MXPA98010243A - Antimicrobial agents for oral hygiene products - Google Patents

Abstract

Oral hygiene compositions include an antimicrobial agent selected from cedarwood oil, chloramphenicol, citronella oil, Glycyrrhiza glabra extract, juicy fruit basil oil, lemon basil oil, and Rosmarinus officinalis oil. Application of these oral hygiene compositions to the oral cavity effectively reduces or prevents the growth of bacteria associated with dental plaque, and with dental caries and/or periodontal diseases such as Actinomyces viscosus, Campylobacter rectus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus sanguis.

Description

ANTIMICROBIAL AGENTS FOR ORAL HYGIENE PRODUCTS FIELD OF THE INVENTION The present invention relates to antimicrobial agents for use in oral hygiene products and methods for using these agents.

BACKGROUND OF THE INVENTION Periodontal disease and dental caries are of great economic and public health interest, worldwide. Now, it is highly recognized that these two oral diseases are caused by bacteria growing in masses on the teeth and in the gingival area. A descriptive term commonly used for these bacterial masses is "dental plaque". In the case of periodontal disease, Schluger et al. (Schluger, Youdelis, Page &Johnson, Periodontal Diseases, second edition, pages 153-262, Lea &Febiger, 1990) report that dental plaque bacteria, which grow in the area where the teeth and tissues are found - gingival, cause an inflammation of the gingiva called u gingivitis. "This is characterized by gingiva ("gum") swollen edematous that reddens and bleeds easily. If the removal of plaque is inadequate, gingivitis may progress to "periodontitis" or EU31HT124 periodontal disease in many individuals. Periodontitis in general is characterized by an inflammation of the tissues around the teeth, which leads to a resorption of the supporting bone. Periodontal disease is the leading cause of tooth loss among adults. Dental caries (cavities) are also caused by bacteria, St-reptococcus mutans is the main etiological agent (McGhee, Michale &Cassell, Dental Microbiology, page 279, Harper &Row, 1982). The prevention of dental plaque or the removal of it has long been the focus of development, with the ultimate goal of inhibiting both caries and periodontal diseases. While the formation of dental plaque can be inhibited to a certain degree by brushing the teeth at frequent intervals, gentle brushing is not sufficient to efficiently prevent the formation of dental plaque or to remove substantially all of the dental plaque that is has formed on the tooth. Since frequent gentle brushing is not sufficient to prevent tooth decay or periodontal disease due to the plaque's pathogenic bacteria, chemical methods using antibacterial agents such as chlorhexidine, benzalkonium chloride, and cetylpyridinium chloride have been proposed. In addition, the use of EU31HT124 natural products for the treatment of teeth and gums is ancient in the art, having been practiced and recommended since the mid-1880s. Since then, many patents have described compositions of oral products containing extracts of natural products. There are numerous natural essential oils available. Many of these oils are described in KIRK OTHMER ENCYCLOPEDIA OF CHEMICAL TECHNOLOGY, 4th ed. , Vol. 17m pp. 603-674, John Wiley & Sons, Inc. Morton Pader, in "Oral Hygiene Products and Practice," Cosmetic Science and Technology Series, vol. 6, pp. 356-373, Marcel Dekker, Inc., discloses sanguinaria extract as an anti-plaque agent with antimicrobial properties. Pader also discloses that volatile oils such as eucalyptol, menthol, thymol, methylsalicylate have varying degrees of antimicrobial activity, and antimicrobial activity has been reported under appropriate test conditions. Pader describes that cinnamon oil is a very weak antiseptic, and that eucalyptus oil and eucalyptol are antiseptic. Pader notes that some essential oils are used in other products primarily for flavor. Among these are cinnamon, cassia, clove, thyme, mint, anise and anethole. Pader also describes that these essential oils have detectable antimicrobial activity. It is known that the compounds: hinokitiol, citral, EU31HT124 geraniol, cocamidopropyl-betaine, berberine and enebrine essence, individually exhibit antimicrobial properties against certain bacteria. US Patent No. 3,940,476 discloses a method for inhibiting the formation of dental plaque, comprising topically applying to the teeth, as an active ingredient, an amount of either or a combination of: (a) allyl isothiocyanate, (b) uranine, (c) obtusaestirene, (d) citral, (e) citronellol, (f) nerol or (g) geraniol. U.S. Patent No. 4,913,895 describes an oral composition that includes a linear polyphosphate or a cyclic polyphosphate and menthol, anethole or mixtures thereof in an aqueous medium. It is reported that the composition has antibacterial effects and prevents the development of stones and periodontal diseases. U.S. Patent No. 4,966,754 discloses that certain essential oils and combinations thereof possess antimicrobial properties against Aspergillus niger, Candida albicans, Staphylococcus aureus and Pseudomonas aeruginosa and, therefore, are useful as preservatives in cosmetic compositions. A mixture of 14 essential oils is described which provides desirable antimicrobial compositions against the indicated microorganisms.

