Compositions and methods for promoting hair growth

Abstract

The present invention provides topical therapeutic compositions and methods of use for promoting hair growth. The present invention utilizes topical compositions comprising microbial growth by-products and pharmacological or naturopathic hair growth promoters. In particular aspects, the microbial growth by-products are biosurfactants.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 325,827, filed March 31, 2022, which is incorporated by reference in its entirety. [Background technology]

[0002] background Human hair is mostly made up of terminal and vellus hair. Terminal hair is a long, coarse, pigmented hair whose bulb, or root, is located deep in the dermis. Vellus hair is a short, fine, thin, non-pigmented hair whose bulb is located in the superficial dermis. At the base of the hair bulb is the papilla, which contains blood vessels and nerves that nourish the hair cells to stimulate growth. The number of papillae is determined at birth, determining the number of hairs present.

[0003] All hair, both in humans and animals, goes through a life cycle that includes three phases: anagen phase (growth phase), catagen phase (catagen phase) and telogen phase (resting phase). The anagen phase is the period of active hair growth. For example, in the case of the scalp, this typically lasts for 3-5 years. The catagen phase is a short transitional period between the anagen and telogen phases. Again, in the case of scalp hair life cycle, the catagen phase lasts approximately 1-2 weeks. The final phase is the telogen phase, during which all growth stops and the hair finally falls out in preparation for new growth. Scalp hair in the telogen phase lasts for approximately 3-4 months before it falls out and new hair starts to grow.

[0004] Some individuals may experience changes in the hair life cycle and / or quality of hair, which may lead to hair loss. For example, taking certain medications, such as chemotherapy drugs, contraceptives, steroids, and antibiotics, can disrupt the normal hair life cycle and cause hair loss. Some of this hair loss may be reversible by stopping the medication, but in some cases, hair loss may become permanent.

[0005] Skin damage and the presence of scar tissue can lead to permanent hair loss, as can genetic and hormonal causes such as alopecia or "male pattern baldness." In alopecia, the scalp begins to lose hair at the temples and top of the head. This incurable condition is mostly limited to men, but women are also affected from time to time.

[0006] As alopecia progresses, the hair itself changes from terminal to vellus hair. Furthermore, when alopecia develops, the proportion of hair in the telogen phase gradually increases, and the proportion of hair in the active growth anagen phase correspondingly decreases. Alopecia is also associated with a severe reduction in hair follicles, for example, a reduction of about one-third or more of hair follicles. Therefore, the combination of the above factors, i.e., the transition from terminal to vellus hair, the increase in the number of telogen phase hairs (some of which are falling out), and the reduction and disappearance of hair follicles, ultimately leads to baldness.

[0007] One known treatment for alopecia is hair transplantation. A plug of skin containing hair is transplanted from a hair-bearing scalp area to the balding area with considerable success; however, this procedure is time-consuming, painful, and expensive. Other non-drug related treatments include, for example, ultraviolet light, massage, psychiatric treatment, revascularization surgery, acupuncture, and exercise therapy. However, none of these are universally accepted as effective.

[0008] The most common treatment approach to alopecia to date is drug therapy.For example, the use of microemulsion creams containing both estradiol and oxandrolone as active ingredients, and the use of organic silicones, have been shown to be useful.In addition, the compounds bimatoprost and minoxidil are useful in stimulating hair growth and / or preventing hair loss.

[0009] Bimatoprost is an eye medication used to treat high intraocular pressure, glaucoma, and to increase eyelash length. Typically, bimatoprost is applied as liquid eye drops.

[0010] Minoxidil is a drug that treats severe hypertension through the dilation of the peripheral vasculature. Dermatologists have recognized that prolonged vasodilation in certain areas of the human body other than the scalp can enhance hair growth even in the absence of vasodilatory medications. For example, increased hair growth around surgical scars is not uncommon. Similarly, arteriovenous fistulas are known to result in conditions of increased vascularity with enhanced hair growth. Initially introduced as an oral medication to treat hypertension, topical solutions and foams containing minoxidil have been introduced to prevent or treat hair loss. Summary of the Invention

[0011] Quick Overview The present invention provides topical therapeutic compositions for promoting skin, scalp and hair health. More specifically, the present invention provides compositions and methods for promoting hair growth and / or treating hair loss using microbial growth by-products. Advantageously, such compositions and methods may be useful for subjects experiencing hair loss for many reasons, including alopecia and / or chemotherapy.

[0012] In certain embodiments, the present invention provides a topical composition for promoting hair growth, the composition comprising one or more biological amphiphilic molecules, hi certain embodiments, the topical composition comprises one or more biological amphiphilic molecules in combination with a pharmacological or naturopathic hair growth promoter.

[0013] In a preferred embodiment, the biological amphiphilic molecule is a biosurfactant, e.g., selected from low molecular weight glycolipids (e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosylerythritol lipids, and trehalose lipids), lipopeptides (e.g., surfactin, iturin, fengycin, arthrofactin, and lichenisin), flavopeptides, phospholipids (e.g., cardiolipin), fatty acid ester compounds, fatty acid ether compounds, and high molecular weight polymers, e.g., lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes.

[0014] The one or more biosurfactants may further include any one or combination of modified forms, derivatives, fractions, isoforms, isomers or subtypes of biosurfactants, including biologically or synthetically modified forms. In certain embodiments, the biosurfactant is a salt form of the biosurfactant.

[0015] In certain specific embodiments, the biosurfactant is a glycolipid, more specifically a sophorolipid (SLP) and / or a mannosylerythritol lipid (MEL). In certain embodiments, a combination of SLP and MEL is utilized.

[0016] In certain embodiments, biosurfactants can function as active ingredients in topical compositions to promote hair growth and / or prevent hair loss.For example, in some embodiments, biosurfactants can enhance the proliferation and / or function of skin fibroblasts and / or accelerate the growth of hair papilla cells and hair.

[0017] In certain embodiments, biosurfactants may function as adjuvants and / or additives that can enhance skin penetration of active and inactive ingredients in the composition, such as pharmacological or naturopathic hair growth promoters, and / or that can enhance formulation of the composition, such as by enhanced emulsification.

[0018] In certain embodiments, the pharmacological hair growth promoter of the composition is minoxidil or bimatoprost.

[0019] In certain exemplary embodiments, the topical compositions of the present invention comprise SLP, MEL and minoxidil. Advantageously, compositions comprising a combination of these ingredients exhibit surprising synergistic effects on promoting hair growth compared to the individual ingredients used alone.

[0020] In some embodiments, the compositions include live or inactivated microorganisms, their growth by-products, and / or culture supernatants of the microorganisms. In certain embodiments, the microorganism is a yeast, such as, for example, Starmerella bombicola, Wickerhamomyces anomalus NRRL Y-68030, Meyerozyma guilliermondii, and / or Pseudozyma aphidis. In certain embodiments, the microorganism is a bacterium, such as, for example, Bacillus coagulans, Bacillus amyloliquefaciens NRRL B-67928, or Bacillus subtilis B4 NRRL B-68031.

[0021] In some embodiments, the topical composition may further comprise a dermatologically acceptable carrier, such as a water-in-oil or oil-in-water emulsion, or an aqueous serum.

[0022] In some embodiments, the topical composition can further include adjuvants and additives typically found in topical skin and hair care compositions, such as: organic solvents, silicones, stabilizers, thickeners, humectants, conditioners, surfactants, colorants, fragrances, pH adjusters, buffers, local anesthetics, anti-biofilm agents, anti-fungals, vitamins, minerals, botanicals, extracts, essential oils, retinoids, anti-comedo agents, moisturizers, sunscreens, and / or other therapeutic and non-therapeutic ingredients known to, for example, heal, repair, rejuvenate, moisturize, protect, and / or improve the skin and scalp while simultaneously stimulating hair growth.

[0023] In certain embodiments, the present invention provides a method for promoting hair growth, wherein the topical composition of the present invention is applied directly to the skin area of ​​a subject where hair loss has occurred and / or hair growth is desired. In some embodiments, the topical composition is formulated and applied as a cream, lotion, ointment, gel, paste, essence, tonic, spray, aerosol, solid bar, eye drops or oil; wherein the formulation is suitable for application to the skin of, for example, the scalp, eyelids, face, limbs or trunk.

[0024] In some embodiments, the method can improve scalp health, prevent hair loss, and / or stimulate hair growth in a subject in need thereof. In some embodiments, the subject is experiencing hair loss as a result of male pattern baldness, chemotherapy, skin injury, burns, scars, or side effects of medication.

[0025] In certain embodiments, the method may further include application of an additional composition to enhance hair growth and / or to complement the activity of the topical composition as part of a hair growth regimen. For example, in one embodiment, the method may include cleansing the scalp or other skin area experiencing hair loss with a cleansing composition comprising a mixture of SLPs and MEL.

[0026] In certain embodiments, the application of topical composition and / or cleanser can also be complemented with the administration of oral compounds such as DHT (dihydrotestosterone) blockers.Subjects with androgenetic alopecia experience hair follicle shrinkage due to the binding of DHT to hair follicles; therefore, in some embodiments, DHT blockers can be useful in preventing hair loss through this mechanism.

