Oral care compositions containing rhamnolipids

Abstract

An oral care composition containing a specific mixture of a single rhamnolipid and a double rhamnolipid is disclosed. The composition has antibacterial efficacy against oral microorganisms, such as Porphyromonas gingivalis, Streptococcus mutans, and Campylobacter rectum. A method of treating or preventing gingivitis in a subject is also disclosed. Also disclosed are methods of preventing microorganisms from growing into a biofilm and / or reducing the number of microorganisms present in the form of a biofilm in the oral cavity of a subject.

Description

BACKGROUND OF THE INVENTION

[0001] The present technology generally relates to oral care compositions comprising biosurfactants such as rhamnolipids. More specifically, the present technology relates to oral care compositions comprising rhamnolipids that are efficacious against oral-related microorganisms, including biofilm-forming bacteria.

[0002] A recent trend is to formulate products using ingredients based on renewable raw materials. Such ingredients are considered "green" or "natural" because they are derived from renewable and / or sustainable sources. Therefore, they are more environmentally friendly than ingredients from fossil fuels or other non-renewable sources. Ingredients with a high biorenewable carbon index (BCI), such as greater than 80, indicate that the ingredient contains carbon primarily from plant, animal, or marine-based sources.

[0003] Rhamnolipids are surface-active glycolipids produced by various bacteria and are an example of a "green" ingredient because they can be prepared by fermentation based on renewable raw materials. It is desirable for oral care compositions to contain actives (such as rhamnolipids) from renewable sources and that also have effective antibacterial properties, so that the oral care composition can be used to reduce oral care-related microorganisms (encapsulated and non-encapsulated) and treat / prevent related biofilms.

[0004] The applicant has determined that the use of rhamnolipids in oral care compositions can meet the above objectives while also furthering the United Nations Sustainable Development Goals ("SDGs"). The oral care compositions of the present technology and their related methods of use can provide efficacy against oral care-related microorganisms and biofilms by using rhamnolipid actives, thereby promoting better health and well-being. Advantageously, rhamnolipids are bio-based renewable source actives that are derived from a bacterial fermentation process that produces biodegradable waste with less environmental impact. These benefits further the Sustainable Development Goal #3 (Good Health and Well-being) and Sustainable Development Goal #12 (Responsible Consumption and Production).

[0005] Brief Description of the Invention

[0006] In one aspect, the present technology provides an oral care composition comprising from about 0.025 to about 99% by weight of a purified rhamnolipid mixture based on the total weight of the composition, wherein the rhamnolipid mixture comprises mono-rhamnolipid and di-rhamnolipid, and the weight ratio of mono-rhamnolipid:di-rhamnolipid is from 10:90 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 15:85 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 20:80 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 80:20, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 70:30, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 60:40, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 50:50, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 45:55, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 60:40, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 50:50, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 45:55, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 48:52, measured by high performance liquid chromatography (HPLC).

[0007] In a related aspect, the present technology provides an oral care composition comprising from 0.025% to 99% by weight of a purified rhamnolipid mixture based on the total weight of the composition, wherein the rhamnolipid mixture comprises at least 10% mono-rhamnolipid (% by weight of total rhamnolipids), alternatively at least 15% mono-rhamnolipid, alternatively at least 20% mono-rhamnolipid, alternatively at least 25% mono-rhamnolipid, alternatively at least 30% mono-rhamnolipid, alternatively at least 35% mono-rhamnolipid, alternatively at least 40% mono-rhamnolipid, determined by high performance liquid chromatography (HPLC).

[0008] In another aspect, the present technology provides an oral care composition comprising from 0.025% to 99% by weight of a purified rhamnolipid mixture based on the total weight of the composition, wherein the rhamnolipid mixture comprises a weight ratio of from about 10:90 to about 47:53, alternatively from about 20:80 to about 47:53, alternatively from about 30:70 to about 47:53, alternatively from about 40:60 to about 47:53, alternatively from about 40:60 to about 45:55, alternatively 43:57 to 45:55 of mono-rhamnolipid and di-rhamnolipid, from about 5% to about 38% by weight, alternatively 29% to about 40% by weight, alternatively from about 29% to about 37.5% by weight of Rha-C10-C10 mono-rhamnolipid salt based on the total weight of the rhamnolipids in the rhamnolipid mixture, and from about 34% to about 80% by weight, 34% to about 45% by weight, alternatively 36% to about 45% by weight of RhaRha-C10-C10 di-rhamnolipid salt based on the total weight of the rhamnolipids, as determined by high performance liquid chromatography (HPLC).

[0009] In some embodiments, the purified rhamnolipid mixture can comprise from about 5% to about 38% by weight of Rha-C10-C10 mono-rhamnolipid based on the total weight of the rhamnolipids, and from about 34% to about 80% by weight of RhaRha-C10-C10 di-rhamnolipid based on the total weight of the rhamnolipids, as measured by high performance liquid chromatography (HPLC).

[0010] In some embodiments, the purified rhamnolipid mixture comprises a weight ratio of 40:60 to 45:55 of mono-rhamnolipid and di-rhamnolipid, the amount of Rha-C10-C10 mono-rhamnolipid is about 29% to 37.5% by weight based on the total weight of the rhamnolipids, and the amount of RhaRha-C10-C10 di-rhamnolipid is about 36% to 45% by weight based on the total weight of the rhamnolipids, as measured by high performance liquid chromatography (HPLC).

[0011] Another aspect of the present technology provides a method of treating or preventing gingivitis in a subject suffering from or susceptible to gingivitis, comprising administering to the subject an effective amount of a composition comprising a rhamnolipid mixture, such as described above, thereby treating or preventing gingivitis in the subject.

[0012] Another aspect of the present technology provides a method for treating or preventing gingivitis in a subject suffering from or prone to gingivitis, comprising administering to the subject an effective amount of a composition comprising a mixture of purified rhamnolipids, thereby treating or preventing gingivitis in the subject, wherein the mixture of rhamnolipids comprises mono-rhamnolipid and di-rhamnolipid, and the weight ratio of mono-rhamnolipid:di-rhamnolipid is from 10:90 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 15:85 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 20:80 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 90:10, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 80:20, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 70:30, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 60:40, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 50:50, alternatively mono-rhamnolipid:di-rhamnolipid is from 30:70 to 45:55, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 60:40, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 50:50, alternatively mono-rhamnolipid:di-rhamnolipid is from 40:60 to 45:55, as measured by high performance liquid chromatography (HPLC).

[0013] In some embodiments, the present technology provides a method for treating or preventing gingivitis in a subject suffering from or prone to gingivitis, as described above, wherein the purified rhamnolipid mixture may comprise from about 5% to about 38% by weight of Rha-C10-C10 mono-rhamnolipid based on the total weight of the rhamnolipids, and from about 34% to about 80% by weight of RhaRha-C10-C10 di-rhamnolipid based on the total weight of the rhamnolipids, as measured by high performance liquid chromatography (HPLC).