EU31HT124 U.S. Patent No. 4,999,184 discloses oily compositions containing certain pyrophosphate salts and reportedly provide an anti-calculation benefit. U.S. Patent No. 5,316,760 describes a mouth care product that contains a combination of Urtica dioica extract and an extract of Juniperus communis. It is described that the combination of these extracts leads to a synergistic reduction of both dental plaque and bleeding and inflammation of the gingiva. The extract of Achillaea millefolium is also described as a suitable additive to the combination of extracts of Urtica dioica and Juniperus communis. U.S. Patent No. 5,472,684 discloses a composition that includes thymol and eugen, and optionally a sesquiterpene alcohol, such as fernesol, which allegedly has antiplaque and antigingivitis effects. The essence of the Australian tea tree, sagebrush essence and eucalyptol is described as improving the anti-plaque and anti-gingivitis activity of mouthrinses formulated from the compositions described. A property that characterizes the efficiency of an antimicrobial agent as antiplaque agent and anticalculus agent is the minimum inhibitory concentration or MIC of the agent. The CIM is the minimum concentration in EU31HT124 micrograms per milliliter of an antimicrobial agent at which no bacterial growth is observed. At concentrations below the MIC, an antimicrobial agent is ineffective in eliminating or inhibiting the growth and reproduction of bacteria. Concentrations above the MIC, the antimicrobial agent is effective in eliminating or inhibiting the growth of reproduction of bacteria. Typically, antimicrobial agents are introduced into the oral cavity at an initial concentration. Almost immediately, the initial concentration begins to decrease due to the dynamics of the oral cavity. Eventually, the concentration of the antimicrobial agent within the oral cavity will fall below the MIC. In this way, it has been an object of those who work in developing antiplaque and anticalculus formulations to use antimicrobial agents that have low MICs. Chlorhexidine has a MIC of approximately one μg / ml and is the standard against which other antimicrobial agents are measured. While chlorhexidine has a desirable MIC, it also exhibits an undesirable taste and has the undesirable side effect of staining teeth.

