AU2019100472A4

AU2019100472A4 - Methods and compositions for treating gingivitis and inflammation and reducing plaque and tartar

Info

Publication number: AU2019100472A4
Application number: AU2019100472A
Authority: AU
Inventors: Kourosh MADDAHI; Hessam Nowzari
Original assignee: Mge Holdings LLC
Current assignee: Mge Holdings LLC
Priority date: 2019-05-02
Filing date: 2019-05-02
Publication date: 2019-06-06
Anticipated expiration: 2027-05-02

Abstract

Abstract Methods of reducing gingivitis and plaque buildup by administering an oral care product (a dentifrice or a mouthwash) that consists essentially of a sea salt, preferably Dead Sea salt, and xylitol. In mouthwash and dentifrice embodiments, the oral care product contains aloe vera leaf juice.

Description

Methods and Compositions for Treating Gingivitis and Inflammation and Reducing Plaque and Tartar

Field

The present invention relates to oral care methods that (i) promote healthy teeth and gums and freshens breath, (ii) reduce gingivitis and inflammation, (iii) reduce build-up of plaque and tartar, and (iv) protect the oral microbiome by neutralizing toxins caused by bacteria commonly found in the oral cavity - namely Streptococcus mutans and Porphyromonas gingivalis

- without killing the bacteria. Oral care compositions that contain natural and naturally.0 derived, non-cytotoxic, non-bactericidal ingredients that repair and protect enamel and gums and reduce cavities without fluoride.

Background

The therapeutic and medicinal benefits of Dead Sea salts have been reported in the scientific literature, typically in connection with diseases of the skin and joints. See, e.g., Uriel Katz et al., .5 Scientific Evidence of the Therapeutic Effects of Dead Sea Treatments: A Systematic Review, Seminars in Arthritis and Rheumatism, Vol. 42, No. 2 (October 2012), pp.186-200, citing Z. EvenPaz, J. Shani, The dead sea and psoriasis: Historical and geographic background, Int J Dermatol, Vo. 28, No. 1 (1989), pp. 1-9 (345 g of mineral per liter (34.5% or 34.5 g/100 mL); Id. citing S. Sukenik et al., The Dead Sea—a unique resort for patients suffering from joint diseases, Harefuah, Vol. 149, No. 3 (2010), pp. (175-179)(180 to 215 g of mineral per liter). Dan Buskila et al., Balneotherapy for Fibromyalgia at the Dead Sea, Rheumatol Int, Vol. 20 (2001), pp. 105-108.

The water of the Dead Sea contains concentrated salts other than NaCI - including, but not limited to, MgCb, CaCb, KCI, and MgBr2. Among the separate ions present in the Dead Sea water are chloride (212.4 g/l), magnesium (40.65 g/l), sodium (39.15 g/l), calcium (16.86 g/l), potassium (7.26 g/l), bromide (5.12 g/l), sulfate (0.47 g/l), and bicarbonate (0.22 g/l). See, e.g.,

I.L. Schamberg, Treatment of psoriasis at the Dead Sea, Int J Dermatol, Vol. 17, No. 6 (1978), pp. 524-525; Paz and Shani, supra.

2019100472 02 May 2019

European Patent Application EP1074245A2 discloses use of mineral salt, in particular Dead Sea salts, as an active ingredient in a mouthwash to assist in combatting bacteria and gum irritation and inflammation.

Essential oils have been used for the treatment of a variety of ailments since ancient times. The safety and efficacy of essential oils in dentistry have been reported in numerous clinical studies.

See, e.g., Namrata Dagli et al., Essential oils, their therapeutic properties, and implication in dentistry: A Review J Int Soc Prev Community Dent. Vol. 5, No. 5 (2015), pp. 335-340.

The present invention meets the long-felt but as yet unmet need for oral care products (in particular, dentifrices and mouthwashes) that effectively reduce gingivitis as well as plaque and .0 tartar build-up, with safe and effective amounts of natural and naturally-derived ingredients.

Compositions of the present invention also meet the need for reducing levels of cavity-causing glucansucrases and neutralizing Leukotoxin A, Gingipains, and lipopolysaccharides from P. gingivalis, all without causing a decrease in the levels of common bacteria in the oral cavity Streptococcus mutans, Porphyromonas gingivalis, Staphylococcus aureus, Fusobacterium .5 nucleatum, subsp. nucleatum Knorr and Fusobacterium nucleatum Strain W1481.