[0014] In some embodiments, the present technology provides a method for treating or preventing gingivitis in a subject suffering from or prone to gingivitis, as described above, wherein the mixture of purified rhamnolipids comprises mono-rhamnolipid and di-rhamnolipid in a weight ratio of 40:60 to 45:55, the amount of Rha-C10-C10 mono-rhamnolipid is about 29% to 37.5% by weight based on the total weight of the rhamnolipids, and the amount of RhaRha-C10-C10 di-rhamnolipid is about 36% to 45% by weight based on the total weight of the rhamnolipids, as measured by high performance liquid chromatography (HPLC).

[0015] Another aspect of the present technology provides a method for preventing the growth of oral bacteria and / or reducing the number of oral bacteria in a subject, whether in planktonic or biofilm form, comprising administering to the subject an effective amount of a composition comprising a rhamnolipid mixture, such as described above, to provide an inhibitory and / or killing effect on the target microorganism.

[0016] Another aspect of the present technology provides a method for preventing the growth of oral bacteria and / or reducing the number of oral bacteria in a subject, whether in planktonic or biofilm form, comprising administering to the subject an effective amount of a composition comprising a rhamnolipid mixture to provide an inhibitory and / or killing effect on the target microorganism, wherein the rhamnolipid mixture comprises mono-rhamnolipid and di-rhamnolipid, and the weight ratio of mono-rhamnolipid to di-rhamnolipid is from about 10:90 to about 47:53, alternatively from about 20:80 to about 47:53, alternatively from about 30:70 to about 47:53, alternatively from about 40:60 to about 47:53, alternatively from about 40:60 to about 45:55, alternatively 43:57 to 45:55 mono-rhamnolipid:di-rhamnolipid, measured by high performance liquid chromatography (HPLC).

[0017] In some embodiments, the present technology provides a method for preventing the growth of oral bacteria and / or reducing the number of oral bacteria in a subject, whether in planktonic or biofilm form, as described above, wherein the rhamnolipid mixture may comprise from about 5% to about 38% by weight of Rha-C10-C10 mono-rhamnolipid based on the total weight of the rhamnolipids, and from about 34% to about 80% by weight of RhaRha-C10-C10 di-rhamnolipid based on the total weight of the rhamnolipids, measured by high performance liquid chromatography (HPLC).

[0018] In some embodiments, the present technology provides a method for preventing the growth of oral bacteria and / or reducing the number of oral bacteria in a subject, whether in planktonic or biofilm form, as described above, wherein the rhamnolipid mixture comprises a weight ratio of mono-rhamnolipid to di-rhamnolipid of 40:60 to 45:55, the amount of Rha-C10-C10 mono-rhamnolipid is about 29% to 37.5% by weight based on the total weight of the rhamnolipids, and the amount of RhaRha-C10-C10 di-rhamnolipid is about 36% to 45% by weight based on the total weight of the rhamnolipids, measured by high performance liquid chromatography (HPLC).

[0019] In another aspect of the present technology, the purified rhamnolipid compositions disclosed herein are associated with certain biological activities. One such property is the ability to reduce the viability and biological activity (e.g., growth) of Porphyromonas gingivalis. For example, in one aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum inhibitory concentration (MIC) against Porphyromonas gingivalis, as determined as described in Example 1 (Antibacterial and Bactericidal Efficacy Tests Against Porphyromonas gingivalis). In this regard, as determined by the method described in Example 1 (Antibacterial and Bactericidal Efficacy Tests Against Porphyromonas gingivalis), the rhamnolipid compositions disclosed herein exhibit an MIC against Porphyromonas gingivalis of from about 21 ppm to about 106 ppm, from about 25 ppm to about 106 ppm, from about 30 ppm to about 106 ppm, from about 35 ppm to about 106 ppm, from about 40 ppm to about 106 ppm, or from about 41 ppm to about 106 ppm.

[0020] In another aspect of the present technology, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum bactericidal concentration (MBC) against Porphyromonas gingivalis, as determined as described in Example 1 (Antibacterial and Bactericidal Efficacy Tests Against Porphyromonas gingivalis). In this regard, as determined by the method described in Example 1 (Antibacterial and Bactericidal Efficacy Tests Against Porphyromonas gingivalis), the rhamnolipid compositions disclosed herein exhibit an MBC against Porphyromonas gingivalis of from about 62 ppm to about 296 ppm, from about 70 ppm to about 296 ppm, from about 80 ppm to about 296 ppm, from about 90 ppm to about 296 ppm, from about 100 ppm to about 296 ppm, from about 110 ppm to about 296 ppm, from about 120 ppm to about 296 ppm, or from about 123 ppm to about 296 ppm.

[0021] In yet another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum inhibitory concentration (MIC) against Streptococcus mutans, as determined as described in Example 2 (Antibacterial and Bactericidal Efficacy Tests Against Streptococcus mutans). In this regard, as determined by the method described in Example 2 (Antibacterial and Bactericidal Efficacy Tests Against Streptococcus mutans), the rhamnolipid compositions disclosed herein exhibit an MIC against Streptococcus mutans of from about 62 ppm to about 889 ppm, from about 70 ppm to about 889 ppm, from about 80 ppm to about 889 ppm, from about 90 ppm to about 889 ppm, from about 100 ppm to about 889 ppm, from about 110 ppm to about 889 ppm, from about 120 ppm to about 889 ppm, or from about 123 ppm to about 889 ppm.

[0022] In another aspect of the present technology, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum bactericidal concentration (MBC) against Streptococcus mutans, as determined as described in Example 2 (Test for antibacterial and bactericidal efficacy against Streptococcus mutans). In this regard, as determined by the method described in Example 2 (Test for antibacterial and bactericidal efficacy against Streptococcus mutans), the rhamnolipid compositions disclosed herein exhibit an MBC against Streptococcus mutans of from about 556 ppm to about 2666 ppm, from about 600 ppm to about 2666 ppm, from about 700 ppm to about 2666 ppm, from about 800 ppm to about 2666 ppm, from about 900 ppm to about 2666 ppm, from about 1000 ppm to about 2666 ppm, from about 1100 ppm to about 2666 ppm, or from about 1111 ppm to about 2666 ppm.

[0023] In yet another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum inhibitory concentration (MIC) against Campylobacter rectus, as determined as described in Example 3 (Test for antibacterial and bactericidal efficacy against Campylobacter rectus). In this regard, as determined by the method described in Example 3 (Test for antibacterial and bactericidal efficacy against Campylobacter rectus), the rhamnolipid compositions disclosed herein exhibit an MIC against Campylobacter rectus of from about 21 ppm to about 98 ppm, from about 25 ppm to about 98 ppm, from about 30 ppm to about 98 ppm, from about 35 ppm to about 98 ppm, from about 40 ppm to about 98 ppm, or from about 41 ppm to about 98 ppm.

[0024] In yet another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum bactericidal concentration (MBC) against Campylobacter rectus, as determined as described in Example 3 (Test for antibacterial and bactericidal efficacy against Campylobacter rectus). In this regard, as determined by the method described in Example 3 (Test for antibacterial and bactericidal efficacy against Campylobacter rectus), the rhamnolipid compositions disclosed herein exhibit an MBC against Campylobacter rectus of from about 21 ppm to about 98 ppm, from about 25 ppm to about 98 ppm, from about 30 ppm to about 98 ppm, from about 35 ppm to about 98 ppm, from about 40 ppm to about 98 ppm, or from about 41 ppm to about 98 ppm.

