Rhamnolipid for scalp health

Abstract

The technologies presented herein generally relate to the formulation of dandruff treatment compositions, and to the formulation and use of rhamnolipid in these compositions. More specifically, the technologies relate to compositions and methods for controlling, treating, or preventing dandruff in subjects affected or susceptible to dandruff. The technologies also relate to compositions and methods for treating excessive Malassezia fungi on the skin of mammals.

Description

[Technical Field]

[0001] This technology generally relates to the use of rhamnolipids to improve and / or promote the maintenance of a healthy scalp. More specifically, this technology relates to compositions and methods for treating and / or preventing the dandruff cycle by applying compositions comprising specific mixtures of rhamnolipids to the human scalp. In certain embodiments, application of the compositions described herein, according to the methods and regimens described herein, inhibits the growth, colonization, and / or reduces the viability of skin-causing microorganisms, such as the lipophilic yeast Malassezia. [Background technology]

[0002] The human scalp is typically characterized by the development of dense terminal hairs and numerous sebaceous glands and microorganisms. This environment contributes to the establishment of specific microenvironments involved in the development of skin pathological conditions, including, for example, seborrheic dermatitis and dandruff, psoriasis, atopic dermatitis, alopecia, and scalp aging.

[0003] Seborrheic dermatitis is a common chronic inflammatory skin disease affecting the scalp, characterized by erythema and oily scales (scaly rash). The non-inflammatory form of this disease affects approximately 50% of the world's population (with dry and desquamating scalp symptoms) and is commonly known as dandruff.

[0004] The dandruff cycle is initiated by an imbalance of endogenous factors, including host immunity, skin microbiome, sebaceous gland activity, and / or epidermal barrier damage. One or more of these conditions can create favorable conditions that lead to Malassezia infection. Malassezia fungi are lipophilic yeasts that account for over 80% of the fungal population found on human skin.

[0005] Malassezia furfur is the primary causative fungal species of dandruff and its associated symptoms and problems. Other species include M. dermatis, M. equi, M. globosa, M. japonica, M. obtusa, M. ovalis (also known as Pityrosporum ovale), M. pachydermatis, M. restricta, M. slooffiae, M. sympodialis, or combinations thereof. Once an infection is established, seborrheic dermatitis develops, leading to damage to the epidermal barrier function, increased sebum secretion, further colonization of Malassezia fungi (and secretion of metabolites), and ultimately an amplification of the inflammatory response. This results in a long-term, sometimes chronic, decline in scalp health and aesthetics. Other conditions such as psoriasis, atopic dermatitis, and accelerated scalp aging have also been reported to be associated with skin damage, oxidative stress, microbial infection, microbiome imbalance, and immunological response mechanisms.

[0006] Current market solutions involve the use of highly irritating and less selective chemical components such as coal tar, zinc pyrithione, selenium sulfide, salicylic acid, or a limited range of antifungal agents, which lose effectiveness over time. One limitation of these conventional chemicals is that while they are effective in suppressing fungal infections and / or removing dandruff from the scalp by exfoliation for aesthetic purposes, these chemicals fail to provide a comprehensive, sustainable, and natural solution to the dandruff cycle.

[0007] Therefore, there is an ongoing need for new compositions, methods, and treatment regimens that promote the maintenance of a healthy scalp. More specifically, there is a need for compositions and methods for treating and / or preventing the dandruff cycle, particularly for inhibiting or preventing the proliferation, colonization, and / or reducing the survival rate of the lipophilic yeast Malassezia.

[0008] Furthermore, formulating products with ingredients based on renewable raw materials is a recent trend. These ingredients are considered "environmentally friendly" or "natural" because they originate from renewable and / or sustainable sources. As a result, they are more environmentally friendly than ingredients derived from fossil fuels or other non-renewable sources. Ingredients with a high Bio-Renewable Carbon Index (BCI), such as over 80, indicate that the ingredient contains carbon primarily from plant, animal, or marine-based sources.

[0009] Rhamnolipid is a surfactant glycolipid produced by various bacterial species and can be prepared by fermentation based on recycled raw materials, making it an example of an "environmentally friendly" ingredient. It would be desirable to provide compositions and methods of use containing active ingredients derived from renewable sources such as rhamnolipid that can be used to promote the maintenance of a healthy scalp. Providing such compositions containing naturally derived rhamnolipid would help achieve sustainability goals by ensuring sustainable consumption through the use of bio-based antimicrobial materials.

[0010] The applicant determined that certain rhamnolipid compositions can satisfy the above objectives while also promoting the UN Sustainable Development Goals ("SDGs"). The rhamnolipid compositions of this technology contribute to better health and well-being by providing equivalent or better efficacy in maintaining a healthy scalp (e.g., in preventing and / or treating dandruff). Advantageously, the rhamnolipid compositions described herein are bio-based, renewablely sourced active materials obtained from a bacterial fermentation process that generates biodegradable waste with minimal environmental impact. These advantages further enhance SDG Goal 3 (Good Health and Well-being) and SDG Goal 12 (Responsible Consumption and Production). [Overview of the Initiative]

[0011] One aspect of this technology relates to compositions and methods for promoting the maintenance of a healthy scalp. More specifically, the technology described herein relates to compositions and methods of use for treating and / or preventing the dandruff cycle. In certain embodiments, application of the compositions described herein, according to the methods and regimens described herein, inhibits the growth, colonization, and / or reduces the survival rate of skin-causing microorganisms, such as lipophilic yeasts of the genus Malassezia.

[0012] A further aspect of this technology relates to a natural and holistic alternative to the highly irritating and non-selective chemicals currently available for the prevention and treatment of dandruff. More specifically, the technology described herein relates to the use of a novel rhamnolipid composition to promote the maintenance of a healthy scalp microbiome, which includes inhibiting (or otherwise controlling) the proliferation, colonization, and / or reducing the viability of lipophilic yeasts of the genus Malassezia, as well as reducing the progression in the dandruff cycle.

[0013] In one aspect of the present technology, a dandruff treatment composition is provided comprising (a) a mixture of rhamnolipid salts, (b) at least one surfactant selected from the group consisting of anionic surfactants, amphoteric surfactants, nonionic surfactants, cationic surfactants, and combinations thereof, and (c) a dermatologically acceptable carrier, which, when applied to the skin of a mammal, exhibits an antifungal effect against Malassezia fungi in the absence of other antifungal active ingredients (i.e., inhibits, prevents growth, and / or otherwise reduces survival rate).

[0014] In another embodiment, a dandruff treatment composition is provided, comprising (a) a mixture of zinc-rhamnolipid salts in an amount of about 0.01% to about 30% by weight, based on the total weight of the composition, and (b) a dermatologically acceptable carrier in an amount totaling 100% by weight. When applied to mammalian skin (e.g., mammalian scalp), the composition exhibits an antifungal effect against Malassezia fungi in the absence of other antifungal active ingredients.

[0015] Further embodiments of the present technology provide a dandruff treatment composition comprising: (a) a mixture of rhamnolipid salts in an amount of about 0.01% to about 30% by weight based on the total weight of the composition; (b) at least one anionic surfactant in an amount of about 1% to about 5% by weight, about 1% to about 10% by weight, about 1% to about 15% by weight, about 1% to about 20% by weight, about 1% to about 30% by weight, or about 1% to about 40% by weight of the composition; (c) at least one amphoteric surfactant in an amount of about 1% to about 5% by weight, about 1% to about 10% by weight, about 1% to about 15% by weight, about 1% to about 20% by weight, about 1% to about 30% by weight, or about 1% to about 40% by weight of the composition; and (d) a dermatologically acceptable carrier in an amount totaling 100%. When applied to mammalian skin (e.g., mammalian scalp), the composition exhibits antifungal activity against Malassezia fungi in the absence of other antifungal active ingredients.

[0016] Furthermore, the rhamnolipid salt mixture contains monorhamnolipid and dirhamnolipid in weight ratios of monorhamnolipid to dirhamnolipid of, for example, 10:90 to 90:10, 40:60 to 60:40, or 40:60 to 48:52. The rhamnolipid salt mixture further contains, for example, about 5% to about 50% by weight, or about 5% to about 38% by weight, of Rha-C10-C10 monorhamnolipid salt, based on the total weight of the rhamnolipid salt mixture, and about 30% to about 80% by weight, or about 34% to about 80% by weight, of Rha-C10-C10 dirhamnolipid salt, based on the total weight of the rhamnolipid salt mixture.