[0027] Advantageously, in some embodiments, the use of a multi-part regimen can provide longer lasting, consistent results for subjects experiencing hair loss. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] Detailed Description The present invention provides topical therapeutic compositions for promoting skin, scalp and hair health.More specifically, the present invention provides compositions and methods for promoting hair growth and / or treating hair loss using microbial growth by-products.Advantageously, such compositions and methods can be useful for subjects experiencing hair loss for many reasons, including alopecia and / or chemotherapy.

[0029] Selected definitions As used herein, a plant "extract" refers to a material obtained by exposing a part of a plant to a solvent and removing the solvent, or by using a variety of chemical, immunological, biochemical, or physical techniques known to those skilled in the art, including, but not limited to, precipitation, steam distillation, centrifugation, filtration, column chromatography, detergent lysis, and cold pressing (or squeezing). Plant extracts include, for example, essential oils. Plant material includes parts of a plant, including fruits, roots, stems, leaves, flowers, or parts thereof.

[0030] As used herein, the term "probiotic" refers to a microorganism that, when administered in adequate amounts, confers a health benefit to the host. In certain embodiments, probiotics are administered to the digestive tract of a subject, for example, to provide a digestive benefit to the subject.

[0031] The terms "naturopathic," "natural," and "naturally-derived" as used in reference to a compound or substance refer to a substance found in nature, meaning that it is produced from earthly processes or by living organisms. A natural product can also be isolated or purified from its natural source and utilized or incorporated into a variety of uses, such as foods, beverages, cosmetics, and supplements. As such, a natural product can be combined with other natural or non-natural products that do not co-exist in nature. A natural product can also be produced by chemical synthesis in a laboratory, so long as no artificial ingredients or materials (i.e., synthetic materials that cannot exist naturally as products of the earth or living organisms) are added.

[0032] As used herein, the terms "cure" and / or "ameliorate" a condition or disorder mean to eradicate, alleviate, improve, or reverse to some extent the signs or symptoms of the condition or disorder, including, but not requiring, a complete cure of the condition or disorder.

[0033] As used herein, "preventing" a condition or disorder means avoiding, delaying, forestalling, or minimizing the onset of certain signs or symptoms of the condition or disorder. Prevention can be, but need not be, absolute or complete, meaning that signs or symptoms may still appear in the future. Prevention can include reducing the severity of the onset of such a condition or disorder and / or inhibiting the progression of a condition or disorder to a more severe condition or disorder.

[0034] According to the present invention, "promoting" hair growth means increasing or accelerating the rate of hair growth and / or creating conditions at the hair loss site that are favorable for increased hair growth. In some embodiments, this can be achieved by changing the life cycle of hair, for example, increasing the anagen or growth phase and / or decreasing the telogen phase. Hair growth can include hair lengthening, hair thickening, increased hair strength, and increased number of hairs. In some embodiments, promoting hair growth also includes preventing and / or slowing hair loss.

[0035] The term "wound" as used herein refers to damage to tissue caused, for example, by a cut, blow, or other impact. In accordance with the present invention, wounds include skin injuries classified as "open wounds", including, for example, cuts, abrasions, ulcers, lesions, abrasions, contusions, punctures, lacerations, burns, lacerations, incisions, gunshot wounds, bites, stab wounds, and abrasions.

[0036] The term "burn" as used herein refers to a wound caused by heat (hot or cold), chemicals (acids, bases, etc.), friction, radiation (sunburn, UV, etc.), or electrical sources. Burns can be "minor" burns, including first degree burns, which cause superficial damage to the outer dermis layer, and second degree burns, which cause damage to the epidermis layer of cells. Symptoms of burns include, for example, stinging, blisters, itching, peeling skin, rashes, redness, and swelling.

[0037] The term "scar" as used herein refers to the mark or growth on the skin where injury, such as wound, burn, sore, surgical incision or puncture, has not healed properly and fibrous connective tissue has developed instead of normal tissue.Scar includes hypertrophic scar, which is an area of ​​tissue raised above the surrounding skin due to overproduction of collagen; keloid, which is another form of excessive scar formation, where tissue forms a large protruding neoplasm; atrophic scar, which is where the underlying structural tissue is lost, resulting in a sunken or depressed appearance (e.g. acne scar); and stretch mark, which is caused by the sudden stretching of skin, such as during pregnancy, growth spurt, or skin renewal.

[0038] The term "subject" as used herein refers to an animal, preferably a mammal. In the context of the present invention, the preferred subject is a human. The subject may be of any gender, age, or developmental stage, such as infant, toddler, adolescent, teenager, young adult, middle-aged, or senior.

[0039] As used herein, "topical" means suitable for local application to the external surface of the skin, i.e., dermal application. In some embodiments, "topical" can include application to the eye, including application to the eyelids.

[0040] As used herein, "dermatologically acceptable," "cosmetically acceptable," and "topically acceptable" are used interchangeably and are intended to mean that a particular ingredient is safe and non-toxic for application to the integument (e.g., skin) at the levels of use. In one aspect, the ingredients of the present compositions are recognized as GRAS (Generally Regarded as Safe).

[0041] As used herein, the terms "therapeutically effective amount," "effective amount," and "effective dose" are used to refer to an amount of something (e.g., compound, composition, time) that can achieve a desired amount of hair growth in a subject. The actual amount will vary depending on many factors, including, but not limited to, the particular condition or disorder causing the hair loss, the severity of the condition, the size, age, and health of the subject, and the method of administration.

[0042] As used herein, a "microbe-based composition" refers to a composition that includes components produced as a result of the growth of a microorganism or other cell culture. Thus, a microbe-based composition may include the microorganism itself and / or by-products of microbial growth (e.g., biosurfactants, solvents and / or enzymes). The cells may be in a vegetative state, a spore state, or a mixture of both. The cells may be in a planktonic form, a biofilm form, or a mixture of both. The cells may be live or inert, intact or lysed. The cells may be removed from the medium in which they were grown, or may be present in a concentration of, for example, at least 1 x 10 per ml of composition. 3 , 1×10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 , 1×10 11 The microbe-based composition may be present at a cell concentration of 1000 to 2000 ng / ml or more. In one embodiment, the microbe-based composition may comprise only the medium in which the cells have been grown, with the cells removed (although in some cases some residual cellular material may remain in the medium). The by-products of growth are present in the medium and may include, for example, metabolic products, cell membrane components, expressed proteins, and / or other cellular components. In one embodiment, the microbe-based composition comprises only microbial growth by-products.

[0043] The present invention further provides a "microbe-based product"; this is the product that will be actually applied to achieve a desired result. The microbe-based product may simply be a microbe-based composition harvested from a microbe culturing process. Alternatively, the microbe-based product may have additional components added to it. These additional components may include, for example, stabilizers, buffers, carriers, and other additives and / or adjuvants suitable for a particular application. The microbe-based product may include a mixture of the microbe-based composition. The microbe-based product may also include one or more components of the microbe-based composition that have been processed in some way, such as, but not limited to, filtration, centrifugation, dissolution, drying, purification, etc.

[0044] "Metabolite" refers to any substance produced by metabolism (e.g., a growth by-product) or a substance necessary to participate in a particular metabolic process. Examples of metabolites include, but are not limited to, enzymes, acids, solvents, alcohol, proteins, carbohydrates, vitamins, minerals, trace elements, amino acids, polymers, and biosurfactants.

[0045] As used herein, the terms "isolated" or "purified" when used in reference to a biological or natural substance, such as a nucleic acid molecule, polynucleotide, polypeptide, protein, organic compound (such as a small molecule), microbial cell / strain, or host cell, means that the substance is substantially free of other compounds (such as cellular material) with which it is naturally associated, i.e., the substance does not exist in nature without these other compounds and / or has different or unique properties compared to those found in the naturally occurring substance.

[0046] In certain embodiments, a purified compound is at least 60% by weight of the compound of interest. Preferably, the preparation is at least 75% by weight, more preferably at least 90%, and most preferably at least 99% or 100% (w / w) of the desired compound. Purity is measured by appropriate standard methods, such as, for example, column chromatography, thin layer chromatography, or high performance liquid chromatography (HPLC) analysis.

[0047] As used herein, "isomer" refers to a molecule that has the same chemical formula as another molecule, but has a unique structure. Isomers can be structural isomers, in which atoms and functional groups are bonded at different positions, and stereoisomers (spatial isomers), in which the bond structure is the same but the geometric arrangement of atoms and functional groups in space is different. MEL isomers can differ, for example, in the bond type and bond position of carbohydrates, fatty acids, and / or acetyl groups.

[0048] As used herein, "surfactant" refers to a surface-active substance, i.e., a compound that reduces the surface tension (or interfacial tension) between two phases. Surfactants act, for example, as detergents, wetting agents, emulsifiers, foaming agents, and / or dispersing agents. "Biosurfactant" refers to a surfactant produced by an organism and / or produced using naturally occurring substrates.

[0049] The transitional term "comprising" is synonymous with "including" or "containing" and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes elements, steps, or ingredients not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to the specified materials or steps and those that do not "materially affect the basic and novel characteristics" of the claimed invention. Use of the term "comprising" contemplates other embodiments "consisting of" or "consisting essentially of" the recited components.

[0050] As used herein, the term "or" is understood to be inclusive unless otherwise stated or clear from context. As used herein, the terms "a," "an," and "the" are understood to be singular or plural unless otherwise stated or clear from context.

[0051] Unless otherwise specified or clear from the context, the term "about" as used herein is understood to mean within normal tolerances in the art, for example, within 2 standard deviations of the mean. "About" may be understood to mean within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value.