[0025] In another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic Log 10 reduction against Porphyromonas gingivalis, as determined as described in Example 7 (the amount of rhamnolipid is 0.1% by weight of the composition and the contact time is 30 seconds), a reduction of from about 3 to about 6, from about 3.5 to about 6, from about 4 to about 6, from about 4.5 to about 6, from about 5 to about 6, or from about 5.5 to about 6.

[0026] In another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic Log 10Reduction, as determined in Example 7 (the amount of rhamnolipid present is 0.05% by weight of the composition and the contact time is 30 seconds), is a reduction of about 2.5 to about 6, about 3 to about 6, about 3.5 to about 6, about 4 to about 6, about 4.5 to about 6, about 5 to about 6, or about 5.5 to about 6.

[0027] In another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic Log 10 Reduction, as determined in Example 7 (the amount of rhamnolipid present is 0.025% by weight of the composition and the contact time is 30 seconds), is a reduction of about 2 to about 6, about 2.5 to about 6, about 3 to about 6, about 3.5 to about 6, about 4 to about 6, about 4.5 to about 6, about 5 to about 6, or about 5.5 to about 6. Detailed Description of the Invention

[0029] Definitions

[0030] The "bioregenerable carbon index" (BCI) refers to the calculation of the percentage of carbon from bioregenerable sources and is calculated based on the following: dividing the amount of bioregenerable carbon by the total amount of carbon in the entire molecule.

[0031] As used herein, "bioregenerable" is defined as being derived from animal, plant, or marine materials.

[0032] As used herein, an "effective amount" means an amount of an active ingredient or composition that, when administered to a subject, is capable of preventing a recognized microorganism in the subject's oral cavity and / or reducing the amount of a recognized microorganism. The actual amount may vary depending on a variety of factors, including but not limited to the microorganism titer, the presence and amount of biofilm, the age and health status of the subject, and the form of administration.

[0033] As defined herein, "rhamnolipid" is a glycolipid having a lipid moiety (comprising one or more generally linear, saturated or unsaturated β-hydroxycarboxylic acid moieties) and a sugar moiety having one or more rhamnose units.

[0034] As defined herein, unless otherwise stated, "Log 10 Reduction" is calculated using the following formula: Log 10 Reduction = Log 10 (untreated) - Log 10 (treated).

[0035] The sugar moiety and the lipid moiety are linked by a β-glycosidic bond between the 1-OH group of the rhamnose moiety of the sugar moiety and the 3-OH group of the β-hydroxycarboxylic acid of the lipid moiety. Accordingly, the carboxylic acid of one carboxylic acid moiety serves as the terminus of the rhamnolipid. When the rhamnolipid contains more than one rhamnose moiety, each rhamnose moiety not linked to the lipid moiety is linked to another rhamnose moiety by a 1-4 β-glycosidic bond. In embodiments where there are two or more β-hydroxycarboxylic acids in the rhamnolipid, the β-hydroxycarboxylic acid moieties are independently selected from each other. In some embodiments, the β-hydroxycarboxylic acid moieties may be the same. In some embodiments, they are different from each other.

[0036] This technology generally relates to oral care compositions comprising specific rhamnolipid mixtures. Although rhamnolipids have been found to have antibacterial activity, the specific rhamnolipid mixtures described herein provide better efficacy against anaerobic bacteria in oral care applications compared to other rhamnolipid mixtures known in the art. The rhamnolipids in the rhamnolipid mixture of this technology may have the following structure (I):

[0037]

[0038] In this formula, R 9 is a hydrogen atom (H) or an aliphatic group, the main chain of which has from 1 to about 46, such as from 1 to about 42, from 1 to about 40, from 1 to about 38, from 1 to about 36, from 1 to about 34, from 1 to about 30, from 1 to about 28, including for example 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 or 28 carbon atoms and from 1 to about 3 (including 2) oxygen atoms. In some embodiments, the main chain of the corresponding aliphatic group bears a terminal carboxylic acid group and / or an internal ester group. As an illustrative example in this regard, R 9 may be of the formula -CH(R 5 )----CH 2 -COOR 6 . In these illustrative moieties, R 5 may be an aliphatic moiety having a main chain length of from 1 to about 19, such as from 1 to about 17, from 1 to about 15, from 1 to about 13, from about 2 to about 13, from about 3 to about 13 or from about 4 to about 13, including for example 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. R 4 in formula (I) is a hydrogen atom (H), or a rhamnopyranosyl moiety. R 6 is a hydrogen atom.

[0039] Unless otherwise specified, the term "aliphatic" refers to straight-chain or branched hydrocarbon chains, which can be saturated, mono-unsaturated or poly-unsaturated, and contain heteroatoms. As used herein, the term "heteroatom" refers to an atom of any element other than carbon or hydrogen. In the present text, unsaturated aliphatic groups contain one or more double bonds (alkenyl moieties). The branches of the hydrocarbon chain can contain straight chains as well as non-aromatic cyclic elements. Unless otherwise specified, the hydrocarbon chain can be of any length and contain any number of branches. Generally, the hydrocarbon (main) chain contains from 1 to about 5, to about 10, to about 15 or to about 20 carbon atoms. Examples of alkenyl moieties are straight-chain or branched hydrocarbon moieties containing one or more double bonds. Alkenyl moieties generally contain from about 2 to about 20 carbon atoms and one or more (e.g., two) double bonds (such as from about 2 to about 10 carbon atoms) and one double bond. Examples of alkyl groups are methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, the n isomers of these groups, isopropyl, isobutyl, isopentyl, sec-butyl, tert-butyl, neopentyl, 3,3-dimethylbutyl. Both the main chain and the branches can also contain heteroatoms, such as N, O, S, Se or Si, or a carbon atom can be replaced by one of these heteroatoms. The aliphatic moiety can be substituted or unsubstituted with one or more functional groups. The substituents can be any functional group, such as but not limited to amino, amido, carbonyl, carboxyl, hydroxyl, nitro, thio and sulfonyl.

[0040] In a more specific embodiment, the rhamnolipid or rhamnolipid salt in the structure has structure (II):

[0041]

[0042] wherein x is 1 or 2; y is 4, 6 or 8; z is 4, 6 or 8; and M is H, or a metal, such as an alkali metal Li, Na or K, an alkaline earth metal Mg or Ca, or a transition metal Mn, Fe, Cu or Zn. In the case of alkaline earth metals and transition metals, multiple rhamnolipid salt moieties can be combined with each metal.

[0043] The mixture of rhamnolipids contains a mixture of mono-rhamnolipids and di-rhamnolipids.

[0044] In some embodiments, the mono-rhamnolipid is present in an amount of 10% to 90%, alternatively about 15% to 90%, alternatively about 20% to about 90%, alternatively about 30% to about 90%, alternatively about 40% to about 90%, alternatively about 10% to about 80%, alternatively about 15% to about 80%, alternatively about 20% to about 80%, alternatively about 30% to about 80%, alternatively about 40% to about 80%, alternatively about 30% to about 70%, alternatively about 30% to about 60%, alternatively about 30% to about 50%, based on the total weight of the rhamnolipid, and is measured by high performance liquid chromatography (HPLC).