[0017] Furthermore, the rhamnolipid salt mixture is available in concentrations of approximately 0.01% to 30% by weight, approximately 0.01% to 20% by weight, approximately 0.01% to 10% by weight, approximately 0.01% to 5% by weight, approximately 0.05% to 30% by weight, approximately 0.05% to 20% by weight, approximately 0.05% to 10% by weight, approximately 0.05% to 5% by weight, and approximately It may be present in the dandruff treatment compositions described herein in amounts of 0.1% to about 30% by weight, about 0.1% to about 20% by weight, about 0.1% to about 10% by weight, about 0.1% to about 5% by weight, about 0.3% to about 20% by weight, about 0.3% to about 10% by weight, about 0.3% to about 5% by weight, about 0.3% to about 3%, about 0.3% to about 2%, or about 0.3% to about 1%.

[0018] Further embodiments of this technology include one or more methods for controlling, treating, or preventing dandruff in subjects who are affected or susceptible to dandruff. These methods include, for example, applying an effective amount of one or more of the dandruff treatment compositions described herein to the hair of the subject.

[0019] Another aspect of this disclosure includes one or more methods for treating an excess of Malassezia fungi on mammalian skin, including on the scalp of mammals. These methods include, for example, applying an effective amount of one or more of the dandruff treatment compositions described herein to the skin of a mammal.

[0020] In other embodiments, the compositions and methods disclosed herein have a favorable effect on fundamental host factors, including microbiome balance. Similarly, the compositions and methods disclosed herein promote healthy immunological responses, such as promoting the maintenance of a healthy epidermal layer and accelerating scalp-related wound healing. [Modes for carrying out the invention]

[0021] The present technology is described in connection with one or more preferred embodiments, and those skilled in the art will understand that the present technology is not limited to only those specific embodiments. On the contrary, the technology described in the present specification includes all alternatives, modifications, and equivalents that may be included within the spirit and scope of the appended claims.

[0022] In general, the present disclosure is directed to compositions and methods for promoting the improvement and / or maintenance of a healthy scalp. The methods disclosed in the present specification include applying the composition to the scalp of a mammal, the composition comprises at least one rhamnolipid, and applying the composition to the scalp of the mammal inhibits or reduces the growth, colonization and / or viability of Malassezia fungi as well as the progression in the dandruff cycle.

[0023] As defined in the present specification, "rhamnolipid" is a glycolipid having a lipid moiety comprising one or more, typically linear, saturated or unsaturated B-hydroxy-carboxylic acid moieties and a sugar moiety of one or more rhamnose units. The term Zn-rhamnolipid or zinc rhamnolipid is a salt of the rhamnolipid described in the present specification, and the salt is zinc.

[0024] The sugar moiety and the lipid moiety are linked via a B-glycosidic bond between the 1-OH group of the rhamnose moiety of the sugar moiety and the 3-OH group of the B-hydroxy-carboxylic acid of the lipid moiety. Thus, the carboxylic acid of one carboxylic acid moiety defines the terminus of the rhamnolipid. When two or more rhamnose moieties are contained in the rhamnolipid, each of the rhamnose moieties not linked to the lipid moiety is linked to another rhamnose moiety via a 1,4B-glycosidic bond. In embodiments where two or more B-hydroxy-carboxylic acids are present in the rhamnolipid, the B-hydroxy-carboxylic acid moieties are selected independently of each other. In some embodiments, the B-hydroxycarboxylic acid moieties may be the same. In some embodiments, they are different from each other.

[0025] The present technology generally relates to compositions for maintaining a healthy scalp, and compositions comprising specific mixtures of rhamnolipids in their salt forms. The rhamnolipids may have the following structure (I):

Chemical Formula

[0026] The term "aliphatic," unless otherwise specified, means a linear or branched hydrocarbon chain, which may be saturated, monounsaturated, or polyunsaturated, and may contain heteroatoms. As used herein, the term "heteroatom" means an atom of any element other than carbon or hydrogen. In this specification, an unsaturated aliphatic group contains one or more double bonds (alkenyl moieties). Branching of a hydrocarbon chain may include linear and non-aromatic cyclic elements. Unless otherwise specified, a hydrocarbon chain may be of any length and may contain any number of branches. Typically, a hydrocarbon (main) chain contains 1 to about 5, about 10, about 15, or about 20 carbon atoms. Examples of alkenyl moieties include linear or branched hydrocarbon moieties containing one or more double bonds. Alkenyl moieties generally contain about 2 to about 20 carbon atoms and one or more, for example, two double bonds, or for example, about 2 to about 10 carbon atoms and one double bond. Examples of alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, n-isomers of these radicals, isopropyl, isobutyl, isopentyl, sec-butyl, tert-butyl, neopentyl, and 3,3-dimethylbutyl. Both the main chain and the branching may further contain heteroatoms such as N, O, S, Se, or Si, or carbon atoms may be substituted by one of these heteroatoms. The aliphatic moiety may be substituted with one or more functional groups or may be unsubstituted. Substituents may be any functional group, such as amino, amide, carbonyl, carboxyl, hydroxyl, nitro, thio, and sulfonyl, but are not limited to these.

[0027] In a more specific embodiment, the rhamnolipid salt in the structure has structure (II): [ka] wherein x is 1 or 2, y is 4, 6 or 8, z is 4, 6 or 8, and M is H, or an alkali metal Li, Na or K, an alkaline earth metal Mg or Ca, or a metal such as a transition metal Mn, Fe, Cu, or Zn. In the case of alkaline earth metals and transition metals, multiple rhamnolipid moieties may associate with each metal.

[0028] The mixture of rhamnolipids preferably comprises mono (x=1) and di (x=2) rhamnolipids, y and z are 6, and M is H, Zn or Na. In some embodiments, the mixture of rhamnolipids is a mixture of zinc-rhamnolipid salts. The monorhamnolipid is C 26 H 48 O9, which may be referred to as Rha-C10-C10 by the formula. The IUPAC name is 3-[3-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxydecanoyloxy]decanoic acid. The dirhamnolipid is C 32 H 58 O 13 , which may be referred to as RhaRha-C10-C10 by the formula. The 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. In general, the mixtures of rhamnolipids disclosed herein comprise various types of monorhamnolipids and dirhamnolipids, and the mixtures expressly encompass all possible combinations of the monorhamnolipids and dirhamnolipids disclosed herein. Further, unless otherwise stated, the amounts of individual monorhamnolipids or dirhamnolipids disclosed herein mean that the respective monorhamnolipids or dirhamnolipids can be present in the mixture of rhamnolipids in the indicated amounts, and the mixtures of rhamnolipids disclosed herein are expressly meant to include all possible combinations of the amounts of the monorhamnolipids and dirhamnolipids disclosed herein. A generally preferred mixture of rhamnolipids is the SEP-RM rhamnolipid composition as described below.

[0029] Rha-C10-C10 may be present in the mixture in amounts of approximately 5% to 50%, 5% to 38%, or 10% to 38%, or 15% to 37.5%, or 20% to 37.5%, or 20% to 50%, or 25% to 45%, or 25% to 37.5%, or 30% to 40%, or 29% to 40%, or 35% to 38%, based on the total weight of rhamnolipid. RhaRha-C10-C10 may be present in the mixture in amounts of approximately 20% to 50% by weight, or approximately 30% to 80%, or approximately 34% to 80%, or approximately 35% to 75%, or approximately 35% to 65%, or approximately 35% to 60%, or approximately 35% to 55%, or approximately 35% to 50%, or approximately 35% to 45%, or approximately 30% to 40%, or approximately 35% to 38%, or approximately 36% to 40% by weight, based on the total weight of rhamnolipid.

