Methods and compositions for controlling acne using rhamnolipids

Abstract

Disclosed is an acne treatment composition containing a mixture of rhamnolipid salts. The composition has the effect of aiming at echinodermis herpes. A method for managing, treating or preventing acne in a subject is also disclosed.

Description

Background Art

[0001] The present technology generally relates to the use of biosurfactants, such as rhamnolipids, for the treatment, control or prevention of acne. More specifically, the present technology relates to a method of treating, controlling or preventing acne by applying a composition comprising a specific rhamnolipid mixture to the skin of a subject suffering from or susceptible to acne. The present technology also relates to a composition comprising a rhamnolipid mixture having efficacy against Cutibacterium acne (formerly known as Proprionibacterium acne).

[0002] The current anti-acne market is dominated by products that meet the FDA's topical anti-acne product OTC monograph. Only four active ingredients are currently approved for use in this category. The two most commonly used actives are salicylic acid and benzoyl peroxide. The remaining two, resorcinol and sulfur, present significant challenges in appearance and formulation that have hampered their inclusion as consumer-grade anti-acne products.

[0003] Benzoyl peroxide and salicylic acid also have certain adverse side effects when used as topical anti-acne products at the monograph level. At usage levels, both ingredients can cause severe skin reactions ranging from mild irritation to severe contact dermatitis and skin peeling. Therefore, the benefits and side effects must be weighed. It would be desirable to have an active ingredient for use in an acne treatment composition that provides equivalent or better anti-Epidermidis acnes activity without the associated side effects that may accompany benzoyl peroxide and salicylic acid.

[0004] Another common treatment option for treating or managing acne is prescription strength, broad spectrum, non-selective antibiotics. In addition to the disadvantage that broad spectrum antibiotics require a doctor's prescription, non-selective antibiotics also reduce the overall microbiome, including potentially beneficial bacteria. It would be ideal to have an over-the-counter active that could specifically target Epidermobacterium acnes and minimize disruption to the overall microbiome.

[0005] 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 derived primarily from plant, animal or marine-based sources.

[0006] Rhamnolipids are interfacially active glycolipids produced by various bacteria and are examples of "green" ingredients since they can be prepared by fermentation based on renewable raw materials. It would be desirable to provide compositions comprising active ingredients derived from renewable sources, such as rhamnolipids, that could be used to control acne.

[0007] Applicants have determined that a specific rhamnolipid salt mixture can meet the above goals while also advancing the United Nations Sustainable Development Goals ("SDGs"). The rhamnolipid salt mixture of the present technology contributes to better health and well-being by providing the same or better efficacy against Epidermobacterium acnes than other common acne treatments without the associated side effects, as well as better efficacy than other rhamnolipids. The rhamnolipid salt mixture is advantageously a bio-based, renewable source of actives obtained from a bacterial fermentation process that produces a biodegradable waste product with less environmental impact. These benefits further promote Sustainable Development Goal #3 (Good Health and Well-Being) and Sustainable Development Goal #12 (Responsible Consumption and Production). Summary of the invention

[0008] In one aspect, the present technology provides an acne treatment composition comprising about 0.01% to about 99% active weight of a purified rhamnolipid mixture based on the total weight of the composition; and at least one dermatologically acceptable carrier, wherein the rhamnolipid mixture comprises salts of monorhamnolipid and dirhamnolipid, the weight ratio of monorhamnolipid: dirhamnolipid is 10:90 to 47:53, the amount of Rha-C10-C10 monorhamnolipid salt is about 5% to about 38% by weight based on the total weight of rhamnolipids in the rhamnolipid mixture, and the amount of RhaRha-C10-C10 dirhamnolipid salt is about 34% to about 80% by weight based on the total weight of rhamnolipids in the rhamnolipid mixture. In some embodiments, the rhamnolipid mixture comprises salts of monorhamnolipid and dirhamnolipid, the weight ratio of monorhamnolipid and dirhamnolipid is 40:60 to 45:55, alternatively 40:60 to 48:52, and the amount of Rha-C10-C10 monorhamnolipid salt is about 29% to about 40% by weight, alternatively about 29% to 37.5%, and the amount of RhaRha-C10-C10 dirhamnolipid salt is about 34% to about 45% by weight, alternatively about 36% to about 45% by weight, based on the total weight of rhamnolipids in the rhamnolipid mixture.

[0009] Another aspect of the present technology provides a method for treating, controlling or preventing acne in a subject suffering from or susceptible to acne, comprising administering to the subject an effective amount of a composition comprising a purified rhamnolipid salt mixture, thereby treating, controlling or preventing acne in the subject, wherein the purified rhamnolipid mixture comprises monorhamnolipid salts and dirhamnolipid salts, the weight ratio of monorhamnolipid: dirhamnolipid is about 10:90 to about 47:53, the amount of Rha-C10-C10 monorhamnolipid salt is about 5% to about 38% by weight, and the amount of RhaRha-C10-C10 dirhamnolipid salt is about 34% to about 80% by weight, based on the total weight of the rhamnolipid mixture. In some embodiments, the purified rhamnolipid mixture comprises salts of monorhamnolipid and dirhamnolipid, the weight ratio of monorhamnolipid and dirhamnolipid is 40:60 to 45:55, alternatively 40:60 to 48:52, and the amount of Rha-C10-C10 monorhamnolipid salt is about 29% to about 40% by weight, alternatively about 29% to 37.5%, and the amount of RhaRha-C10-C10 dirhamnolipid salt is about 34% to about 45% by weight, alternatively about 36% to about 45% by weight, based on the total weight of rhamnolipids in the rhamnolipid mixture. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 Figure 2 shows the Log of Epidermobacterium acnes after treatment with rhamnolipid test samples and control samples. 10 Reduced graph.

[0011] Figure 2 Figure 2 is a graph showing enumeration of Epidermidis acnes after treatment with rhamnolipid test and comparison samples as well as control samples.