[0045] In some embodiments, the mono-rhamnolipid is present in an amount of 10% to about 47%, alternatively about 15% to about 45%, alternatively about 20% to about 45%, alternatively about 30% to about 45%, alternatively about 40% to about 45%, alternatively about 43% to about 45%, 10% to about 48%, alternatively about 15% to about 48%, alternatively about 20% to about 48%, alternatively about 30% to about 48%, alternatively about 40% to about 48%, alternatively about 43% to about 48%, alternatively about 10% to about 47%, alternatively about 15% to about 47%, alternatively about 20% to about 47%, alternatively about 30% to about 47%, alternatively about 40% to about 47%, alternatively about 43% to about 47%, based on the total weight of the rhamnolipid.

[0046] The di-rhamnolipid is present in an amount of about 52% to about 90% by weight, alternatively about 52% to about 85%, alternatively about 52% to about 80%, alternatively about 52% to about 70%, alternatively about 52% to about 60%, alternatively about 52% to about 57%, alternatively about 53% to about 90% by weight, alternatively about 53% to about 85%, alternatively about 53% to about 80%, alternatively about 53% to about 70%, alternatively about 53% to about 60%, alternatively about 53% to about 57%, alternatively about 54% to about 85%, alternatively about 55% to about 80%, alternatively about 55% to about 70%, alternatively about 55% to about 60%, alternatively about 55% to about 57% by weight, based on the total weight of the rhamnolipid, and is measured by high performance liquid chromatography (HPLC). The ratio of mono-rhamnolipid:di-rhamnolipid can be from about 10:90 to about 48:52, alternatively about 47:53, alternatively from about 40:60 to about 45:55. In some embodiments, the ratio of mono-rhamnolipid:di-rhamnolipid can be from 43.5:56.5 to 45:55, alternatively from 43:57 to 45:55 or from 43:57 to 48:52.

[0047] The rhamnolipid mixture preferably comprises mono- (where x = 1) and di- (where x = 2) rhamnolipids, where y and z are 6 and M is H or Na. The mono-rhamnolipid can be referred to as Rha-C10-C10, with the molecular formula C 26 H 48 O 9 . Its IUPAC name is 3-[3-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxydecanoyloxy]decanoic acid. The di-rhamnolipid can be referred to as RhaRha-C10-C10, with the molecular formula C 32 H 58 O 13 . Its IUPAC name is 3-[3-[4,5-dihydroxy-6-methyl-3-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid. Rha-C10-C10 can be present in the mixture in the following amounts: about 5% to 38%, alternatively about 10% to about 38%, alternatively about 15% to about 37.5%, alternatively about 20% to about 37.5%, alternatively about 25% to about 37.5%, alternatively about 29% to about 37.5%, alternatively about 35% to about 37% by weight, based on the total weight of the rhamnolipids, as measured by high performance liquid chromatography (HPLC). RhaRha-C10-C10 can be present in the mixture in the following amounts: about 34% to about 80%, alternatively about 35% to about 75%, alternatively about 35% to about 65%, alternatively about 35% to about 60%, alternatively about 35% to about 55%, alternatively about 35.5% to about 50%, alternatively about 36% to about 45%, alternatively about 36% to about 38% by weight, based on the total weight of the rhamnolipids in the rhamnolipid mixture, as measured by high performance liquid chromatography (HPLC).

[0048] In addition to Rha-C10-C10 and RhaRha-C10-C10, the rhamnolipid mixture may further comprise RhaRha-C10-C12 in an amount of about 5% to about 15%, alternatively about 9% to about 12%, alternatively about 10% to about 12.5% by weight based on the total weight of the rhamnolipids, and Rha-C10-C12 in an amount of about 0.2% to about 6% by weight, alternatively about 2% to about 5%, alternatively about 3.5% to about 5% by weight based on the total weight of the rhamnolipids, each measured by high performance liquid chromatography (HPLC). The rhamnolipid mixture may further comprise, for example, RhaRha-C10-C12:1 in an amount of about 0.2% to about 5% by weight, alternatively 1% to about 4% by weight based on the total weight of the rhamnolipids; RhaRha-C8-C10 in an amount within the range of about 0.2% to about 5% by weight, alternatively about 1% to about 4% by weight based on the total weight of the rhamnolipids; Rha-C8-C10 in an amount within the range of about 0.2% to about 5% by weight, alternatively about 1% to about 4% by weight based on the total weight of the rhamnolipids; or any combination thereof, each measured by high performance liquid chromatography (HPLC).

[0049] Rhamnolipids can be produced by rhamnolipid-producing microorganisms capable of synthesizing / producing rhamnolipids under suitable conditions. Such microorganisms include, but are not limited to, bacteria, particularly bacteria of the phyla Actinobacteria, Firmicutes, and Proteobacteria. Rhamnolipids are of natural origin and thus have a BCI of 100. In certain embodiments, the rhamnolipid-producing microorganism used for producing rhamnolipids is Pseudomonas aeruginosa.

[0050] Methods for culturing rhamnolipid-producing bacteria and methods for producing rhamnolipids by fermentation are known in the art, such as U.S. Patent No. 11,142,782 and U.S. Patent No. 10,144,943, the entire contents of which are incorporated herein by reference. Methods for purifying rhamnolipids are also known in the art, such as U.S. Patent No. 9,884,883 and U.S. Patent No. 10,829,507, the entire contents of which are incorporated herein by reference. The rhamnolipids in the rhamnolipid mixture used in the present technology are separated from the fermentation medium, washed, deodorized, decolorized, and neutralized to form purified rhamnolipid salts.

[0051] The content of the rhamnolipid component in the oral care composition can be 0.025% to 99% by weight, alternatively 0.050% to 99% by weight, alternatively 0.075% to 99% by weight, alternatively 0.25% to 99% by weight, alternatively 0.5% to 99% by weight, alternatively 0.1% to 99% by weight based on the total weight of the composition.

[0052] The oral care composition of the present technology can be in liquid form and contains at least one carrier suitable for oral care such that the total percentage of the composition reaches 100% (taking into account any additives that may be present in addition to rhamnolipids). As will be understood by at least those skilled in the art, various carriers, vehicles, diluents, etc. are suitable for the practice of the present technology. Therefore, it should also be understood that, for the purposes of the present technology and in describing its various formulations, applications, uses, and compositions, the terms "carrier", "vehicle", and "diluent" should be considered non-exhaustive and interchangeable.

[0053] Water is a suitable carrier and can be deionized water, hard water, soft water, distilled water, tap water, or a combination thereof. Water can be used alone as a carrier or in combination with other carriers suitable for oral care applications.

