CZ384898A3 - Antimicrobial preparations 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

This invention relates to antimicrobial agents for use in oral hygiene products and methods of using such agents.

State of the art

Periodontal disease and dental caries are both health and economic problems worldwide.

It is now known with certainty that both of these oral diseases are caused by bacteria that grow in large numbers on the teeth and in the periodontal region. The term commonly used to describe these bacterial formations is dental plaque.

In the case of periodontal disease, Schluger et al. describes, see Schluger, Yuodelis, Psge and Johnson, Periodontal Disease','

2nd ed. ₀ , pp. 153-262, Lea and Febiger, 1990, that plaque bacteria, growing in the area where the teeth meet the gum tissue, cause gum inflammation, so-called gingivitis. The _disease is characterized by swelling and edema of the gums, which is felt in

They turn red and pale. If plaque removal is inadequate, gingivitis can progress to periodontitis in many individuals. Periodontal disease is usually characterized by chronic inflammation of the tissue around the teeth, leading to resorption of the supporting bone. Periodontal disease leads to tooth loss in adults. Dental caries (cavities) are also caused by bacteria, with Streptococcus mutans being the main etiological agent, see McGhee, Michalek _G Cassell, Dental Microbiology, p.

279, Harper and Row, 1982.

Prevention or removal of dental plaque has long been a focus of interest in order to inhibit both dental caries and periodontal disease. While plaque formation can be inhibited to some extent by frequent tooth brushing, it is important to note that plaque is a major cause of dental caries.

-2 intervals, brushing alone is insufficient to effectively prevent the formation of dental plaque or to remove essentially all of it that has formed on the teeth.

Since tooth brushing alone is not sufficient to prevent or remove plaque, chemical methods have been proposed using antibacterial agents such as chlorhexidine, benzalkonium chloride, and cetylpyridinium chloride. The use of natural products for tooth brushing is a relatively old practice, dating back to about 1880. Since then, numerous patented methods have been described for oral compositions and similar products containing extracts from natural products. Numerous natural oils are mentioned, and many of them are described, see Kirk-Othmer Encyclopedia of Chemical Technology, 4th ed. vol. 17, pp. 603-674, J. Wiley and Sons, Inc. Uorton Pader describes in Oral Hygiene Products and Practice, Cosmetic Science and Technology Series, vol. 6, pp. 356-373 (Marcel Dekker, lne) extracts of Sanguinaria as agents effective against plaque with antimicrobial properties. Pader also describes volatile oils _such as eucalyptol, menthol, thymol, methyl salicylic acid ester with varying degrees of antimicrobial activity as well as antiplatelet activity as indicated under appropriate test conditions and procedures. Pader describes that cinnamon oil is a very weak antiseptic and both eucalyptus oil and eucalyptol are antiseptics. Pader notes that some of the oils are used in other products mainly for their fragrance. These include cinnamon oil, cassia oil, clove oil, frankincense oil, peppermint oil, anise oil and anethole. Pader also states that some of the oils listed have detectable antimicrobial activity.

For example, it is known that cocamidopropylbetsin, hinokitiol and berhsrin, as well as essential oils, citral, geraniol and juniper berry oil, increase antimicrobial efficacy against some bacteria.

U.S. Patent No. 3,940,476 describes a method of inhibiting the formation of dental plaque, comprising topically applying to the teeth as an active ingredient either alone or in combination the following substances:

* · · · « · · _3_

a) allyl isothiocyanate,

b) uranine,

c) obtusastyrene,

d) citral

é) citronellol, £j nerol, or

g) geraniol.

U.S. Patent No. 4,913,895 describes an oral composition comprising a linear polyisophosphate or a cyclic polyphosphate and menthol, anethole or mixtures thereof in an aqueous medium. The composition is said to have antibacterial effects and to prevent the development of plaque and periodontal disease.

US Patent No. 4,966,754 discloses that certain essential oils and combinations thereof exhibit antimicrobial properties against Aspergillus niger, Candida _albicans , Staphylococcus aureus, and Pseudomonas aeruginosa and are therefore suitable as preservatives in cosmetic compositions. A mixture of 14 essential oils is described which provides the necessary antimicrobial properties against the aforementioned microorganisms.

U.S. Patent No. 4,999,184 describes an oily composition containing certain salts of pyrophosphoric acid and is claimed to provide protection against bacterial growth.

U ₀ S.patent document 5 316 7čO describes a product with regard to oral care, containing a combination of nettle (Urtica dioica) and juniper (Juniperus communis) extracts. It is described that the combination of these extracts is associated with a synergistic reduction of both dental plaque and bleeding or inflammation of the mucosa. It also describes the extract of common yarrow (Achillea milíefolium) as a suitable additive and a combination of nettle (Urtica dioica) and juniper (Juniperus communis) extracts.