[0052] The recitation of a list of chemical groups in a definition of a variable herein includes a definition of that variable as a single group or combination of the listed groups. The recitation of an embodiment of a variable or aspect herein includes that embodiment as a single embodiment or in combination with other embodiments or portions thereof.

[0053] Any composition or method provided herein can be combined with any one or more of the other compositions and methods provided herein.Other features and advantages of the present invention will be apparent from the following description of its preferred embodiments and from the claims.All documents cited herein are incorporated herein by reference.

[0054] Topical Hair Growth Composition In certain embodiments, the present invention provides topical compositions for promoting hair growth, the compositions comprising one or more biological amphiphilic molecules in combination with a pharmacological or naturopathic hair growth-promoting agent.

[0055] In a preferred embodiment, the biological amphiphilic molecule is a biosurfactant, e.g., selected from low molecular weight glycolipids (e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosylerythritol lipids, and trehalose lipids), lipopeptides (e.g., surfactin, iturin, fengycin, arthrofactin, and lichenisin), flavolips, phospholipids (e.g., cardiolipin), fatty acid ester compounds, fatty acid ether compounds, and high molecular weight polymers, e.g., lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes.

[0056] Biosurfactants are amphiphilic substances, meaning that they contain both polar (hydrophilic) and non-polar (hydrophobic) groups. Due to their amphiphilic structure, biosurfactants reduce the surface and interfacial tension between liquid, solid, and gas molecules.

[0057] Moreover, biosurfactants accumulate at interfaces, resulting in the formation of aggregated micellar structures in solution. The ability of biosurfactants to form pores and destabilize biological membranes allows for their use, for example, as antibacterial and antifungal agents, as well as delivery agents for other compounds across membranes, etc. Moreover, biosurfactants are biodegradable, have low toxicity, and can be produced using low-cost renewable resources. They can inhibit microbial adhesion to various surfaces, prevent biofilm formation, and exhibit strong emulsifying and demulsifying properties.

[0058] The one or more biosurfactants may further comprise any one or combination of modified forms, derivatives, fractions, isoforms, isomers or subtypes of biosurfactants, including biologically or synthetically modified forms. In one embodiment, the one or more biosurfactants are present in the composition at a critical micelle concentration (CMC).

[0059] In some embodiments, the amphipathic molecule is utilized in a crude form, where the molecule is present in the growth medium (e.g., broth) in which the amphipathic molecule-producing microorganism is cultured and is recovered therefrom without purification. The crude form can, for example, contain at least 0.001%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 99% of the amphipathic molecule in the growth medium. In another embodiment, the amphipathic molecule is purified from the product of the culture.

[0060] In certain embodiments, biosurfactant can function as an active ingredient in topical compositions to promote hair growth and / or prevent hair loss.For example, in some embodiments, biosurfactant can enhance the proliferation and / or function of skin fibroblasts and / or accelerate the growth of hair papilla cells and hair.In some embodiments, biosurfactant according to the present invention can also kill pathogens present in skin / scalp, regulate the immune system of skin, kill melanocytes to allow replacement cell growth, and reduce oxidative stress.

[0061] In certain embodiments, biosurfactants may function as adjuvants and / or additives that can enhance skin penetration of active and inactive ingredients in the composition, such as pharmacological or naturopathic hair growth promoters, and / or that can enhance formulation of the composition, such as by enhanced emulsification.

[0062] In a preferred embodiment, the biosurfactant is a sophorolipid (SLP) and / or a mannosylerythritol lipid (MEL), both of which are glycolipid biosurfactants produced by certain yeasts. Glycolipids generally contain a mono- or oligosaccharide group or a glycerol group that can be acetylated or alkylated, attached to a sphingolipid, and one or more fatty acids.

[0063] SLPs are produced by yeasts of the Starmerella clade, such as Candida apicola and Starmerella bombicola. In some embodiments, Wickerhamomyces anomalus is the SLP producer (e.g., W. anomalus NRRL Y-68030). SLPs consist of the disaccharide sophorose linked to a long-chain hydroxy fatty acid. Such SLPs are partially acetylated 2-O-β-D-glucopyranosyl-D-glucopyranose units β-glycosidically linked to 17-L-hydroxyoctadecanoic acid or 17-L-hydroxy-Δ9-octadecenoic acid. Hydroxy fatty acids are generally 16 or 18 carbon atoms and may contain one or more unsaturated bonds. The carboxyl group of the fatty acid may be free (acidic or open) or internally esterified at the 4″ position (lactone form).

[0064] SLPs have environmental compatibility, high biodegradability, low toxicity, high selectivity and specific activity over a wide range of temperature, pH and salinity conditions. Furthermore, in some embodiments, the small micellar size of SLPs can help the micelles and the compounds encapsulated therein to move through nanoscale pores and spaces (e.g., between epithelial cells and within biofilm matrices), which can be advantageous for applications in the fields of cosmetics and dermatology. In certain embodiments, the micellar size of SLPs is less than 100 nm, less than 50 nm, less than 20 nm, less than 15 nm, less than 10 nm, or less than 5 nm.

[0065] In a particular embodiment, the SLPs according to the invention are represented by general formula (1) and / or general formula (2) and have different fatty acid chain lengths (R 3 ), and in some cases R 1 and / or R 2 It is obtained as a collection of more than 30 structural analogues, each of which has acetylation or protonation at the base. TIFF2025511022000001.tif42134

[0066] In the general formula (1) or (2), R 0R can be either a hydrogen atom or a methyl group. 1 and R 2 are each independently a hydrogen atom or an acetyl group. 3 is a saturated aliphatic hydrocarbon chain or an unsaturated aliphatic hydrocarbon chain having at least one double bond, and may have one or more substituents.

[0067] Non-limiting examples of the substituents include halogen atoms, hydroxyl groups, lower (C1-6) alkyl groups, halo lower (C1-6) alkyl groups, hydroxy lower (C1-6) alkyl groups, halo lower (C1-6) alkoxy groups, etc. 3 Generally, it has 11 to 20 carbon atoms, preferably 13 to 17 carbon atoms, and more preferably 14 to 16 carbon atoms.

[0068] In a particular embodiment, the SLP according to the invention is a salt form of the SLP represented by the general formula (3), wherein R 4 = a metal ion, for example Na, K, Mg, Cu or Ca.

[0069] Advantageously, SLPs have several benefits as active or inactive ingredients in topical compositions, including, for example: emulsifying oil-in-water or water-in-oil mixtures; reducing inflammation and oxidative stress; removing damaged keratinocytes from the upper layers of the skin; enhancing wound healing through antibacterial and anti-inflammatory effects; promoting fibroblast metabolism; improving collagen synthesis and toning and remodeling the skin; stimulating the synthesis of leptin by adipocytes, helping to reduce the subcutaneous adiposity that causes cellulite; inhibiting the activity of elastase, reducing the appearance of wrinkles; exfoliating the stratum corneum to reduce skin blemishes and inhibit melanin production; combating microbial dandruff, acne and body odor; and / or reducing inflammatory conditions such as dermatitis, eczema, and psoriasis.

[0070] In certain preferred embodiments, the SLPs comprise a mixture of lactone-type and linear SLPs, e.g., a mixture of SLPs according to general formulas (1) and (2). In some embodiments, the ratio of lactone-type SLPs to linear SLPs is between 50:50 and 10:90, between 40:60 and 20:80, or about 30:70 lactone-to-linear.

[0071] In one embodiment, the glycolipid is MEL. MEL contains either 4-OBD-mannopyranosyl-meso-erythritol or 1-OBD-mannopyranosyl-meso-erythritol as the hydrophilic portion and a fatty acid group and / or an acetyl group as the hydrophobic portion. One or two hydroxyls, typically at C4 and / or C6 of the mannose residue, may be acetylated. Additionally, there may be one to three esterified fatty acids with chain lengths of 8-12 carbons or more.

[0072] MEL and MEL-like substances (e.g., mannose-based substances) are produced primarily by Pseudozyma and Ustilago spp., although the MEL structures produced by each species vary greatly. Certain mannose-based substances with MEL-like properties can also be produced by the yeast Meyerozyma guilliermondii.

[0073] MEL is non-toxic and stable over a wide range of temperatures and pH. Furthermore, MEL can be used without additional preservatives.

[0074] MEL can be produced in over 93 different combinations, grouped into five major categories: MEL A, MEL B, MEL D, triacetylated MEL A, and triacetylated MEL B / C. These molecules can be modified, either synthetically or naturally. For example, MEL can contain chains of different carbon lengths, or different numbers of acetyl and / or fatty acid groups.

[0075] MEL molecules and / or modified forms thereof according to the invention may include, for example, triacylated, diacylated, monoacylated, triacetylated, diacetylated, monoacetylated and non-acetylated MEL, as well as stereoisomers and / or structural isomers thereof.

[0076] Other mannose-based substances / MEL-like substances exhibiting similar structures and similar properties can also be used in accordance with the present invention, such as mannosyl-mannitol lipids (MML), mannosyl-arabitol lipids (MAL), and / or mannosyl-ribitol lipids (MRL).