EU31HT124 SUMMARY OF THE INVENTION In one aspect the present invention is an oral hygiene composition that includes an antimicrobial agent selected from cedar wood essence, chloramphenicol, citronella essence, Glycyrrhiza glabra extract, juicy fruit basil essence, essence of lemon basil, and oil of Rosmarinus officinalis. According to the present invention, oral hygiene compositions including these antimicrobial agents are effective in inhibiting and preventing the growth of bacteria present in oral cavities such as Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus. sanguis. Oral hygiene compositions formed in accordance with the present invention are characterized in that the antimicrobial agent is present in an amount sufficient to retard the growth of the bacteria or to kill the bacteria. Antimicrobial agents can be combined with components typically found in oral hygiene products such as dentifrices. These components include abrasives, humectants, binders and surfactants. Other dentifrice components include flavoring agents and thickening agents. According to the present invention, the EU31HT124 oral hygiene compositions and the products of the present invention can be used in the method for practicing oral hygiene which includes the step of contacting an oral cavity with an antimicrobial agent selected from cedar wood oil, chloramphenicol, Citronella oil, Glycyrrhiza glabra extract, juicy fruit basil essence, lemon basil essence, and Rosmarinus officinalis oil.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT As used herein, the following terms have the following meanings: "Cedarwood oil" refers to extracts of complete, volatile oil, derived primarily from the heart of the Juniperus virginiana tree or Juniperus ashei. The constituents of the complete oil extract include tujopsen, cedrol, alpha-copaene, alpha-cedrene, beta-cedrene and iddrol. The CAS number for cedar wood oil is 8000-27-9. "Chloramphenicol" refers to 2,2-dichloro-N- [2-hydroxyl-1- (hydroxymethyl) -2- (4-nitrophenyl) -ethyl] acetamide. Chloramphenicol is derived from Streptomyces venezuelae or by organic synthesis. The CAS number for chloramphenicol is 56-75-7. "Essence of citronella" refers to an essence Commercially available EU31HT124 produced by steam distillation of either Cymbopogon nardus or Cymbopogon winterianus. The CAS number for citronella essence is 8000-29-1. "Glycyrrhiza glabra extract", also known as sweet stick extract, refers to the crude powder extract of Glycyrrhiza glabra. Several varieties that include typical G. and G. glandulif ra exist. The extract of Glycyrrhiza includes glycyrrhizic acid and glycyrrhetinic acid as constituents. The complete extract is available from commercial sources or can be collected by solvent extraction, such as extraction with ethanol described below. ?? Juicy fruit basil essence "refers to the complete extract of a sectioned variety of basil with a component of juicy fruits Juicy fruit basil is a crop of Ocimum basilicum L. w Essence of lemon basil" refers to the extract of full, volatile oil, from a selected variety of basil with a "citral component." Lemon basil is a culture of Ocimum basilicum L. with a high citral content. "Lemon essence" refers to the complete oil extract , volatile, from a variety of fresh citrus trus lemon peel.