U ₀ S. patent documents 472-684 describe a composition containing thymol and eugenol and optionally one of the sesquiterpene alcohols such as farnesol, which according to the findings is characterized by antiplatelet effects, as well as antigingivitis effects. Australian tea tree oil and eucalyptol are described as supporting components against platelet formation and against gingivitis in rinsing the mouth with the said • fc • fc « · • fc

11« · · fc · compos.

.IN

One of the properties that characterizes an antimicrobial agent as effective against the formation of trails and clumps is the minimum inhibitory concentration (hereinafter MIC) of the agent. The MIC is the minimum concentration in micrograms per milliliter at which bacterial growth is not observed. At concentrations higher than the MIC, the antimicrobial agent is effective in the sense that it kills bacteria or inhibits their growth and reproduction.

Typically, antimicrobial agents are introduced into the oral cavity at a certain initial concentration. Almost immediately, the initial concentration begins to decrease in response to the dynamics of the oral space. Eventually, the concentration of the antimicrobial agent in the oral cavity drops below the HIC value. Thus, the aim of this work was to develop an anti-platelet and anti-clumping formulation using an antimicrobial agent with a low MIC value.

The HIC value of chlorhexidine is approximately 1 µg/ml and is a standard against other antimicrobial agents measured. Although chlorhexidine has the required HIC value, it has an unpleasant taste and undesirable side effects, such as tooth discoloration.

Essence and invention

According to one aspect, the present invention provides an oral hygiene composition comprising an antimicrobial agent selected from the group consisting of cedarwood oil, chloramphenicol, lemongrass oil, licorice extract in

(Glycyrrhiza glabra), basil oil from juicy fruit, basil oil from lemons and rosemary oil (Rosmarinus officinalis). According to the present invention, oral hygiene compositions containing these antimicrobial agents are effective in inhibiting and preventing the growth of bacteria present in the oral cavity, such as Actinomyces viscosus, Pusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans and Streptococcus sanguis. Oral compositions prepared in accordance with the process of the present invention are characterized in that they contain antimicrobial agents in an amount sufficient to inhibit the growth of bacteria or kill them.

• ·

-5« · »

Antimicrobial agents can be combined with devices typically used in the dental and oral hygiene field, such as toothpastes. The combination may include abrasives, humectants, binders, and surfactants. Other preparations in connection with toothpastes include flavors and preservatives.

Accordingly, in accordance with the present invention, the oral hygiene compositions and products of the present invention may be used in oral hygiene that involves contacting the oral cavity with an antimicrobial agent selected from the group consisting of cedarwood oil, chloramphenicol, lemon tree oil, licorice extract (Glycyrrhiza glabra), extracts and oils of citronella fruit, lemon tree oil, and rosemary oil (Rosmarinus officinalis).

Detailed description of the invention

As the following names are used, p3k means:

"Cedarwood oil" means the oily extracts of the wood of the juniper (Juniperus virginiana or Juniperus ashei). The constituents of such an oily extract are thujopsen, cedrol, -kopaen, oL-cedren^ £>-cedren and widdrol.

The CAS number of cedarwood oil is 8000-27-9.

Chloramphenicol is 2,2-dichloro-E ^T -(.2-hydroxy-1-(hydroxymethyl)-2-(4-nitrophenyl)-ethyl)-acotamide. The origin can be either Streptomyces venezuelae or organic synthesis,

The CAS number of chloramphenicol is 56-75-7.

Citronella oil is a commonly available oil as a product of steam distillation using either Cymbopogon nardus or Gymbopogon winterianus.

CAS number in this case: 8000-29-1,

Licorice (Glycyrrhiza glabra) extract, also known as licorice root extract, refers to the crude powdered extract of licorice. There are several varieties, such as G.typica and G _e glandulifera. Licorice (Glycyrrhiza glabra) contains glycyrrhizic and glycyrrhetinic acids in the extract. The extract as a whole is commercially available or can be obtained by solvent extraction, such as ethanol, as described below.

Basil oil with fruit juice refers to the total extract of basil and components of the nature of fruit juices. It is *

-6Lemon basil oil is a volatile oily extract from a selected type of basil with a citral component. Lemon basil oil is a product of the cultivation of the Onimum bascilicum basil with a high citral content®

Lemon oil is the total oily extract of the fresh peel of the lemon Citrus limon. It is also known by other similar names ₀

CAS-Numbers: 8008-5&-8.

Rosemary oil refers to the total oil extract of rosemary (Rosmarinus officinalis). It is also known as Rosemary _oil .

CAS-number?: .04604-14-8 _o

All of the substances listed above are commercially available.