Summary

The present invention is directed to oral care products - toothpastes, mouthwashes, and dental floss - comprised of (i) a sea salt, preferably, Dead Sea salt, at a concentration of from 0.5% to 3.0%; and (ii) xylitol, and uses of such products to reduce plaque and gingival inflammation.

Mouthwash and dentifrice embodiments of the invention also contain aloe vera leaf juice.

Oral care products of the present invention do not contain any of the following: fluoride; artificial colors, dyes or flavors; paraben or formaldehyde preservatives; bleaching agents (i.e., peroxides); sodium lauryl sulfate or sodium laureth sulfate; petroleum-derived glycerin; and genetically modified organisms (GMOs).

Oral care products of the present invention are (xx) non-cytotoxicity within the framework of ISO 10993-5 Biological Evaluation of Medical Devices - Tests For In Vitro Cytotoxicity, and (yy) non-bactericidal, each as described in greater detail below

Detailed Description

2019100472 02 May 2019

Non-cytotoxicity of oral care products of the present invention is confirmed within the framework of ISO 10993-5:2009 based on the widely-used Trypan blue exclusion test. (Trypan blue is a colorant which stains dead cells, i.e., cells with loss of membrane integrity.) More particularly, Balb/c 3T3 clone A31 cells (ATCC CCL 163; 86th passage) are seeded in multi-well plates (24 wells, each 15.5 mm in diameter) at the starting density of 30,000 cells/cm² in culture medium - Dulbecco's Modified Eagle Medium (DMEM) - supplemented with 10 % (v/v) fetal calf serum (FCS). No antibiotics are used. Cultures are incubated at 37°C in a humidified .0 atmosphere containing 5% (v/v) CO₂, for 24 hours, and are examined with a microscope to verify a sub-confluent monolayer with less than de minimus alteration in morphology. Culture medium is withdrawn and replaced with a solution of one of the following: oral care products of the present invention at 5,000 pg/mL, as well as dilutions 1,500 pg/mL, 500 pg/mL and 150 pg/mL; a negative control (phenol, Chemical Abstract Service No. 108-95-2, 0.64mg/mL); and .5 a positive control (DMEM supplemented with 10 % (v/v) FCS and 1% antibiotics (v/v). A positive control is defined as statistically significant (30% or greater) inhibition of cell growth as compared to the negative control.

Wells are incubated at 37°C in a humidified atmosphere containing 5 % (v/v) CO₂, for a 24-hour period. Photomicrographs are taken (320x magnification) showing the cell layer in contact with 20 the negative control, the positive control and the oral care product of the present invention.

Morphology and cell density of the cultures are observed. At the end of the incubation period, culture medium is removed. Cells are detached for two minutes using 250 pL trypsin (0.05 % (w/v) in Hank's balanced solution Ca++ and Mg++ free supplemented with 1 mM EDTA. 250 pL of a Trypan blue solution at 0.2 % (w/v) in 0.15 M NaCI and 10 % (v/v) FCS are added. Cells are 25 incubated for two minutes. Living cells (uncolored) are counted using a hemocytometer. Cell morphology and cell density of medium treated with 5,000 pg/mL of the oral care product of the present invention are observed to be comparable to those of negative control; neither

2019100472 02 May 2019 shows statistically significant (30% or greater) inhibition of cell growth. In contrast, cells treated with the positive control show greater than 50% inhibition on cell growth.

Oral care products of the present invention are “non-bactericidal” by which is meant that when administered at a concentration of 5 mg/ml in a 10 ml solution also containing 0.1 ml of artificial saliva (AFNOR NF S91-141, Pickering Laboratories, Inc. Mountain View, California) with 1 x 10⁵ to 1 x 10⁶ colony forming units (“CFUs”) of a bacteria selected from the group of S. mutans, P. gingivalis,

S. aureus, Fusobacterium nucleatum subsp. nucleatum Knorr (ATCC® 25586D-5™), and Fusobacterium nucleatum Strain W1481, the bacterial count (i.e., number of CFUs) does not decrease.

.0 By the term dentifrice is meant a preparation for cleansing and polishing the teeth, that may, and preferably does provide one or more therapeutic benefits (as described below). As will be understood by the skilled artisan, a dentifrice (also referred to in the art and in this application as a toothpaste) may be formulated as a paste, gel or powder and is preferably applied with a toothbrush.

