Production and use of ozone oxidation products of oleic-acid sophorolipids

Abstract

The subject invention provides a method for improving the solubility of azelaic acid (AzA), wherein the method comprises oxidizing a linear sophorolipid to produce AzA and a short chain, truncated sophorolipid. The truncated sophorolipid can bring the AzA into solution at higher levels and lower pH than when AzA is produced using oleic acid starting materials and methods that are known in the art.

Description

[0001] DESCRIPTION

[0002] PRODUCTION AND USE OF OZONE OXIDATION PRODUCTS OF OLEIC-ACID SOPHOROLIPIDS

[0003] CROSS-REFERENCE TO RELATED APPLICATION

[0004] This application claims priority to U.S. Provisional Patent Application No. 63 / 583,091, filed September 15, 2023, which is incorporated herein by reference in its entirety.

[0005] BACKGROUND OF THE INVENTION

[0006] Azelaic acid (AzA) is used as a starting material for producing industrial products, including polyesters, plasticizers and lubricants. Azelaic acid is also a component of hair- and skincare products. For example, Finacea® (azelaic acid) Gel (Bayer Healthcare Pharmaceuticals Inc. Whippany, N.I.) is prescribed for topical treatment of mild to moderate rosacea. Other uses for azelaic acid as an ingredient include, for example, treatment of acne and skin hyperpigmentation. Chemically, azelaic acid is a Cg aliphatic saturated dicarboxylic acid, which is most commonly produced via ozonolysis of oleic acid, followed by dioxygen oxidation.

[0007] Despites its potential uses, formulating products containing azelaic acid remains a challenge due to its low water solubility at pH less than 8 (< 2 mg / mL). Active levels of azelaic acid in personal care products range from, e.g., 3-10%, while some pharmaceutical products such as Finacea®, contain, e.g., 15% azelaic acid. Formulators must use either a complex combination of lipophilic solvents and surfactants or high pH to achieve these formulation loading levels; however, consumer demand for lower toxicity, more bio-based skincare, healthcare and personal care products is on the rise.

[0008] Accordingly, methods are needed for improving the solubility of azelaic acid in all pH ranges, which reduce and / or replace the use of harsh, synthetic additives with potential for negative impacts on health and the environment.

[0009] BRIEF SUMMARY OF THE INVENTION

[0010] The subject invention provides methods for improving the formulation of consumer goods, including pharmaceuticals, cosmetics and personal care products. More specifically, the subject invention provides methods for improving the solubility of azelaic acid (AzA) for its use in such products in a broad range of pH levels. Advantageously, the methods can help reduce and / or eliminate the need for harsh, synthetic surfactants and / or solvents to achieve a desired level of solubilized AzA in lower pH formulations.

[0011] In certain embodiments, the subject invention provides a method for improving the solubility of AzA, wherein the method comprises utilizing a sophorolipid (SLP) as a starting material in the production of AzA (as opposed to using oleic acid). Through oxidation (preferably two-step oxidation), the SLP is converted into AzA and a short chain, truncated (C9) SLP (tSLP). FIG. 1.

[0012] Advantageously, the tSLP can bring the AzA into solution at higher concentrations and at lower pH (i.e., < pH 8) than when AzA is produced using oleic acid starting materials and methods that are known in the art. Improved solubility can improve the function of the AzA in a formulation as intended, as well as improve the overall appearance and clarity of the solution containing the AzA. Notably, the AzA solution will appear clear or transparent rather than white or cloudy.

[0013] Furthermore, the tSLP side product can provide additional benefits in an end product formulation. For example, the tSLP can have similar utility to traditional C18 linear SLP, but providing improved water solubility, improved bioavailability for other active components, improved solvency for other active components and improved biodegradability.

[0014] Other aspects of the subject invention are directed to improved formulations comprising an AzA and / or tSLP produced according to the subject invention, as well as methods of producing and / or formulating a pharmaceutical, cosmetic and / or personal care product by mixing the AzA and / or tSLP with one or more active and / or inert ingredients that are typical for such compositions (e.g., carriers, pH modifiers, emollients, moisturizers, antimicrobials, and the like). Advantageously, in some embodiments, the compositions produced according to the subject invention can exhibit improved bioavailability and dermal penetrability of active compounds.

[0015] BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 depicts a reaction scheme in which a linear sophorolipid is subjected to primary and secondary oxidation to yield a truncated SLP and azelaic acid.

[0017] DETAILED DESCRIPTION OF THE INVENTION

[0018] As used herein, “surfactant” means a compound that lowers the surface tension (or interfacial tension) between phases. Surfactants act as, e.g., detergents, wetting agents, emulsifiers, foaming agents, and / or dispersants. A “biosurfactant” is a surface-active substance produced by a living cell and / or using naturally-derived substrates.