[0077] Advantageously, MEL has several benefits as an active or inactive ingredient in topical compositions, including, for example: reducing skin inflammation; preventing cell damage resulting from the use of synthetic surfactants such as SDS; stimulating hair bulb cell and hair growth; repairing and / or strengthening damaged hair (especially MEL-A and MEL-B); enhancing the viability of fibroblasts and dermal papilla cells (especially MEL-A); reducing sweating and aiding the skin's moisture barrier function; and / or reducing the melanin content of age spots.

[0078] In certain preferred embodiments, the topical compositions of the present invention comprise a mixture of SLPs and MELs.

[0079] In preferred embodiments, the concentration of SLPs in the total topical composition is about 100-500 ppm, about 125-375 ppm, about 150-300 ppm, about 180-250 ppm, or about 200 ppm.

[0080] In preferred embodiments, the concentration of MEL (or MEL-like substance) in the total topical composition is 0.01-100 ppm, about 1-75 ppm, about 2-50 ppm, about 3-25 ppm, about 5-10 ppm, about 4-6 ppm, about 5 ppm or less.

[0081] In one embodiment, the biosurfactant may include one or more lipopeptides, such as, for example, surfactin, iturin, fengycin, arthrofactin, viscosin, amphicin, syringomycin, and / or lichenisin.

[0082] In a specific embodiment, the lipopeptide biosurfactant is surfactin. Lipopeptides are produced by various probiotic and non-pathogenic bacteria, such as Bacillus natto, Bacillus coagulans, Bacillus subtilis, Bacillus amyloliquefaciens, lactic acid bacteria, etc. In some embodiments, lipopeptides are produced by certain strains of the genus Bacillus, such as B. amyloliquefaciens NRRL B-67928 or B. subtilis B4 NRRL B-68031.

[0083] In certain embodiments, the compositions according to the invention further comprise one or more pharmacological or naturopathic hair growth promoters, such as, for example, minoxidil, bimatoprost, finasteride, dihydrotestosterone, biotin, fibroblast growth factors, keratinocyte growth factors, basic fibroblast growth factors, platelet derived growth factors, spironolactone, vitamins and minerals (e.g., B vitamins, vitamin E, vitamin A, vitamin C, vitamin D, iron, folic acid, zinc), retinol, collagen peptides, omega-3, -6 and -9 fatty acids, plant extracts (e.g., lemon (Citrus limon L.), strawberry (Fragaria ananassa L.), rye (Secale cereale L.), epimedium spp., Geranium sibiricum L., olive (Olea europaea L.), tea plant (Camellia sinensis, Arabica coffee tree (Coffea arabica), Safflower (Carthamus tinctorius L.), Ginseng (Panax ginseng Mayer), Houttuynia cordata Thunb., Sophora glavescens Aiton, Illicium anisatum L., Illicium verum Hook. f., Barley (Hordeum vulgare L.), Lotus fruit (Nelumbinis semen), Japanese cypress (Chamaecyparis obtuse), Polygonum multiflorum, Alder tree (Alnus sibirica), Winter mallow (Malva verticillate), Chinese linden tree (Magnolia officinalis), Angelica gigas gigas), Sappan (Caesalpinia sappan), Paper berry (Broussonetia papyrifera), Oriental oak (Thuja orientalis), Sweet potato (Ipomoea batatas), Japanese laurel (Ishige sinicola), Plum (Prunis miraKoehne, Saussurea lappa, Cornus officinalis, Anemarrhena asphodeloides, Salvia plebeian, Undariopsis peterseniana, Pueraria thomsonii, Platycladus orientalis, Polygonum multiforum, Rosemary (Rosmarinus officinalis), Avicennia marina, Ecklonia cava, Saw palmetto (Sabal serrulatum), and bioactive compounds (e.g., sinapic acid, icariin, corilagin, gallic acid, oleuropein, caffeine, hydroxysafflor yellow A, linoleic acid, β-sitosterol, quercitrin, L-marquiain, medicarpin, shikimic acid, procyanidins (including B2 and B3), ginsenoside Rb1, anthraquinones, flavonoids, tannins, saponins, a- Terpinyl acetate, sabinene, isobornyl acetate, limonene, β-D-glucoside, emodin, oregonin, myristoleic acid, liposomal honokiol, decursin, 3-deoxysappanchalcone, 7-hydroxycoumarin, protocatechuic acid, ferulic acid, protocatechuic acid, epicatechin, β-sitosterol, kaempferol, isoquercetin, octaphlorethol A A), vitamin E, oleic acid, costunolide, morroniside, timosaponin BII, phenolic acids, terpenes, terpenoids, saponin, myricitrin, isoquercitrin, quercitrin, myricetin, afzelin, quercetin, kaempferol, amentoflavone, hinokiflavone, ginsenoside Ro, rhyscion, avicequinone C, diecol, etc.

[0084] In certain embodiments, the pharmacological or naturopathic hair growth promoter is a compound that can stimulate resting hair follicles (telogen phase) to growing hair follicles (anagen phase) (e.g., prolong the anagen phase).

[0085] In some embodiments, the pharmacological or naturopathic hair growth promoting agent is a compound that can promote the expression of growth factors, reduce inflammatory cytokines, activate Wnt / β-catenin signaling, promote Hedgehog pathway signaling (Shh), inhibit 5α-reductase, and / or improve cell cycle progression.

[0086] In a particular embodiment, the pharmacological hair growth promoter is minoxidil.

[0087] In certain embodiments, the pharmacological or naturopathic hair growth promoter is present in the topical composition at a concentration of 0.001% to 90%, 0.01% to 50%, 0.05% to 10%, or about 5% by weight of the total composition.

[0088] In certain exemplary embodiments, the topical compositions of the present invention comprise SLP, MEL, and minoxidil. Advantageously, compositions comprising such a combination of ingredients exhibit a surprising synergistic effect on promoting hair growth compared to each ingredient used alone.

[0089] In certain embodiments, in addition to having a therapeutic effect in itself, the biosurfactant can increase the percutaneous absorption of minoxidil, which can help to enhance benefits such as changes in hair growth phase and hair thickness. In certain embodiments, the biosurfactant can form micelles, liposomes, or vesicles around minoxidil to enhance the absorption of minoxidil through the skin. In certain embodiments, the biosurfactant can bind to minoxidil to facilitate the absorption of minoxidil through the skin.

[0090] In a specific example, the composition comprises 200 ppm SLPs, 5 ppm MEL, and 5% minoxidil, where the SLPs are in a 70:30 ratio of linear SLPs to lactone SLPs.

[0091] In some embodiments, the topical compositions of the present invention comprise live or inactivated microorganisms capable of producing growth by-products useful for hair growth, skin healing and rejuvenation, and / or supernatants obtained from the culture of such microorganisms. In certain embodiments, the microorganisms are yeasts, such as Starmerella bombicola, Wickerhamomyces anomalus (e.g., NRRL Y-68030), Meyerozyma (Pichia) guilliermondii, and / or Pseudozyma aphidis. In one specific embodiment, the composition comprises live or inactivated Wickerhamomyces anomalus.