.5 Dentifrice embodiments of the present invention are administered by dispensing at least about 0.25 grams of a dentifrice of the present invention (commonly described in the scientific literature and in recommendations from dental professionals and toothpaste manufacturers as a pea sized amount), and more preferably at least 0.50 grams of dentifrice, and brushing the teeth for at least 30 seconds, preferably for at least 60 seconds, and still more preferably for at 20 least 120 seconds, at least once per day, in the evening prior to going to sleep. In certain preferred embodiments, the dose of toothpaste per administration ranges from about 0.4 to about 0.6 grams.

By brushing is meant placing the bristles of a toothbrush in contact with the teeth, preferably at an angle of about 45 degrees to the gum line (where the gums and teeth meet), and moving 25 the bristles in gentle, short strokes along the outer surfaces (cheek side), the inside surfaces (tongue side) and the chewing surfaces of all teeth. The strokes may be in a back-and-forth motion (side-to-side, or up-and-down) or a circular motion. After brushing, the dentifrice is expectorated.

2019100472 02 May 2019

Still more preferably, dentifrice embodiments of the present invention are administered by brushing the teeth for at least 30 seconds, preferably for at least 60 seconds, and still most preferably for at least 120 seconds, twice daily - once in the evening prior to going to sleep; and once in the morning after breakfast.

By the term mouthwash is meant a solution that is swished, preferably vigorously, around the mouth, and then expectorated, thereby cleaning the mouth and making the breath smell pleasant.

Mouthwash embodiments of the present invention are administered by swishing about 15-20 ml (about 0.5-0.75 fluid ounces) in the mouth, for a period of time sufficient to contact the .0 teeth, the gums, the roof of the mouth and the tongue. Preferably, mouthwash is swished for at least 30 seconds, preferably for at least 60 seconds, at least once per day, in the evening prior to going to sleep.

Still more preferably, mouthwash embodiments of the present invention are administered for at least 30 seconds, preferably for at least 60 seconds, most preferably for at least 120 seconds, .5 at least once per day twice daily -once in the evening, after brushing prior to going to sleep;

and once in the morning after brushing after breakfast.

In especially preferred embodiments, a person practicing a method of the present invention (e.g., a consumer or a patient) brushes his/her teeth with a dentifrice of the invention for at least 30 seconds, more preferably for at least 60 seconds, and then swishes a mouthwash of the 20 invention in his/her mouth for a period of at least 30 seconds, preferably at least 60 seconds - a period of time sufficient for the mouthwash to contact the teeth, the gums, the roof of the mouth and the tongue.

Persons practicing a method of the present invention are instructed to abstain from drinking or eating for at least 30 minutes after administering the oral care compositions of the invention;

and, since oral care compositions of the invention are not intended for ingestion, to expectorate the oral care composition after use (i.e., brushing in the case of a dentifrice, or swishing in the case of a mouthwash).

2019100472 02 May 2019

Dental floss is a cord of thin filaments used to remove food and plaque from between teeth.

Flossing means the mechanical cleansing of proximal tooth surfaces with dental floss, subgingivally and supragingivally.

Preferred mouthwash embodiments of the present invention are alcohol free - a term understood by the person having ordinary skill in the art as free of ethyl alcohol. Alcohol free mouthwashes may contain other alcohols, including menthol or glycerol.

As used in the present application, an essential oil is a mixture of terpenic hydrocarbons, especially monoterpenes and sesquiterpenes, and oxygenated derivatives such as aldehydes, ketones, epoxides, alcohols, and esters.

.0 Numbers used in describing quantities of ingredients and/or reaction conditions are to be understood as being modified in all instances by the term about.

Unless otherwise indicated, percentages, parts and ratios are to be understood as based upon the total weight of the composition.

Numerical ranges are meant to include numbers within the recited range, and combinations of .5 subranges between, the given ranges. For example, a range from 1-5, includes 1, 2, 3, 4 and 5, as well as subranges such as 2-5, 3-5, 2-3, 2-4, 1-4, etc.

At least one means one or more, and also includes individual components as well as mixtures/combinations.

Sea salt is a mixture of inorganic salts from sea water or from inland bodies of salt water. Sea salt may be in the form of a precipitate (on the bottom of a marsh or salt pan or flat) or crystals that float on the surface of the water (known as fleur de sei).