EU31HT124 is also known as lemon essence or Ci trus lemon essence. The CAS number for the lemon essence is 8008-56-8. "Rosmarinus officinalis oil" refers to the extract of complete oil, from one the flowering tips of Rosmarinus officinalis. The essence of Rosmarinus officinalis is also known as the rosemary extract, or the essence extract of Rosmarinus officinalis. The CAS number for the essence of Rosmarinus officinalis is 84604-14-8. All of the above is available from commercial sources. Minimum inhibitory concentration or MIC "refers to the minimum concentration in micrograms per milliliter of an antimicrobial agent at which no growth of bacteria is observed." Concentrations below the MIC, the antimicrobial agent is ineffective in killing or inhibiting growth and reproduction of the bacteria Concentrations above the MIC, the antimicrobial agent is effective in removing or inhibiting the growth and reproduction of the bacteria An oral hygiene composition formed in accordance with the present invention includes a selected antimicrobial agent. from wood oil EU31HT124 cedar, chloramphenicol, citronella oil, Glycyrrhiza glabra extract, juicy fruit basil essence, lemon basil essence, and Rosmarinus officinalis oil. These oral hygiene compositions can be incorporated into oral hygiene products formulated in accordance with the present invention, such as dentifrices, mouthwashes and mouthwashes. Preferred oral hygiene compositions include an antimicrobial agent selected from cedarwood oil, chloramphenicol, Glycyrrhiza glabra extract. Particularly preferred for incorporation into an oral hygiene composition formulated in accordance with the present invention are the antimicrobial agents selected from cedar wood oil and Glycyrrhiza glabra extract. Preferred antimicrobial agents are selected because these agents are surprisingly effective in retarding the growth and / or preventing the growth of representative gram-positive and gram-negative oral pathogenic bacteria such as Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus sanguis. The following examples illustrate the effectiveness of the antimicrobial agents of the present invention against these particular bacteria. Preferably, the EU31HT124 antimicrobial agent is effective against more than one of the bacteria indicated above and preferably against all the bacteria mentioned above. The particular amount of the antimicrobial agent present in compositions formed in accordance with the present invention is not limited to any particular value, provided that the amount present is effective to retard the growth of the bacteria and / or prevent the growth of the bacteria, it is say, in an amount that is greater than the MIC of the antimicrobial agent with respect to particular bacteria. As illustrated in the examples that follow, the antimicrobial agents of the present invention exhibit MICs ranging from about 3.1 to about 156 against representative oral pathogens, Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus sanguis. Dentifrices or toothpastes are, in general, a thick suspension of a polishing, abrasive material in an aqueous wetting system. Typically, dentifrices include: an abrasive to remove the thin film, a humectant (s) to provide a vehicle for flavor, a abrasive, thickening agent (s) to structure and stabilize the dentifrice, an agent EU31HT124 surfactant mainly to supply foam during use, a fluoride to prevent the formation of cavities and a flavoring to make the taste of the product pleasant. Many abrasives are available for dentifrices, examples include: silica xerogel, silica precipitate, dicalcium phosphate, dicalcium phosphate dihydrate, alumina trihydrate, calcium pyrophosphate, calcium carbonate and insoluble sodium metaphosphate. Examples of suitable humectants include sorbitols, glycerin, and polyethylene glycols. Silica aerogels, pyrogenic silica, silica precipitates, carboxymethylcellulose, carboxyvinyl polymers, xanthan gum and carrageenan, are examples of materials that are suitable as thickeners. Exemplary surfactants include sodium lauryl sulfate and dodecylbenzene sulfonate. Numerous flavoring agents are commercially available with those that provide mint or other refreshing flavors, such as cinnamon which is commonly used. Oral rinses or mouthwashes are in general, pourable, flavored aqueous emulsions, which in most cases have incorporated a EU31HT124 antimicrobial. Typical components of an oral rinse include flavoring agents to make the product pleasant to use and to emphasize the therapeutic or refreshing qualities, surfactant agent (s) to maintain stable dispersion flavor, humectant (s) to improve the sensation in the mouth, thickening agent and an active agent. Frequently, a surfactant is used to impart light foaming properties to the oral rinse. Dentifrices and oral rinses that incorporate the compositions for inhibiting the growth of the bacteria formed in accordance with the present invention are formulated in a conventional manner with the antimicrobial agents which are presented in an amount ranging from about 0.001 wt% to about 5.0 % by weight based on the total weight of the toothpaste. Preferably, the antimicrobial agents are present in an amount ranging from about 0.01% by weight to about 2.5% by weight. The specific components used in dentifrices and oral rinses incorporating the antimicrobial agents of the present invention are not limited to those discussed above; however, the selected components do not preferably have an antagonistic effect on the antimicrobial properties of the selected antimicrobial agent.EU31HT124 The following examples illustrate the effectiveness of the oral hygiene compositions of the present invention against the bacteria present in the oral cavity, the MIC of the antimicrobial agents in the compositions against these bacteria, the dentifrice formulations including antimicrobial compositions of the present invention. , and the consumer preference for these tooth formulations, and the in vivo effectiveness data for these toothpaste formulations.

EXAMPLE 1 Determination of the minimum inhibitory concentration of antimicrobial agents The following example illustrates how the antimicrobial agents useful in accordance with the present invention retard or prevent the growth of dental plaque bacteria present in the oral cavity. In addition, the example illustrates the lower concentration of various antimicrobial agents that will inhibit the visible in vitro growth of a particular bacterium. The assay used a microtitre plate to dilute the antimicrobial agent at varying concentrations in order to determine the MIC. Table 1 below provides a listing of the antimicrobial agents used in this example and EU31HT124 abbreviations for them.