[0012] Figure 3 To show the Log of Epidermobacterium acnes after treatment with rhamnolipid test sample and comparison sample and control sample 10 Reduced graph.

[0013] Figure 4 Figure 2 is a graph showing enumeration of Epidermidis acnes after treatment with rhamnolipid test and comparison samples as well as control samples.

[0014] Figure 5 To show the Log of Epidermobacterium acnes after treatment with rhamnolipid test sample and comparison sample and control sample 10 Reduced graph. DETAILED DESCRIPTION

[0015] definition

[0016] "Biorenewable Carbon Index" (BCI) refers to the calculation of the percentage of carbon derived from biorenewable resources and is based on dividing the number of biorenewable carbons by the number of total carbons in the entire molecule.

[0017] For the purposes of this article, “biorenewable” is defined as derived from animal, plant or marine materials.

[0018] The terms "active", "% active" and "% active by weight" refer to the amount of active ingredient, without taking into account the amount of water or other solvents that may be present in the ingredient.

[0019] "Antimicrobial agent" refers to an agent that is effective in controlling the growth of, reducing and / or killing microorganisms (such as bacteria, viruses, fungi, yeast, algae, cyanobacteria, etc.).

[0020] As used herein, "effective amount" refers to the amount of an active ingredient or composition that, when administered to a subject, is capable of controlling, treating or preventing acne caused by Epidermobacterium acnes. The actual amount may vary depending on a variety of factors, including but not limited to the severity of acne, the age and health of the patient, and the form of administration.

[0021] As defined herein, a "rhamnolipid" is a glycolipid having a lipid portion comprising one or more (typically linear) saturated or unsaturated β-hydroxycarboxylic acid moieties and a sugar portion of one or more units of rhamnose.

[0022] 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. Thus, the carboxylic acid of one carboxylic acid moiety defines the terminus of the rhamnolipid. When more than one rhamnose moiety is included in the rhamnolipid, each of the rhamnose moieties that are not linked to the lipid moiety is linked to another rhamnose moiety by a 1,4-β-glycosidic bond. In embodiments where two or more β-hydroxycarboxylic acids are present in the rhamnolipid, the β-hydroxycarboxylic acid moieties are selected independently of one another. In some embodiments, the β-hydroxycarboxylic acid moieties may be the same. In some embodiments, they are different from one another.

[0023] The present technology generally relates to acne treatment compositions comprising a specific mixture of purified rhamnolipids in salt form. Although acidic protonated forms of rhamnolipids have shown antimicrobial activity against Epidermidis herpes simplex, a bacterium known to cause acne conditions, purified rhamnolipids in neutralized salt form have not shown similar antimicrobial activity due to the acidic nature of rhamnolipids. However, the specific mixture of rhamnolipid salts in a pH neutral environment described herein surprisingly provides better efficacy against Epidermidis herpes simplex than other rhamnolipid mixtures known in the art when applied to the skin. The rhamnolipid may have the following structure (I):

[0024]

[0025] In this formula, R 9 is a hydrogen atom (H) or an aliphatic group having a backbone of 1 to about 46, such as 1 to about 42, 1 to about 40, 1 to about 38, 1 to about 36, 1 to about 34, 1 to about 30, 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 1 to about 3 (including 2) oxygen atoms. In some embodiments, the backbone of the corresponding aliphatic group carries a terminal carboxylic acid group and / or an internal ester group. As an illustrative example in this regard, R 9 It can be of the formula -CH(R 5 )----CH2-COOR 6 In these illustrative structural parts, R 5 It can be an aliphatic moiety wherein the backbone has a length of 1 to about 19, such as 1 to about 17, 1 to about 15, 1 to about 13, about 2 to about 13, about 3 to about 13, or about 4 to about 13, including, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. 4 is a hydrogen atom (H), or a rhamnopyranosyl moiety. 6 It's a hydrogen atom.

[0026] Unless otherwise specified, the term "aliphatic" refers to a straight or branched hydrocarbon chain, which can be saturated or monounsaturated or polyunsaturated and include heteroatoms. As used herein, the term "heteroatom" refers to an atom of any element other than carbon or hydrogen. Herein, unsaturated aliphatic groups contain one or more double bonds (alkenyl moieties). The branch of the hydrocarbon chain can contain straight chains and non-aromatic cyclic elements. Unless otherwise specified, the hydrocarbon chain can be any length and include any number of branches. Typically, the hydrocarbon (main) chain contains 1 to about 5, to about 10, to about 15 or to about 20 carbon atoms. The example of the alkenyl moiety is a straight or branched hydrocarbon moiety containing one or more double bonds. The alkenyl moiety typically contains about 2 to about 20 carbon atoms and one or more (e.g., two) double bonds (such as about 2 to about 10 carbon atoms) and a double bond. Examples of alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, isomers of these groups, isopropyl, isobutyl, isopentyl, sec-butyl, tert-butyl, neopentyl, 3,3-dimethylbutyl. Both the main chain and the side chain may also contain heteroatoms, such as N, O, S, Se or Si, or the carbon atom may be substituted by one of these heteroatoms. The aliphatic moiety may or may not be substituted with one or more functional groups. The substituent may be any functional group, such as, but not limited to, amino, amide, carbonyl, carboxyl, hydroxyl, nitro, sulfenyl and sulfonyl.

[0027] In a more specific embodiment, the rhamnosyl lipid salt in the structure has the following structure (II):

[0028]

[0029] 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 may be associated with each metal.

[0030] Rhamnolipid mixtures include mixtures of monorhamnolipids and dirhamnolipids.Monorhamnolipids can be present in the following amounts: about 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%, 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%, based on the total weight of the rhamnolipid in the mixture.

[0031] The dirhamnolipid may be 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. The amount of monorhamnolipid and dirhamnolipid in the rhamnolipid mixture described herein is determined by HPLC. The ratio of monorhamnolipid: dirhamnolipid may be from about 10:90 to about 48:52, alternatively about 47:53, alternatively about 40:60 to about 45:55. In some embodiments, the ratio of monorhamnolipid: dirhamnolipid may range from 43:57 to 45:55 or 43:57 to 48:52.