Minimum inhibitory concentration (MIC) means the minimum concentration in micrograms per milliliter of an antimicrobial agent at which no further growth of bacteria is observed. At a concentration below the MIC, the antimicrobial agent is ineffective in killing, inhibiting, or reproducing bacteria. At a concentration above the MIC, the antimicrobial agent is effective in killing or inhibiting the growth or reproduction of bacteria.

Oral hygiene compositions prepared in accordance with the present invention contain an antimicrobial agent from the group of cedarwood oil, chloramphenicol, citronella oil, extract in

licorice, basil oil with fruit juice, basil lemon oil and rosemary oil. These oral hygiene compositions can be incorporated into oral hygiene products prepared according to the present invention, such as toothbrushes, mouthwashes and mouthwashes.

Preferred oral hygiene compositions comprise an antimicrobial agent selected from the group consisting of cedarwood oil, chloramphenicol and licorice extract. Particularly preferred oral hygiene compositions in accordance with the present invention comprise an antibacterial agent selected from the group consisting of cedarwood oil and licorice extract. Preferred antibacterial agents are selected because such agents are surprisingly effective in slowing the growth and/or inhibiting the growth of exemplary gram-positive and gram-negative oral pathogenic bacteria such as Actinomyces viscosus, Pusocterium nucleatum, Porphytomonas gingivalis, Streptococcus mutans and Streptococcus Sanguis. The examples which follow demonstrate the effectiveness of the antibacterial agents of the present invention particularly against the bacteria just mentioned. In the manufacture, an antibacterial agent is used which is effective against more than one of the bacteria as mentioned hereinabove, and preferably against all of the bacteria mentioned hereinabove.

The amount of antimicrobial agent present in the composition of the present invention is not limited in any way, provided that the amount present in the composition is sufficient to effectively delay and/or prevent bacterial growth. That is, it must be an amount greater than the MIC value of the antibacterial agent with respect to a particular bacterium.

As demonstrated in the examples that follow, the antibacterial agents of the present invention exhibit MIC values ranging from about 3.1 to about 156 against representative oral pathogens such as Actinomyces viscosus, Pusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Streptococcus sanguis.

Toothpowders or toothpastes are usually a concentrated mixture of abrasive polishing materials in an aqueous wetting medium. Typically, toothpastes contain a conventional abrasive agent to remove staining deposits, a humectant or humectants to provide flavor to the vehicle, abrasives and thickeners to adjust the texture and stabilize the toothpowder, surfactants to create foam during use, fluorides to prevent tooth decay, and flavorings to give the product an acceptable taste.

Numerous abrasive agents are suitable for use in toothpastes, and examples include silica xerogel, precipitated silica, dicalcium phosphate, its dihydrate, alumina trihydrate, calcium pyrophosphate, calcium carbonate, and insoluble sodium metaphosphate.

-8• · • · · • · · • · « « « * » » «·*· · • · · · · ««·«««« «« tt

Examples of suitable humectants include sorbitol, glycerol and polyethylene glycols.

Suitable thickeners include: silica aerogels, fumed or precipitated silica, carboxymethylcellulose, carboxyvinyl polymers, xanthan gum and carrageenan.

Examples of suitable surfactants include sodium laurylsulfuric acid and dodecylbenzenesulfonic acid and its salts.

Numerous flavoring agents are commercially available, including those that include mint and other refreshing flavors such as _cinnamon oil.

Oral rinses or mouthwashes are usually an aqueous, mobile emulsion with flavors, to which in most cases an antimicrobial agent is added. Typical components of water rinses also include flavoring agents so that the product can be used with pleasure, emphasizing the therapeutic and refreshing qualities, surfactants are present to maintain the flavor in a stable dispersion, humectants to improve the overall mouthfeel, thickeners and active agents, and a surfactant is often added to achieve a light foaming of the preparation.

Toothpastes and mouthwashes containing antimicrobial agents for inhibiting the growth of bacteria in the oral cavity in accordance with the present invention are prepared by conventional methods, with the antimicrobial agent being present in an amount ranging from about 0.001% (w/w) to about 5.0% (w/w), based on the total weight of the formulation. Preferably, the antimicrobial agent is present in an amount ranging from about 0.01% (w/w) to about 2.5% (w/w). The specific ingredients used in the antimicrobial agent-containing toothpastes _and mouthwashes of the present invention are not limited to those listed above. _However , it is preferred that the selected ingredients do not have antagonistic effects on the antimicrobial properties of the antimicrobial agent.

Further examples demonstrate the effectiveness of the oral hygiene compositions of the present invention against bacteria present in the oral cavity.