[0030] In addition to Rha-C10-C10 and RhaRha-C10-C10, the rhamnolipid mixture may contain RhaRha-C10-C12 in amounts of approximately 5% to 15% by weight, or approximately 10% to 15% by weight, or approximately 8% to 14% by weight, or approximately 9% to 13% by weight, or approximately 10% to 12% by weight, based on the total weight of the rhamnolipid, and Rha-C10-C12 in amounts of approximately 0.2% to 6% by weight, or approximately 1% to 5% by weight, or approximately 2% to 5% by weight, or approximately 3% to 5% by weight, or approximately 4% to 5% by weight, based on the total weight of the rhamnolipid. The rhamnolipid mixture may also contain RhaRha-C10-C12:1 in amounts ranging from about 0.2% to about 5% by weight, or about 1% to about 4% by weight, or about 3% to about 5% by weight, based on the total weight of the rhamnolipid; RhaRha-C8-C10 in amounts ranging from about 0.2% to about 5% by weight, or about 1% to about 5%, or about 1% to about 4%, or about 2% to about 5%, or about 3% to about 4% by weight, based on the total weight of the rhamnolipid; and RhaC8-C10 in amounts ranging from about 0.2% to about 5% by weight, or about 1% to about 5%, or about 1% to about 4%, or about 2% to about 5%, or about 3% to about 4% by weight, based on the total weight of the rhamnolipid. The rhamnolipid mixture may also contain RhaRha-C12-12 in amounts of approximately 0.1% to 0.5% by weight, or approximately 0.2% to 0.4% by weight, or approximately 0.2% to 0.3% by weight, based on the total weight of the rhamnolipid.

[0031] The rhamnolipid mixture may include a mixture of monorhamnolipid and dirhamnolipid. Monorhamnolipid may be present in amounts of about 10% to about 90% by weight, or about 10% to about 80%, or about 10% to about 70%, or about 10% to about 60%, or about 10% to about 50%, or about 15% to about 50%, or about 20% to about 60%, or about 20% to about 50%, or about 30% to about 50%, or about 40% to about 50%, or about 45% to about 50%, or about 44% to about 50%, or about 45% to about 48%, or about 42% to about 48% by weight, based on the total weight of rhamnolipid in the mixture. Diramnolipid may be present in amounts of approximately 10% to 90% by weight, or approximately 20% to 70% by weight, or approximately 30% to 60% by weight, or approximately 40% to 60% by weight, or approximately 45% to 55% by weight, or approximately 50% to 85% by weight, or approximately 50% to 80% by weight, or approximately 50% to 70% by weight, or approximately 50% to 60% by weight, or approximately 52% to 58% by weight. The weight ratio of monoramnolipid to diramnolipid can be approximately 10:90 to 90:10, or approximately 15:85 to 85:15, or approximately 20:80 to 80:20, or approximately 25:75 to 75:25, or approximately 30:70 to 70:30, or approximately 35:65 to 65:35, or approximately 40:60 to 60:40, or approximately 40:60 to 45:55, or 45:55 to 55:45, or 47:53 to 53:47, or approximately 40:60 to 50:50, or approximately 40:60 to 48:52.

[0032] The terms "activity," "activity%," and "activity by weight%" refer to the amount of the active ingredient, regardless of the amount of water or other solvents that may be present with the ingredient.

[0033] As used herein, “effective amount” means the amount of an active ingredient or composition that, when administered to mammalian skin (e.g., mammalian scalp), can accelerate or otherwise promote the prevention and / or treatment of dandruff. This includes inhibiting or reducing the growth, colonization, and / or survival rate of Malassezia fungi, as well as reducing their progression in the dandruff cycle.

[0034] A monoramnolipid may contain one or more monoramnolipid-monolipid congeners, for example, Rha-C8-:2, Rha-C8, Rha-C10, Rha-C12:2, Rha-C12, Rha-C14:2, or a combination thereof. A monoramnolipid may also contain one or more monoramnolipid-dilipid congeners, for example, Rha-C8-C8, Rha-C8-C10:1, Rha-C10:1-C8, Rha-C8-C10, Rha-C10-C8, Rha-C10-C10:1, Rha-C10-C10, Rha-C8-C12, Rha-C12-C8, Rha-C10-C12:1, Rha-C12:1- This includes C10, Rha-C10-12, Rha-C12-C10, Rha-C10-C14:1, Rha-C12-C12:1, Rha-C10-C14, Rha-C12-C12, Rha-C12-C14, Rha-C14-C14, Rha-C14-C16, Rha-C16-C16, Rha-C10-C10-CH3, Decenoyl-Rha-C10-C10, or combinations thereof.

[0035] Diramnolipid may contain one or more diramnolipid-monolipid congeners, such as Rha-Rha-C8, Rha-Rha-C10, Rha-Rha-C12:1, Rha-Rha-C12, Rha-Rha-C14, or combinations thereof. Diramnolipid may also contain one or more diramnolipid-dilipid congeners, for example, Rha-Rha-C8-C8, Rha-Rha-C8-C10, Rha-Rha-C10-C8, Rha-Rha-C10-C10:1, Rha-Rha-C10-C10, Rha-Rha-C8-C12:1, Rha-Rha-C12:1-C8, Rha-Rha-C10-C12:1, Rha-Rha-C12:1-C10, Rha-Rha-C10-C12, Rha-Rha-C12-C10, Rha-Rha-C1 This includes 0-C14:1, Rha-Rha-C12-C12:1, Rha-Rha-C12:1-C12, Rha-Rha-C12-C12, Rha-Rha-C12-C14, Rha-Rha-C14-C12, Rha-Rha-C14-C14, Rha-Rha-C14-C16, Rha-Rha-C16-C14, Rha-Rha-C16-C16, Rha-Rha-C14-C14-C14, Rha-Rha-C10-C10-CH3, Decenoyl-Rha-Rha-C10-C10, or combinations thereof.

[0036] Rhamnolipid can be produced from rhamnolipid-producing microorganisms capable of synthesizing / producing rhamnolipid under favorable conditions. Such microorganisms include, but are not limited to, bacteria of the phylum Pseudomonadota, Actinobacteria, Fimicutes, and Proteobacteria. Rhamnolipid is of natural origin and therefore has a BCI of 100. In certain embodiments, the rhamnolipid-producing microorganism for producing rhamnolipid is Pseudomonas aeruginosa. Methods for culturing rhamnolipid-producing bacteria and methods for producing rhamnolipid from fermentation are known in the art, for example, from U.S. Patent No. 11,142,782 and U.S. Patent No. 10,144,943, both of which are incorporated herein by reference in their entirety. Methods for purifying rhamnolipid are also known in the art, for example, from U.S. Patent No. 9,884,883 and U.S. Patent No. 10,829,507, both of which are incorporated herein by reference in their entirety.

[0037] In addition to rhamnolipid salts, the dandruff treatment compositions of this technology may include at least one surfactant selected from the group consisting of anionic surfactants, amphoteric surfactants, nonionic surfactants, cationic surfactants, and combinations thereof. In some embodiments, at least one surfactant is a combination of an anionic surfactant and an amphoteric surfactant. Examples of surfactants that can be used in dandruff treatment compositions include sulfonated alkylbenzenes, sulfonated methyl esters, sulfonated alphaolefins, paraffin sulfonates, alkyl sulfates, alkyl alkoxy sulfates, alkyl alkoxy carbonates, alkyl sulfonates, alkyl alkoxy sulfonates, acyl lactic acid, alkyl isethionate, alkyl sarcosinate, alkyl betaine, alkylamidopropyl betaine, alkyl sultaine, alkylamine oxides, and quaternary ammonium compounds. Specific surfactants that can be used in dandruff treatment compositions include sodium laureth sulfate, sodium lauryl sulfate, cocamidopropyl betaine, sodium C14-16 olefin sulfonate, ammonium laureth sulfate, ammonium lauryl sulfate, sodium methyl 2-sulfolaurate, disodium 2-sulfolaurate, cetyl betaine, cocamidopropyl hydroxysultaine, sodium lauroyl sarcosinate, glyceryl caprylate / caprate, behenamidopropyl dimethylamine, brassichamidopropyl dimethylamine, behentrimonium methosulfate, behentrimonium chloride, cetrimonium chloride, stearamidopropyl dimethylamine, their salts, and combinations thereof.