[0019] Biosurfactants are a structurally diverse group of surface-active substances consisting of two parts: a polar (hydrophilic) moiety and non-polar (hydrophobic) group. Due to their amphiphilic structure, biosurfactants can, for example, increase the surface area of hydrophobic water-insoluble substances, increase the water bioavailability of such substances, and change the properties of bacterial cell surfaces. Biosurfactants can also reduce the interfacial tension between water and oil and, therefore, lower the hydrostatic pressure required to move entrapped liquid to overcome the capillary effect. Biosurfactants accumulate at interfaces, thus reducing interfacial tension and leading to the formation of aggregated micellar structures in solution. The formation of micelles provides a physical mechanism to mobilize, for example, oil in a moving aqueous phase.

[0020] The ability of biosurfactants to form pores and destabilize biological membranes also permits their use as antibacterial, antifungal, and hemolytic agents to, for example, control pests and / or microbial growth.

[0021] Typically, the hydrophilic group of a biosurfactant is a sugar (e.g., a mono-, di-, or polysaccharide) or a peptide, while the hydrophobic group is typically a fatty acid. Thus, there are countless potential variations of biosurfactant molecules based on, for example, type of sugar, number of sugars, size of peptides, which amino acids are present in the peptides, fatty acid length, saturation of fatty acids, additional acetylation, additional functional groups, esterification, polarity and charge of the molecule.

[0022] These variations lead to a group of molecules comprising a wide variety of classes, including, for example, glycolipids (e.g., sophorolipids, rhamnolipids, cellobiose lipids, mannosylerythritol lipids and trehalose lipids), lipopeptides (e.g., surfactin, iturin, fengycin, arthrofactin and lichenysin), flavolipids, phospholipids (e.g., cardiolipins), fatty acid ester compounds, and high molecular weight polymers such as lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes. Each type of biosurfactant within each class can further comprise subtypes having further modified structures.

[0023] Like chemical surfactants, each biosurfactant molecule has its own HLB value depending on its structure; however, unlike production of chemical surfactants, which results in a single molecule with a single HLB value or range, one cycle of biosurfactant production typically results in a mixture of biosurfactant molecules (e.g., subtypes and isomers thereof).

[0024] The phrases “biosurfactant” and “biosurfactant molecule” include all forms, analogs, orthologs, isomers, and natural and / or anthropogenic modifications of any biosurfactant class (e.g., glycolipid) and / or subtype thereof (e.g., sophorolipid).

[0025] A particular biosurfactant group of interest herein is the sophorolipid group. Sophorolipids are glycolipid biosurfactants produced by fermentation of microbes, such as Starmerella spp. yeasts and / or Candida spp. yeasts, e.g., Starmerella (Candida) bombicola (e.g., ATCC 22214), Candida apicola, Candida batistae, Candida floricola, Candida riodocensis, Candida stellate and / or Candida knot, using carbon sources such as pure fatty acids, fatty acid mixtures, pure fatty acid esters, mixtures of fatty acid esters, triglycerides along with carbohydrate sources such as com syrup, dextrins and glucose.

[0026] The production of sophorolipids with the use of renewable substrates and different microbial species, as well as the variation in culture parameters (incubation time, stirring speed, pH of the medium and added nutrients), allow for the acquisition of compounds with distinct structural and physical properties. This makes it possible to produce a wide variety of compounds that can elicit different physical, chemical, biochemical, and biophysical properties.

[0027] Sophorolipids are amphiphilic molecules containing a hydrophilic carbohydrate group (sophorose) and a hydrophobic fatty acid. Sophorose is a disaccharide that consists of two glucose molecules linked via a B-1,2 bond. The sophorose in sophorolipids can be acetylated at the 6’- and / or 6 ’’-positions. The fatty acid in sophorolipids is glycosidically attached to the sophorose molecule through its hydroxyl group, and its terminal carboxylic acid group is either free (as shown in Fig. 1) or internally esterified (i.e., lactonic form), generally at the 4”-position. Most commonly, the fatty acids consist of 16 or 18 carbon atoms with one degree of unsaturation, but the fatty acid structure varies depending on the carbon sources provided during the production (e.g., degree of unsaturation, chain length) and the microorganisms conducting the fermentation. For example, it can consist of chain lengths from C12 to C22 having various degrees and positions of unsaturation.

[0028] Further, the degree of lactonization of sophorolipids and acetylation of the sophorose portion may be influenced by, inter alia, the carbon source fed during sophorolipid production. For example, it has been found that sophorolipids derived from rapeseed, sunflower, and palm oils rich in Cl 8:0 and Cl 8: 1 fatty acids are formed with higher levels of diacetylated lactones than sophorolipids produced from the corresponding fatty acid ester feedstocks.