[0032] The rhamnolipid mixture preferably comprises mono (where x=1) and di (where x=2) rhamnolipids, wherein y and z are 6 and M is H or Na. The monorhamnolipid may be referred to as Rha-C10-C10 and has the formula C 26 H 48 O9. The IUPAC name is 3-[3-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxydecanoyloxy]decanoic acid. Rhamnolipid can be referred to as RhaRha-C10-C10, with the formula C 32 H 58 O 13The 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 an amount of 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 rhamnolipid. RhaRha-C10-C10 can be present in the mixture in an amount of about 34% to about 80% by weight based on the total weight of the rhamnolipid, 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%.

[0033] In addition to Rha-C10-C10 and RhaRha-C10-C10, the rhamnolipid mixture may also contain RhaRha-C10-C12 and Rha-C10-C12, the amount of RhaRha-C10-C12 being about 5% to about 15% by weight, alternatively about 9% to about 12%, alternatively about 10% to about 12.5% ​​based on the total weight of the rhamnolipid, and the amount of Rha-C10-C12 being about 0.2% to about 6%, alternatively about 2% to about 5%, alternatively about 3.5% to about 5% by weight, based on the total weight of the rhamnolipid. The rhamnolipid mixture can also contain RhaRha-C10-C12:1, RhaRha-C8-C10 and Rha-C8-C10, the amount of RhaRha-C10-C12:1 is about 0.2% to about 5% by weight, alternatively 1% to about 4% by weight, based on the total weight of rhamnolipids, the amount of RhaRha-C8-C10 ranges from about 0.2% to about 5% by weight, alternatively about 1% to about 4% by weight, based on the total weight of rhamnolipids, and the amount of Rha-C8-C10 ranges from about 0.2% to about 5% by weight, alternatively about 1% to about 4% by weight, based on the total weight of rhamnolipids.

[0034] In yet another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum inhibitory concentration (MIC) against Epidermidis acnes as determined as shown in Example 3 (MIC / MBC Test). In this aspect, the rhamnolipid compositions disclosed herein exhibit a MIC against Propionibacterium acnes of about 21 ppm to about 98 ppm, about 21 ppm to about 90 ppm, about 21 ppm to about 80 ppm, about 21 ppm to about 70 ppm, about 21 ppm to about 50 ppm, about 40 ppm to about 98 ppm, about 40 ppm to about 90 ppm, about 40 ppm to about 80 ppm, about 40 ppm to about 70 ppm, or about 40 ppm to about 50 ppm, as determined according to the method described in Example 3 (MIC / MBC Test).

[0035] In another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic minimum bactericidal concentration (MBC) against Epidermidis acnes as determined as shown in Example 3 (MIC / MBC Test). In this regard, the rhamnolipid compositions disclosed herein exhibit an MBC against Epidermidis acnes of about 61 ppm to about 296 ppm, about 61 ppm to about 200 ppm, about 61 ppm to about 150 ppm, about 61 ppm to about 125 ppm, about 75 ppm to about 296 ppm, about 75 ppm to about 200 ppm, about 75 ppm to about 150 ppm, about 75 ppm to about 125 ppm, about 100 ppm to about 296 ppm, about 100 ppm to about 200 ppm, about 100 ppm to about 150 ppm, about 100 ppm to about 125 ppm, about 120 ppm to about 296 ppm, about 120 ppm to about 200 ppm, about 120 ppm to about 150 ppm, or about 120 ppm to about 125 ppm.

[0036] In another aspect, the rhamnolipid compositions disclosed herein exhibit a characteristic percentage reduction in Epidermidis acnes colony forming units (CFU) per mL using a significantly reduced amount of rhamnolipid and contact time, as shown in Example 7 (time-dependent comparative test). In this aspect, the rhamnolipid compositions disclosed herein exhibit a percentage reduction in Epidermidis acnes colony forming units (CFU) per mL of 70% to about 99.999%, alternatively 80% to about 99.999%, alternatively 85% to about 99.999%, alternatively 90% to about 99.999%, alternatively 95% to about 99.999%, or alternatively 99% to about 99.999%, as determined according to the method set forth in Example 7 (using a reduced amount of rhamnolipid of 0.003% by weight (based on the total weight of the composition), and a reduced contact time of 15 minutes). Furthermore, in this regard, the rhamnolipid compositions disclosed herein exhibit from about 0.52 to about 5.0 log 10 Reduce, alternatively from about 0.70 to about 5 log 10 Reduce, alternatively from about 0.82 to about 5 log 10 Reduce, alternatively from about 1.0 to about 5 log 10 Reduce, alternatively from about 1.3 to about 5 log 10 Reduce, alternatively from about 2.0 to about 5 log 10 Reduced reduction in Epidermoepidermidis acnes colony forming units (CFU) per mL as determined according to the method described in Example 7 (using a reduced amount of rhamnolipid of 0.003% by weight (based on the total weight of the composition), and a reduced contact time of 15 minutes).

[0037] Rhamnolipid can be produced by rhamnolipid-producing microorganisms with the ability to synthesize / produce rhamnolipid under suitable conditions. Such microorganisms include but are not limited to bacteria, particularly Pseudomonas (Pseudomonadota), Actinobacteria (Actinobacteria), Firmicutes (Fimicutes) and Proteobacteria (Proteobacteria) bacteria. Rhamnolipid is naturally derived, and therefore has a BCI of 100. In a particular embodiment, the rhamnolipid-producing microorganism for producing rhamnolipid is Pseudomonas aeruginosa. The method of cultivating rhamnolipid-producing bacteria and the method of producing rhamnolipid from 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. The method of purifying rhamnolipid is 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.