-9f space, report MIC values of antimicrobial agents in compositions against bacteria, describe toothpastes and mouthwashes containing antimicrobial compositions of the present invention, consumer benefits of such dental care compositions, and in vivo efficacy data of such hygiene compositions

Examples of embodiments of the invention

Example 1

Determination of minimum inhibitory efficacy and concentration of antimicrobial agents.

FROM

Another example illustrates how antimicrobial agents useful in accordance with the present invention retard or prevent the growth of, with respect to dental plaque, bacteria present in the oral cavity. The example further provides the lowest concentrations of various antimicrobial agents that visibly inhibit the growth of the corresponding bacteria in vitro.

The test uses a microtiter plate to dilute an antimicrobial agent to various concentrations to determine the MIC.

FROM

Table 1 lists the antimicrobial agents used in this example and their abbreviations:

Table 1

FROM

Antimicrobial agents and their abbreviations

Before cedarwood oil (RG1) chloramphenicol (CR1) citronella oil (CTR1) licorice oil (GLY) Glgcyrrhiza glabra basil oil with fruit juice (JPB1) lemon oil (LMB1) lemon oil (LM01) rosemary oil (R0P1) Rosmarinus officinalis

The bacterial culture was incubated overnight at 37°C.

x by diluting the antimicrobial agent as described below, • · < · • · • ·< · i e ι

-10 the bacterial culture was pelleted (at 2000 rpm) into a pellet which was resuspended in phosphate buffer. The inoculum was normalized spectrophotometrically to an optical density at 550 nanometers between 0.18-0.22, which is equivalent to 5.0 ♦ 10 ⁷ colony forming units (CPU per milliliter). The inoculum was then set aside until: the dilution of the antimicrobial agent was complete, ,

A sterile 96-well polystyrene plate was used to dilute the antimicrobial agents, each containing 100 microliters of distilled water. 100 microliters of antimicrobial agent was added to the first well of each column. Stock solutions of antimicrobial agents were prepared using methanol as a solvent to keep the agents in solution. This resulted in a half dilution of the stock solution. From these wells, 100 microliters were transferred to the next well of the column, and so on to the next column. Each transfer represented a half dilution of the concentration of the previous well. After the dilution of the antimicrobial agent was complete, 80 microliters of growth medium specific for the bacteria under study were added to each well. Specific growth media for the bacteria are listed in Table 2 _.

Next, 20 microliters of inoculum were added to each well. The result in the first well of each column was a final dilution, corresponding to a quarter of the stock solution. The remaining wells were always half the dilution of the previous well with each transfer _by

The plate and 96 wells were incubated under conditions that varied depending on the microorganism. The incubation conditions are listed in Table 2.

Aerobic bacteria were incubated at room temperature and anaerobic bacteria were incubated in an atmosphere of 10% hydrogen gas, 5% carbon dioxide, and the remainder nitrogen gas. After 48 hours of incubation, microbial growth on the incubated plate was measured using an optical density (OD) spectrometer. A well containing • « 0

-110 0 0 0 ’ « 4 «· ·0 the lowest achievable dilution with a spectrophotometric reading below 0.05 OD (i.e. no detectable microbial growth) was considered exemplary for an antimicrobial agent. The η.ilC value of the agent was determined by calculation from the concentration of the starting stock solution and the resulting dilution in a 96-well plate.

The specific bacteria inoculated into a 96-well plate are listed in Table 2 along with the growth medium and incubation conditions for that microorganism.

Table 1 and 2

Microorganism/growth medium/incubation conditions microorganism

ATCC o. Growth medium

Campylobacter rectus 33238

Actinomyces viscosus (AV) 19246

Puso bacteriura nucleatum (FN) 10933

Porphyromonas gingivslis (FG) 33277

Streptococcus mutsns (SI.·) 25175 Streptococcus sanguis (SS)49295

Comment:

CR~

TSB ¹

TSB

PG" ,1

1.

4.

Incubation conditions hours/37°C/anaerobic ” aerobic anaerobic

Π

IT

1T

II tl u

Π aerobic υ

Tryptic, soy, brew, 3.0% w/v, yeast extract 0.1% and 999 milliliters of distilled water.

Tryptic soy mash, 3.0% w/v, yeast extract 0.5%, L-cysteine 0.05%, hemir 0.0005%, menadione 0.00002%, 990 milliliters of purified water.

Tryptic soy infusion, - 3.0% w/v, yeast extract 0.5%, pontone 1%, L-cysteine extract, hemin 0.0005%, menadione 0.00002%, 990 milliliters distilled water). Liver-heart infusion infusion 0.74% w/v, yeast extract 0.01%, sodium formic acid 0.2%, sodium fumaric acid 0.01%, hemin 0.005% and 990 milliliters distilled water

-12Β·** ♦ to • ·* from

In the following tables, antimicrobial agents are indicated by the abbreviations from Table 1, and then five microorganisms are listed in Table 2 with reference to the abbreviations in Table 2.