[0038] The dandruff treatment composition may further contain one or more optional components, such as additives and co-compounds. Examples of additives include stabilizers (e.g., trihydroxystearin, ethylene glycol distearate polymer, and mixtures thereof), rheological modifiers (e.g., sodium chloride and sodium xylene sulfonate), emollients, skin conditioning agents, emulsifiers / suspending agents, fragrances, pigments, dyes, herbal extracts, vitamins, builders, enzymes, pH adjusters, preservatives (e.g., one or more compounds selected from butylene glycol, pentylene glycol, hexylene glycol, 1,2-hexanediol, caprylyl glycol, decylen glycol, and glyceryl caprylate), foaming agents having one or more volatile materials (e.g., isobutane, propane, and 1,3,3,3-tetrafluoropropene), and antimicrobial agents. Specific examples of additives include, but are not limited to, polyglycerol alkyl esters, glyceryl esters, glycol esters, benzoic acid esters, vitamins such as vitamin A, vitamin E, or pantothenic acid, or any combination thereof. In some embodiments, one or more additives may include monoglycerides, diglycerides, or combinations thereof.

[0039] Optional co-components may include materials for improving hair shine and / or reducing hair damage. For example, the dandruff treatment compositions disclosed herein may be formulated with compounds that remove aliphatic calcium carboxylate salts from hair, including silicones, siloxanes, natural oils, mineral oils, natural or synthetic waxes, or calcium chelating agents (e.g., diethylenetriaminepentaacetic acid (DTPA), etidronic acid, and mixtures thereof).

[0040] In addition, the dandruff treatment compositions disclosed herein may be formulated with high-melting-point aliphatic compounds, such as aliphatic alcohols including cetyl alcohol, stearyl alcohol, behenyl alcohol, or brassica alcohol. In yet another embodiment, the dandruff treatment compositions disclosed herein may be formulated with 2-pyridinol-N-oxide materials, including unsubstituted 2-pyridinol-N-oxide, substituted 2-pyridinol-N-oxide, and their salts and tautomers. For example, the dandruff treatment compositions disclosed herein may be formulated with 1-hydroxy-4-methyl-6-(2,4,4-trimethylpentyl)-2-pyridone monoethanolamine salt.

[0041] The dandruff treatment compositions disclosed herein may also be formulated with a gel network. The gel network comprises a lamellar structure (e.g., an aggregate of microsheets and / or vesicles). The gel network generally comprises a lamellar solid crystalline phase containing at least one fatty acid and at least one secondary component (e.g., at least one secondary surfactant or additional fatty amphiphilic substance, and water or other suitable solvent).

[0042] In some embodiments, the compositions disclosed herein may also be formulated with one or more additional antifungal or anti-dandruff components. Such components include zinc pyrithione, sulfur, selenium sulfide, coal tar, and soluble anti-dandruff active ingredients, for example, one or more compounds selected from the group of azoles (e.g., crimbazole, ketoconazole, itraconazole, econazole, and erbiol), hydroxypyridines (e.g., piroctone olamine, cyclopirox, rilopirox, and MEA-hydroxyoctyloxypyridinone), keratolytic agents (e.g., salicylic acid and other hydroxy acids), strobilurins (e.g., azoxystrobin and metal chelating agents, e.g., 1,10-phenanthroline, and hinokitiol).

[0043] The dandruff treatment composition of this technology also contains at least one carrier suitable for personal care or dermatological care, so that the total percentage of the composition is 100% by weight. 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, or a combination thereof.

[0044] The dandruff treatment compositions described herein may have a predetermined pH range. For example, the dandruff treatment compositions described herein may have a pH in the range of 4.5 to 8.0, 5.0 to 7.5, 5.5 to 7.5, or 6.0 to 7.5.

[0045] The dandruff treatment compositions described herein can be formulated as solutions, aqueous emulsions, gels, pastes, or creams.

[0046] In one embodiment, the present technology provides an anti-dandruff composition and a method of use, the composition comprising a mixture of rhamnolipid salts in an amount of 0.01% to 95% by weight, or 0.01% to 30% by weight, or 0.01% to 20% by weight, or 0.01% to 10% by weight, wherein the mixture of rhamnolipid salts is in a weight ratio of 10:90 to 90:10 monorhamnolipid:diramnolipid, or 15:85 to 90:10 monorhamnolipid:diramnolipid, or 20:80 to 90:10 monorhamnolipid:diramnolipid, or 30:70 to 90:10 monorhamnolipid:diramnolipid, or 40:60 to 90:10 monorhamnolipid: Diramnolipid, or monoramnolipid at 30:70-80:20:diramnolipid, or monoramnolipid at 30:70-70:30:diramnolipid, or monoramnolipid at 30:70-60:40:diramnolipid, or monoramnolipid at 30:70-50:50:diramnolipid, or monoramnolipid at 30:70-45:55:diramnolipid, or monoramnolipid at 40:60-60:40:diramnolipid, or monoramnolipid at 40:60-50:50:diramnolipid, or monoramnolipid at 40:60-45:55:diramnolipid, including monoramnolipid and diramnolipid.

[0047] In another aspect, the present technology provides an anti-dandruff composition and a method of use, the composition comprising a mixture of rhamnolipid salts in an amount of about 0.1% to about 99% by weight (or about 0.05% to about 99% by weight, based on the total weight of the composition), wherein the mixture of rhamnolipid salts is in a ratio of about 10:90 to about 47:53, or about 20:80 to about 47:53, or about 30:70 to about 47:53, or about 40:60 to about 47:53, or about 40:60 to about 45:55, or It comprises mono-rhamnolipid and di-rhamnolipid in a weight ratio of 43:57 to 45:55, Rha-C10-C10 mono-rhamnolipid salt in an amount of approximately 5% to 50% by weight, or approximately 5% to 38% by weight, based on the total weight of rhamnolipid in the mixture of rhamnolipids, and Rha-C10-C10 di-rhamnolipid salt in an amount of approximately 30% to 80% by weight, or approximately 34% to 80% by weight, based on the total weight of rhamnolipid in the mixture of rhamnolipids.

[0048] Further embodiments of the present technology provide a method for controlling, treating or preventing dandruff in subjects affected or susceptible to dandruff, or a method for treating excessive Malassezia fungi on the skin of mammals, the method comprising administering an effective amount of a composition comprising a mixture of rhamnolipid salts to the skin of a mammal, the mixture of rhamnolipid salts being in a weight ratio of 10:90 to 90:10 monorhamnolipid:diramnolipid, or 15:85 to 90:10 monorhamnolipid:diramnolipid, or 20:80 to 90:10 monorhamnolipid:diramnolipid, or 30:70 to 90:10 monorhamnolipid:diramnolipid, or 40:60 to 90:10 mono Rhamnolipid:diramnolipid, or monoramnolipid:diramnolipid at 30:70-80:20, or monoramnolipid:diramnolipid at 30:70-70:30, or monoramnolipid:diramnolipid at 30:70-60:40, or monoramnolipid:diramnolipid at 30:70-50:50, or monoramnolipid:diramnolipid at 30:70-45:55, or monoramnolipid:diramnolipid at 40:60-60:40, or monoramnolipid:diramnolipid at 40:60-50:50, or monoramnolipid:diramnolipid at 40:60-45:55, including monoramnolipid and diramnolipid. The composition exhibits antifungal activity against fungi of the genus Malassezia. In certain embodiments, the Malassezia fungus is M. furfur. Other Malassezia fungi include, for example, M. dermatis, M. equi, M. restricta, M. globose, M. ovalis (also known as Pityrosporum ovale), M. pachydermatis, M. restricta, M. sympodialis, M. obtusa, M. slooffiae, or combinations thereof.

[0049] In some embodiments, the technology provides a method for treating and / or preventing dandruff or excessive Malassezia fungi on mammalian skin, as described above, wherein the rhamnolipid mixture comprises monorhamnolipid and dirhamnolipid in a weight ratio of 40:60 to 45:55 or 40:60 to 48:52, with about 20% to about 40% by weight of Rha-C10-C10 monorhamnolipid and about 30% to 50% by weight of Rha-C10-C10 dirhamnolipid, based on the total weight of rhamnolipid in the rhamnolipid mixture.