[0038] The rhamnolipid salt mixture can be used alone as the sole active ingredient in the acne treatment composition. When used alone, the rhamnolipid mixture can be based on the total weight of the composition, based on an active weight of about 0.003% to about 99%, alternatively about 0.003% to about 25%, alternatively about 0.003% to about 10%, alternatively about 0.003% to about 6%, alternatively about 0.003% to about 0.1% by active weight, alternatively about 0.01% to about 99% by active weight, alternatively about 0.02% to about 25%, alternatively about 0.1% to about 10%, alternatively about 0.2% to about 6% by active weight. The rhamnolipid salt mixture can also be used in combination with another anti-acne active ingredient such as salicylic acid, benzoyl peroxide, resorcinol, resorcinol monoacetate, sulfur or a combination thereof. When used with another co-active, the rhamnolipid mixture can be based on the total weight of the composition, by active weight from about 0.003% to about 95%, alternatively from about 0.003% to about 25%, alternatively from about 0.003% to about 10%, alternatively from about 0.003% to about 6%, alternatively from about 0.003% to about 0.1%, alternatively from about 0.01% to about 95%, alternatively from about 0.02% to about 25%, alternatively from about 0.1% to about 10%, alternatively from about 0.2% to about 6% by active weight. The combination of a rhamnolipid salt mixture with another co-active ingredient can help alleviate the irritation potential of the co-active without reducing or inhibiting its activity. Combining a rhamnolipid salt mixture with another co-active ingredient can also allow for a reduction in another co-active ingredient, which may also help reduce the overall irritation potential.

[0039] The acne treatment composition can be formulated into any therapeutic form commonly used for dermatological applications. For example, the composition can be in the form of an aqueous solution, suspension, cream, lotion, gel, paste, spray, cream, foam or emollient, or impregnated on a pad or wipe.

[0040] The acne treatment composition of the present technology also includes at least one carrier suitable for personal or dermatological care to bring the total percentage of the composition to 100%. As at least one skilled in the art will understand, various carriers, vehicles, diluents, etc. are suitable for the practice of the present technology. Therefore, it should also be understood that with respect to the present technology and when describing its various formulations, applications, uses and compositions, the terms "carrier", "vehicle" and "diluent" should be considered non-exhaustive and interchangeable.

[0041] 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 personal care, such as alcohols, such as ethanol, isopropanol or benzyl alcohol; glycols, such as propylene glycol or polyethylene glycol. Other carriers may include, but are not limited to, solvents, emulsifiers or solubilizers.

[0042] When the treatment form is a cream, gel or paste, the acne treatment composition may include, but is not limited to, plant gums, starches, celluloses, waxes, silicones, silica or clays as carrier ingredients. When the treatment form is a spray, the composition may include a propellant.

[0043] In addition to rhamnolipid actives and carriers, the acne treatment compositions of the present technology may also include optional ingredients known in the art. Such other components or additives may include, but are not limited to, surfactants, pH regulators, skin conditioners, antioxidants, preservatives, fragrances, pigments, dyes, and other excipients.

[0044] The acne treatment compositions of the present technology can have a pH ranging from about 4.0 to about 8.5, alternatively from about 5.0 to about 8.0, desirably 5.5 to 7.0.

[0045] The acne treatment composition of the present technology can be used to treat, control or prevent acne by applying the composition to the skin of a subject in an amount effective to treat, control or prevent acne. "Application" can refer to any commonly used application method, such as but not limited to applying a cream or gel containing the acne treatment composition to the skin surface and leaving the cream or gel on the skin; spraying a liquid containing the acne treatment composition on the skin surface and wiping the skin or leaving the spray on the skin; wiping the skin with a wet wipe impregnated with the acne treatment composition and leaving the composition on the skin; applying a pad impregnated with the acne treatment composition and leaving the pad on the skin surface; or an aqueous or non-aqueous liquid cleanser intended to treat the skin surface.

[0046] The dosage form and treatment regimen of the acne treatment composition using the present technology can vary with the type and intensity of the disease. For example, the type of acne can include non-inflammatory acne (e.g., comedonal acne) or inflammatory acne (e.g., papulopustular acne or nodular acne). The severity of acne can also be divided into: mild (e.g., less than 20 acnes, or less than 15 inflammatory lesions, or a total lesion count of less than 30); moderate (e.g., 20-100 acnes, or 15-50 inflammatory lesions, or a total lesion count of between 30-125); or severe (e.g., greater than 5 nodules, or a total inflammation count of greater than 50, or a total lesion count of greater than 125). In one or more embodiments, one, two, three, four or more daily dosage regimens can be used according to the treatment method of the present technology. Depending on the patient's condition and response, the daily dosing regimen can continue for one, two, three, four, five, six or more weeks.

[0047] The acne treatment composition of the present technology comprises a specific mixture of rhamnosyl lipid salts and a carrier, and has efficacy against Epidermobacterium acnes. In some embodiments, the composition shows a >4.6 Log after 1 hour as determined according to the industry standard American Society for Testing and Materials (ASTM) E1054. 10 reduction (>99.997% reduction) and >2.95Log after 15 minutes 10The acne treatment composition provides a minimum inhibitory concentration (MIC) of 41 ppm or less and a minimum bactericidal concentration (MBC) of 123 ppm or less against Epidermidis acnes as determined according to industry standards (Clinical and Laboratory Standards Institute (CLSI) M7, M11 and M26). The rhamnolipid mixture in salt form also provides additional benefits. For example, because the rhamnolipid salt is at a neutral pH, it does not cause side effects such as redness, peeling, dryness or other potential effects that acidic anti-acne actives such as salicylic acid and benzoyl peroxide may cause. In addition, the rhamnolipid mixture has the potential to provide overall skin health, ranging from improving barrier function and moisture content to rebalancing the skin's resident flora, which helps inhibit the growth of Epidermidis acnes, thereby alleviating acne conditions.

[0048] Those skilled in the art will recognize that modifications may be made to the present technology 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.