[0054] The oral care composition of the present technology can contain optional ingredients known in the art. Such other components or additives can include pH regulators, hydrotropic agents or other solubilizers for obtaining and maintaining a clear single-phase composition, emulsifiers, binders, excipients, stabilizers, chelating agents, gelling agents, buffers, surfactants, lubricants, flavoring components, sweeteners, colorants, vitamins, fluoride sources, preservatives, and other functional ingredients. In some embodiments, for example, the oral care composition can contain a fluoride source such as stannous fluoride, sodium fluoride, potassium fluoride, sodium monofluorophosphate, potassium monofluorophosphate, ammonium fluoride, ammonium monofluorophosphate, fluorinated amines, etc. In some embodiments, the oral care composition can also contain an abrasive. The abrasive can be, for example, a silica abrasive such as hydrated silica, precipitated silica, or silica gel, alternative abrasive materials such as phosphates, or a combination thereof. In some embodiments, the oral care composition can also contain a humectant such as sorbitol, glycerin, xylitol, polyethylene glycol, propylene glycol, or a combination thereof. In some embodiments, the oral care composition can contain a bicarbonate such as sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate, or a combination thereof. In some embodiments, the oral care composition can also contain hydrogen peroxide for tooth whitening.

[0055] In some embodiments, the oral care composition can comprise a foaming agent, such as high molecular weight polyethylene glycol (PEG). In one aspect, the rhamnolipids of the present technology can be formulated (or otherwise used) as a natural foaming agent, having properties similar to those of synthetic foaming agents. For example, the rhamnolipids of the present technology have foaming properties similar to those of sodium lauryl ether sulfate (SLES) in rinse-off applications (e.g., as shown using the standardized shaking foam method). Since the foaming mechanism in oral care has significantly greater shear forces in smaller and more confined areas, the foaming properties of the rhamnolipids disclosed herein can translate as a substitute for natural foaming agents in oral care. In some embodiments, the oral care composition can also comprise a desensitizing agent.

[0056] In some embodiments, particularly where the oral care composition is a mouthwash, the oral care composition can comprise an alcohol, such as ethanol.

[0057] In some embodiments, particularly where the oral care composition is toothpaste, the oral care composition can comprise a binder. Examples of suitable binders include carboxymethyl cellulose, hydroxyethyl cellulose, hydroxyethyl carboxymethyl cellulose, natural and synthetic gums, polyvinylpyrrolidone, colloidal carboxyvinyl polymer, algal colloids, and the like.

[0058] In some embodiments, the oral care composition can comprise a flavoring agent or flavorant. Suitable flavoring agents include wintergreen oil, peppermint oil, and spearmint oil.

[0059] The pH value of the oral care composition of the present technology can range from about 5 to about 12; alternatively from about 6 to about 8. As the rhamnolipids become protonated, the pH value decreases from 6 to 5, and the solubility of the individual rhamnolipid component in water decreases.

[0060] The oral care composition and / or formulation of the present technology can be in a variety of forms, such as aqueous solutions, gels, pastes, sprays, creams, homogeneous or heterogeneous mixtures, or powders. The end uses of the composition include, but are not limited to, dentifrices, oral rinses, mouthwashes, toothpastes, dental powders, dissolvable strips, lozenges, chewing gums, wipes, or any other oral care product known in the art. The oral care composition can be used to inhibit and / or reduce the appearance of plankton and / or the formation of biofilms (e.g., dental plaque), and has shown antibacterial and bactericidal efficacy against oral bacteria, including Porphyromonas gingivalis, Streptococcus mutans, and Campylobacter rectus.

[0061] In some embodiments, the oral care composition can be used to treat and / or prevent dental plaque, biofilm, and / or tartar formation; gingivitis, tooth decay, periodontal disease, or any other oral health condition associated with the presence or accumulation of oral bacteria. The frequency of administration of the oral care composition can be configured based on the specific type of the product, the strength of the product (the dose provided per use), the oral health condition of the user, and the like. In some embodiments, for example, the user care composition can be administered once a day, twice a day, three times a day, once every two days, once every three days, and so on.

[0062] The oral care composition can be administered prophylactically, i.e., to prevent dental plaque, biofilm, and / or tartar formation, gingivitis, tooth decay, periodontal disease, or any other oral health condition associated with the presence or accumulation of oral bacteria in an individual, or as part of a treatment plan, i.e., to treat dental plaque, biofilm, and / or tartar formation, gingivitis, tooth decay, periodontal disease, or any other oral health condition associated with the presence or accumulation of oral bacteria in an individual suffering from one or more oral health conditions. For prophylactic treatment, the individual may not be suffering from the oral health condition that the oral care composition is intended to prevent. However, in some embodiments, the individual may be particularly predisposed or susceptible to the oral health condition that the oral care composition is intended to prevent.

[0063] When used to treat dental plaque, biofilm, and / or tartar formation, gingivitis, tooth decay, periodontal disease, or any other oral health condition associated with the presence or accumulation of oral bacteria in an individual suffering from one or more oral health conditions, the dosing regimen can continue for one week, two weeks, three weeks, four weeks, five weeks, six weeks, or more weeks, depending on the patient's condition and response. At the end of the treatment, the oral care composition or a variant thereof (e.g., a composition containing a lower dose of the active ingredient) can continue to be used as a prophylactic treatment.

[0064] Those skilled in the art will recognize that the technology can be modified without departing from the spirit or scope of the invention. The invention is further illustrated by the following examples, which should not be construed as limiting the spirit or scope of the invention to the specific procedures or compositions described therein. Examples

[0065] Example 1: Antibacterial and Bactericidal Efficacy Test Against Porphyromonas gingivalis

[0066] Using the composition containing only rhamnolipid active ingredient disclosed herein, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against the representative oral care-related microorganism Porphyromonas gingivalis were determined.

[0067] As used herein, the MIC is the concentration of rhamnolipid at which no growth of the target bacteria is detected relative to the control after sufficient incubation in nutrient broth medium. The MIC is determined according to industry standards (Clinical and Laboratory Standards Institute (CLSI) M7, M11, and M26), where samples are serially diluted, typically 1:2 or 1:3, in a nutrient medium suitable for the growth of the test microorganism and are sufficiently pre-reduced to remove oxygen before anaerobic testing. The pre-reduction is carried out by achieving indirect atmospheric sequestering of oxygen through the reaction of hydrogen on a palladium catalyst. Subsequently, the samples are exposed to a test challenge microorganism at a final concentration of at least 1.0E+06 CFU / mL. After an initial incubation period where the positive control (nutrient medium, microorganism, and water instead of the active ingredient) is sufficient to develop mature and measurable growth, the presence / absence of growth in the samples is visually evaluated by turbidity assessment. A sample that does not become turbid compared to the positive control and is comparable to the negative control marks the concentration of inhibition.

[0068] As used herein, the MBC is the lowest concentration of an antimicrobial composition required to kill a specific target bacteria. The MBC is determined according to industry standards CLSI M7, M11, and M26. For test samples that show bacteriostatic effects in the MIC test, their bactericidal properties are further detected by the following operations: aliquots taken from each test concentration are neutralized and plated onto an appropriate agar medium to determine the presence of survivors. Samples showing a ≥3.0 Log 10 reduction (≥99.9% reduction) compared to the control are determined as the bactericidal concentration.

[0069] A representative strain tested is Porphyromonas gingivalis (Pg) ATCC 33277.

[0070] The representative rhamnolipid compositions tested include:

[0071]

[0072] The efficacy of these rhamnolipid samples against Porphyromonas gingivalis ATCC 33277 was tested, and the minimum inhibitory concentration (MIC) values and minimum bactericidal concentration (MBC) values shown are presented in Table 1.

