invention include lysozyme, derived from egg white, which contains a single chain of polypeptide cross-linked with four disulfide bridges having a molecular weight of 14,600 daltons. The enzyme can show antibacterial properties by facilitating the hydrolysis of bacterial cell walls by segmenting the glycosidic bond between carbon number 1 of Nacethylmuramic acid and carbon number 4 of N-acetyl-D-glucosamine, which in vivo these two carbohydrates are polymerized to form the cell wall polysaccharide. Additionally, pectinase, an enzyme that is present in most plants, facilitates the hydrolysis of the pectin polysaccharide to give sugars and galacturonic acid.

The antibacterial agents useful in the practice of the present invention include all known cationic agents. Particularly useful are quaternary ammonium compounds and related compounds. Suitable compounds include benzethonium chloride, or diisobutylphenoxyethoxyethyl dimethyl benzyl ammonium chloride, and cetyl pyridinium chloride.

Other cationic quaternary ammonium antibacterial compounds useful and desirable in the practice of the present invention include those in which one or two of the substituents in quaternary nitrogen have a carbon chain length (typically an alkyl group) of about 8 to 20, typically 10 to 18 carbon atoms, while the remaining substituents have a lower number of carbon atoms (typically an alkyl or benzyl group), such as 1 to 7 carbon atoms, typically methyl or ethyl groups. Dodecyl trimethyl ammonium bromide, benzyl dimethyl stearyl ammonium chloride, cetyl pyridinium chloride (CPC) and 5-amino-1,3-bis (2-ethylhexyl) -5-methyl hexa quaternary hydroxy pyrimidine are typical quaternary ammonium antibacterial agents. The invention contemplates preferred pyridinium components, such as cetyl (C-16), stearyl (C-18) and myristyl (C-18) pyridinium and salts thereof formed with halide or other anion. A more illustrative list of useful antibacterial agents is provided in US Pat. No. 5,776,435 to Gaffar et al., incorporated herein by reference.

Preferred abrasive agents for use in the implementation of the present invention include silica materials and in particular silica and precipitated amorphous silica gels having an oil absorption rate of less than 100 cc / 100 g of silica, and preferably in the range of about 45 cc / 100 g to less than about 70 cc / 100 g of silica.

The composition may include a vehicle or paste and preferably comprises an aqueous phase, in which a humectant is contained. The humectant is preferably glycerol, sorbitol, xylitol and / or propylene glycol of molecular weight in the range of 200 to 1,000, but other humectants and mixtures thereof can also be used. The humectant concentration typically amounts to a total of about 5 to about 70% by weight of the oral composition.

The oral treatment compositions of the present invention may contain a variety of optional ingredients. As described below, such optional ingredients may include, but are not limited to, thickening agents, surfactants, a source of fluoride ions, a synthetic anionic polycarboxylate, a flavoring agent, tartar agents and coloring agents.

The dentifrice composition of the present invention may also contain ingredients that stabilize enzymes in a dentifrice environment. These stabilizers protect the enzyme against inactivation by chelating the metal impurities present in the dentifrice composition, which have a propensity to denature the active site of the enzyme by protecting the enzyme against oxidation. Chelating agents include ethylene diamine tetraacetic acid (EDTA) and

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sodium gluconate in concentrations between 0.01% and 1%, preferably between 0.1 and 0.5%. Agents that stabilize the enzyme against oxidation include sodium bisulfite, metal gallates, sodium stannate and ascorbic acid, in concentrations between about 0.03 and about 2.0%, preferably between about 0.1 and about 0.75%.

Synthetic anionic polycarboxylates can also be used in the dentifrice compositions of the present invention as an efficacy enhancing agent for any antibacterial, anti-tartar or other active agent within the dentifrice composition. Such anionic polycarboxylates are generally employed in the form of their free acids or preferably in the form of alkali metal salts (eg potassium and preferably sodium) or water soluble, partially neutralized or, more preferably, fully neutralized ammonium salts. 1: 4 to 4: 1 copolymers of anhydride or maleic acid with another ethylenically unsaturated polymerizable monomer, preferably methyl vinyl ether / maleic anhydride having a molecular weight (MW) of about 30,000 to about 1,800,000, and most are preferred preferably from about 30,000 to about 700,000. Examples of these copolymers are available from the GAF Corporation under the trade name Gantrez (eg AN 139 (M.W. 500,000), AN 119 (M.W. 250,000); S-97 Pharmaceutical Quality (M.W. 700,000), AN 169

(M.W. 1,200,000 - 1,800,000), and AN 179 (M.W. greater than 1,800,000)), among which the preferred copolymer is S-97 Pharmaceutical Quality (M.W. 700,000).