[0049] Example

[0050] Example 1: Efficacy against Epidermobacterium herpetiformis

[0051] A study was conducted to determine the antimicrobial activity of a rhamnolipid mixture of the present technology alone against Epidermidis acnes, available from the American Type Culture Collection (ATCC) (Manassas, VA) as ATCC 6919. Test samples were prepared by mixing 6.0% by weight of the rhamnolipid mixture into an aqueous matrix. Two samples were prepared, one using a rhamnolipid mixture processed by a non-solvent purification process and the other using a rhamnolipid mixture processed by a solvent extraction process. The rhamnolipid mixture in each sample contained mono-rhamnolipid and di-rhamnolipid in a weight ratio ranging from 43:57 to 45:55. The efficacy of the test samples was compared to salicylic acid in deionized water at a concentration of 0.3%, using deionized water and buffer as controls. The results are shown in Table 1.

[0052] Table 1

[0053]

[0054]

[0055] *Solubility limit in water at 25°C

[0056] The results in Table 1 show that the rhamnolipid mixture proved effective in killing E. acnes and showed comparable efficacy to salicylic acid.

[0057] Example 2: Efficacy of Skin Analogs Against Epidermidis herpes simplex

[0058] A study was conducted to determine the antimicrobial activity of the rhamnolipid mixture of the present technology against Epidermidis acnes using skin analogs as a growth substrate for Epidermidis acnes. The samples tested are shown in Table 2:

[0059] Table 2

[0060]

[0061] The rhamnolipid mixture was processed by a non-solvent purification process and had a weight ratio of mono-rhamnolipid to di-rhamnolipid in the range of 43:57 to 45:55. The skin analog substrate was prepared by embedding primary adult dermal fibroblasts in a fibrin matrix to produce a dermis equivalent (DE). DE was cultured to allow the fibroblasts to remodel the matrix. Primary neonatal keratinocytes were applied to the DE surface and cultured under liquid for 48 hours. Labskin was cultured at an air-liquid interface until a stratified epidermis was formed.

[0062] Epidermidis herpes microorganisms (NCTC 737) were incubated anaerobically at 37°C for 4 days in a fortified Clostridium agar medium containing furazolidone. Approximately 1.1 x 10 8 CFU mL -1 Labskin was inoculated with an initial inoculum of Epidermobacterium acnes bacteria. Ten microliters of the inoculum were used to colonize each Labskin unit, and the Labskin was incubated at 37°C for 24 hours. After 24 hours, five Labskin units were left untreated, and the remaining Labskin units were treated with 11 microliters of the control or test article. All Labskin units were then incubated at 37°C for 24 hours. A biopsy sample of 8 mm in diameter was aseptically removed from the center of each Labskin unit, and the number of viable microorganisms was determined by recovering the appropriate culture medium.

[0063] The technical replicate counts obtained from the selective solid medium were averaged and used to calculate colony forming units per square centimeter (CFU cm -2 ). By calculating each CFU cm -2 Log of value 10 The values ​​are averaged to calculate Log 10 Log is calculated using the following equation 10 difference:

[0064] Log10 Difference = Log 10 (Processing)-Log 10 (Unprocessed)

[0065] The results are Figure 1 Displayed graphically in . Figure 1 The results in show that the rhamnolipid test sample significantly reduced the population of E. acnes at both concentrations tested, although the higher concentration (0.2%) of the rhamnolipid sample had a greater effect on the population of E. acnes. The 0.2% rhamnolipid sample was also slightly more effective than the acne gel control, and almost as effective as the salicylic acid control, although the 0.2% rhamnolipid concentration was 10 times lower than the concentrations of the acne gel and salicylic acid.

[0066] Example 3: MIC / MBC Test

[0067] A study was conducted to evaluate the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of a rhamnolipid mixture of the present technology (Example 3 composition) and a commercially available rhamnolipid product (Comparative composition) containing a rhamnolipid mixture having a monorhamnolipid: dirhamnolipid ratio of about 4:96 against Epidermidis herpes ATCC 6919. The rhamnolipid mixture in the Example 3 composition was processed using a non-solvent purification process and had a weight ratio of monorhamnolipid to dirhamnolipid ranging from 43:57 to 45:55. The Example 3 composition and the Comparative composition are compositions of only rhamnolipid actives diluted in deionized water.

[0068] As used herein, MIC is the concentration of the active substance in which the growth of target bacteria is not detected after sufficient incubation in nutrient broth medium relative to the control. MIC is determined according to industry standards (Clinical and Laboratory Standards Institute (CLSI) M7, M11 and M26), wherein the sample is typically diluted 1:2 or 1:3 in a nutrient medium suitable for the growth of the test microorganism, and fully pre-reduced to remove oxygen before anaerobic testing. Pre-reduction is performed by indirect atmospheric isolation of oxygen using a hydrogen reaction on a palladium catalyst. Subsequently, the sample is exposed to a test challenge microorganism with a final concentration of at least 1.0E+06CFU / mL. After an initial incubation period sufficient to develop measurable growth in the positive control group (a test container containing a nutrient medium and microorganisms; free of active ingredients), the presence / absence of growth in the sample is visually evaluated by turbidity evaluation. The sample concentration that does not become turbid compared to the positive control and is comparable to the negative control marks the concentration of the inhibitory effect.

[0069] As used herein, MBC is the minimum concentration of an antibacterial composition required to kill a specific target bacterium. MBC is determined according to industry standards CLSIM7, M11, and M26. Test samples that show inhibitory effectiveness in MIC tests are further tested for bactericidal properties by neutralizing aliquots extracted from each test concentration and plated onto appropriate agar media to determine the presence of survivors. After 48 hours, a ≥3.0 Log 10 The samples with reduced concentration (≥99.9% reduction) were determined as bactericidal concentrations. The results are shown in Table 3.

[0070] Table 3

[0071]

[0072] The results in Table 3 show that the rhamnolipid mixture of the present technology is more effective against Epidermidis herpes simplex than the comparative composition. The Example 3 composition exhibited a MIC of 41 ppm, which is lower than the 123 ppm of the comparative composition. The Example 3 composition also exhibited a lower MBC at 123 ppm compared to the MBC of the comparative composition at 370 ppm.