[0050] The rhamnolipid compositions disclosed herein are associated with specific biological properties. One such property is the ability to reduce the growth of Malassezia versicolor. In some embodiments, the rhamnolipid compositions disclosed herein exhibit a characteristic reduction in Malassezia versicolor colony-forming units (CFUs) per agar plate (compared to a control without rhamnolipid and performed according to the method described in Example 2, "Solid Agar Embedding Antifungal Efficacy Test Method" below). For example, the rhamnolipid compositions disclosed herein may show a reduction in the biological activity (i.e., growth) of Malassezia versicolor, as determined according to the method described in Example 2, of about 25% to about 99%, about 50% to about 99%, about 75% to about 99%, about 25% to about 75%, about 25% to about 50%, or about 25% to about 40%.

[0051] In other embodiments, the rhamnolipid compositions disclosed herein exhibit characteristic reduction rates of Malassezia colony-forming units (CFUs) per mL (compared to a control without rhamnolipid and carried out as described in the 96-well plate liquid method of Example 3 below). For example, the rhamnolipid compositions disclosed herein show that when the initial load of Malassezia (i.e., microbial challenge) is about 5E+05 (CFU / mL), the reduction rate of Malassezia biological activity (i.e., growth) determined according to the method of Example 3 is about 40% to about 70%, about 40% to about 65%, about 50% to about 70%, or about 55% to about 65%. The rhamnolipid compositions disclosed herein may result in a reduction in the biological activity (i.e., growth) of Malassezia versicolor, as determined according to the method described in Example 3, when the initial load (i.e., microbial challenge) of Malassezia versicolor is about 5E+04 (CFU / mL), and the reduction may be about 40% to about 80%, about 40% to about 75%, about 50% to about 80%, about 60% to about 80%, or about 65% to about 75%. When the initial load (i.e., microbial challenge) of Malassezia versicolor is about 5E+03 (CFU / mL), the reduction may be about 50% to about 90%, about 50% to about 85%, about 60% to about 90%, about 70% to about 90%, or about 70% to about 85%. The rhamnolipid compositions disclosed herein may result in a reduction in the biological activity (i.e., growth) of Malassezia versicolor, as determined according to the method described in Example 3, when the initial load (i.e., microbial challenge) of Malassezia versicolor is about 6E+02 (CFU / mL), and / or when the initial load (i.e., microbial challenge) is about 3E+02 (CFU / mL), the reduction may be about 50% to about 97%, about 70% to about 97%, about 80% to about 97%, about 85% to about 97%, or about 90% to about 97%.

[0052] In some embodiments, the rhamnolipid compositions disclosed herein also exhibit a specific rate of reduction in the biological activity (i.e., growth) of Malassezia versicolor when formulated with anionic, cationic, amphoteric, or nonionic co-compounds. For example, when formulated with at least one surfactant selected from the group consisting of anionic surfactants, amphoteric surfactants, nonionic surfactants, cationic surfactants, and combinations thereof, the rhamnolipid compositions disclosed herein may exhibit a rate of reduction in the biological activity (i.e., growth) of Malassezia versicolor, as determined according to the method described in Example 4 (Selective Formulation Example / Test using 96-well plate liquid method), when the initial load (i.e., microbial challenge) of Malassezia versicolor is about 2.5E+05 (CFU / mL), of about 70% to about 100%, about 80% to about 100%, about 90% to about 100%, or about 95% to about 100%.

[0053] Items of the present invention The present invention further relates to the following items.

[0054] 1. A dandruff treatment composition comprising a rhamnolipid salt mixture in an amount of about 0.01% to about 30% by active weight, based on the total weight of the composition, wherein the rhamnolipid salt mixture comprises monorhamnolipid and diramnolipid having a monorhamnolipid:diramnolipid weight ratio of about 40:60 to about 60:40. The rhamnolipid salt may be zinc-rhamnolipid. The rhamnolipid composition also comprises, based on the total weight of rhamnolipids present in the composition, about 29% to about 40% by weight of C10-C10 monorhamnolipid, about 35% to about 50% by weight of C10-C10 dirhamnolipid, about 0.2% to about 5% by weight of C8-C10 monorhamnolipid, about 0.2% to about 5% by weight of C8-C10 dirhamnolipid, about 2% to about 5% by weight of C10-C12 monorhamnolipid, and about 8% to about 14% by weight of C10-C12 dirhamnolipid. The composition also further comprises at least one surfactant selected from the group consisting of anionic surfactants, amphoteric surfactants, nonionic surfactants, cationic surfactants, and combinations thereof, and at least one dermatologically acceptable carrier in an amount that gives 100% of the total weight of the composition.

[0055] 2. The above-mentioned composition wherein at least one surfactant is one or more of the following: sodium laureth sulfate, sodium lauryl sulfate, cocamidopropyl betaine, sodium C14-16 olefin sulfonate, ammonium laureth sulfate, ammonium lauryl sulfate, sodium methyl 2-sulfolaurate, disodium 2-sulfolaurate, cetyl betaine, cocamidopropyl hydroxysultaine, sodium lauroyl sarcosinate, glyceryl caprylate / caprate, behenamidopropyl dimethylamine, blascicamidopropyl dimethylamine, behentrimonium methosulfate, behentrimonium chloride, cetrimonium chloride, or stearamidopropyl dimethylamine, for example, a combination of sodium laureth sulfate, sodium lauryl sulfate, and cocamidopropyl betaine, where, based on the weight of the composition, the amount of sodium laureth sulfate is about 1% to about 2% by weight, and the amount of sodium lauryl sulfate is about 1% to about 2% by weight. The amount is in weight percent, and the amount of cocamidopropyl betaine is about 0.5% to about 2% by weight, or a combination of sodium lauryl sulfate and cocamidopropyl betaine, where, based on the weight of the composition, the amount of sodium lauryl sulfate is about 2% to about 4% by weight and the amount of cocamidopropyl betaine is about 0.5% to about 2% by weight, or a combination of sodium C14-16 olefin sulfonate and cocamidopropyl betaine, where, based on the weight of the composition, the amount of sodium C14-16 olefin sulfonate is about 2% to about 4% by weight and the amount of cocamidopropyl betaine is about 0.5% to about 2% by weight, or a combination of ammonium laureth sulfate, ammonium lauryl sulfate, and cocamidopropyl betaine, where, based on the weight of the composition, the amount of ammonium laureth sulfate is about 1% to about 3% by weight, the amount of ammonium lauryl sulfate is about 1% to about 3% by weight and the amount of cocamidopropyl betaine is about 0%.A composition comprising 5% to about 2% by weight of one of the following: or a combination of methyl 2-sulfolaurate sodium, 2-sulfolaurate disodium, cetylbetaine, and cocamidopropyl hydroxysultaine, wherein, based on the weight of the composition, the combined amounts of methyl 2-sulfolaurate sodium and 2-sulfolaurate disodium are about 1% to about 2% by weight, the amount of cetylbetaine is about 1% to about 2% by weight, and the amount of cocamidopropyl hydroxysultaine is about 0.5% to about 3%, or about 1% to about 2% by weight. One or more surfactants may be, based on the weight of the composition, a total amount of sodium methyl 2-sulfolaurate and disodium 2-sulfolaurate in an amount of about 1% to about 5% by weight, or sodium lauroyl sarcosinate in an amount of about 1% to about 5% by weight, or glyceryl caprylate / caprate in an amount of about 0.1% to about 1.0% by weight, or sodium lauryl sulfate in an amount of about 2% to about 5% by weight, or sodium C14-16 olefin sulfonate in an amount of about 2% to about 5% by weight, or [Examples]

[0056] The technology and its advantages described herein will be better understood by referring to the following examples. These examples are provided to illustrate specific embodiments of the technology. By providing these examples, the inventors do not limit the scope and spirit of the technology.

[0057] Example 1 - Preparation of a rhamnolipid composition After fermentation, the entire culture containing rhamnolipid undergoes several purification steps. The specific combination of steps depends on the rhamnolipid active ingredient and purity requirements for the target application. The specific rhamnolipid mixtures used herein are purified by a solvent extraction process. As used herein, the acronym SEP-RM refers to Solvent Extraction Purified-Rhamnolipid Mixture (or composition).