When present, the anionic polycarboxylate is used in effective amounts to achieve the desired potentiation of the efficacy of any of the antibacterial, anti-tartar or other active agent agents within the oral treatment composition. Generally, anionic polycarboxylates are present in a form of composition for exemplary toothpaste oral treatment between about 0.05% and about 4% by weight, preferably from about 0.5% to about 2.5% by weight.

Various other materials can be incorporated into the oral treatment compositions of the present invention, including numbing agents (such as potassium nitrate), bleaching agents, preservatives, silicones, and chlorophyll compounds. These additives, when present, are incorporated into the oral treatment composition in amounts that substantially do not adversely affect the desired properties or characteristics.

EXAMPLE 1 - Dentifrice formulation containing non-stick enzyme.

Enzymes are formulated in a silica-based formulation. Table 1 gives the exemplary dentifrice formulations. Table 1 provides the exemplary dentifrice formulations. All values are percentages by weight, unless otherwise indicated. The key ingredients of the formulation are enzymes, ficin, papain and crilase, at 0.226% by weight, a mixed silica abrasive system for cleaning greater than 25% by weight, and a mixed surfactant system of 0.5% SLS / 1.5 % pluronic acid 1% betaine, all by weight. Also included is a peppermint and peppermint flavoring agent that is stable with enzymes. The compositions are each chosen to provide attributes (such as foam, taste, mouthfeel and aesthetics) without compromising the activity of the enzymes. For enzyme concentration, a dose response study was conducted to select effective enzyme levels before selecting clinical formulations. Based on in vitro studies, clinical products were prepared at the OPTC (Bucal Process Technology Center) under GMP conditions.

Table 1: Formulations of dentifrices containing enzymes.

INGREDIENT

Ficina Papain Crilase

70% Sorbitol

24,374 24,374 24,374

99.5% Glycerinasynthetic - USP

20.0 20.0 20.0

Purified water

17.0 17.0 17.0

Dental type silica (Sylodent XWA 650 - USP)

17.0 17.0 17.0

Dental type silica (Zeodent 115)

8,000 8,000 8,000

Polyethylene Glycol 600 (PEG-12) NF

3,000 3,000 3,000

Synthetic amorphous precipitated silica No.2 - Zeodent 165

2,500 2,500 2,500

29% sodium lauryl sulfate

0.5 0.5 0.5

Poloxomer 407

1,500 1,500 1,500

CMC Sodium 2000S - 12 USP

1,200 1,200 1,200

Natural and artificial mixed peppermint

1,100 1,100 1,100

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Enzyme

FicinaPapaínaCrilasa EXAMPLE II: Enzyme characterization.

30% Cocamidopropyl betaine

1,000 1,000 1,000

Sodium monofluorophosphate - USP

0.760 0.760 0.760

Tetrasodium pyrophosphate - Fine (FCC)

0.500 0.500 0.500

USP sodium saccharin

0.500 0.500 0.500

Xanthan gum - NF

0.400 0.400 0.400

Poly OXWSR-N 750

0.100 0.100 0.100

Blue poly 50

0,300 0,300 0,300

Blue solution

0.040 0.040 0.040

Ficina

0.226 0 0

Papain

0 0.226 0

Crilase

0 0 0.226

TOTAL

100 100 100

Total protein concentration

Total protein content was determined using a non-interfering assay of Genoteca total protein concentration. The total protein concentration of the crilase powder is approximately 30 to 40%, and 60 to 70% for papain EDC (Enzyme Development Corporation) and approximately 90% or more protein concentration for ficin.

Activity test

Protease activity was evaluated using a generic Panvera protease assay at 40 ° C and at neutral pH. The activity was established in relation to papain on the basis of equal weight of pure dust samples. Protease crilase enzymes are a mixture of eight different proteases; then, the activity of any one of the eight was not discernible by the present essay. The trial helped determine the dose for dose response studies. In clinical formulations, enzymes were dosed on the basis of equal weight. Table 2 gives the relative activity of pure enzymes.