TABLE 1 Antimicrobial agents and abbreviations for the same cedar wood oil (RC1) chloramphenicol (CR1) citronella essence (CTR1) extract of Glycyrrhiza glabra (GLY) essence of juicy fruit basil (JFB1) essence of lemon basil (LMB1) ) essence of lemon (LM01) essence of Rosmarinus officinalis (R0F1) A bacterial culture was incubated at 37 ° C. Prior to the dilution of the antimicrobial agents as described below, the bacterial culture was stirred at 2,000 rpm in a pellet and redispersed in a buffered phosphate solution. The inoculum was normalized with a spectrophotometer at an optical density of 550 nanometers from 0.18-0.22 (equivalent to 5.0 x 107 colony forming units, CFU per milliliter). The inoculum was set aside, until the completion of the dilution of the antimicrobial agent. A 96-well, sterile polystyrene plate was used to dilute the agent combinations EU31HT124 antimicrobial. Using the aseptic technique, 100 microliters of distilled water was placed in each test cavity. In the first cavity in each column, 100 microliters of the antimicrobial agent was added. Concentrated solutions of the antimicrobial agents were prepared with methanol for a solvent to bring the agents into solution. This resulted in a half dilution of the concentrated solution. Then 100 microliters of these cavities were transferred to the next cavity in the column, and so on, down in each column. Each transfer achieved a half dilution of the concentration in the appropriate cavity. After the dilution of the antimicrobial agent was finished, 80 microliters of the growth medium specific for the bacteria under study was added to each cavity. The specific growth media for a given bacterium are set forth in Table 2. Then, 20 microliters of inoculum were then added to each cavity. This resulted in the first cavity having a final dilution of a quarter based on the concentration of the concentrated dilutions. The remaining cavities were a dilution to the half of the previous cavity, in each transfer. The 96-well plate was incubated under conditions that varied based on microorganisms EU31HT124 particular. The incubation conditions are set forth in Table 2. The aerobic bacteria were incubated under normal environmental conditions and the anaerobic bacteria were incubated under an atmosphere of 10% hydrogen gas, 5% carbon dioxide gas, and the rest gas of nitrogen. After 48 hours of incubation, the incubated plate was read for microbial growth with a spectrophotometer, through optical density (OD). The cavity containing the lowest dilution possible with a spectrophotometer reading below 0.05 OD (ie, no detectable microbial growth) was considered representative for the antimicrobial agent. The MIC for the agent was determined taking into account the concentration of the concentrated start solution and the resulting dilutions in the 96-well plate. The specific bacteria inoculated in the 96-well plate are set forth in Table 2 below together with the growth medium and the incubation conditions for that microorganism.

TABLE 2 Microorganism Conditions / Growth Medium / Incubation Microorganism No. of ATCC Conditions Medium Incubation Growth EU31HT124 1. Tryptic soy broth 3.0% by weight by volume, yeast extract 0.1%, and 999 milliliters of distilled water. 2. Tryptic soy broth 3.0% by weight by volume, yeast extract 0.5%, L-cysteine 0.05%, Hemin 0.0005%, Menadione 0.00002%, and 990 milliliters of distilled water. 3. Tryptic soy broth 3.0% by weight to volume, yeast extract 0.5%, peptone 1.0%, extract of L-cysteine, Hemin 0.0005%, Menadione 0.00002%, and 990 milliliters of distilled water. 4. Brain and heart infusion broth 0.74% by weight by volume, 0.01% yeast extract, 0.2% sodium formate, 0.03% sodium fumarate, 0.005% heme and 990 milliliters EU31HT124 of distilled water.

In the following Tables, the antimicrobial agents were identified with respect to the abbreviations set forth in Table 1. In addition, the five microorganisms set forth in Table 2 are referred to by the abbreviations set forth in Table 2.

TABLE 3 CIM (μg / ml) for antimicrobial agents for bacteria indicated This example illustrates the minimum inhibitory concentration of the indicated antimicrobial agents and the EU31HT124 capacity and ability of antimicrobial agents to inhibit the growth of specific bacteria.