Dead Sea salt is a mixture of natural hygroscopic minerals and micronutrients found in the Dead Sea and is comprised of sodium chloride, potassium chloride, calcium chloride and magenesium bromide. S. Halevy et al., J. Eur. Acad. Dermatol. Venereol., Vol. 9, pp. 237-242 (1997). Dead 25 Sea salt also contains separate ions: Br-, Rb+, Li+, Fe3+, AI3+, SO42-, Sr2+ and Mn2+. See, e.g.,

2019100472 02 May 2019

Dead Sea salt is present in oral care products of the invention at a concentration of less than about 3%, preferably a concentration of from about 0.1% to 2%, still more preferably a concentration of from about 0.5% to about 1.5%.

Another preferred sea salt suitable for use in the oral care products of the present invention is Himalayan salt, which is harvested from the Punjab Region of Pakistan, and is comprised of sodium chloride (about 95-98%), with about 2 to 3% polyhalite (potassium, calcium, magnesium, sulfur, oxygen, hydrogen), fluoride, iodine, and smaller amounts of other trace .0 minerals.

Xylitol is a pentahydric alcohol and is present in oral care products of the invention at a concentration of at least about 7.5%, more preferably at least about 10%, or at a dose of about 0.1 g/brushing or rinsing.

Aloe vera leaf juice useful in mouthwash and toothpaste embodiments of the present invention .5 preferably contains (i) glycosides at a concentration of at least about 1%, preferably at least about 2%, and still more preferably at about 3%, as well as (ii) at least two, preferably three, anti-inflammatory agents selected from the group of anthraquinones, sterols, auxins and gibberellins and (iii) and immunomodulatory muccopolysachharides, preferably Acemannan.

Oral care products of the present invention contain one or more essential oils selected from the 20 group consisting of spearmint oil, Wintergreen oil, and peppermint oil.

Spearmint oil is the volatile oil obtained from the leaves of Mentha viridis (also known as Mentha spicata).

Wintergreen oil is the volatile oil obtained from the leaves of Gaultheria procumbens.

Peppermint oil is a volatile oil obtained from the whole plant Mentha piperita.

In one set of preferred embodiments, oral care products of the present invention include at

2019100472 02 May 2019 least one essential oil in the genus Mentha, selected from Mentha piperita (peppermint) oil and

Mentha viridis (spearmint) leaf oil.

In yet another preferred embodiment, oral care products of the present invention include Gaultheria procumbens (wintergreen) leaf oil.

In one even more preferred embodiment, oral care products of the present invention include peppermint oil and one of wintergreen oil or spearmint oil.

In another even more preferred embodiment, oral care products of the present invention include wintergreen oil and one of peppermint oil or spearmint oil.

In a still further even more preferred embodiment, oral care products of the present invention .0 include spearmint oil and one of peppermint oil or wintergreen oil.

In especially preferred embodiments, the oral care products of the present invention contain spearmint oil, peppermint oil, and wintergreen oil.

Menthol, an alcohol that can be isolated from peppermint or other mint oils, can also be used in oral care products of the present invention.

.5 Oral care products of the present invention also preferably include oils of one or both of Ocimum basilicum (basil) or Eugenia caryophyllus (clove flower).

In certain preferred embodiments of the invention, basil oil is present at a concentration of up to about 0.5%.

In certain preferred embodiments of the invention, clove oil is present at a concentration of at least about .005%. In other preferred embodiments of the invention, clove oil is present at a concentration of at least about 0.01%. In certain of these preferred embodiments, clove oil is preferably at a concentration of up to about 0.02%.

Other essential oils that may be included in oral care products of the present invention include Melaleuca alternifolia (tea tree) leaf oil, the oil distilled from the leaves of the Melaleuca alternifolia, and Zingiber officinale (ginger) root oil, which is obtained from the dried rhizomes

2019100472 02 May 2019 of Zingiber officinale.

Dentifrice embodiments ofthe present invention may include mild abrasives (to remove debris and residual surface stains), humectants (to prevent water loss in the toothpaste), thickening products, also known in the art as binders (to stabilize the toothpaste formula), flavoring products (for taste) and detergents (to create foaming action).

Mild abrasives suitable for use in the toothpaste embodiments of the present invention include calcium carbonate, dehydrated silica gels, hydrated aluminum oxides, magnesium carbonate, phosphate salts and silicates. Silica, also called silicone dioxide, bentonite clay and hydrated silica are minerals. Some toothpastes ofthe present invention preferably contain hydrated .0 silica.