[0092] In certain embodiments, the microorganism is a bacterium, such as Lactobacillus spp., Bifido spp., Lactococcus spp., Streptococcus spp., and Bacillus spp., such as Bacillus acidiceler, B. acidicola, B. acidiproducens, B. acidocaldarius, B. acidoterrestrisr, B. aeolius, B. aerius, B. aerophilus, B. agaraldherens, B. agaradhaerens, B. agri, B. aidingensis, B. akibai, B. alcalophilus, B. algicola, B. alginolyticus, B. alkalidiazotrophicus, B. alkalinitrilicus, B. alkalisediminis, B. alkalitelluris, B. altitudinis, B. alveayuensis, B. alvei, B. amyloliquefaciens, B. B. a. subsp. amyloliquefaciens, B. a. subsp. plantarum, B. mylolyticus, B. andreesenii, B. aneurinilyticus, B. anthracia, B. aquimalis, B.aquimaris, B. arenosi, B. arseniciselenatis, B. arsenicus, B. aurantiacus, B. arvi, B. aryabhattai, B. asahii, B. atrophaeus, B. axarquiensis, B. azotofixans, B. azotoformans, B. badius, B. barbaricus, B. bataviensis, B. beisingensis beijingensis, B. benzoevorans, B. beringensis, B. berkeleyi, B. beveridgei, B. bogoriensis, B. boroniphilus, B. borstelensis, B. brevis migula, B. butanolivorans, B. canaveralius, B. carboniphilus, B. cecembensis, B. cellulosilyticus, B. centrosporus, B. cereus cereus, B. chagannorensis, B. chitinolyticus, B. chondroitinus, B. choshinensis, B. chungangensis, B. cibi, B. circulans, B. clarkii, B.B. clausii, B. coagulans, B. coahuilensis, B. cohnii, B. composti, B. curdlanolyticus, B. cycloheptanicus, B. cytotoxicus, B. daliensis, B. decisifrondis, B. decolorationis, B. deserti, B. dipsosauri, B. drentensis, B. edaphicus, B. ehimensis ehimensis, B. eiseniae, B. enclensis, B. endophyticus, B. endoradicis, B. farraginis, B. fastidiosus, B. fengqiuensis, B. firmus, B. flexus, B. foraminis, B. fordii, B. formosus, B. fortis, B. fumarioli, B. funiculus, B. fusiformis, B. galactophilus galactophilus, B. galactosidilyticus, B. galliciensis, B. gelatini, B. gibsonii, B. ginsengi, B. ginsengihumi, B. ginsengisoli, B. globisporus, B.globisporus, B. g. subsp. globisporus, B. g. subsp. marinus, B. glucanolyticus, B. gordonae, B. gottheilii, B. graminis, B. halmapalus, B. haloalkaliphilus, B. halochares, B. halodenitrificans, B. halodurans, B. halophilus, B. halosaccharovorans, B. halosaccharovorans, B. hemicellulosilyticus, B. hemicentroti, B. herbersteinensis, B. horikoshii, B. horneckiae, B. horti, B. huizhouensis, B. humi, B. hwajinpoensis, B. idriensis, B. indicus, B. infantis, B. infernus, B. insolitus, B. invicta invictae, B. iranensis, B. isabeliae, B. isronensis, B. jeotgali, B. kaustophilus, B. kobensis, B. kochii, B. kokeshiiformis, B. koreensis, B. korlensis, B.B. kribbensis, B. krulwichiae, B. laevolacticus, B. larvae, B. laterosporus, B. lautus, B. lehensis, B. lentimorbus, B. lentus, B. licheniformis, B. ligniniphilus, B. litoralis, B. locisalis, B. luciferensis, B. luteolus, B. luteus luteus, B. macauensis, B. macerans, B. macquariensis, B. macyae, B. malacitensis, B. mannanilyticus, B. marisflavi, B. marismortui, B. marmarensis, B. massiliensis, B. megaterium, B. mesonae, B. methanolicus, B. methylotrophicus, B. megranus migulanus, B. mojavensis, B. mucilaginosus, B. muralis, B. murimartini, B. mycoides, B. naganoensis, B. nanhaiensis, B. nanhaiisediminis, B. nealsonii, B. neidei, B.neidei, B. neizhouensis, B. niabensis, B. niacini, B. novalis, B. oceanisediminis, B. odysseyi, B. okhensis, B. okuhidensis, B. oleronius, B. oryzaecorticis, B. oshimensis, B. pabuli, B. pakistanensis, B. pallidus, B. panascisori panacisoli, B. panaciterrae, B. pantothenticus, B. parabrevis, B. paraflexus, B. pasteurii, B. patagoniensis, B. peoriae, B. persepolensis, B. persicus, B. pervagus, B. plakortidis, B. pocheonensis, B. polygoni, B. polymyxa, B. popilliae, B. pseudoalkalinephilus, B. pseudalcalophilus, B. pseudofirmus, B. pseudomycoides, B. psychrodurans, B. psychrophilus, B. psychrosaccharolyticus, B. psychrotolerans, B. pulvifaciens, B.B. pumilus, B. purgationiresistens, B. pycnus, B. qingdaonensis, B. qingshengii, B. reuszeri, B. rhizosphaerae, B. rigui, B. ruris, B. safensis, B. salarius, B. salexigens, B. saliphilus, B. schlegelii, B. sediminis, B. serenatarsenatis selenatarsenatis, B. selenitireducens, B. seohaeanensis, B.shacheensis, B. shackletonii, B. siamensis, B. silvestris, B. simplex, B. siralis, B. smithii, B. soli, B. solimangrovi, B. solisalsi, B. songklensis, B. sonorensis, B. sphaericus sphaericus, B. sporothermodurans, B. stearothermophilus, B. stratosphericus, B. subterraneus, B. subtilis, B. s. subsp. inaquosorum, B. s. subsp. spizizenii, B. s. subsp. subtiliss. subtilis, B. taeanensis, B. tequilensis, . B. thermantarcticus, B. thermoaerophilus, B. thermoamylovorans, B. thermocatenulatus, B. thermocloacae, B. thermocopriae, B. thermodenitrificans, B. thermoglucosidasius, B. thermolactis, B. thermoleovorans, B. thermophilus, B. thermoruber, B. thermosphaericus, B. thiaminolyticus, B. thiaminolyticus, B. thioparans, B. thuringiensis, B. tianshenii, B. trypoxylicola, B. tusciae, B. validus, B. vallismortis, B. vedderi, B. velezensis, B. vietnamensis, B. vireti, B. vulcani, B. wakoensis, B. weihenstephanensis, B. xiamensis, B. xiamenensis, B. xiaoxiensis, B. zhanjiangensis, B. amyloliquefaciens NRRL B-67928, B. subtilis B4 NRRL B-68031, and / or Bacillus coagulans (e.g., BC-30).

[0093] In some embodiments, the composition comprises a culture supernatant of a microorganism in place of a pharmacological or naturopathic hair growth promoter. For example, in one specific embodiment, the composition comprises the supernatant of SLP, MEL, and a Bacillus species, such as Bacillus coagulans.

[0094] Additional microbial growth by-products useful according to the present invention include mannoproteins, β-glucans, enzymes, and other metabolic products that have bioemulsifying and surface / interfacial tension reducing activity.

[0095] In some embodiments, the topical cosmetic composition may contain a therapeutically effective amount of enzymes and / or proteins produced by microorganisms. For example, about 0.001% to about 20%, about 0.01% to about 15%, or about 0.05% to about 10% by weight of one or more enzymes and / or proteins. These include, but are not limited to, exo-β-1,3-glucanases, chitinases, esterases, lipases, glycosidases, amylases, and proteases that are beneficial for improving skin health.

[0096] In some aspects, the topical therapeutic composition may further comprise a dermatologically acceptable carrier or vehicle.

[0097] The carrier or vehicle may include, for example, water; saline; saline solution; ointments; creams; oil-in-water emulsions; water-in-oil emulsions; silicone-in-water emulsions; water-in-silicone emulsions; wax-in-water emulsions; water-oil-water triple emulsions; microemulsions; gels; vegetable oils; mineral oils; ester oils, such as octyl palmitate, isopropyl myristate and isopropyl palmitate; ethers, such as dicapryl ether and dimethyl isosorbide; alcohols, such as ethanol and isopropanol; fatty alcohols, such as cetyl alcohol, cetearyl alcohol, stearyl alcohol and behenyl alcohol; isoparaffins, such as isooctane, isododecane, isopropyl alcohol ... cane (IDD) and isohexadecane; silicone oils such as cyclomethicone, dimethicone, dimethicone crosspolymer, polysiloxane and their derivatives, preferably organic modified derivatives including PDMS, dimethicone copolyol, dimethiconol, and amodimethiconol; hydrocarbon oils such as mineral oil, petrolatum, isoeicosane, and polyolefins such as (hydrogenated) polyisobutene; polyols such as propylene glycol, glycerin, butylene glycol, pentylene glycol, hexylene glycol, caprylyl glycol; waxes such as beeswax, carnauba, ozokerite, microcrystalline wax, polyethylene wax, and botanical wax; or any combination or mixture thereof. The aqueous vehicle includes one or more solvents miscible with water, such as lower alcohols such as ethanol, isopropanol, etc. The vehicle can comprise from about 1% to about 99%, from 10% to about 85%, from 25% to 75%, or from 50% to about 65% by weight of the composition.

[0098] The term "oil" as used herein includes silicone oil unless otherwise specified. The emulsion may contain an emulsifier, such as a nonionic, anionic or amphoteric surfactant, or a gallant, typically in an amount of about 0.001% to about 5% by weight.

[0099] In some embodiments, the topical composition may further comprise additional adjuvants and additives commonly found in skin and hair care compositions, such as organic solvents, stabilizers, silicones, thickeners, emollients, sunscreens, moisturizers, fragrances, or others described herein. The amount of each component, whether active or inactive, is that amount conventionally used in the cosmetic field to achieve its intended purpose, and typically ranges from about 0.0001% to about 25%, or from about 0.001% to about 20% of the composition, although amounts outside these ranges may also be used. The nature and amount of these components must be compatible with the manufacture and function of the composition of the present disclosure.

[0100] In one embodiment, the composition can include additional skin actives, including, but not limited to, keratolytic agents, desquamating agents, keratinocyte proliferation promoters, collagenase inhibitors, elastase inhibitors, depigmenting agents, anti-inflammatory agents, steroids, anti-acne agents, antioxidants, and advanced glycation end products (AGE) inhibitors, to name just a few.

[0101] In one embodiment, the composition can include anti-aging ingredients, such as, but not limited to, botanicals (e.g., Butea frondosa extract); phytol; phytonic acid; phospholipids; silicones; petrolatum; triglycerides; omega fatty acids; retinoids; hydroxy acids (including alpha-hydroxy acids and beta-hydroxy acids), salicylic acid and alkyl esters of salicylic acid; exfoliants (e.g., glycolic acid, 3,6,9-trioxaundecanedioic acid, etc.), estrogen synthase stimulating compounds (e.g., caffeine and derivatives); compounds capable of inhibiting 5α-reductase activity (e.g., linoleic acid, linoleic acid, finasteride, and mixtures thereof); and barrier function enhancers (e.g., ceramides, glycerides, cholesterol and its esters, alpha-hydroxy and omega-hydroxy fatty acids and their esters).

[0102] In one embodiment, the composition may include an exfoliant. Suitable exfoliants include, for example, alpha-hydroxy acids, beta-hydroxy acids, oxa-acids, oxadiacids, and derivatives thereof (such as esters, anhydrides, salts, etc.). Suitable hydroxy acids include, for example, glycolic acid, lactic acid, malic acid, tartaric acid, citric acid, 2-hydroxyalkanoic acid, mandelic acid, salicylic acid, and derivatives thereof. An example of an exfoliant is glycolic acid. When present, the exfoliant may comprise about 0.001% to about 20% by weight of the composition.