EXAMPLE 2 Dentifrice Formulations This example describes the dentifrice formulations comprising the antimicrobial agents of the present invention. Formulations 18-88 do not include the antimicrobial agents according to the present invention.

TABLE 4 Formulation 5-82: 0.1% Cedarwood Oil, (P / P) EU31HT124 TABLE 5 Formulation 5-81: 0.1% Cedarwood Oil, (P / P) EU31HT124 TABLE 6 Formulation 5-108: 0.4% Cedarwood Oil, (P / P) EU31HT124 TABLE 7 Formulation 18-87: 0.5% Glycyrrhiza glabra extract, (P / P) TABLE 8 Formulation 18-90: cedar wood oil 0.1%, (P / P) EU31HT124 TABLE 9 Formulation 18-25: Hinokitiol 0.1%, EU31HT124 TABLE 10 Formulation 18-88: Control Carbomer refers to polymer composed of acrylic acid cross-linked with allyl sucrose available as CARBOPOL® 940. CARBOPOL® 940 is commercially available EU31HT124 from B.F. Goodrich. SYLODENTR 750 and 700 are silica gels (silicon dioxide) and SYLODENTR 15 is a silica gel (silicon dioxide). SYL0DENTR is commercially available from W.R. Grace & Co, - Conn. Davison Chemical Division. The formulations described above were formed in a vacuum mixer by adding the deionized water and dispersing the carbomer while vacuum is applied. When the carbomer was well dispersed, sodium hydroxide was added. In another vessel, the 70% sorbitol solution and the Poloxamer were heated and mixed. The Poloxamer solution was then added to the vacuum mixer and mixed in the carbomer mixture. The xanthan gum was then added to the glycerin, and the resulting solution was added to the vacuum mixer and incorporated into the mixture. The salts were then added to the vacuum mixer, followed by silicas that were mixed slowly. The active agents, flavor, sodium lauryl sulfate and coloring agents were then added and allowed to mix until they were well incorporated. This example illustrates various formulations of dentifrice incorporating the antimicrobial agents of the present invention EU31HT124 EXAMPLE 3 Consumer Acceptance This example illustrates consumer acceptance of dentifrice formulations incorporating an antimicrobial agent according to the present invention. The formulations set forth in Tables 4-9 were evaluated for consumer acceptance. For comparison purposes, several commercially available dentifrices were also evaluated. These included Viadent Gel (Formulation 5-35), available from Viadent, Inc.; Crest Regular Blue Paste (Formulation 5-97), available from Proctor & Gamble Company; and Listerine Teal Gel (Formulation 5-99), available from Warner Wellcome. The study of preference was carried out with a group of approximately 20 people. A tube of toothpaste formulation was given to each person to use during the course of a week. At the end of the week, participants filled out a questionnaire, the results of which are summarized in Table 11 below. The score interval was 1-10 with one that is poor and with 10 that is good. The scores were counted and averaged.

EU31HT124 TABLE 11 Consumer assessment This example illustrates how the complete satisfaction of the consumer for the formulations with the active agents according to the present invention is EU31HT124 greater than full consumer satisfaction for certain commercially available products.