[0073] Example 4: Comparative Efficacy Testing Against Epidermidis herpes simplex Using Skin Analogs

[0074] A study was conducted to compare the efficacy of the rhamnolipid mixture of the present technology and comparative treatments against Epidermidis acnes using a skin analog as a growth substrate for Epidermidis acnes. The samples tested are shown in Table 4:

[0075] Table 4

[0076]

[0077]

[0078] *Revolution Skincare 2% Salicylic Acid BHA Anti-Blemish Serum

[0079] The exemplary rhamnolipid mixtures each had a weight ratio of monorhamnolipid to dirhamnolipid ranging from 43:57 to 45:55. The competing rhamnolipid mixture had a weight ratio of monorhamnolipid to dirhamnolipid of 4:96.

[0080] A skin analog substrate ("Labskin") was prepared by embedding primary adult dermal fibroblasts in a fibrin matrix to create a dermis equivalent (DE). DE was cultured to allow the fibroblasts to remodel the matrix. Primary neonatal keratinocytes were applied to the DE surface and cultured under liquid for 48 hours. Labskin was cultured at an air-liquid interface until a stratified epidermis was formed.

[0081] Epidermidis herpes simplex microorganisms (NCTC 737) were incubated anaerobically at 37°C for 4 days in a fortified Clostridium agar medium containing furazolidone. 8 CFU mL -1 Labskin was inoculated with an initial inoculum of Epidermobacterium acnes bacteria. Ten microliters of the inoculum were used to colonize each Labskin unit, and the Labskin was incubated at 37°C for 24 hours. After 24 hours, five Labskin units were left untreated, and the remaining Labskin units were treated with 11 microliters of the control or test article. All Labskin units were then incubated at 37°C for 24 hours. A biopsy sample of 8 mm in diameter was aseptically removed from the center of each Labskin unit, and the number of viable microorganisms was determined by recovering the appropriate culture medium.

[0082] The technical replicate counts obtained from the selective solid medium were averaged and used to calculate colony forming units per square centimeter (CFU cm -2 ). Figure 2 The enumeration of E. acnes bacteria for the control and test samples is shown graphically. The graphical results show that the dPBS sample caused a slight decrease in the average number of E. acnes bacteria compared to "untreated", while the positive control (acnecide gel) and each test sample caused a significant decrease in the average number of E. acnes bacteria.

[0083] By calculating each CFU cm -2 Log of value 10 The values ​​are averaged to calculate Log 10 Log is calculated using the following equation 10 difference:

[0084] Log10 difference = Log10 (treated) - Log10 (untreated)

[0085] Log 10 The difference results are displayed graphically in Figure 3 Log 10 Differential analysis allows for an overall comparison of treated samples. This analysis can be used to determine biologically significant changes in treated samples relative to "untreated" controls. A significant difference is defined as ±0.5 Log 10 CFU cm -2 . Figure 3 The results in the graph in show that the dPBS control did not significantly reduce the population of E. acnes, while the acnecide gel, salicylic acid, and each rhamnolipid treated samples significantly reduced the population of E. acnes. Figure 3 The results in also show that the exemplary rhamnolipids of the present technology were all more effective (greater reduction in the population of E. acnes) than the competing rhamnolipid mixture, and that the 0.5% and 0.8% concentrations of the exemplary rhamnolipids had greater reductions in the population of E. acnes than any treatment except Acnecide gel. Figure 3 A dose response effect of the exemplary solvent-purified rhamnolipids can also be observed in the Figure 4A, where higher concentrations have a greater effect on the E. acnes population.

[0086] Example 5: Comparison of the efficacy of rhamnolipid, salicylic acid and benzoyl peroxide

[0087] A study was conducted to compare the efficacy of the rhamnolipid mixture of the present technology with salicylic acid and benzoyl peroxide, two common over-the-counter anti-acne actives. The samples tested are shown in Table 5. The samples containing salicylic acid and benzoyl peroxide were also mixed using a tip ultrasonicator to ensure that the actives were fully dissolved or well distributed.

[0088] Table 5

[0089]

[0090]

[0091] The skin analog substrate (Labskin) was prepared as described in Example 4. Epidermobacterium acnes microorganisms were cultured as described in Example 4 and inoculated into each Labskin unit as described in Example 4. The Labskin units were incubated at 37°C for 24 hours and treated 24 hours later with 11 microliters of control (dPBS), vehicle, or test article. All Labskin units were then incubated at 37°C for 24 hours, after which a biopsy sample of 8 mm in diameter was aseptically removed from the center of each Labskin unit and the number of viable microorganisms was determined by recovering the appropriate culture medium.

[0092] The technical replicate counts obtained from the selective solid medium were averaged and used to calculate colony forming units per square centimeter (CFU cm -2 ). Figure 4 The enumeration of E. acnes for the control, vehicle, and test samples is shown graphically. The graphical results show that the vehicle sample showed a slight increase in the mean number of E. acnes, while the 0.5% w / w salicylic acid test sample showed a slight decrease in the mean number of E. acnes compared to the dPBS control. All other test samples showed a clear or significant decrease in the mean number of E. acnes.

[0093] Log between dPBS control and vehicle and test samples10 The difference was calculated as described in Example 4, and Log 10 The difference results are displayed graphically in Figure 5 middle. Figure 5 The graphical results in show a slight increase in E. acnes recovered from the vehicle sample, and a moderate decrease in E. acnes recovered from the 0.5% w / w salicylic acid sample. Samples containing 2% w / w salicylic acid and 2% w / w salicylic acid plus 0.5% w / w rhamnolipids showed a clear decrease in the number of E. acnes recovered, while the rhamnolipid and benzoyl peroxide samples alone and in combination showed a significant decrease in the recovered population of E. acnes. Figure 5 The results in also show that samples of the present technology containing rhamnolipids at a concentration of 0.5% w / w are more effective (greater reduction in the population of Epidermobacterium acnes) than test samples containing salicylic acid, and have comparable efficacy to the 2.5% w / w benzoyl peroxide test sample, even though the rhamnolipid concentration was 5 times lower than that of the 2.5% w / w benzoyl peroxide sample.