[0058] For example, the SEP-RM rhamnolipid compositions used herein are prepared according to at least the following processing steps: (1) fermentation of suitable rhamnolipid-producing bacteria, (2) biomass separation, (3) sterilization, (4) purification (e.g., filtration), (5) acidification, (6) bleaching, (7) washing, (8) solvent extraction, (9) neutralization and dilution, and (10) final finishing steps(s). As discussed below, the solvent extraction process / step is intended to result in a higher purity rhamnolipid mixture / composition for personal care applications, as described herein. Additional solvent extraction steps are required, for example, to achieve a lighter color and milder odor profile required for the personal care market.

[0059] At the end of fermentation, the total culture typically contains rhamnolipid along with fermentation biomass and other by-products. To separate the biomass solids, the culture can be centrifuged. The resulting centrifuged culture is then subjected to sterilization (e.g., high-temperature sterilization), and subsequently, the centrifuged and sterilized culture is purified by filtration to remove suspended solids.

[0060] Further purification is achieved by treating the purified culture medium with acid, which converts the rhamnolipid into a water-insoluble form that settles at the bottom and separates from the bulk aqueous phase. This highly concentrated acidified rhamnolipid is then isolated (referred to as acidified concentrated clarified culture medium (ACCB)). The ACCB is then treated with a bleaching agent and subsequently washed with water to remove residual bleaching agent and other water-soluble impurities. At this point, the decolorized and washed ACCB is concentrated (e.g., with approximately 45% or more active ingredients, or approximately 35% to 55% active ingredients) and purer (e.g., with a purity of approximately 75% or more, or approximately 65% ​​to 85% purity).

[0061] Solvent Extraction Process – To achieve better purity, color, and odor, the decolorized acidified concentrated purified culture medium (ACCB) undergoes a solvent extraction process. This is generally carried out by dissolving the decolorized and washed ACCB in an organic solvent, preferably ethyl acetate. The typically formed lag layer is separated from the bulk solution. Activated carbon is then added to the rhamnolipid solution in ethyl acetate for further decolorization and deodorization. The slurry is filtered, and the resulting solution is stripped under vacuum to remove the ethyl acetate. The highly concentrated ACCB obtained as a residue is then neutralized and diluted (e.g., to about 20% to about 30% active ingredients, preferably about 25% active ingredients) to obtain crude SEP-RM. As a final purification step, the crude SEP-RM is washed with ethyl acetate to extract the defoamer, and the final product is optically clear SEP-RM with about 25% active ingredients and about 85% purity.

[0062] Rhamnolipid, a zinc salt, was prepared by adding an equimolar amount of ZnCl2 to the active component of rhamnolipid (such as SEP-RM) in an aqueous solution and mixing at ambient temperature for approximately 20 minutes. The solution was then centrifuged to separate it into layers. The upper layer was decanted, and the lower layer was washed with approximately 20 g of DI water and centrifuged. The washing procedure was repeated, the lower layer was collected, and dried in a vacuum oven at 50°C to obtain Zn-rhamnolipid salt.

[0063] Monoramnolipid:diramnolipid ratio and congener distribution The monorhamnolipid:diramnolipid ratio and congener distribution of the SEP-RM rhamnolipid composition of this disclosure can be determined using HPLC (high-performance liquid chromatography). As will be understood by those skilled in the art, HPLC separates rhamnolipid congeners by chain length and the number of rhamnose moieties (more polar congeners elute first). Fractions are collected and analyzed by mass spectrometry to identify each peak. The congener distribution is reported as an area percentage (the active ingredient is calculated in comparison to a known standard).

[0064] The HPLC homologue distribution of the standard SEP-RM rhamnolipid composition of this disclosure is shown below. [Table 1]

[0065] Monorhamnolipid may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts of about 35% to about 55% by weight, or about 38% to about 52% by weight, or about 40% to about 50% by weight, or about 42% to about 48% by weight, or about 42% to about 46% by weight, or about 43% to about 45% by weight, or about 44% by weight, based on the total weight of rhamnolipid in the mixture.

[0066] Diramnolipid may be present in the SEP-RM rhamnolipid composition of this disclosure in an amount of about 45% to about 65% by weight, or about 50% to about 60% by weight, or about 52% to about 58% by weight, or about 53% to about 57% by weight, or about 54% to about 56% by weight, or about 55% by weight, based on the total weight of rhamnolipid in the mixture.

[0067] The weight ratio of monoramnolipid to diramnolipid in the SEP-RM ramnolipid composition disclosed herein may be approximately 35:65 to approximately 65:35, or approximately 38:62 to approximately 62:38, or approximately 40:60 to approximately 60:40, or approximately 42:58 to approximately 58:42, or approximately 43:57 to approximately 57:43, or approximately 41:59 to approximately 44:56, or approximately 42:58 to approximately 44:56, or approximately 42:58, or approximately 43:57, or approximately 44:56, or approximately 50:50.

[0068] Monorhamnolipid C8-C10 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 1% to about 6% by weight, or about 2% to about 5% by weight, or about 3% to about 4% by weight, or about 3% by weight, or about 4% by weight, based on the total weight of rhamnolipids present in the composition.

[0069] Monorhamnolipid C10-C10 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 20% to about 50% by weight, or about 25% to about 45% by weight, or about 30% to about 40% by weight, or about 32% to about 38% by weight, or about 34% to about 38% by weight, or about 35% to about 37% by weight, or about 36% to about 37% by weight, or about 36% by weight, or about 37% by weight, based on the total weight of rhamnolipids present in the composition.

[0070] Monorhamnolipid C10-C12 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 1% to about 8% by weight, or about 2% to about 6% by weight, or about 3% to about 5% by weight, or about 4% to about 5% by weight, or about 4% by weight, or about 5% by weight, based on the total weight of rhamnolipids present in the composition.

[0071] Diramnolipid C8-C10 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 1% to about 6% by weight, or about 2% to about 5% by weight, or about 3% to about 4% by weight, or about 3% by weight, or about 4% by weight, based on the total weight of rhamnolipid present in the composition.

[0072] Diramnolipid C10-C10 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 20% to about 50% by weight, or about 25% to about 45% by weight, or about 30% to about 40% by weight, or about 32% to about 38% by weight, or about 34% to about 38% by weight, or about 35% to about 38% by weight, or about 36% to about 38% by weight, or about 37% by weight, or about 38% by weight, based on the total weight of rhamnolipid present in the composition.

[0073] Diramnolipid C10-C12.1 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 1% to about 7% by weight, or about 2% to about 6% by weight, or about 3% to about 5% by weight, or about 3% to about 4% by weight, or about 3% by weight, or about 4% by weight, based on the total weight of rhamnolipid present in the composition.

[0074] Diramnolipid C10-C12 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 8% to about 14% by weight, or about 9% to about 13% by weight, or about 10% to about 12% by weight, or about 11% to about 12% by weight, or about 11% by weight, or about 12% by weight, based on the total weight of rhamnolipid present in the composition.

[0075] Diramnolipid C12-C12 congeners may be present in the SEP-RM rhamnolipid composition of this disclosure in amounts ranging from about 0.1% to about 0.5% by weight, or about 0.2% to about 0.4% by weight, or about 0.2% to about 0.3% by weight, or about 0.25% to about 0.3% by weight, or about 0.2% by weight, or about 0.25% by weight, or about 0.25% by weight, or about 0.25% by weight, based on the total weight of rhamnolipid present in the composition.

[0076] The SEP-RM rhamnolipid compositions provided herein can be processed with or without a bleaching step (which occurs after acidification and before any solvent extraction step). For the SEP-RM rhamnolipid compositions disclosed herein, the monorhamnolipid:diramnolipid ratio and the associated congener distribution are not affected by whether or not a bleaching step is included in the purification process.