Table 3 /

Value i JIC (ug/ml) of antimicrobial agents for labeled bacteria Reagent AV CR PG SM SS PCI 31.3 31.3 31.3 7.6 31.3 15.6 Chl 3.1 — 3.1 6.3 3.1 3.1 CTR1 62.5 12 5 31.3 31.3 62.5 31.3 GLf 15.6 15.6 15.6 7.8 15.6 7.8 JFB1 156.3 156.3 156.3 62.5 156.3 156.3 LMB1 12 5 12 5 62.5 31.3 62.5 62.5 LM01 312.5 125 62.5 31.3 125 125 R0P1 125 125 62.5 62.5 125 125 In this example j are uvc prices me normal inhibitory

The concentration of the antimicrobial agents and the ability of the agents to inhibit the growth of specific bacteria.

Example 2

This example describes tooth protection preparations containing anximaxcrobi^ini cimcla cle of this vy na z

pravgk 16-88 do not contain an antimicrobial agent in accordance with this invention.

/Ί τ,;

in agreement

T a b l e 1 a n d 4

Formulation 5-82: 0.1% cedarwood oil (w/w)

Folder % by weight sorbitol 70% 37.00 poloxamer 407 (PLUROSIC P127) 9.50 ^oda^r. deprived ions 24.50 carbomer 940 (CARBOPOL 940) 0.30

• 0 • » ·

-13» * 0 · • · · · · · · cont.table 4

Component % wt.

sodium hydroxide 0.20 xanthan gum 0.40 glycerol 4.75 sodium fluoride 0.25 sodium derivative of sscharin 0.30 sylodent 750 (silicon dioxide) 9.50 sylodent 15 (silicon dioxide) 9.50 cedar wood oil 0.50 flavor 0.30 FDsnd C blue ??1 1% solution 0.10 titanium dioxide 0.90 sodium laurylsulfuric acid 1.40

Table 5

Formulation 5-81: 0.1% cedarwood oil (w/w)

Component % wt.

sorbitol 70% 58.63 sodium carboxymethylcellulose 0.35 ion-free water 3.00 polyethylene glycol 600 5.00 glycerol 10.00.

sodium fluoride 0.22 soony esnvaL saccharin 0.30 sodium salt of benzoic acid 0.50 sylodent 700 (silicon dioxide) 15.00 aerosil 200 (silicon dioxide) 3.00 flavor 0.80 ethanol 1.50

0.10

PDandC blue Ό- 1 1% solution 0.10 sodium lauryl sulfate 1.50 « · *« · * 1 « # 4 ·· ·*

-14csóar wood ΐ ε b u 1 k ε 6

Formulation 5-108: 0.4% oil w/w

OlozRs sorbitol 70% deionized water csrbomer 340 (CAR30P0L 940) sodium hydroxide xanthan gum sodium fluoride sodium saccharin derivative SYlODSiíT 750 (silicon dioxide) SiLODBhT 15 (silicon dioxide cedarwood oil pner/j.to v e eim 1 o titanium dioxide sodium lauryl sulfate % wt.

50.00

24.63

0.30

0.20

0.5000

0.22

0.55

10.00

10.00

0.40

0.60

1.00

Addition 18-37:

T a, b u 1 0.5% oil k a 7 . , from cedar wood (wt/r wt.

(otherwise)

Component

sorbitol 70% 50.00 deionized water 24.53 carbomer 940 (CAR30P0L 940) 0.30 pine hydroxide' 0.2 0 xabthan gum 0.50 sodium fluoride 0.22 sodium saccharin derivative Ά (— Γ’ o, 00 SYLoDEIÍT 750 (silicon dioxide) 10.00 Sylodent 15 (silicon dioxide) K-,00 cinnamon extract 0.50 in flavoring agent 0.80 titanium dioxide 1.00 sodium laurylsulfuric acid 1.40

-15Table δ

Preparation 15-90: 1% ceáro' n/k^otn oil /he tree Ingredient % mass n„ sorbitol 70% 50.00 ions, water removed 2 4.03 carbomer 940 (CŮR30POL 940) 0.30 sodium hydroxide 0.20 xanthan gum 0.50 sodium fluoride 0.22 sodium saccharin derivative 0.55 SýLODENT 750 (silicon dioxide) 10.00 SYLGOBIT 15 (silicon dioxide) 10.00 oil with cn of crushed wood 1.00 in flavoring agent 0.30 titanium dioxide 1.00 sodium salt of Isuryl sulfuric acid 1.40