[0094] Example 6: Time-dependent comparison test

[0095] A study was conducted to compare the time-dependent efficacy of a rhamnolipid mixture of the present technology (composition of Example 6), a commercially available rhamnolipid product (comparative composition), and salicylic acid against Epidermidis herpes simplex (ATCC#6919). The rhamnolipid mixture in the composition of Example 6 was processed using a non-solvent purification process and had a weight ratio of monorhamnolipid to dirhamnolipid ranging from 48:52 to 45:55. The comparative composition contained a rhamnolipid mixture having a monorhamnolipid: dirhamnolipid ratio of about 4:96. The test samples were prepared by diluting rhamnolipid and salicylic acid in sterile water. The concentrations of rhamnolipid actives in the rhamnolipid test samples were 0.003wt% active, 0.01wt% active, and 0.03wt% active. The concentrations of salicylic acid actives in the test samples were 0.03wt%, 0.1wt%, and 0.3wt%. Prior to the start of the study, the samples were fully pre-reduced to remove oxygen by indirect atmospheric isolation of oxygen using a hydrogen reaction on a palladium catalyst. The test samples were inoculated with a culture of at least 1.0E+06 CFU / mL of Epidermidis acnes and incubated at 37.0±2.0°C, with aliquots taken at 15 minutes and 60 minutes. The sample aliquots were neutralized by diluting in an oxygen reduction buffer containing flow cytometry dyes, SYTO 9, and propidium iodide. The samples were analyzed using flow cytometry, where the cells were characterized by morphology and viability compared to a deionized water control. The results are shown in Table 6.

[0096] Table 6

[0097]

[0098] 1 The pH of the comparative test sample was adjusted to be comparable to that of the Example 7 composition test sample.

[0099] 2 Greater than or equal to the maximum log reduction limit of detection.

[0100] The results in Table 6 show that the Example 6 composition has better efficacy than the comparative composition at an equal concentration of 0.003% percent active after 15 minutes. The results also show that the Example 6 composition has better efficacy at a concentration of 0.003% than salicylic acid at a concentration of 0.30% after 15 minutes.

[0101] Example 7: Efficacy of the formulation product

[0102] A study was conducted to compare the time-dependent efficacy of a formulated product containing a rhamnolipid mixture of the present technology (Example 7 composition) and commercially available acne treatment products: Neutrogena Stubborn Acne AM Treatment (benzoyl peroxide acne treatment) and Clean & Clear Advantage Acne Spot Treatment (salicylic acid acne treatment) against Epidermobacterium acnes (ATCC #6919). The Example 7 composition comprises an acne base composition and a rhamnolipid mixture of the present technology at different concentrations. The acne base composition comprises propylene glycol in an amount ranging from 57.67% to 60% by weight, xanthan gum in an amount ranging from 1.92% to 2% by weight, a mixture of benzyl alcohol / benzoic acid / dehydroacetic acid in an amount ranging from 0.72% to 0.75% by weight, and water up to a total of 100% by weight of the total weight of the composition. The rhamnolipid mixture is subjected to a non-solvent purification process and has a weight ratio of monorhamnolipid to dirhamnolipid in the range of 48:52 to 45:55.

[0103] The formulated product and commercial product samples were tested undiluted at product active concentrations of 0.0% (base composition only), 0.1% and 1.0% rhamnolipid for the Example 7 composition, 2.5% for benzoyl peroxide treatment, and 2.0% for salicylic acid treatment. The formulated product and commercial product samples were also diluted in 75% deionized water to active concentrations of 0.025% and 0.25% (for the Example 7 composition), 0.625% (for benzoyl peroxide treatment), and 0.50% (for salicylic acid treatment). The product samples were inoculated with at least 1.0E+06 CFU / mL of a culture of Epidermidis acnes and incubated at 37.0±2.0°C for 15 minutes. The test sample aliquots were neutralized by diluting in an oxygen reduction buffer containing flow cytometry dyes, SYTO 9, and propidium iodide. The samples were analyzed using flow cytometry, where the cells were characterized by morphology and viability compared to a deionized water control. The results of the undiluted preliminary test samples are shown in Table 7, and the results of the diluted test samples are shown in Table 8.

[0104] Table 7

[0105]

[0106]

[0107] 1 The test samples were non-significantly (<0.1%) diluted by E. acnes.

[0108] 2 Greater than or equal to the maximum log reduction limit of detection.

[0109] The results in Table 7 show that the product composition formulated in Example 7 performed comparably to a national benchmark containing 2.5% benzoyl peroxide (pH 5.0) and 2.0% salicylic acid (pH 3.8) at both 0.10% and 1% active weight concentrations of rhamnolipid.

[0110] Table 8

[0111]

[0112] 1 The samples were diluted to receive 75% of the total volume with sterile deionized water.

[0113] 2 Greater than or equal to the maximum log reduction limit of detection.

[0114] The results in Table 8 show that the composition of Example 7 diluted to a rhamnolipid concentration of 0.25% performed as well as the national benchmark containing 0.625% benzoyl peroxide (pH 5.0) and 0.5% salicylic acid (pH 3.8).

[0115] The embodiments and examples described herein are illustrative and do not limit the currently described technology in any way. The scope of the present technology described in this description is the complete scope defined or implied by the claims. In addition, unless otherwise stated, any references pointed out in the detailed description of the invention part of this application are incorporated herein by reference in their entirety.

[0116] The present technology is now described in sufficient, clear and concise terms to enable those skilled in the art to practice the same. It should be understood that the foregoing describes preferred embodiments of the present technology and that modifications may be made therein without departing from the spirit or scope of the present technology as set forth in the appended claims. In addition, the examples provided are not exhaustive but are intended to illustrate several embodiments that fall within the scope of the claims.