Table 2: Activity of several enzymes by protease assay.

CPU activity / g (+/- 3)

23 25 47

EXAMPLE III: Study of anti-union efficacy - Artificial mouth

To test the efficacy of the active compounds in formulas for toothpastes, the artificial mouth model was used. This is described in a general way in Herles, S., S. Olsen, et al. (1994). "Chemostat flow cell system: an in vitro model for the evaluation of antiplate agents." J Dent Res 73 (11): 1748-55. The artificial mouth model is a flow system that simulates the human mouth. Saliva-coated hydroxyapatite discs (salivacoated hydroxyapatite: SHAP) served as artificial teeth, and a culture of bacteria consisting of the main oral bacteria in humans was flowed through the system at a rate consistent with human saliva in vivo. Eight discs were then placed per treatment in the chemostatic flow cell and the oral bacteria culture was allowed to flow through the discs overnight (20 to 24 hours) to see if the active substances applied to the surfaces of the disc prevented biofilm formation After 24 hours the SHAP discs were removed and the amount of bacteria was determined. The ABS610 values of the solutions were then measured and analyzed. This result gave an estimate of the amount of bacteria that were attached to the SHAP discs. From this value, the percentage of non-stick efficacy of the pastes is evaluated in relation to a corresponding negative control. Two concentrations of protease were tested for each of the three proteases, namely, 0.226 percent by weight and 0.065 percent by weight. The artificial mouth anti-adhesion experiment suggests that at the two concentrations tested for each of the three proteases, on average, all three had an efficacy above that of the negative control pastes, which did not contain

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None of the enzymes. However, based on statistical analysis, the efficacy of crilase was not demonstrated. Papain and ficin showed efficacy at a concentration of 0.226%. Ficin showed similar efficacy at the two concentrations 0.065% and 0.226%. Based on these results, GMP lots were prepared for the proof of concept clinical studies. In vitro results indicated that ficin was better than papain, which was in turn better than the corresponding placebo paste.

EXAMPLE IV: Clinical evaluation in vivo.

Two clinical studies were conducted with humans to verify the non-stick efficacy of the products in relation to the corresponding negative control paste. The clinical procedure for measuring non-stick efficacy with the ultimate benefit of plaque reduction is indicated below.

A. Determination of the modified gingival margin plate index (Modified Gingival Margin Plaque Index: MGMPI) - Test products 0.226% ficin and corresponding negative control.

Fifteen (15) internal panelists were recruited and enrolled in this clinical study. Fourteen (14) panelists completed this three-week study. A panelist was excluded from the study because of a mild illness that required the use of antibiotics. Panelists gave the dental clinic a mouth exam and a medical history review. All acceptable panelists received prophylaxis (dental cleaning) and began a one-week wash with Colgate Great Regular. During the treatment phase of the clinical study, the panelists informed the dental clinic on their assigned morning. They were subjected to tartar removal and prophylaxis to remove all dental stones and plaque. The panelists then used 1.5 g of the assigned toothpaste and brushed for 60 seconds and then rinsed for 5 seconds with 10 ml of water. They were instructed to refrain from all oral hygiene for 24 hours at the end of which they returned to the dental clinic rinsed with solution and scored their plates. At the end of the scoring of the panels the panelists resumed normal oral hygiene (brushing the entire mouth twice a day) using the washing product. The results are shown below.

Punctuation

from the license plate Toothpaste with  enzyme Placebo toothpaste

(MGMPI)

0.226% ficina) correspondent

24 hour average score

14.55 ± 8.50 * 30.38 ± 17.99

* Statistically significant (p <0.05) with respect to placebo.

A t-test was used to determine the differences between products (p <0.05). The toothpaste with enzyme (0.226% ficina) is statistically different from the corresponding placebo. The ficin-containing toothpaste showed a clinical reduction of plaque by a non-adhesive mode of action.

B. Determination of the plaque index of the modified gingival margin (MGMPI) - Test products 0.226% papain and corresponding negative control.

This clinical procedure was identical to that described above for ficin, except for the fact that 14 panelists were enrolled in this study, all of whom completed the study.

Score of

 the plate Pasta  from  teeth with enzyme Placebo toothpaste

(MGMPI)

0.226% papain) correspondent

24 hour average score

17.07 ± 7.03 * 30.38 ± 17.95

* Statistically significant (p <0.05) with respect to placebo.