EXAMPLE 4 Bacterial Reduction in the Gingival Edge in vivo This example illustrates how the dentifrice formulations that include the compositions of the present invention reduce the levels of bacteria at the gingival margin. In this example, the brushing habits of the subjects remained constant. On the day of sampling, the subjects had not brushed their teeth during 6-18 (overnight) before the test. The level of bacteria was determined by recovering samples in vivo and counting the number of bacterial colonies formed on agar after 48 hours. Subjects were 18-65 years of age, had a minimum of 20 teeth, and had normal salivary flow. Subjects with systemic diseases that affect oral tissues, such as diabetes, blood cell abnormalities, Down syndrome, or known HIV infection, were rejected; with glosistis, moderate severe gingivitis, periodontitis or other oral infections, or were taking an antibiotic Systemic EU31HT1 or had done so two weeks before the start of the test. Each of the formulations was rated for antimicrobial efficacy in approximately 20 subjects. The subjects were asked to keep the microbial plate overnight, for the morning when the samples were collected by "individual brushing" of a formulation. In the baseline, a sample of the margin plate or gingival margin was taken with a sterile cotton and placed in one milliliter of sterile, phosphate buffered saline (PBS). The cottons were gently rubbed along the gingival margin of the mandibular buccal surface between the incisor teeth. The subjects were then brushed immediately after the baseline sample with a toothpaste formulation using an electric toothbrush for two minutes. At intervals of one to two hours, after the completion of the brushing, samples of the gingival plate were recovered with cottons as described above. The clinical design was a cross study of several periods. By cross-study of several periods it is meant an experiment in which subjects are subjected to a first treatment, then they "cross over" to receive a second, then a third and finally a fourth treatment. The experimental design allows each subject EU31HT124 serve as your own control. A two-day "wash" period was used among the different antibacterial test formulations. The gingival samples were placed in the test tubes and remained at room temperature until it was processed in the space of 1 hour. Samples in the test tube were serially diluted in sterile PBS with a Spiral Plater (Spiral Systems, Cincinnati, Ohio) and plated on blood agar supplemented with 5% sheep blood (BBL, Becton Dickinson, Cockeysville) . The plate samples were then incubated aerobically at 37 ° C for 48. The total aerobic colony forming units were counted. The bacterial counts were transformed into units of log10, the average and standard deviations for each test condition were generated. The data were paired by subject for relevant comparisons and analyzed by the Wilcoxon sign-rank test, non-parametric, for level of significance. We hypothesized that the test depended on statistics using the multi-period crossover design of the clinical study. The data is presented later in Table 12. Formulations 18-25, 18-87, and 18-90 included the components set forth in Tables 9, 7, and 8, respectively. Formulation 18-88 included the EU31HT124 components set forth in Table 10 (no antibacterial agent) and used with a control for comparison purposes against formulations 18-25, 18-87 and 18-90.

TABLE 12 Effect of dentifrice formulations containing antimicrobial agent on bacterial levels in vivo.

Statistical results: Test significance against control (formulation 18-88) at 2 hours.

EU31HT124 The equal pairs of Wilcoxon sign-rank with the data transformed by log10. ** p < 0.01, ** p < 0.05, * p < 0.1 NA = not applicable This example illustrates how toothpaste formulations that include antimicrobial agents reduce gingival bacterial levels in vivo.

EXAMPLE 5 Ethanol extraction of Glycyrrhiza glabra 25 grams of powdered plant material of Glycyrrhiza glabra was combined with 250 grams of a mixture of 250 grams of a 95: 5 ethanol / water mixture. The mixture was stirred overnight at room temperature. The solids were removed from the stirred mixture with a Whatman filter number 4 in a Buchner funnel. The additional removal of the solids was achieved with a Whatman filter number 5 in a Buchner funnel. The additional solids were removed with a Whatman 1 micrometer filter in a Buchner funnel. A vacuum filtration apparatus and a 0.2 micron filter were used to clean the solution in a final time. The clean solution was then concentrated to a solid using a rotoevaporizer. Approximately 2.5 grams of an oxide-colored solid was collected as the crude extract of Glycyrrhiza glabra.

EU31HT124 While the preferred embodiment of the invention has been illustrated and described, it will be appreciated that various changes may be made therein without departing from the spirit and scope of the invention.