Claims

1. An acne treatment composition comprising: (a) a purified rhamnolipid salt mixture in an amount of about 0.003% to about 99% by weight based on the total weight of the composition, wherein the purified rhamnolipid salt mixture comprises monorhamnolipid and dirhamnolipid, the weight ratio of monorhamnolipid: dirhamnolipid is 10:90 to 47:53, the amount of Rha-C10-C10 monorhamnolipid salt is about 5% to about 38% by weight based on the total weight of the rhamnolipid salt mixture, and the amount of RhaRha-C10-C10 dirhamnolipid salt is about 34% to about 80% by weight based on the total weight of the rhamnolipid salt mixture; and (b) at least one dermatologically acceptable carrier, and optionally one or more additives, in amounts up to a total of 100% by weight of the composition.

2. The acne treatment composition according to claim 1, wherein the weight ratio of monorhamnolipid to dirhamnolipid is 40:60 to 47:53, preferably 42:58:47:

53.

3. The acne treatment composition according to claim 1 or claim 2, wherein the amount of Rha-C10-C10 monorhamnolipid salt is from about 29% to about 37.5% by weight based on the total weight of the rhamnolipid salt mixture.

4. The acne treatment composition according to any one of claims 1 to 3, wherein the amount of RhaRha-C10-C10 di-rhamnolipid salt is from about 36% to about 45% by weight based on the total weight of the rhamnolipid mixture.

5. The acne treatment composition according to any one of claims 1 to 4, wherein the amount of RhaRha-C10-C12 di-rhamnolipid salt is from about 5% to about 15% by weight based on the total weight of the rhamnolipid mixture.

6. The acne treatment composition according to any one of claims 1 to 5, wherein the amount of RhaRha-C10-C12:1 dirhamnolipid salt 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.

7. The acne treatment composition according to any one of claims 1 to 6, wherein the amount of RhaRha-C8-C10 dirhamnolipid salt 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.

8. The acne treatment composition according to any one of claims 1 to 7, wherein the amount of Rha-C10-C12 monorhamnolipid salt is from about 0.2% to about 6%, preferably from about 3.5% to about 5% by weight based on the total weight of the rhamnolipid mixture.

9. The acne treatment composition according to any one of claims 1 to 8, wherein the amount of Rha-C8-C10 monorhamnolipid salt 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 acne treatment composition of any one of claims 1-9, wherein the amount of rhamnolipid salt in the composition is from about 0.003% to about 6% by weight of the composition.

11. The acne treatment composition according to any one of claims 1 to 10, wherein the composition comprises one or more additives.

12. The acne treatment composition of claim 11, wherein the one or more additives include one or more of surfactants, pH adjusters, skin conditioners, antioxidants, preservatives, fragrances, pigments, and dyes.

13. The acne treatment composition of any one of claims 1-12, wherein the dermatologically acceptable carrier comprises water.

14. The acne treatment composition of any one of claims 1-13, wherein the dermatologically acceptable carrier comprises alcohols, glycols, solvents, emulsifiers, solubilizers, vegetable gums, starches, celluloses, waxes, silicones, silica, and clays.

15. The acne treatment composition according to any one of claims 1 to 14, wherein the composition exhibits an inhibitory effect on Epidermidis acnes.

16. The acne treatment composition according to any one of claims 1 to 15, wherein the composition exhibits a bactericidal effect against Epidermidis acnes.

17. The acne treatment composition of any one of claims 1-16, wherein the composition has a pH of 4.0 to 8.

5.

18. The acne treatment composition of any one of claims 1-17, wherein the composition is an aqueous solution, suspension, cream, lotion, gel, paste, spray, cream, foam, emollient, pad or wipe.

19. An acne treatment composition according to any one of claims 1 to 18, wherein the composition comprises a co-active ingredient in combination with the rhamnolipid salt mixture.

20. The acne treatment composition of claim 19, wherein the co-active ingredient is salicylic acid, benzoyl peroxide, resorcinol, resorcinol monoacetate, or sulfur, or any combination thereof.

21. A method of controlling, treating or preventing acne in a subject suffering from or susceptible to acne comprising applying to the skin of the subject an effective amount of the acne treatment composition of any one of the preceding claims.

22. A method for controlling, treating or preventing acne in a subject suffering from or susceptible to acne, comprising applying to the skin of the subject an effective amount of a composition comprising a purified rhamnolipid salt mixture, thereby controlling, treating or preventing acne in the subject, wherein the purified rhamnolipid salt mixture comprises monorhamnolipid salts and dirhamnolipid salts, the weight ratio of monorhamnolipid: dirhamnolipid is 10:90 to 47:53, the amount of Rha-C10-C10 monorhamnolipid salt is from about 5% to about 38% by weight based on the total weight of the rhamnolipid mixture, and the amount of RhaRha-C10-C10 dirhamnolipid salt is from about 34% to about 80% by weight based on the total weight of the rhamnolipid salt mixture.

23. The method of claim 22, wherein the purified rhamnolipid salt mixture is present in the composition in an amount of 0.003% to 6.0% by weight, based on the total weight of the composition.

24. The acne treatment composition of any one of claims 1-20, wherein the purified rhamnolipid exhibits a percentage reduction in Epidermobacterium acnes colony forming units (CFU) per mL of about 70% to about 99.999%.

25. The acne treatment composition of any one of claims 1-20, wherein the purified rhamnolipid exhibits a relative humidity of about 0.52 to about 5.0 cm -2 Log of colony forming units (CFU) of Epidermobacterium acnes 10 reduce.

26. The acne treatment composition of any one of claims 1-20, wherein the purified rhamnolipid exhibits a minimum inhibitory concentration (MIC) against Epidermidis acnes of about 21 ppm to about 98 ppm.

27. The acne treatment composition of any one of claims 1-20, wherein the purified rhamnolipid exhibits a minimum bactericidal concentration (MBC) against Epidermobacterium acnes of about 61 ppm to about 296 ppm.

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