[0077] Example 2 - Solid agar-embedded antifungal efficacy test method To evaluate the efficacy of the rhamnolipid compositions disclosed herein against Malassezia furfur, a solid agar embedding test method was used. In this method, the test sample was diluted in undiluted nutrient agar matrix and dispensed into Petri dishes at 20 mL per dish. The test species, Malassezia furfur ATCC 14521 (i.e., M. furfur), was supplied to each dish at different levels using fully cultured and prepared cultures. The nutrient agar matrix / Malassezia furfur mixture was maintained at 45±2°C, thoroughly mixed, and solidified by cooling to 20-25°C. The plates were then incubated at 30±2°C for up to 7 days until sufficient growth occurred in the control. The absence of growth compared to the control is characteristic of antifungal activity.

[0078] The nutrient agar matrices used included: potato dextrose agar (PDA) (BD 213400, a general medium for fungal species), and modified Dixon agar (MD) (ATCC medium 2693, consisting of proteins, carbon, oil, and acid preferred by Malassezia). The test samples used contained rhamnolipid compositions prepared using solvent extraction as described above (e.g., SEP-RM / lot A having 26.9% active ingredient, 47.2% mono-rhamnolipid (RL), and 52.8% di-RL, and SEP-RM / lot B having 25.7% active ingredient, 47.6% mono-RL, and 52.4% di-RL).

[0079] The test results are as follows: [Table 2]

[0080] Example 3 - 96-well plate liquid method

[0081] To evaluate the efficacy of the rhamnolipid compositions disclosed herein against Malassezia versicolor, a 96-well plate liquid culture method was used. In this method, samples were diluted in a low-intensity nutrient broth liquid medium, and up to 300 μL per well was dispensed into the plate wells. Malassezia versicolor was supplied to each well set at different levels using well-grown and prepared cultures. Resazurin, a blue fluorescent dye used as a redox indicator in cell viability and growth assays, was added to each sample and control well. The mixtures were kept at 20–25°C and thoroughly mixed. The plates were then incubated at 30±2°C for up to 3 days until mature biological activity was detected from the control, first visually (resazurin changes color upon metabolic treatment) and then quantitatively (by using a fluorescence-enabled plate reader). The tests were performed using 0.3x concentration Modified Dixon (MD), which is associated with the scalp and is a nutrient matrix favored by microorganisms.

[0082] The test samples used contained rhamnolipid compositions prepared using solvent extraction as described above (SEP-RM / Lot A, containing 26.9% active ingredient, 47.2% mono-rhamnolipid (RL), and 52.8% di-RL, and SEP-RM / Lot B, containing 25.7% active ingredient, 47.6% mono-RL, and 52.4% di-RL). Rhamnolipid was added to each test mixture at an active ingredient concentration of 1.0%.

[0083] The test results are shown below. [Table 3]

[0084] Example 4 - Selected Formulation Example The efficacy of the rhamnolipid compositions disclosed herein against Malassezia was evaluated in the presence of anionic, amphoteric, or nonionic co-compounds. The tests were performed using the 96-well plate liquid culture method described above, at the activity concentrations described below in specific test samples. The test samples used contained rhamnolipid compositions (SEP-RM / Lot B having 47.6% mono-RL and 52.4% di-RL) prepared using solvent extraction as described above. Rhamnolipid was added to each test mixture at an active ingredient concentration of 1.0%, as shown below.

[0085] As outlined in the table below, rhamnolipid (SEP-RM / Lot B, containing 25.7% active ingredient, 47.6% mono-RL, and 52.4% di-RL) was mixed in water with various co-compounds (the %(w / w) figures are based on the total weight of the composition): [Table 4] [Table 5] [Table 6] [Table 7] Table 8 Table 9 Table 10 Table 11 Table 12 Table 13 Table 14 Table 15 Table 16 Table 17 Table 18

[0086] To evaluate efficacy, Malassezia versicolor was added to the rhamnolipid formulation described above, according to the 96-well plate liquid culture method described above. Individual samples were diluted in a low-intensity nutrient broth liquid medium, and up to 300 μL was dispensed into plate wells at a time. Malassezia versicolor was supplied to each set of wells using well-grown and prepared cultures at a level of 2.5E+05 cfu / ml. Resazurin, a blue fluorescent dye used as a redox indicator in cell viability and growth assays, was added to each sample and control well. The mixture was kept at 20–25°C and thoroughly mixed. The plate was then incubated at 30±2°C for up to 3 days until mature biological activity was detected from the control, first visually (resazurin changes color upon metabolic treatment) and then quantitatively (by using a fluorescence-compatible plate reader). The tests were performed using 0.3x concentration Modified Dixon (MD), which is related to the scalp and is a nutrient matrix favored by microorganisms.

[0087] As described below, the rhamnolipid prepared above (SEP-RM / Lot B, having 47.6% mono-RL and 52.4% di-RL) demonstrated efficacy at 1% (w / w based on the total weight of the composition) (47% inhibition against a 2.5E+05 cfu / mL M. furfur challenge). The SEP-RM / Lot B rhamnolipid performed significantly better than the comparative sample (a commercially available rhamnolipid product with a mono:di-rhamnolipid ratio of 4:96), which showed only 19% inhibition (against a 2.5E+05 cfu / mL M. furfur challenge). When the co-compound was added to 1% SEP-RM / lot B (i.e., the sample described above) containing 47.6% mono-RL and 52.4% di-RL, the performance increased to 100% inhibition, which is the maximum detectable value (for a M. furfur challenge of 2.5E+05 cfu / ml).

[0088] The test results are shown below. [Table 19]

[0089] Domestic brand anti-dandruff products were also tested according to the protocol described above (this example and Example 3). As shown below, the efficacy of the SEP-RM rhamnolipid formulation (mentioned above) is consistent with the performance of three domestic brand anti-dandruff products (0.3x concentration) and quaternary ammonium compounds (1839-220) at 0.3x RTU. [Table 20]

[0090] Example 5: Zn-rhamnolipid activity To evaluate the efficacy of the Zn-rhamnolipid compositions disclosed herein against M. furfur, the 96-well plate liquid culture method described in Example 3 was used. The test samples contained a mixture of rhamnolipids prepared using solvent extraction as described above and converted to Zn-rhamnolipid salts. The Zn-rhamnolipid mixture was added as an active ingredient at a concentration of 1.0% to either water or a base shampoo composition containing 10% by weight of sodium lauryl sulfate, 3% by weight of cocamidopropyl betaine, and 3% by weight of sodium laureth sulfate, and a total of 100% by weight of water as active ingredients. Comparative samples were prepared from a domestic brand (H&S) dandruff shampoo product containing 1% zinc pyrithione, as well as from Zn-rhamnolipid samples prepared from commercially available rhamnolipid compositions containing approximately 3% monorhamnolipid and 97% dirhamnolipid converted to Zn-rhamnolipid salts as described above. Water alone, the base composition alone, and an aqueous zinc pyrithione solution were used as controls. The sample dilutions and test results are shown below. [Table 21]

[0091] Example 6 - Zn-rhamnolipid combined with an additional antifungal agent The efficacy of Zn-rhamnolipid in combination with different antifungal active ingredients in water or base shampoo compositions described in Example 5 was evaluated using the 96-well plate liquid culture method described in Example 3. The antifungal active ingredients tested were salicylic acid and zinc pyrithione. The active ingredient concentrations and test results of the tested samples are shown below. [Table 22]

[0092] This technology is described herein in complete, clear, and concise terms so that those skilled in the art can implement it. The foregoing describes preferred embodiments of the technology and should be understood to be modifiable without departing from the spirit or scope of the technology as set forth in the appended claims. Furthermore, these embodiments are provided for illustrative purposes only, not exhaustively, of some embodiments within the claims.

Claims

1. A composition for treating dandruff, (a) A mixture of rhamnolipid salts in an amount of about 0.01% to about 30% by weight, based on the total weight of the composition, wherein the mixture of rhamnolipid salts comprises monorhamnolipid and diramnolipid in a weight ratio of monorhamnolipid:diramnolipid of 10:90 to 90:10, based on the total weight of the mixture of rhamnolipid salts, about 5% to about 50% by weight, Rha-C10-C10 mono-rhamnolipid salt, and about 30% to about 80% by weight, RhaRha-C10-C10 diramnolipid salt, based on the total weight of the mixture of rhamnolipid salts. (b) At least one surfactant selected from the group consisting of anionic surfactants, amphoteric surfactants, nonionic surfactants, cationic surfactants, and combinations thereof, (c) comprising a dermatologically acceptable carrier in an amount totaling 100% by weight, The composition reduces Malassezia fungi present on mammalian skin in the absence of other antifungal active ingredients.