Tg lump 9 Preparation 18"25: 0.1% hino kitiol ,at Folder In % : ό t n. i o n u ii tired in o q 3 92.02 carbomer 540 (Gá.RBOPOL 940) 1.00 sodium hydroxide 0.75 sodium saccharin derivative 0.50 sodium fluoride 0.23 to blow 0, bO hinokitiol 0.10 ethanol 3.50 sodium laurylsulfuric acid 1.10 T a b l e 1 10 Preparation 18-88: control Folder mass n. sorbitol 70% 17.70 poloxamer 407 (PLURONIC F127) 9.50 deionized water 25.50

·· · ·

• · • » »

-16p round and bulky 10

Ingredient carbomsr 940 (CARBOPOL 940) sodium hydroxide and gum glycerol sodium iluoride sodium saccharin derivative

SYLODERT 750 (silicon dioxide)

3YL0DENT 15 (silicon dioxide) ritonium oxide sodium lauryl sulfate ρ nmoon. 0.30 0.20 0.40 4.75 0.25 0.40 9.50 9.50 1.00 1.00 fCarbomer is a polymer of acrylic acid cross-linked with allyl derivatives of s ε cho r o sy, available as CARBOPOL 940. This is commonly available from 3Goodrich, SYLCDSIIT 750 and 700 are silica gels (silicon oxides) and SYLODBIiT 15 is also available. SYLCDRRT is available from W.R.Grace and Co., Conn® Davison Chem.Bivision.

The formulations described hereinabove are obtained in a vacuum mixer by adding deionized water and dispersing the carbomer under vacuum. After the carbomer is well dispersed, sodium hydroxide is added. In another container, 70% sorbitol and poloxamer are mixed and heated, this mixture is then added to the vacuum mixer and the procarbomer. The xanthan gum is mixed with a mixer with a stirring rod, which is also added to the vacuum mixer. Then the salts are added to the vacuum mixer, then the silicas are slowly mixed in. Finally, the active agents, flavors, sodium lauryl sulfate and coloring agents are added, followed by thorough mixing.

This example describes several toothpaste-type preparations incorporating antimicrobial agents according to the present invention.

Example 3

Suitability from a customer perspective This example describes the suitability of dental products,

Of containing an antimicrobial agent in accordance with the present invention

The preparations, as listed in Tables 4-9, were prepared 9 9 · 1

9 « *9 *·

-17 evaluations from the consumer's point of view. For comparison, several commercially available preparations were also evaluated. These include: Viadent Gel (preparation 5-35), a product of Viadent, Inc.; Crest Regular Blue Paste (preparation 5-97), a product of the Procter and Gamble Company; and Listerine Seal Gel (preparation 5-99), a product of Warner Wellcome.

Prelerence studies have been conducted by the Icumen for about a year

people. Each of the axes 00 cyia c one tube dental preparation in to to use of b Ι·:;:; th day. pcs end of th days of the week they were uc a s i π í c i to that Π1 IV j V J & X li iij p research j are short nuey in that lump 11 here further. Scope of assessment was 1 to 10 seconds η -r>i r/ a 1:.;, 1 means weak above sec and 10 means good luck dress. Result the were section ny and u their diameter. T a b at 1 k and 11 Benefit night dreams come true consumer Preparation 5-82 5-81 5-103 5 -35 5-97 5-95 color 6, OR 8.00 6.75 2.72 6.64 7,- > appearance o, 4-s 7.90 6.90 3.16 6.62 6.75 < _ 1 !_Á U 5.80 6.20 7.15 3.72 6.27 4.00 taste, additionally 6.80 5.85 6.05 3.38 6.09 and what aroma 7.19 6.73 7.05 4.33 6.32 4.55 cleaning 7.38 5.35 i, Oo 3.94 6.41 6.10 foam 6.42 5.10 7.55 3.66 6.05 5.85 consistency 7.23 5.95 7.70 4.00 6.14 5.75 z dry mouth 8.14 6.90 7.10 3.89 7.31 5.70 feeling another her trs 7.00 6.16 7.05 3 3 ^Q 5.50 5.15 overall mouthfeel 7.33 6.16 7.05 3.11 6.09 /! /’ “r, satisfies overall 6.90 6.10 6.90 3.05 ς c = 4.20 about this example yes, I understand not that total peace and ness customer in case of preparation svků with active component according to it's invention is higher is compared to oelkovo at peace women student from

in the case of some commonly available products.

Reducing the excess of gum bacteria "in vivo" This example demonstrates that jsk dental preparations containing

Example 4

-18 compositions according to this invention reduce the level of bacteria, namely their excess in the gums.