[0073] Table 1

[0074]

[0075] Using common microbiological methods known as the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), the rhamnolipid mixtures tested showed improved bacteriostatic and bactericidal efficacy against Porphyromonas gingivalis ATTC 33277.

[0076] Example 2: Antibacterial and Bactericidal Efficacy Testing of Streptococcus mutans

[0077] The MIC and MBC values of the representative rhamnolipid compositions of the present disclosure against the oral care-related microorganism Streptococcus mutans were also determined.

[0078] The representative strain tested was Streptococcus mutans ATCC 25175. The rhamnolipid composition tested was Sample 1 used for Porphyromonas gingivalis above. The comparative composition was the same comparative sample used for Porphyromonas gingivalis above.

[0079] As shown in Table 2, the rhamnolipid compositions of Sample 1 and the comparative sample showed MIC values of 123 ppm and 1111 ppm, respectively. The rhamnolipid compositions of Sample 1 and the comparative sample also showed MBC values of 1111 ppm and 3333 ppm, respectively. Both compositions were tested against Streptococcus mutans ATCC 25175.

[0080] Table 2

[0081]

[0082] Using common microbiological methods called minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), the rhamnolipid mixtures tested showed improved antibacterial and bactericidal efficacy against Streptococcus mutans ATCC 25175.

[0083] Example 3: Antibacterial and Bactericidal Efficacy Testing of Campylobacter rectus

[0084] The MIC and MBC values of the representative rhamnolipid compositions of the present disclosure against the oral care-related microorganism Campylobacter rectus were also determined.

[0085] The representative strain tested was Campylobacter rectus ATCC 33238. The rhamnolipid composition tested was Sample 1 used for Porphyromonas gingivalis above. The comparative composition was the same comparative sample used for the Porphyromonas gingivalis example above.

[0086] As shown in Table 3, the rhamnolipid compositions of Sample 1 and the comparative sample showed MIC values of 41 ppm and 123 ppm, respectively. The rhamnolipid compositions of Sample 1 and the comparative sample also showed MBC values of 41 ppm and 123 ppm, respectively. The efficacy of both compositions against Campylobacter rectus (Cr) ATCC 33238 at 1.9E+07 CFU / mL was tested in Modified Reinforced Clostridial Broth (RMCB) at pH 6.8 ± 0.2 after 28 hours of exposure.

[0087] Table 3

[0088]

[0089] Using common microbiological methods known as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), the tested rhamnolipid mixtures showed improved antibacterial and bactericidal efficacy against Campylobacter rectus ATCC 33238.

[0090] The above examples demonstrate that the rhamnolipid mixtures of the present disclosure have antibacterial and bactericidal effects against oral care-related microorganisms, including, for example, Porphyromonas gingivalis, Streptococcus mutans, and Campylobacter rectus. Surprisingly, the above examples also show that increasing the weight percentage of mono-rhamnolipid, i.e., increasing the ratio of mono-rhamnolipid to di-rhamnolipid to be higher than the 4:96 ratio found in comparative commercially available rhamnolipid products, can result in improved antibacterial and / or bactericidal effects against oral care-related microorganisms, including, for example, Porphyromonas gingivalis, Streptococcus mutans, and Campylobacter rectus.

[0091] Example 4: Qualitative dissociation test of Porphyromonas gingivalis biofilm

[0092] This test method evaluates the ability of the test compound to prevent the growth and establishment of Porphyromonas gingivalis biofilm.

[0093] The representative strain tested was Porphyromonas gingivalis (Pg) ATCC 33277. The rhamnolipid composition tested was Sample 3 for Porphyromonas gingivalis in Table 1 above.

[0094] First, an established 1:10 culture of Porphyromonas gingivalis was prepared in a nutrient medium. Then, 180 μL of the test culture and 20 μL of a sterile deionized water control or one of the following were dispensed into a 96-well microtiter plate:

[0095] i. A solution containing rhamnolipid at an active substance level of 2.6%;

[0096] ii. A solution containing rhamnolipid at an active substance level of 1.6%;

[0097] iii. A solution containing rhamnolipid at an active substance level of 0.6%;

[0098] More specifically, four wells of the plate were filled with 180 μL of the culture and 20 μL of the 2.6% active solution, four wells of the plate were filled with 180 μL of the culture and 20 μL of the 1.6% active substance solution, and four wells of the plate were filled with 180 μL of the culture and 20 μL of the 0.6% active substance solution. Each set of four wells was accompanied by a set of four adjacent control wells, each containing 180 μL of the culture and 20 μL of sterile deionized water.

[0099] The lid was placed on the microtiter plate, and then the plate was incubated at a temperature of about 37 ± 2 °C for about 72 ± 2 hours.

[0100] At the end of the incubation period, each well was stained with 0.4% crystal violet, and the results were evaluated by the following staining score:

[0101] 0 – Colorless

[0102] 1 – Very light staining

[0103] 2 – Medium light staining

[0104] 3 – Medium staining

[0105] 4 – Medium dark

[0106] 5 – Dark.

[0107] Based on visual observation, it was concluded that in the presence of the active rhamnolipid mixture, the growth of Porphyromonas gingivalis produced an observed score of 2 (medium light staining), compared to a control observed score of 5 (dark). Therefore, based on the results measured by this qualitative method, it was concluded that the introduction of the rhamnolipid mixture dissociated the established Porphyromonas gingivalis biofilm.

[0108] Example 5: Quantitative dissociation test of Porphyromonas gingivalis biofilm

[0109] This test method evaluates the ability of a test compound to dissolve, remove, and / or kill an already formed Porphyromonas gingivalis biofilm.

[0110] The representative strain tested was Porphyromonas gingivalis (Pg) ATCC 33277.

[0111] Rhamnolipid composition products at 0.165% by weight and 0.325% by weight were evaluated to assess the ability to dissociate an already formed Porphyromonas gingivalis biofilm. This test was conducted on common oral care organisms at a set contact time suitable for industrial use. First, the test organisms were grown in tryptic soy broth medium (ATCC medium 2722) supplemented for 3 days, then 270 μl of the grown culture was transferred to a sterile 96-well plate and aged for 48 ± 2 hours to allow the culture to form a mature biofilm on the well plate walls and reach a high titer.

[0112] At the start of the test, aspirate the culture medium from the inoculation wells, taking care not to disrupt the biofilm. Add 270 μl of sterile deionized water to the evaluation wells and 30 μl of sterile deionized water to the initial 270 μl of sterile deionized water wells to complete a 1:10 dilution, which is evaluated as a control for the selected contact time. After the contact time exposure, aspirate all the contents, still taking care not to disrupt the biofilm present on the well walls. Wash the wells twice with sterile deionized water to help quench the active substances. Repeat the exposure process for each test product (0.165% by weight product and 0.325% by weight product) for evaluation. To complete the 1:10 dilution for exposure, add 270 μl of sterile deionized water to the inoculation wells and 30 μl of the test product to the initial sterile deionized water wells, evaluate the sample efficacy for the selected contact time, and then wash twice to quench.