EU31HT124

Claims (27)

1. CLAIMS 1. A composition, comprising: an antimicrobial agent selected from the group consisting of: cedarwood oil, chloramphenicol, citronella oil, Glycyrrhiza glabra extract, juicy fruit basil essence, lemon basil essence , lemon essence and oil of Rosmarinus officinalis.
2. 2 . The oral hygiene composition according to claim 1, wherein the antimicrobial agent is selected from the group consisting of: cedar wood oil, chloramphenicol and Glycyrrhiza glabra extract. The oral hygiene composition according to claim 2, wherein the antimicrobial agent is selected from the group consisting of: cedarwood oil and Glycyrrhiza glabra extract. The composition according to claim 1, wherein the antimicrobial agent is present in an amount effective to retard the growth of oral pathogenic bacteria. The composition according to claim 4, wherein the oral pathogenic bacteria are selected from the group consisting of: Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, EU31HT124 Streptococcus mutans and Streptococcus sanguis. The composition according to claim 1, wherein the antimicrobial agent is present in an amount effective to kill the bacteria. The composition according to claim 6, wherein the bacteria are selected from the group consisting of: Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans and Streptococcus sanguis. The composition according to claim 1, wherein the antimicrobial agent is present in an amount effective to inhibit the formation of dental plaque. The composition according to claim 1, wherein the antimicrobial agent is present in an amount greater than the minimum inhibitory concentration of the agent. 10. A dentifrice comprising: an antimicrobial agent selected from the group consisting of: cedarwood oil, chloramphenicol, citronella oil, Glycyrrhiza glabra extract, juicy fruit basil essence, lemon basil essence, essence of lemon and oil of Rosmarinus officinalis; an abrasive; EU31HT124 a humectant; a binder; and a surfactant. The dentifrice according to claim 10, wherein the antimicrobial agent is selected from the group consisting of: cedar wood oil, chloramphenicol and Glycyrrhiza glabra extract. The dentifrice according to claim 11, wherein the antimicrobial agent is selected from the group consisting of: cedarwood oil and Glycyrrhiza glabra extract. The dentifrice according to claim 10, wherein the antimicrobial agent is present in an amount effective to retard the growth of oral pathogenic bacteria. The dentifrice according to claim 13, wherein the bacteria are selected from the group consisting of: Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans and Streptococcus sanguis. 15. The dentifrice according to claim 10, wherein the antimicrobial agent is present in an amount effective to kill the oral pathogenic bacteria. 16. The dentifrice according to claim 15, in EU31HT124 where the oral pathogenic bacteria is selected from the group consisting of: Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans and Streptococcus sanguis. 17. The dentifrice according to claim 10, wherein the antimicrobial agent is present in an amount effective to inhibit the formation of dental plaque. 18. The dentifrice according to claim 10, wherein the antimicrobial agent is present in an amount greater than the minimum inhibitory concentration of the agent. 19. A method to inhibit or prevent the growth of oral pathogenic bacteria, comprising the step of: contacting an oral cavity with an antimicrobial agent selected from the group consisting of: cedar wood oil, chloramphenicol, citronella oil, Glycyrrhiza glabra extract, juicy fruit basil essence, essence of lemon basil, lemon essence, Rosmarinus officinalis oil. The method according to claim 19, wherein the antimicrobial agent is selected from the group consisting of: cedar wood oil, chloramphenicol and Glycyrrhiza glabra. EU31HT124 21. The method according to claim 18, wherein the antimicrobial agent is selected from the group consisting of: cedar wood oil and Glycyrrhiza glabra. The method according to claim 19, wherein the step of contacting further comprises contacting the oral cavity with an amount of the antimicrobial agent effective to retard the growth of the oral pathogenic bacteria. The method according to claim 22, wherein the oral pathogenic bacteria selected from the group consisting of: Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus sanguis. The method according to claim 19, wherein the step of contacting further comprises contacting the oral cavity with an amount of the antimicrobial agent effective to kill the oral pathogenic bacteria. 25. The method according to claim 24, wherein the oral pathogenic bacteria are selected from the group consisting of: Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus sanguis. 26. The method according to claim 19, in EU31HT124 wherein the step of contacting further comprises contacting the oral cavity with an amount of the antimicrobial agent effective to inhibit the formation of the dental plaque. 27. The method according to claim 19, wherein the step of contacting further comprises contacting the oral cavity with an amount of the antimicrobial agent greater than the minimum inhibitory concentration of the antimicrobial agent. EU31HT124