In this example, the subjects' brushing habits were maintained. On the day of data collection, subjects did not brush their teeth for 16 to 13 hours (overnight) before testing, and bacterial levels were evaluated by in vivo sample preparation and bacterial colony counting, resulting in

The age of the test subjects was 13 to 65 years, at least 20 teeth, normal salivary flow. Those with chronic diseases affecting the gum tissues, such as diabetes, blood cell abnormalities, Down's syndrome, or known HIV infection, were excluded, as were those with glossitis, gingivitis, periodontitis, or other oral infections, or if they were systematically administered antibiotics, or if they had been 2 weeks before the start of the tests.

Each of the products was tested for antimicrobial efficacy on about 20 people. The subjects were asked to keep their mouths covered the night before testing, with a simple tooth brushing test the next morning.

Initially, a sample of excess gingival plaque was collected with a sterile cotton swab and placed in 1 ml of phosphate-buffered saline (PBS). The cotton swabs were gently moved along the gingival surface of the mandible using tweezers. Immediately after sample collection, the subjects brushed their teeth using a dental preparation and an electric toothbrush for 2 minutes. At intervals of one hour and two hours after the completion of brushing, the same procedure as described above was used to

Gum plaques were removed using a cotton swab.

The clinical picture is a cross-section of a study lasting a longer period. This means a study in which the test subjects were used for the first exposure to the studied preparation, then monitored for the second, third and even fourth time. The experimental result allows each test subject to self-control. A washout period of two days was always used between testing the different antibacterial preparations.

in

The gum samples were then transferred to test tubes • · ··« ···9

-19s tsrage left;; at room temperature until processing within an hour. Samples in tubes were?/ serially diluted with sterile PBS solution (see previous page) using Spiral Platsr, Spiral Systems, Cincinnati, Ohio, USA and placed on blood agar supplemented with 5h sheep blood (BBL, Becton Dickinson, Cockeyville, LJarylanc, USA). Then the samples were incubated aerobically at 37° C. for 48 hours. Bacteria were counted with respect to all aerobic colony forming units.

The bacterial count was converted to log standard deviation values under all test conditions, the data were combined taking into account the prevailing

The sigmoid colon and ε were analyzed using the nonparametric Uilcoxon sign-rank test ¹¹ with a view to determining the level of significance. Hypothesis testing was based on statistical data using a multi-period cross-section of clinical studies.

The results are shown in Table 12,

Formulations 18-25, 18-87 and 18-90 contained the ingredients listed in Tables 9, 7 and 8. Formulation 18-88 contained the ingredients listed in Table 10 (without antibacterial agent) and was used for control and comparative purposes in comparison with Formulations 18-25, 13-87 and 18-90.

-2012 effect of dental preparations containing antibacterial agents on bacterial levels in vivo

Product Active _ingredient w/w (1.10 CFU/ml) Reduction Significant Number of bacteria time level tested-

for a start 2 hours against control th A' 18-25 hinokitiol 0.1 53.11 4.47 49.8% ++ 17 18-87 extract from licorice 0.5 3 3.81 2.14 7 υ, Op· +++ 17 16-90 cedar wood 1.0 31.62 3.10 64.5% +++ 17 18-88 control 33.88 8.91 ΪΪΡ IIP 17 Statistical results : T estova effect against the control simple row (18-88) in 2 hours iny ”Vileozon sign-rank matched psirs conversion to uda is log _1Q M4 p below 0.01 ++ p below 0.05 4 p below 0.1 liP - unusable In this example, proven drugs with content- hem and of antimicrobial agents el according to this inventions destroy in vivo

I ri·—.«-ΊΚ'ΙΛ 1-. -·. I « X -s 1Z| — ·.

XC UXUW UU.OLO ¹ 7 JL·!! U Í5 KU ^r JX IX» . vs. - ·, <— ^! said Mr

Extraction of St. John's wort (Glycyrrhiza glabrs) with ethanol

250 g _of powdered St. John's wort material) with ethanol and water 95:5. The mixture is stirred at room temperature for one night, the solid mixture is filtered off using a Buchner funnel with portions of filter paper.

-21Whatman

No. 4. Further separation of the solids follows on a Buchner funnel with Whatman No. 5 filter paper, with any further solids on a Buchner funnel using Whatman 1 miti$nstr filter paper. For final purification, a vacuum filtration device with a 0.2 micron filter is used. The clear solution is then concentrated to a solid using a rotary evaporator and about 2.5 g of a rust-colored solid is isolated as the crude St. John's wort extract.

Although preferred embodiments of the invention have been described and illustrated, it is obvious that numerous variations are possible without departing from the scope of the invention.