2. A composition for treating dandruff, (a) A mixture of zinc-rhamnolipid salts in an amount of about 0.01% to about 30% by weight, based on the total weight of the composition, wherein the mixture of zinc-rhamnolipid salts comprises monorhamnolipid and diramnolipid in a weight ratio of monorhamnolipid:diramnolipid of 10:90 to 90:10, based on the total weight of the mixture of zinc-rhamnolipid salts, about 5% to about 50% by weight, based on the total weight of the mixture of zinc-rhamnolipid salts, and about 30% to about 80% by weight, based on the total weight of the mixture of zinc-rhamnolipid salts, (b) comprising a dermatologically acceptable carrier in an amount totaling 100% by weight, (c) A composition that reduces Malassezia fungi present on the skin of mammals.

3. The composition according to claim 2, wherein the composition further comprises at least one surfactant selected from the group consisting of anionic surfactants, amphoteric surfactants, nonionic surfactants, cationic surfactants, and combinations thereof.

4. The composition according to any one of claims 1 to 3, wherein the amount of the rhamnolipid salt is about 0.1% to about 10% by weight, 0.1% to about 5% by weight, about 0.3% to about 5% by weight, about 0.3% to about 3% by weight, about 0.3% to about 2% by weight, or about 0.3% to about 1% by weight of the composition.

5. The composition according to claim 1 or claim 3 to 4, wherein at least one surfactant is present in an amount of about 2% to about 40% by weight of the composition.

6. The composition according to claim 1 or any one of claims 3 to 5, wherein the at least one surfactant comprises an anionic surfactant in an amount of about 1% to about 40% by weight of the composition.

7. The composition according to any one of claims 1 or 3 to 6, wherein the at least one surfactant comprises an amphoteric surfactant in an amount of about 1% to about 40% by weight, about 1% to about 30% by weight, about 1% to about 20% by weight, or about 1% to about 10% by weight of the composition.

8. The composition according to any one of claims 1 to 7, wherein the mixture of rhamnolipid salts comprises monorhamnolipid and diramnolipid in a weight ratio of 40:60 to 60:40 monorhamnolipid:diramnolipid, 45:55 to 55:45 monorhamnolipid:diramnolipid, or 47:53 to 53:47 monorhamnolipid:diramnolipid, based on the total weight of the mixture of rhamnolipid salts.

9. The composition according to any one of claims 1 to 8, wherein the mixture of rhamnolipid salts comprises, based on the total weight of the mixture of rhamnolipid salts, about 20% to about 60% by weight, about 30% to about 50% by weight, about 40% to about 50% by weight, or about 45% to about 48% by weight of monorhamnolipid.

10. The composition according to any one of claims 1 to 9, wherein the mixture of rhamnolipid salts comprises about 20% to about 70% by weight, about 30% to about 60% by weight, about 40% to about 60% by weight, or about 45% to about 55% by weight, or about 50% to about 55% by weight, based on the total weight of the mixture of rhamnolipid salts.

11. The composition according to any one of claims 1 to 10, wherein the mixture of rhamnolipid salts comprises Rha-C10-C10 monorhamnolipid salt in an amount of about 20% to about 50% by weight, about 25% to about 45% by weight, about 30% to about 40% by weight, or about 35% to about 38% by weight, based on the total weight of the mixture of rhamnolipid salts.

12. The composition according to any one of claims 1 to 11, wherein the mixture of rhamnolipid salts comprises RhaRha-C10-C10 diramnolipid salt in an amount of about 20% to about 50% by weight, about 25% to about 45% by weight, about 30% to about 40% by weight, or about 35% to about 38% by weight, based on the total weight of the mixture of rhamnolipid salts.

13. The composition according to any one of claims 1 to 12, wherein the mixture of rhamnolipid salts comprises Rha-C8-C10 monorhamnolipid salt in an amount of about 1% to about 10% by weight, about 1% to about 5% by weight, about 2% to about 5% by weight, or about 3% to about 4% by weight, based on the total weight of the mixture of rhamnolipid salts.

14. The composition according to any one of claims 1 to 13, wherein the mixture of rhamnolipid salts comprises RhaRha-C8-C10 diramnolipid salt in an amount of about 1% to about 10% by weight, about 1% to about 5% by weight, about 2% to about 5% by weight, or about 3% to about 4% by weight, based on the total weight of the mixture of rhamnolipid salts.

15. The composition according to any one of claims 1 to 14, wherein the mixture of rhamnolipid salts comprises Rha-C10-C12 monorhamnolipid salt in an amount of about 1% to about 10% by weight, about 1% to about 5% by weight, about 2% to about 5% by weight, about 3% to about 5% by weight, or about 4% to about 5% by weight, based on the total weight of the mixture of rhamnolipid salts.

16. The composition according to any one of claims 1 to 15, wherein the mixture of rhamnolipid salts comprises RhaRha-C10-C12 diramnolipid salt in an amount of about 1% to about 20% by weight, about 5% to about 15% by weight, about 10% to about 15% by weight, or about 10% to about 12% by weight, based on the total weight of the mixture of rhamnolipid salts.

17. The composition according to any one of claims 1 to 16, wherein the composition comprises one or more additives.

18. The composition according to claim 17, wherein the one or more additives include one or more of the following: pH adjusters, skin conditioning agents, antioxidants, preservatives, fragrances, pigments, and dyes.

19. The composition according to claim 17, wherein one or more of the additives include monoglycerides, diglycerides, or combinations thereof.

20. The composition according to claim 1 or any one of claims 3 to 19, wherein the at least one surfactant is selected from the group consisting of sulfonated alkylbenzene, sulfonated methyl ester, sulfonated alphaolefin, paraffin sulfonate, alkyl sulfate, alkyl alkoxy sulfate, alkyl alkoxy carbonate, alkyl sulfonate, alkyl alkoxy sulfonate, acyl lactylate, alkyl isethionate, alkyl sarcosinate, alkyl betaine, alkylamidopropyl betaine, alkyl sultaine, alkylamine oxide, quaternary ammonium compounds, salts thereof, and combinations thereof.

21. The composition according to any one of claims 1 to 20, wherein the dermatologically acceptable carrier comprises water.

22. The composition according to any one of claims 1 to 21, wherein the dermatologically acceptable carrier comprises one or more alcohols, glycols, or solvents.

23. The composition according to any one of claims 1 to 22, wherein the composition further comprises at least one antifungal active component selected from the group consisting of zinc pyrithione, sulfur, selenium sulfide, salicylic acid, coal tar, ketoconazole, and piroctone olamine.

24. The composition according to any one of claims 1 to 23, wherein the Malassezia species is M. furfur, M. dermatis, M. equi, M. globesa, M. japonica, M. obtusa, M. ovalis (also known as Pityrosporum ovale), M. pachydermatis, M. restricta, M. slooffiae, M. sympodialis, or a combination thereof.

25. The composition according to any one of claims 1 to 24, wherein the composition has a pH in the range of 4.5 to 8.

0.

26. The composition according to any one of claims 1 to 25, wherein the composition is a solution, gel, paste, or cream.

27. A method for controlling, treating, or preventing dandruff in a person who is affected by or susceptible to dandruff, comprising applying an effective amount of the dandruff treatment composition according to any one of claims 1 to 26 to the scalp and / or hair of the person.

28. A method for controlling, treating, or preventing an excess of Malassezia fungi on the skin of a mammal, comprising applying an effective amount of the dandruff treatment composition according to any one of claims 1 to 27 to the skin of the mammal.

29. The method according to claim 28, wherein the Malassezia species is M. furfur, M. dermatis, M. equi, M. globesa, M. japonica, M. obtusa, M. ovalis (also known as Pityrosporum ovale), M. pachydermatis, M. restricta, M. slooffiae, M. sympodialis, or a combination thereof.

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