[0103] In one embodiment, the composition may include one or more antioxidants. Suitable antioxidants include, for example, compounds with phenolic hydroxyl functionality, such as ascorbic acid and its derivatives / esters; beta-carotene; catechin; curcumin; ferulic acid derivatives (e.g., ethyl ferulate, sodium ferulate); gallic acid derivatives (e.g., propyl gallate); lycopene; reductic acid; rosmarinic acid; tannic acid; tetrahydrocurcumin; tocopherol and its derivatives, such as tocopherol acetate; uric acid; or any mixture thereof. Other suitable antioxidants are those with one or more thiol functional groups (--SH), either reduced or non-reduced, such as glutathione, lipoic acid, thioglycolic acid, and other sulfhydryl compounds. Antioxidants may be inorganic, such as bisulfite, metabisulfite, sulfite, or other inorganic salts and acids containing sulfur. The antioxidants, individually or collectively, may comprise from about 0.001% to about 10% (w / w), or from about 0.01% to about 5% (w / w) of the total weight of the composition.

[0104] Non-biological surfactants can also be added to the formulation. Examples of surfactants include, but are not limited to, alkyl sulfates, alkyl ether sulfates (e.g., sodium / ammonium lauryl sulfate and sodium / ammonium laureth sulfate), amphoterics (e.g., amphoacetates and amphopropionates), sulfosuccinates, alkyl polyglucosides, betaines (e.g., cocamidopropyl betaine (CAPB)), sultaines, sarcosinates, isethionates, taurates, ethoxylated sorbitan esters, alkanolamides, and amino acid-based surfactants.

[0105] Viscosity modifiers may also be added to the composition and include, for example, cocamide DEA, oleamide DEA, sodium chloride, cellulosic polymers, polyacrylates, ethoxylated esters, alcohols, glycols, xylene sulfonates, polysorbate 20, alkanolamides, and cellulose derivatives (e.g., hydroxypropyl methylcellulose and hydroxyethyl cellulose).

[0106] Additionally, polymers can be added including, for example, xanthan gum, guar gum, polyquaternium-10, PEG-120 methyl glucose dioleate, PEG-150 distearate, PEG-150 polyglyceryl-2 tristearate, and PEG-150 pentaerythrityl tetrastearate.

[0107] A sunscreen or combination of sunscreens may be included to protect the skin and scalp from both UVA and UVB rays. Sunscreens that may be used in the composition include avobenzone, cinnamic acid derivatives (such as octyl methoxycinnamate), octyl salicylate, oxybenzone, octocrylene, titanium dioxide, zinc oxide, or any mixture thereof. The sunscreen may be present at about 1 wt% to about 30 wt% of the total weight of the composition.

[0108] The composition may include a pH adjuster (e.g., citric acid, ethanolamine, sodium hydroxide, etc.) to be formulated within a wide range of pH levels. In one embodiment, the pH of the topical composition ranges from 1.0 to 13.0. In some embodiments, the pH of the topical composition ranges from 2.0 to 12.0. Other pH ranges suitable for the composition include 3.5 to 7.0, or 7.0 to 10.5. To bring the pH of the composition within the desired range, suitable pH adjusters such as sodium hydroxide, citric acid, and triethanolamine may be added.

[0109] The composition may optionally contain other ingredients, additives or adjuvants known to those skilled in the art, including, but not limited to, the following: skin penetration enhancers; emollients (e.g., isopropyl myristate, petrolatum, volatile or non-volatile silicone oils such as methicone and dimethicone, ester oils, mineral oils, and fatty acid esters); humectants (e.g., glycerin, hexylene glycol, caprylyl glycol); skin plumpers (e.g., glycerin, hexylene glycol, caprylyl glycol); plumper) (e.g., palmitoyl oligopeptides, collagen, collagen and / or glycosaminoglycan (GAG) enhancers); anti-inflammatory agents (e.g., aloe vera, bioflavonoids, diclofenac, salicylic acid); chelating agents (e.g., EDTA or its salts, e.g., disodium EDTA); vitamins (e.g., tocopherol and ascorbic acid); vitamin derivatives (e.g., ascorbyl monopalmitate, tocopherol acetate, vitamin E palmitate); thickening agents (e.g., vegetable gums such as hydroxyalkylcellulose, carboxymethylcellulose, carbomer, and xanthan gum); gelling agents (e.g., ester-terminated polyesteramides); structuring agents; proteins; immune modulators (e.g., corticosteroids and nonsteroidal immune modulators).

[0110] Other ingredients that can be included are film formers, moisturizers, minerals, viscosity and / or rheology adjusting agents, insect repellents, skin cooling compounds, skin protectants, lubricants, preservatives, pearls (lusters), chromalite, mica, conditioners, antiallergic agents, antimicrobial agents (e.g. antifungal agents, antiviral agents, antibacterial agents), disinfectants, drugs, light stabilizers, surface smoothing agents, light diffusing agents, and exfoliation promoters.Details about these and other suitable cosmetic ingredients can be found in the "International Cosmetic Ingredient Dictionary and Handbook" 10th Edition (2004) published by CTFA (Cosmetic, Toiletry, and Fragrance Association) on pages 2177-2299, which is incorporated herein by reference in its entirety.The amount of these various substances is that which is conventionally used in the cosmetic or pharmaceutical fields, and may, for example, constitute about 0.01% to about 20% of the total weight of the composition.

[0111] The compositions may be formulated as suspensions, emulsions, hydrogels, multiphase solutions, vesicular dispersions, or other known forms of topical compositions.

[0112] In certain embodiments, a topical composition can be formulated to be applied, for example, by a pen, tube, bottle, brush, stick, sponge, cotton swab, wet paper towel (wipe), spray, dropper, hand, or finger.

[0113] The compositions may be formulated into a variety of product forms, such as, for example, lotions, creams, serums, sprays, aerosols, liquid cakes, solid bars, ointments, essences, gels, pastes, patches, pencils, powders, wet paper towels, soaps, shampoos, conditioners, sticks, foams, mousses, elixirs, or concentrates. In a preferred embodiment, the compositions are formulated to be particularly suitable for topical administration to the scalp.

[0114] How to Promote Hair Growth In certain embodiments, the present invention provides methods for promoting hair growth, wherein a topical composition of the present invention is applied directly to an area of ​​the skin of a subject where hair loss is occurring and / or hair growth is desired, such as the scalp, eyelids, face, limbs, or torso.

[0115] In some embodiments, this method can improve scalp health, prevent hair loss, and / or stimulate hair growth in a subject in need thereof.In some embodiments, the subject experiences hair loss as a result of male pattern baldness, chemotherapy, skin injury, burns, scars, or side effects of medication.

[0116] In some embodiments, "applying" the composition may mean painting, spraying, or otherwise contacting the composition to skin or hair. Reference to "skin" herein is meant to include the scalp in addition to other external parts of the body. In some embodiments, the composition is rubbed or massaged into the skin. In some embodiments, the composition is left on the skin to be fully absorbed; while in other embodiments, the composition is applied to the skin for a therapeutically effective period of time and then washed or removed from the skin, for example with water or a cloth.

[0117] In certain embodiments, the topical composition is applied 0-10 times per day, preferably at least once per day or at least once every other day. In some embodiments, the topical composition is applied daily or every other day for an indefinite period of time, e.g., at least 1, 2, 3 weeks, or more, to achieve and / or maintain a desired level of hair growth.

[0118] In one embodiment, the composition is administered to 1 cm 2 of skin. 2 from about 0.001 to about 100 mg / cm, more commonly from about 0.01 to about 20 mg / cm 2 , or about 0.1 to about 10 mg / cm 2100 mg / kg, although more or less may be used depending on the size of the skin area to be treated.

[0119] In certain embodiments, the method may further include application of an additional composition to enhance hair growth and / or to complement the activity of the topical composition as part of a hair growth regimen. For example, in one embodiment, the method may include cleansing the scalp or other skin area experiencing hair loss with a cleansing composition comprising a mixture of SLPs and MEL.

[0120] The additional cleansing step can be performed, for example, once a day to once a week. In certain embodiments, cleansing is performed immediately prior to application of the topical composition. In certain embodiments, cleansing is performed minutes to hours before or after application of the topical composition, for example, 5 minutes to 12 hours before or after.

[0121] In certain embodiments, the cleansing composition comprises about 150-300 ppm, or about 200 ppm, of SLPs (70:30 ratio of linear SLPs to lactone-type SLPs). In certain embodiments, the cleansing composition comprises about 1-10 ppm, or about 5 ppm, of MEL.

[0122] The cleansing composition may include additional ingredients, such as carriers, moisturizers, fragrances, colorants, etc., as described above in connection with topical compositions for promoting hair growth, etc. In certain aspects, the cleansing composition is a shampoo, conditioner, or tonic for hair.

[0123] In certain embodiments, the application of topical compositions and / or cleansers can also be complemented with the administration of oral compounds, such as DHT (dihydrotestosterone) blockers or 5α-reductase inhibitors. DHT blockers include, for example, alphatradiol, dutasteride, epristeride, and finasteride.