A t-test was used to determine the differences between products (p <0.05). The toothpaste with enzyme (0.226% papain) is statistically different from the corresponding placebo. The dentin containing papain showed a clinical reduction of plaque by a non-adhesive mode of action. Comparing the results for the two clinical studies, it is also concluded that dentin containing ficin showed directionally better non-stick efficacy than dentin containing papain.

The GMP produced batches were aged for 6 weeks at 49 ° C. The enzymatic activity of each of the enzymes remained almost unchanged compared to the initial activity figures.

EXAMPLE V: Dentifrice formulation containing antiadhesive enzyme and antibacterial agent.

The procedure of Example I above is repeated except that the amount of water is reduced and replaced with a corresponding amount of the example antibacterial agent, CPC. Consequently, the water was reduced to 16.7 percent by weight and the CPC was included in an amount of 0.3 percent by weight for each of the three formulations, as in Table 1.

Claims (21)

one.

 A composition for oral treatment comprising: an anti-adhesive agent comprising a non-stick protease enzyme wherein the protease enzyme comprises at least one cysteine protease, a cationic antibacterial agent; and an anionic surfactant that is incompatible with the anti-adhesive agent and the cationic antibacterial agent, and is secreted from the anti-adhesive agent and the cationic antibacterial agent.

2.

 The composition according to claim 1, wherein the at least one cysteine protease comprises at least one of the ficine, papain and crilase proteases.

3.

 The composition according to claim 2, wherein the cysteine protease is ficin.

Four.

The composition according to any of the preceding claims, wherein the anionic surfactant comprises an anionic alkyl sulfate surfactant.

5.

 The composition according to claim 4, wherein the anionic surfactant comprises sodium lauryl sulfate.

6.

 The composition according to any of the preceding claims, wherein the cationic antibacterial agent comprises a quaternary ammonium compound.

7.

 The composition according to claim 6, wherein the cationic antibacterial agent comprises an alkyl pyridinium halide.

8.

 The composition according to claim 7, wherein the cationic antibacterial agent comprises cetyl pyridinium chloride.

9.

 The composition according to claim 8, wherein the cationic antibacterial agent is cetyl pyridinium chloride and the antiadhesive agent is ficin.

10.

 The composition according to any of the preceding claims, wherein, on the basis of 100 parts by weight of the composition for oral treatment, the anti-adhesive agent is present in an amount of up to about 10 parts.

eleven.

 The composition according to claim 10, wherein, on the basis of 100 parts by weight of the oral treatment composition, the anti-adhesive agent is present in an amount of at least about 0.01 parts.

12.

 The composition according to claim 11, wherein, on the basis of 100 parts by weight of the oral treatment composition, the anti-adhesive agent is present in an amount of at least about 0.05 parts.

13.

 The composition according to claim 11, wherein, on the basis of 100 parts by weight of the oral treatment composition, the non-stick agent is present in an amount of about 0.03 parts to about 0.5 parts.

14.

 The composition according to any of the preceding claims, wherein, on the basis of 100 parts by weight of the composition for oral treatment, the cationic antibacterial agent is present in an amount of at least about 0.01 parts.

fifteen.

 The composition according to claim 14, wherein, on the basis of 100 parts by weight of the composition for oral treatment, the cationic antibacterial agent is present in an amount of at least about 0.05 parts.

16.

 The composition according to claim 14, wherein, on the basis of 100 parts by weight of the composition for oral treatment, the cationic antibacterial agent is present in an amount of about 0.03 to about 0.30 parts.

17.

 The composition according to any of the preceding claims, which also comprises a different enzyme, in addition to the non-stick protease enzyme, wherein the different enzyme is chosen from the group of amylases, lipases and other proteases.

18.

 The composition according to any of the preceding claims, wherein in an anionic surfactant is secreted by encapsulation in a vehicle that experiences rupture in the oral cavity.

19.

 The composition according to claim 18, wherein the vehicle is a microsphere.

twenty.

 The composition according to claim 18, wherein the vehicle is a gel matrix.

twenty-one.

 The composition according to claim 1, wherein the cationic antibacterial agent is secreted from the anionic surfactant by encapsulation in a vehicle that experiences rupture in the oral cavity.