[0113] After exposure and washing, dry the plate in an oven for approximately 30 minutes. Add 270 μl of 0.1% crystal violet to each evaluation well and let stand for approximately 15 minutes. Immerse the plate in deionized water, then flip and blot dry on a paper towel, performing this 3 times independently, changing the water between each immersion. Let the plate air dry for approximately 10 minutes, then add 270 μl of 33% glacial acetic acid to each treated well to resuspend the bound dye to the adherent cells. Measure the OD absorbance at 590 nm and measure the % dissociation relative to the positive control absorbance value.

[0114] As shown in Table 4, after a 30-second contact time, the 0.325% by weight rhamnolipid composition produced 80% effective dissociation against the common oral care organism Porphyromonas gingivalis. The rhamnolipid composition produced comparable dissociation efficacy results.

[0115] Table 4

[0116]

[0117] Example 6: Flavorant Solubilization

[0118] The ability of the rhamnolipid mixture to solubilize oil-based flavorants, such as peppermint oil flavorants, was tested. The rhamnolipid composition tested was Sample 3 for Porphyromonas gingivalis described above.

[0119] In the absence of synthetic solubilizers or other solubilizing agents, the rhamnolipid mixture was used as a potential solubilizing agent component for flavor oils, and its functionality was determined through a series of dilutions. The rhamnolipid composition was compared with polysorbate 20, which is a synthetic solubilizer and also the industry standard for flavor oil solubilization. Common peppermint flavorants, such as Peppermint #244456 and Peppermint #244455 from BelleAire Creations, were used as flavor oils.

[0120] The rhamnolipid composition was added to each flavor oil in increasing amounts to determine the ratio of solubilizer component to flavor oil to obtain a clear premix. Once the clear premix was obtained, it was added to deionized water to ensure a clear and transparent final solution.

[0121] The results showed that the solubilization ratio of polysorbate 20 was 1:6 (flavor oil: polysorbate 20), while the solubilization ratio of the rhamnolipid mixture was 1:13 (flavor oil: rhamnolipid mixture). In both cases, the final solution of the premix in deionized water was clear and transparent.

[0122] It is believed that the 1:13 ratio obtained with the rhamnolipid mixture is sufficient for the rhamnolipid mixture to be an acceptable solubilizer for oil-based flavorants such as mint flavorants. This ability of the rhamnolipid composition may be particularly important in certain embodiments or composition types, as compositions that are semi-transparent to opaque in appearance may not be aesthetically pleasing and thus commercially undesirable.

[0123] Example 7: Prevention and Time-to-Kill Efficacy Testing of Porphyromonas gingivalis

[0124] The efficacy of a representative rhamnolipid composition of the present disclosure against Porphyromonas gingivalis was evaluated. The test was conducted at a set contact time suitable for the end use. The selected test organism was grown in the aforementioned medium for 3 days and then subcultured again for an additional 48 ± 2 hours to allow the culture to mature and reach a high titer.

[0125] At the start of the test, a 270 ul volume of the prepared microbial titer was pipetted into a 96-well plate. 30 uL of sterile deionized water was added to prepare a control sample. For the test sample, 30 uL was used instead. After the contact time exposure, a 1:10 dilution was applied and / or the active agent was neutralized with a neutralizing agent. The sample was further diluted and stained with viability dyes (Syto 9 and propidium iodide) and analyzed by flow cytometry. The Log 10 Reduction was calculated using the flow cytometry data.

[0126] As shown in Table 5, compared to the comparative rhamnolipid composition (at the same concentration and contact time), the rhamnolipid compositions of the present disclosure (rhamnolipid compositions of the present disclosure at concentrations of 0.1%, 0.05%, and 0.025% by weight, and when formulated into a finished product (having 0.1%, 0.05%, and 0.025% by weight of rhamnolipid based on the total weight of the composition)) showed (at each concentration) a greater reduction in the Porphyromonas gingivalis titer (within 30 seconds of contact time), and was comparable to the reduction of the tested national brand mouthwash sample (i.e., Crest Pro-Health Advanced Multi-Protection containing 0.02% active ingredient sodium fluoride).

[0127] The formulated product B (FP-B) in Table 5 contains poloxamer 407 (Polyoxamer 407), sodium benzoate, citric acid, and an active ingredient (i.e., rhamnolipid sample 3). The formulated product C (FP-C) in Table 5 is a control and contains the same materials as the corresponding FP-B, except for rhamnolipid sample 3 and citric acid.

[0128] To prepare each of the FP-B compositions (Table 5) tested below, a first FP-B bulk composition was prepared, which contained: 0.29% poloxamer 407 (% by weight, based on the total weight of the composition); 0.19% sodium benzoate (% by weight, based on the total weight of the composition); 0.06% citric acid (% by weight, based on the total weight of the composition); and 1.0% rhamnolipid sample 3 (% by weight, based on the total weight of the composition). To prepare the individual FP-B samples tested below, the FP-B bulk composition was diluted 1 / 10 (for FP-B with 0.1% active ingredient), 1 / 20 (for FP-B with 0.05% active ingredient), and 1 / 40 (for FP-B with 0.025% active ingredient).

[0129] To prepare each of the corresponding FP-C compositions (Table 5) tested below, an FP-C bulk composition was prepared, which contained: 0.30% poloxamer 407 (% by weight, based on the total weight of the composition); and 0.20% sodium benzoate (% by weight, based on the total weight of the composition). To prepare the individual FP-C samples tested in Table 6, the FP-C bulk composition was diluted 1 / 10 (for the FP-C control corresponding to FP-B with 0.1% active ingredient), 1 / 20 (for the FP-C control corresponding to FP-B with 0.05% active ingredient), and 1 / 40 (for the FP-C control corresponding to FP-B with 0.025% active ingredient).

[0130] Table 5

[0131]

[0132]

[0133] The embodiments and examples described herein are illustrative and do not limit the technology described herein in any way. The scope of the technology described in this specification is the full scope defined or implied by the claims. Additionally, any references cited in the detailed description of the invention in this application are hereby incorporated by reference in their entirety, unless otherwise noted.

[0134] The technology has now been described in sufficient, clear, and concise terms such that those skilled in the art to which the technology pertains can practice the technology. It should be understood that the preferred embodiments of the technology have been described above and that modifications can be made thereto without departing from the spirit or scope of the technology as set forth in the appended claims. Additionally, the examples provided are not exhaustive but are used to illustrate several embodiments that fall within the scope of the claims.

Claims

1. An oral care composition, comprising: (a) a rhamnolipid mixture in the following amounts: 0.1% to 99% by weight, alternatively 0.05% to 99% by weight, alternatively 0.025% to 99% by weight, based on the total weight of the composition, wherein the rhamnolipid mixture comprises mono-rhamnolipid and di-rhamnolipid, and the weight ratio of mono-rhamnolipid:di-rhamnolipid is from 10:90 to 90:10, preferably mono-rhamnolipid:di-rhamnolipid is from 40:60 to 60:40, more preferably mono-rhamnolipid:di-rhamnolipid is from 40:60 to 50:50, wherein the Rha-C10-C10 mono-rhamnolipid accounts for about 5% to about 38% by weight of the total weight of the rhamnolipid mixture, and the RhaRha-C10-C10 di-rhamnolipid accounts for about 34% to about 80% by weight of the total weight of the rhamnolipid mixture; and (b) at least one orally acceptable carrier, and optionally one or more additives, in an amount up to 100% of the total weight of the composition.