[0124] DHT is an androgen (male hormone) derived from testosterone in the male body, and is necessary for the development and maintenance of male sexual characteristics, such as muscle tissue and facial hair.Testosterone is converted to DHT through 5α-reductase, which is stored in the oil gland of hair follicle.In some cases, male pattern baldness subjects experience hair follicle shrinkage due to excessive DHT binding to hair follicle; therefore, in some embodiments, DHT blockers may be useful in preventing hair loss through this mechanism.

[0125] In some embodiments, the oral DHT blocker is administered daily to the subject, with the dosage determined by a physician or pharmacist.

[0126] Advantageously, in some aspects, the use of a multi-part regimen can provide longer lasting, consistent results for subjects experiencing hair loss.

[0127] Microbial growth and production of microbial growth by-products The present invention provides methods for culturing microorganisms and producing microbial metabolic products and / or other microbial growth by-products. As used herein, "fermentation" refers to the growth of cells under controlled conditions. Growth can be aerobic or anaerobic.

[0128] In one aspect, the present invention provides materials and methods for producing biomass (e.g., viable cellular material), extracellular metabolites (e.g., small molecules and secreted proteins), residual nutrients and / or intracellular components (e.g., enzymes and other proteins).

[0129] The microbial growth vessel used in accordance with the present invention may be an industrial fermenter or culture reactor. In one embodiment, the vessel may be equipped with or connected to functional controllers / sensors to measure important factors in the culture process, such as pH, oxygen, pressure, temperature, agitator shaft power, humidity, viscosity, microbial density and / or metabolite concentration.

[0130] In further embodiments, the container can also be used to monitor the growth of the microorganisms within the container (e.g., measuring cell count and growth phase). Alternatively, daily samples can be taken from the container and cell counts can be performed using techniques known in the art, such as dilution plating.

[0131] In one embodiment, the method includes supplementing the culture with a nitrogen source. The nitrogen source can be, for example, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium phosphate, ammonia, urea, and / or ammonium chloride. These nitrogen sources can be used alone or in combination of two or more.

[0132] The method can provide oxygen to growing cultures. One embodiment utilizes slow air movement to remove low oxygen-containing air and introduce oxygenated air. The oxygenated air can be ambient air, replenished daily by mechanisms such as an impeller that mechanically agitates the liquid, or an air sparger that provides air bubbles to the liquid to dissolve oxygen into the liquid.

[0133] The method may further comprise supplementing the culture with a carbon source.Carbon sources are generally carbohydrates, such as glucose, sucrose, lactose, fructose, trehalose, mannose, mannitol, and / or maltose; organic acids, such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid, malonic acid, and / or pyruvic acid; alcohols, such as ethanol, propanol, butanol, pentanol, hexanol, isobutanol, and / or glycerol; fats and oils, such as soybean oil, rice bran oil, olive oil, corn oil, sesame oil, and / or linseed oil; and the like.These carbon sources can be used alone or in combination of two or more.

[0134] In one embodiment, the medium contains growth factors and micronutrients for the microorganism. This is particularly suitable for growing microorganisms that do not have the ability to produce all the vitamins they require. The medium may also contain inorganic nutrients, including trace elements such as iron, zinc, copper, manganese, molybdenum and / or cobalt. In addition, sources of vitamins, essential amino acids, and trace elements can be included, for example, in the form of flour or meal, such as corn flour, or in the form of extracts, such as yeast extract, potato extract, beef extract, soybean extract, banana peel extract, or in purified form. Amino acids, such as those useful for protein biosynthesis, can also be included.

[0135] In one embodiment, inorganic salts may also be included. Usable inorganic salts may be potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, magnesium chloride, iron sulfate, iron chloride, manganese sulfate, manganese chloride, zinc sulfate, lead chloride, copper sulfate, calcium chloride, calcium carbonate, and / or sodium carbonate. These inorganic salts may be used alone or in combination of two or more.

[0136] In some embodiments, the culture method may further include adding additional acid and / or antimicrobial agent into the liquid medium before and / or during the culture process.Antimicrobial agents or antibiotics are used to protect the culture from contamination.In addition, antifoaming agents may be added to prevent foam formation and / or accumulation during culture.

[0137] The pH of the mixture must be appropriate for the microorganism of interest. Buffers and pH adjusters, such as carbonates and phosphates, can be used to stabilize the pH near a preferred value. If metal ions are present in high concentrations, it may be necessary to use chelating agents in the liquid medium.

[0138] The methods and apparatus for culturing microorganisms and producing microbial by-products can be carried out in batch, semi-continuous, or continuous processes.

[0139] In one embodiment, the method for culturing the microorganism is carried out at about 5° C. to about 100° C., preferably 15° C. to 60° C., more preferably 25° C. to 50° C. In a further embodiment, the culture can be carried out continuously at a constant temperature. In another embodiment, the culture can be subjected to temperature changes.

[0140] In one embodiment, the equipment used in the method and culture process is sterile. The culture equipment, such as the reactor / vessel, may be separate from, but connected to, a sterilization unit, such as an autoclave. The culture equipment may also be equipped with a sterilization unit for in situ sterilization before inoculation begins. Air may be sterilized by methods known in the art. For example, ambient air may be passed through at least one filter before being introduced into the vessel. In other embodiments, the medium may be pasteurized or, optionally, no heat may be applied at all, in which case low water activity and low pH may be utilized to inhibit bacterial growth.

[0141] In one embodiment, the invention provides a method for producing microbial metabolites, including, for example, biosurfactants, enzymes and / or other proteins, by culturing a microbial strain of the invention under conditions suitable for growth and production of the metabolites. The metabolite content of the resulting culture can be, for example, at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0142] The biomass content of the fermentation broth can be, for example, from 5 g / l to 180 g / l or more, or from 10 g / l to 150 g / l.

[0143] Microbial growth by-products produced by the microorganism of interest may be retained within the microorganism or may be secreted into the liquid medium. In another aspect, the method of producing a microbial growth by-product may further include a step of concentrating and purifying the microbial growth by-product of interest. In a further aspect, the liquid medium may include a compound that stabilizes the activity of the microbial growth by-product.

[0144] In one embodiment, the entire microbial culture is removed when the culture is completed (e.g., when a desired cell density or concentration of a particular metabolite in the medium is reached). In this batch process, an entirely new batch is started once the first batch has been harvested.

[0145] In another embodiment, only a portion of the fermentation product is removed at any one time. In this embodiment, the biomass containing the viable cells remains in the vessel as inoculum for a new culture batch. The removed composition may be cell-free broth or may contain cells. In this way, a quasi-continuous system is created.

[0146] Advantageously, this method does not require complex equipment or high energy consumption. The microorganisms of interest can be cultivated on-site on a small or large scale and used in admixture with the culture medium. Similarly, the metabolic products of the microorganisms can be produced in large quantities where they are needed.

[0147] The microorganisms may be, for example, bacteria, yeast and / or fungi. These microorganisms may be natural or genetically modified. For example, the microorganisms may be transformed with specific genes to exhibit specific properties. The microorganisms may also be mutants of the desired strain. As used herein, "mutant" refers to a strain, genetic variant or subtype of a reference microorganism; a mutant has one or more genetic variations (e.g., point mutations, missense mutations, nonsense mutations, deletions, duplications, frameshift mutations or repeat expansions) compared to the reference microorganism. Procedures for generating mutants are well known in the field of microbiology. For example, UV mutagenesis and nitrosoguanidine are widely used for this purpose.

[0148] In certain embodiments, the microorganisms can produce amphiphilic molecules, enzymes, proteins and / or biopolymers. In particular, microbial biosurfactants can be produced by a variety of microorganisms, such as bacteria, fungi, yeasts, and the like, including, for example, Agrobacterium spp. (e.g., A. radiobacter); Arthrobacter spp.; Aspergillus spp.; Aureobasidium spp. (e.g., A. pullulans); Azotobacter (e.g., A. vinelandii, A. chroococcum); Azospirillum spp. (e.g., A. brasiliensis); brasiliensis); Bacillus spp. (e.g. B. subtilis, B. amyloliquefaciens, B. pumillus, B. coagulans, B. cereus, B. licheniformis, B. falmus, B. laterosporus, B. megaterium); Blakeslea; Candida spp. (e.g. C. albicans, C. rugosa, C. tropicalis, C. lipolytica, C. torulopsis); Clostridium spp. (e.g. C. butyricum, C. tyrobutyricum, C. tyrobutyricum, C. acetobutyricum, and C. beijerinckii); Campylobacter spp.; Cornybacterium spp.; Cryptococcus spp.; Debaryomyces spp. (e.g. D. hansenii)hansenii);Entomophthora spp.;Flavobacterium spp.;Gordonia spp.;Hansenula spp.;Hanseniaspora spp. (e.g. H. uvarum);Issatchenkia spp.;Kluyveromyces spp.;Meyerozyma spp. (e.g. M. guilliermondii);Mortierella spp.;Mycorrhiza spp.;Mycobacterium spp.;Nocardia spp. spp.); Pichia spp. (e.g. P. anomala, P. guilliermondii, P. occidentalis, P. kudriavzevii); Phycomyces spp.; Phythium spp.; Pseudomonas spp. (e.g. P. aeruginosa, P. chlororaphis, P. putida, P. florescens, P. fragi, P. syringae, P. syringae); Pseudozyma spp. (e.g. P. aphidis); Ralslonia spp. (e.g. R. eulropha); Rhodococcus spp. (e.g. R. erythropolis); Rhodospirillum spp. (e.g. R. rubrum); Rhizobium spp.; Rhizopus spp.; Saccharomyces spp. (e.g. S. cerevisiae);cerevisiae, S. boulardii sequela, S. torula; Sphingomonas spp. (e.g. S. paucimobilis); Starmerella spp. (e.g. S. bombicola); Thraustochytrium spp.; Torulopsis spp.; Ustilago spp. (e.g. U. maydis); Wickerhamomyces spp. (e.g. W. anomalus); Williopsis spp.; and / or Zygosaccharomyces spp. (e.g. Z. bailiii). Thank you.