Humectants that may be, and preferably are, ingredients in toothpastes ofthe present invention include glycerin, preferably vegetable glycerine, propylene glycol, and sorbitol.

Glycerin, a sugar alcohol that can be synthesized or obtained from natural sources, is an especially preferred humectant used in toothpastes ofthe invention.

.5 Non-limiting examples of thickening products that may be, and preferably are included in toothpaste embodiments ofthe present invention include gums and colloids. Preferred colloids are of marine origin, even more preferably seaweeds.

Two preferred gums are xanthan gum and biosaccharide gum-1; both are polysaccharides derived from the fermentation of carbohydrates. Xanthan gum is derived from glucose or corn 20 syrup. Biosaccharide gum-1 is derived from sorbitol.

Carrageenan, a polysaccharide hydrocolloid obtained from edible red seaweeds in the Gigartinaceae or Solieriaceae families, may be, and preferably is, present in toothpastes of the invention.

Methods ofthe present invention reduce the amount of plaque by at least 10%, preferably at least 20%, more preferably by at least 30%. Plaque levels can be measured, for example, by the Quigley-Hein Plaque Index (defined by G. A. Quigley and J. W. Hein in 1962 and modified by S.

2019100472 02 May 2019

Turesky, N. D. Gilmore, and I. Glickman in 1970), a system for scoring the amount of plaque buildup on teeth that is well-known to the person having ordinary skill in the art (PHOSITA) of dentistry.

More particularly, the PHOSITA scores the amount of plaque on each facial or lingual surface of each tooth that is examined, t third molars. Scores ranging from 0 to 5 are assigned as follows: 0 - no plaque; 1 - separate flecks of plaque at the cervical margin of the tooth; 2 - a thin continuous band of plaque (up to one mm) at the cervical margin of the tooth; 3 - a band of plaque wider than one mm but covering less than one-third of the crown of the tooth; 4 plaque covering at least one-third but less than two-thirds of the crown of the tooth; 5 - plaque .0 covering two-thirds or more of the crown of the tooth. The Quigley-Hein Plaque Index does not take evaluate third molars and does not take into account plaque buildup on dental restorations (i.e., bridges, crowns, veneers, inlays, and onlays made of porcelain or composite resin). The overall Quigley-Hein Plaque Index (for the entire mouth) is determined by dividing the total score by the number of surfaces examined.

.5 Methods of the present invention reduce gingival inflammation based on the Loe-Silness gingival index preferably at least 25%, more preferably by at least 50%, even more preferably by at least 75%. The Loe-Silness gingival index is a system for scoring gingival inflammation that is well-known to the PHOSITA of dentistry. A score ranging from 0-3 is assigned, where 0.1-1.0 is indicative mild inflammation; 1.1-2.0 indicates moderate inflammation; and 2.1-3.0 indicates .0 severe inflammation. Overall Gl is obtained by adding the values of each tooth and dividing by the number of teeth examined.

Reduction in gingival inflammation can also be expressed in terms of the redness coordinate in the CIE 1976 (L*a*b*) system, in which L* value represents comparative lightness / darkness (lower L* being indicative of darker), and a* and b* values represent chromaticity coordinates (red-green and blue-yellow, respectively). See A.R. Robertson, The CIE 1976 colordifference formulae, Color Res. Appl. Vol. 2, pp. 7-11 (1977). Each of the L*, a* and b* can be plotted in three-dimensional space to characterize a color in absolute terms. The magnitude of the difference between two colors, as perceived by the human eye, is proportional to the

2019100472 02 May 2019 distance between two points defining the two colors on the three-dimensional plot. The difference between the two colors, the Euclidean distance (ΔΕ), is defined by the following equation: AE*ab = [(AL*⁾² + (Aa*)² + (Ab*)2]'^/2.

Methods of the present invention may also be practiced by flossing with a dental floss comprised of filaments that are coated with a sea salt, preferably Dead Sea salt, and xylitol. Preferably, the filaments are coated with spearmint.

Examples

The following examples illustrate compositions and methods of practicing of the present invention in some of its embodiments; the examples should not be construed as limiting the .0 scope of the invention. Other embodiments will be apparent to one skilled in the art from consideration of the specification and examples. It is intended that the specification, including the examples, is considered exemplary only without limiting the scope and spirit of the invention.