2. The oral care composition according to claim 1, wherein the rhamnolipid mixture comprises mono-rhamnolipid and di-rhamnolipid in a weight ratio of about 10:90 to about 47:

53.

3. The oral care composition according to any one of the preceding claims, wherein the amount of the Rha-C10-C10 mono-rhamnolipid is about 29% to about 37.5% by weight based on the total weight of the rhamnolipid mixture.

4. The oral care composition according to any one of the preceding claims, wherein the amount of the RhaRha-C10-C10 di-rhamnolipid is about 36% to about 45% by weight, preferably about 36% to about 38% by weight, based on the total weight of the rhamnolipid mixture.

5. The oral care composition according to any one of the preceding claims, wherein the amount of the RhaRha-C10-C12 di-rhamnolipid is about 5% to about 15%, preferably about 10% to about 12.5% by weight based on the total weight of the rhamnolipid mixture.

6. The oral care composition according to any one of the preceding claims, wherein the amount of the RhaRha-C10-C12:1 di-rhamnolipid is about 0.2% to about 5%, preferably about 1% to about 4% by weight based on the total weight of the rhamnolipid mixture.

7. The oral care composition according to any one of the preceding claims, wherein the amount of the RhaRha-C8-C10 di-rhamnolipid is about 0.2% to about 5%, preferably about 1% to about 4% by weight based on the total weight of the rhamnolipid mixture.

8. The oral care composition according to any one of the preceding claims, wherein the amount of the Rha-C10-C12 mono-rhamnolipid is about 0.2% to about 6%, preferably about 3.5% to about 5% by weight based on the total weight of the rhamnolipid mixture.

9. The oral care composition according to any one of the preceding claims, wherein the amount of the Rha-C8-C10 mono-rhamnolipid is from about 0.2% to about 5%, preferably from about 1% to about 4% by weight based on the total weight of the rhamnolipid mixture.

10. The oral care composition according to any one of the preceding claims, wherein the amount of rhamnolipid in the composition is from about 0.1% to about 25% by weight.

11. The oral care composition according to any one of the preceding claims, wherein the composition comprises one or more additives.

12. The oral care composition according to claim 11, wherein the one or more additives include one or more of surfactants, thickeners, preservatives, flavoring agents, coloring agents, humectants, buffers, chelating agents, sweetening agents, fluoride sources, and abrasives.

13. The oral care composition according to any one of the preceding claims, wherein the orally acceptable carrier comprises water.

14. The oral care composition according to any one of the preceding claims, wherein the composition exhibits an inhibitory effect against Porphyromonas gingivalis and has a minimum inhibitory concentration (MIC) of from about 21 ppm to about 106 ppm.

15. The oral care composition according to any one of the preceding claims, wherein the composition exhibits a bactericidal effect against Porphyromonas gingivalis and has a minimum bactericidal concentration (MBC) of from about 62 ppm to about 296 ppm.

16. The oral care composition according to any one of the preceding claims, wherein the composition exhibits an inhibitory effect against Streptococcus mutans and has a minimum inhibitory concentration (MIC) of from about 62 ppm to about 889 ppm.

17. The oral care composition according to any one of the preceding claims, wherein the composition exhibits a bactericidal effect against Streptococcus mutans and has a minimum bactericidal concentration (MBC) of from about 556 ppm to about 2666 ppm.

18. The oral care composition according to any one of the preceding claims, wherein the composition exhibits an inhibitory effect against Campylobacter rectus and has a minimum inhibitory concentration (MIC) of from about 21 ppm to about 98 ppm.

19. The oral care composition according to any one of the preceding claims, wherein the composition exhibits a bactericidal effect against Campylobacter rectus and has a minimum bactericidal concentration (MBC) of from about 21 ppm to about 98 ppm.

20. The oral care composition according to any one of the preceding claims, wherein when the amount of rhamnolipid present is 0.1% by weight of the composition and the contact time is 30 seconds, the composition exhibits a Log 10 reduction of 3.0 to 6 in Porphyromonas gingivalis.

21. The oral care composition according to any one of the preceding claims, wherein when the amount of rhamnolipid present is 0.05% by weight of the composition and the contact time is 30 seconds, the composition exhibits a Log 10 reduction of 2.5 to 6 in Porphyromonas gingivalis.

22. The oral care composition according to any one of the preceding claims, wherein when the amount of rhamnolipid present is 0.025% by weight of the composition and the contact time is 30 seconds, the composition exhibits a Log 10 reduction of 2.0 to 6 in Porphyromonas gingivalis.

23. The oral care composition according to any one of the preceding claims, wherein the pH of the composition is from 6.0 to 8.

0.

24. The oral care composition according to any one of the preceding claims, wherein the composition contains a flavoring agent and wherein the rhamnolipid mixture is present in an amount effective to dissolve the flavoring agent.

25. The oral care composition according to claim 24, wherein the composition does not contain a synthetic flavoring solubilizer.

26. The oral care composition according to any one of claims 24 - 25, wherein the flavoring agent is selected from: wintergreen oil, spearmint oil, peppermint oil, or any combination thereof.

27. An oral care composition according to any one of claims 24 - 26, wherein the rhamnolipid is present in an amount of 0.025% to 99% by weight based on the total weight of the composition.

28. An oral care composition according to any one of the preceding claims, wherein the rhamnolipid is present in an amount effective to act as a natural foaming agent.

29. An oral care composition according to claim 28, wherein the composition is free of synthetic foaming agents.

30. An oral care composition according to any one of claims 28 - 29, wherein the composition is a toothpaste.

31. An oral care composition according to any one of claims 1 - 29, wherein the composition is an oral rinse, mouthwash, toothpaste, tooth powder, dissolvable strip, lozenge or wipe.

32. A method of treating or preventing gingivitis in a subject suffering from or prone to gingivitis, comprising administering to the subject an effective amount of an oral care composition according to any one of the preceding claims, thereby treating or preventing gingivitis in the subject.

33. A method of treating or preventing gingivitis in a subject suffering from or prone to gingivitis, comprising administering to the subject an effective amount of an oral care composition according to any one of the preceding claims, thereby treating or preventing gingivitis in the subject; and wherein the rhamnolipid mixture comprises mono - rhamnolipid and di - rhamnolipid, and the weight ratio of mono - rhamnolipid:di - rhamnolipid is from 10:90 to 90:10, preferably mono - rhamnolipid:di - rhamnolipid is from 40:60 to 60:40, more preferably mono - rhamnolipid:di - rhamnolipid is from 40:60 to 50:50, wherein Rha - C10 - C10 mono - rhamnolipid accounts for about 5% to about 38% by weight of the total weight of the rhamnolipid mixture, and RhaRha - C10 - C10 di - rhamnolipid accounts for about 34% to about 80% by weight of the total weight of the rhamnolipid mixture; wherein the treating or preventing gingivitis comprises preventing or reducing the growth of Porphyromonas gingivalis.

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