0 • 0 ·« *00

-22· «0 0

Claims (21)

Composition for inhibiting or preventing the growth of oral pathogenic bacteria, characterized in that it comprises an antibacterial agent selected from the group consisting of cedar oil, chloramienicol, citronella oil, St. John's wort extract, basil oil of fruity fruit , lemon and vein oil, lemon oil or rosemary oil. Claim 1, characterized in that o is selected from the group consisting of: 2a, chlorphenicol, and extract of 3. The composition of the invention comprises an antimicrobial agent selected from the group consisting of an oil 3lky 3 _e The composition according to claim 2, characterized in that it contains% antimikrobiQ agent permission from the group consisting of cedarwood oil and Glycyrrhiza glabra extract. The composition of claim 1, comprising the antimicrobial agent in an amount sufficient to delay the growth of oral oathogenic bacteria. The composition of claim 4, wherein the oral bacteria are those selected from the group consisting of Actinomyces viscosus, Csmpylobacter rectus, Pusobacterium nucleatum, Porphyromons gingivslis, Streptococcus mutans and Streptococcus ssnguis. 6 ₀ A composition according to claim 1, wherein the?, <Contains an antimicrobial agent in an 'nosství sufficient to kill bacteria, Composition according to claim 6, characterized in that the bacteria are selected from the group consisting of Actinomyces viscosus, Campylobacter rectus, Pugobacterium nucleatua, Porphyroaonas gingivalis, Streptococcus mutans and Strsptosoccus sanguis. A composition according to claim 1, characterized in that z < it contains an antimicrobial agent in an amount sufficient to inhibit platelet formation. • 00 • 0 0 100. A composition according to claim 1, characterized in that it comprises an antimicrobial agent in an amount v.y is the minimum inhibitory concentration of the agent. 10. Toothpaste containing an antimicrobial agent selected from the group consisting of cedarwood oil, chloramphenicol, lemon seed oil, liquorice extract, basil oil of fat fruit, basil lemon oil, lemon oil and rosemary oil; humectant, binder and surfactant. Dental oasta li. The chiorernfenico agent of claim 10, wherein the antimicrobial is selected from the group consisting of cedar oil from St. John's wort. The toothpaste of claim 11, wherein the antimicrobial agent is selected from the group consisting of cedarwood oil and St. John's wort extract. The toothpaste of claim 10, wherein the antimicrobial agent is present in an amount sufficient to contain the oral pathogenic bacterium. Toothpaste according to claim 13, wherein the bat 14. is selected from the group consisting of Actinomyces viscosus, Gampylobsctsr rectuc, Pusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Stroptococcud sanguis. 15 Dec reagent j - - 4 - T - v v -. - ·, r ‘i n p <2 U i 1U C L1 L- X U i i l / G i iti A dental sow according to claim 10, wherein the dermal bacterial for killing oral is present in a plurality of ways. Toothpaste according to claim 15, wherein the oral pathogenic bacterium is selected from the group consisting of Actinomyces viscosus, Campylobscter rectus, Pusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans and Streptococcus sanguis. X The toothpaste of claim 10, wherein the antimicrobial agent is present in an amount sufficient to inhibit platelet formation. «Fl - -” ·· »fl v -2418. The skirt of claim 10, wherein the antimicrobial agent is present in an amount greater than the minimum inhibitory concentration of the Agent ®. 13. A method of inhibiting or preventing the growth of oral pathogenic bacteria, comprising the step of contacting the oral cavity with an antimicrobial agent permitted by a group consisting of cedar oil, chloramphenicol, citronella oil, extract and St. John's wort fruit, plowed with olrj and rosemary oil. ba lemon oil, lemon oil 21. 20. The method of claim 13, wherein the imicrobial agent is an oleaginous group consisting of wood, chloramphenicol and St. John's wort extract. Method according to claim 18, characterized in that: The antimicrobial agent is allowed from the sxupine consisting of cedarwood oil and shag extract. 22. The method of claim 13, further comprising the step of contacting the oral cavity with an amount of an anti-microbial agent sufficient to delay the growth of the oral pathogenic bacterium. 23. The method of claim 22, wherein the oral pathogenic bacterium is selected from the group consisting of Actinomyces viscosus, Campylbacter rectus, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutnns and Streptococcus sanguis, 24. The method of claim 13, wherein the contacting step comprises further blunting the oral cavity with an amount of an antimicrobial agent sufficient to kill the pathogenic bacterium. 25. The method of claim 24, wherein the oral pathogenic bacterium is selected from the group consisting of Actinomyces viscosus, Campylobacter rectus, Fusobgcterium nucleatum, Forphyromons gingivalis, Streptococcus mutans and Streptococcus sanguis®. 26. The method of claim 19, wherein the contacting step comprises the further step of contacting the oral cavity with an amount of antimicrobial agent sufficient to inhibit platelet formation. 27. The method of claim 19, wherein the contacting step further comprises contacting the oral cavity with an amount of antibacterial agent that is greater, Z than the minimum inhalation concentration of the ontibskteridine reagent is ₀