Some of the examples illustrate preferred embodiments of the invention. Variations of these .5 preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, unless otherwise indicated herein or otherwise clearly contradicted by context, the inventions include all modifications and equivalents of the 20 subject matter disclosed and recited in the claims appended hereto as permitted by applicable law.

Dentifrice Examples

Dentifrices of the present invention are formulated for use by the general population - persons not having sensitivity or xerostomia - and contain the following ingredients at the following 25 concentrations:

Dead Sea salt is present in toothpastes of the invention at a concentration of less than about

2019100472 02 May 2019

3%, preferably from about 0.5% to 2%, most preferably from about 0.75% to about 1.5%.

Xylitol is present in toothpastes of the present invention at a concentration of at least 10% or at a dose of 0.1 g/brushing.

Aloe vera leaf juice is present in toothpastes formulated for use by the general population at a concentration of at least about 10%, preferably at least about 20%, and most preferably at least about 40%.

Mentha piperita (peppermint) oil is present in toothpastes of the invention at a concentration of less than 1%, preferably less than about .075%, more preferably at a concentration of less than about 0.05%, and even more preferably at a concentration of about 0.03%.

.0 Mentha viridis (spearmint) oil is present in toothpastes of the invention, preferably at a concentration of up to about 4%, preferably up to about 2%, more preferably up to about 1%, and still more preferably less than about 0.5%.

Gaultheria procumbens (wintergreen) leaf oil is present in toothpastes of the invention, preferably at a concentration of up to about 1%, preferably from about 0.25% to about 0.5%.

.5 In some preferred toothpaste embodiments, the weight ratio of wintergreen oil to peppermint oil is about 10:1.

In other preferred toothpaste embodiments, the weight ratio of spearmint oil to peppermint oil is about 3:1.

In still other preferred toothpaste embodiments, the weight ratio of wintergreen oil to spearmint oil is about 3:1.

In one preferred mouthwash embodiment, the weight ratio of wintergreen to peppermint is from about 6:1 to about 5:1.

In another preferred mouthwash embodiment, the weight ratio of spearmint to peppermint is about 5:3.

2019100472 02 May 2019

In still another preferred mouthwash embodiment, the weight ratio of wintergreen to spearmint is about 4:1.

Basil oil (also known as Ocimum tenuiflorum oil) is present in toothpastes of the invention, preferably at a concentration of up to about .05%.

Clove oil is present in toothpastes of the invention, preferably at a concentration of up to about 0.05%.

Glycerin is present in toothpastes formulated for use by the general population at a concentration of from about 2.5% to about 20%, preferably from about 5.0% to about 15%.

Carrageenan may be, and preferably is, present in toothpastes of the invention, preferably at a .0 concentration of at least about 0.05%. preferably about 0.1%. Even more preferably, carrageenan is food-grade.

Xanthan gum may be, and preferably is present in toothpastes of the invention, preferably at a concentration of at least about 0.10.

Titanium dioxide may be present in certain toothpastes of the invention; when present, .5 titanium dioxide is present at a concentration of up to about 0.6%.

Hydrated silica may be, and preferably is, present in toothpastes of the invention, at a concentration of from about 10% to about 25%.

Toothpastes of the present invention contain a foaming anionic other than sodium lauryl sulfate, preferably sodium methyl cocoyl taurate or sodium lauroyl sarcosinate. In certain 20 preferred embodiments, sodium methyl cocoyl taurate is present in toothpastes of the invention at a concentration of up to about 2%.

Mouthwash Examples

Dead Sea salt is present in mouthwashes formulated for use by the general population at a concentration of from about .5% to about 5%, preferably from about .75% to about 3%, and 25 most preferably from about 1% to about 2%.

2019100472 02 May 2019

Xylitol is present in mouthwashes formulated for use by the general population at a concentration of from about 5% to about 30%, preferably from about 7% to about 15%, and most preferably from about 8% to about 12%.

Aloe vera leaf juice is present in mouthwashes formulated for use by the general population at 5 a concentration of from about 10% to about 90%, preferably from about 20% to about 85%, and most preferably from about 50% to about 70%.

Clove flower oil is present in mouthwashes formulated for use by the general population at a concentration of from about .005% to about .075%, preferably from about .01% to about .04%, and most preferably from about .01% to about .03%.

.0 Basil oil is preferably present in mouthwashes formulated for use by the general population at a concentration of from about .005% to about .5%, preferably from about .01% to about .2%, and most preferably from about .02% to about .1%.