[0149] In one embodiment, the method utilizes yeasts such as, for example, W. anomalus, Pseudozyma aphidis, Starmerella bombicola, Pichia kudriabzevi, or Pichia guilliermondii (Meierozyma guilliermondii). These yeasts are efficient producers of a variety of amphiphilic molecules, including glycolipids, enzymes, and other useful metabolites. In a specific embodiment, the method utilizes W. anomalus.

[0150] Other microbial strains can also be used in accordance with the present invention, including, for example, other strains capable of accumulating significant amounts of amphipathic molecules. Additional metabolic products useful in accordance with the present invention include mannoproteins, β-glucans, and other molecules with bioemulsifying and surface / interfacial tension reducing properties.

[0151] Preparation of microbial-based products One of the microbial-based products of the present invention is simply the fermentation broth containing the microorganisms and / or microbial metabolic products and / or residual nutrients produced by the microorganisms. The product of the fermentation can be used directly without extraction or purification.

[0152] However, extraction and purification can be easily accomplished using standard extraction and / or purification methods or techniques described in the literature. For example, in certain embodiments, the microbial-based product simply comprises the by-products of microbial growth, either in crude or purified form. In certain embodiments, the by-products are biosurfactants produced by the microorganisms grown according to the present invention.

[0153] The microorganisms and / or broth resulting from microbial growth can be removed from the growth vessel, transported, for example, via piping, and used immediately.

[0154] In other embodiments, the composition (microorganism, broth, or microorganism and broth) can be placed in a suitable size container, for example, taking into account the intended use, the intended application method, the size of the fermentation tank, and the means of transportation from the microorganism growth facility to the location of use. Thus, the container in which the microorganism-based composition is placed can be, for example, 1 gallon to 1,000 gallons or more. In other embodiments, the container is 2 gallons, 5 gallons, 25 gallons, or more.

[0155] In certain embodiments, the compositions of the present invention have one or more of the following advantages, e.g., compared to biosurfactants alone: ​​high concentrations of mannoproteins as part of the outer surface of the yeast cell wall (mannoproteins are highly effective bioemulsifiers that can reach an emulsification index of up to 80%); the presence of the biopolymer β-glucan (emulsifier) ​​in the yeast cell wall; the presence of biosurfactants in the culture, which can reduce both surface and interfacial tension; and the presence of metabolic products (e.g., lactic acid, ethanol, etc.).

[0156] When the microbial-based composition is harvested from the growth vessel, additional ingredients can be added when the harvested product is containerized and / or piped (or transported for use). Additives can be, for example, buffers, carriers, other microbial-based compositions produced in the same or different facility, viscosity modifiers, preservatives, nutrients for microbial growth, tracking agents, solvents, biocides, other microorganisms, and other ingredients specific to the intended use.

[0157] Other suitable additives that may be included in the formulations according to the invention include substances that are routinely used in such preparations. Examples of such additives include surfactants, emulsifiers, lubricants, buffers, solubility regulators, pH adjusters, preservatives, stabilizers, and UV-resistant agents.

[0158] In one embodiment, the composition can further comprise a buffering agent, comprising organic acid and amino acid or their salts.Suitable buffering agents include citrate, gluconate, tartrate, malate, acetate, lactate, oxalate, aspartate, malonate, glucoheptonate, pyruvate, galactarate, glucarate, tartronate, glutamate, glycine, lysine, glutamine, methionine, cysteine, arginine, and mixtures thereof.Phosphate and phosphorous acid or their salts can also be used.Although it is suitable to use synthetic buffering agents, it is preferred to use natural buffering agents such as the above-mentioned organic acid and amino acid or their salts.

[0159] In further embodiments, the pH adjuster includes potassium hydroxide, ammonium hydroxide, potassium carbonate or bicarbonate, hydrochloric acid, nitric acid, sulfuric acid, or mixtures thereof.

[0160] In one aspect, additional components can be included in the formulation, such as aqueous preparations of salts such as sodium bicarbonate or sodium carbonate, sodium sulfate, sodium phosphate, sodium biphosphate, and the like.

[0161] Advantageously, according to the invention, a microorganism-based product can include the medium in which the microorganism was grown. The product can be, for example, at least 1%, 5%, 10%, 25%, 50%, 75%, or 100% growth medium by weight. The amount of biomass in the product can be, for example, from 0% to 100% by weight, including all percentages therebetween.

[0162] Optionally, the product can be stored before use. Preferably, the storage time is short. Thus, the storage time can be less than 60 days, 45 days, 30 days, 20 days, 15 days, 10 days, 7 days, 5 days, 3 days, 2 days, 1 day, or 12 hours. In a preferred embodiment, when live cells are present in the product, the product is stored at a cool temperature, for example, below 20°C, 15°C, 10°C, or 5°C. Meanwhile, biosurfactant compositions can be stored at ambient temperature.

[0163] References TIFF2025511022000003.tif44159

Claims

1. A topical composition comprising one or more biosurfactants, a pharmacological or naturotherapy hair growth promoter, and optionally a dermatologically acceptable carrier, wherein the one or more biosurfactants are glycolipids selected from sophorolipids (SLPs) and mannosylerythritol lipids (MELs).

2. A topical composition according to claim 1, comprising a mixture of SLP and MEL, and The SLP comprises a mixture of linear SLP and lactone-type SLP in a ratio of 70:30; or The mixture contains 100 to 250 ppm of SLP; or The mixture contains 1 to 10 ppm of MEL; or The pharmacological hair growth stimulant is minoxidil or bimatoprost; or The mixture contains 5% minoxidil. Topical composition.

3. Contains 200 ppm SLP, 5 ppm MEL, and 5% minoxidil. The SLP comprises a mixture of linear SLP and lactone-type SLP in a ratio of 70:

30. The topical composition according to claim 1.

4. It further comprises one or more dermatological adjuvants and / or additives selected from organic solvents, silicones, pH adjusters, chelating agents, gelling agents, proteins, vitamins, emollients, oils, hydroxy acids, exfoliants, viscosity modifiers, polymers, minerals, insect repellents, lubricants, preservatives, botanicals, essential oils, clearing agents, non-biological surfactants, antioxidants, thickeners, softeners, sunscreens, moisturizers, colorants, and fragrances, and The composition is formulated as a lotion, cream, gel, ointment, liquid, wipe, soap, shampoo, conditioner, tonic, mousse, foam, or spray, and the formulation is suitable for direct application to the skin, scalp, and hair. The topical composition according to claim 1.

5. The topical composition according to claim 1, further comprising live or inactive cells of a microorganism selected from the group consisting of Wickerhamomyces anomalus NRRL Y-68030, Starmerella bombicola, Meyerozyma guilliermondii, Pseudozyma aphidis, Bacillus coagulans, Bacillus amyloliquefaciens NRRL B-67928, and B. subtilis B4 NRRL B-68031.

6. A topical composition for use in a method for promoting hair growth, comprising SLP, MEL, and a pharmacological or naturotherapy hair growth stimulant, The method comprises the step of applying the topical composition to a skin area of ​​a target where hair growth is desired, wherein the application of the topical composition results in faster hair growth compared to any one of the SLP, MEL, or pharmacological or natural hair growth stimulants alone. The aforementioned topical composition.

7. The SLP comprises a mixture of linear SLP and lactone-type SLP in a ratio of 70:30; or The local composition contains 100 to 250 ppm of SLP; or The topical composition contains 1 to 10 ppm of MEL; or The pharmacological hair growth stimulant is minoxidil or bimatoprost; or The topical composition contains 5% minoxidil; or The topical composition comprises 200 ppm SLP, 5 ppm MEL, and 5% minoxidil; or The SLP comprises a mixture of linear SLP and lactone-type SLP in a 70:30 ratio, and the SLP comprises a mixture of linear SLP and lactone-type SLP in a 70:30 ratio; or The method further comprises administering an orally administered DHT blocker to the subject once daily, wherein the DHT blocker is alfatradiol, dutasteride, epristeride, or finasteride. The topical composition according to claim 6.

8. The subject is experiencing hair loss as a result of alopecia, chemotherapy, skin injury, burns, or drug side effects; or The target skin area to which the topical composition is applied is the scalp, eyelids, face, limbs, or torso; or Apply the composition once a day; or The composition is rubbed into the skin area so that it is absorbed into that area; or The composition is left on the skin area for a therapeutically effective period of time, and then the composition is washed off or otherwise removed from the skin. The topical composition according to claim 6.

9. The topical composition according to claim 6, further comprising the step of cleansing the skin area with a cleansing composition comprising a mixture of SLP and MEL, wherein the cleansing composition is applied 5 minutes to 12 hours before or after the application of the topical composition.

10. The topical composition according to claim 1, further comprising a microbial culture supernatant.