Peppermint oil is present in mouthwashes formulated for use by the general population at a concentration of from about .005% to about .12%, preferably from about .01% to about .1%, .5 and most preferably from about .02% to about .09%.

Spearmint oil is present in mouthwashes formulated for use by the general population at a concentration of from about .01% to about 1%, preferably from about .02% to about .17%, and most preferably from about .05% to about .15%.

Wintergreen oil is present in mouthwashes formulated for use by the general population at a 20 concentration of from about .03% to about 1%, preferably from about .05% to about .5%, and most preferably from about .1% to about .45%.

Clinical Testing Example: Reduction in Plaque Presence and Gingival Health

Ten test subjects were recruited to participate in a double-blinded clinical study. Over a period of twenty days, participants followed three different oral care regimens - each a study arm 25 lasting three days: (i) administering a mouthwash of the present invention, (ii) administering a control mouthwash, and (iii) not administering any mouthwash.

2019100472 02 May 2019

Three days prior to the beginning the first study arm, subjects abstained from oral hygiene - no teeth brushing, no use of mouthwash, no chewing gum. For the duration of each study arm, subjects abstained from brushing the lower anterior teeth.

Three clinical endpoints were recorded before and after each of three study arms: (i) plaque index (PI), based on the Turesky Modification of Quigley-Hein Index; (ii) gingival inflammation (Gl) based on the Loe-Silness gingival index; and (iii) sulcus bleeding (mSBI).

Each study arm was followed twice daily - before going to sleep and in the morning after eating breakfast. In study arms (i) and (ii) participants vigorously swished with the assigned mouthwash for at least one minute.

.0 Between the first and second study arm, and again between the second and third study arm, participants entered a one-week wash out period, during which time they brushed with CREST’ PRO-HEALTH™ toothpaste (Procter and Gamble, Cincinnati, OH) using a new ORAL B® PROFLEX™ toothbrush (Procter and Gamble, Cincinnati, OH) for each washout period of the study. Use of other oral hygiene measures was not permitted, including chewing gum. Additionally, in .5 order to minimize potential crossover effects, the sequence of mouthwash usage was randomized so that 3 subjects used the mouthwash of the invention as the first study arm; 3 subjects used the control mouthwash as the first study arm; and 4 subjects used no mouthwash at all as the first study arm.

Plaque was scored on a scale of 0 to 5 according to the Turesky modification of the Quigley20 Hein Plaque Index. Gingival health was evaluated using the Loe-Silness gingival index and the Sulcus Bleeding Index.

Using a 2-sided T-test and standard statistical approaches for crossover studies, the endpoint and baseline values for each parameter were compared. None of the data showed any evidence of a significant carryover effect (i.e., from one study arm to the next arm). For all of the clinical indices measured, the mouthwash of the invention effects did not differ significantly from those of the control mouthwash (p>0.164). Both mouthwashes removed plaque and maintained gingival health more effectively than when no mouthwash was used (p<0.05).

2019100472 02 May 2019

More particularly, three days of using the mouthwash of the invention or the control mouthwash reduced plaque presence and gingival inflammation to a very similar degree. Plaque indices were reduced by approximately 30% after 3-day use of either mouthwash, whereas plaque levels increased by approx. 25% when no mouthwash was used. Gingival indices and plaque buildup were significantly reduced after use of either mouthwash, whereas they increased significantly in the absence of a mouthwash regimen.

1. A method of reducing gingival inflammation and reducing plaque build-up by

Claims

Claims administering to a subject an effective amount of an oral care product comprised of (a) Dead Sea salt, at a concentration of from about 0.5% to about 3.0%; and (b) xylitol at a concentration of at least 5%;

(c) at least one essential oil selected from the group of peppermint oil, spearmint oil and Wintergreen oil; and (d) optionally, an essential oil of basil or clove flower.
2. The method of claim 1 wherein the oral care product is a dentifrice and contains vegetal .0 glycerin and at least one surfactant, wherein the at least one surfactant (a) is coconut derived and (b) is not sodium lauryl sulfate or sodium laureth sulfate.
3. The method of claim 1 where the oral care product is administered to the subject at least once daily for at least 30 seconds.
4. The method of claim 3 wherein plaque build-up is reduced by at least 10% as measured .5 on the Quigley-Hein Plaque Index.
5. The method of claim 3 wherein gingival inflammation by at least 25% as measured on the Loe-Silness gingival index.