[0029] Thus, sophorolipids are frequently produced as a mixture of related molecules. Differences among the related molecules arise mainly from: their fatty acid structure (degree of unsaturation, chain length, position(s) of unsaturation and position of hydroxylation); whether they are produced in the free or lactonic form, the acetylation pattern; the presence of stereoisomers; and / or whether the glycosidic bond on the fatty acid is at the co-position (e.g., terminal) or co-1 position (subterminal).

[0030] As used herein, the term “sophorolipid,” “sophorolipid molecule,” “SLP” or “SLP molecule” includes all forms, and isomers thereof, of SLP molecules, including, for example, acidic (linear) SLP and lactonic SLP. Further included are mono-acetylated SLP, di-acetylated SLP, esterified SLP, SLP with varying hydrophobic chain lengths, SLP with fatty acid-amino acid complexes attached, and other, including those that are and / or are not described within in this disclosure. SLP can be represented by General Formula (A) and / or General Formula (B), and are obtained as a collection of multiple structural homologues: where R’ and R1independently represent saturated hydrocarbon chains or single or multiple, in particular single, unsaturated hydrocarbon chains having 8 to 20, in particular 12 to 18 carbon atoms, more preferably 14 to 18 carbon atoms, which can be linear or branched and can comprise one or more hydroxy groups, R2and R2independently represent a hydrogen atom or a saturated alkyl functional group or a single or multiple, in particular single, unsaturated alkyl functional group having 1 to 9 carbon atoms, more preferably 1 to 4 carbon atoms, which can be linear or branched and can comprise one or more hydroxy groups, and R3, R3, R4and R4independently represent a hydrogen atom or -COCH3. R5can be, but is not limited to, — H or — OH.

[0031] SLP have environmental compatibility, high biodegradability, low toxicity, high selectivity and specific activity in a broad range of temperature, pH and salinity conditions. Additionally, in some embodiments, SLP can be advantageous due to their small micelle size, which can help facilitate the movement of the micelle, and compounds enclosed therein, through nanoscale pores and spaces. In certain embodiments, the micelle size of a SLP 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. As used herein, “applying” a composition or product refers to contacting it with a target or site such that the composition or product can have an effect on or in that target or site.

[0032] As used herein, an “isolated” or “purified” nucleic acid molecule, polynucleotide, polypeptide, protein or organic compound such as a small molecule (e.g., those described below), is substantially free of other compounds, such as cellular material, with which it is associated in nature. A purified or isolated polynucleotide (ribonucleic acid (RNA) or deoxyribonucleic acid (DNA)) is free of the genes or sequences that flank it in its naturally-occurring state. A purified or isolated polypeptide is free of the amino acids or sequences that flank it in its naturally-occurring state. An isolated microbial strain means that the strain is removed from the environment in which it exists in nature. Thus, the isolated strain may exist as, for example, a biologically pure culture, or as spores (or other forms of the strain) in association with a carrier.

[0033] In certain embodiments, purified compounds are at least 60% by weight the compound of interest. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 98%, by weight the compound of interest. For example, a purified compound is one that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 98%, 99%, or 100% (w / w) of the desired compound by weight. Purity is measured by any appropriate standard method, for example, by column chromatography, thin layer chromatography, or high-performance liquid chromatography (HPLC) analysis.

[0034] A “metabolite” refers to any substance produced by metabolism or a substance necessary for taking part in a particular metabolic process. A metabolite can be an organic compound that is a starting material, an intermediate in, or an end product of metabolism. Examples of metabolites include, but are not limited to, enzymes, acids, solvents, alcohols, proteins, vitamins, minerals, microelements, amino acids, biopolymers and biosurfactants.

[0035] Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 20 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, as well as all intervening decimal values between the aforementioned integers such as, for example, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to sub-ranges, “nested sub-ranges” that extend from either end point of the range are specifically contemplated. For example, a nested sub-range of an exemplary range of 1 to 50 may comprise 1 to 10, 1 to 20, 1 to 30, and 1 to 40 in one direction, or 50 to 40, 50 to 30, 50 to 20, and 50 to 10 in the other direction.

[0036] As used herein a “reduction” means a negative alteration, and an “increase” means a positive alteration, wherein the negative or positive alteration is at least 0.001%, 0.01%, 0.1%, 0.5%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%. The transitional term “comprising,” which is synonymous with “including,” or “containing,” is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. By contrast, the transitional phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim. The transitional phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. Use of the term “comprising” contemplates other embodiments that “consist” or “consist essentially of’ the recited component(s).

[0037] Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms “a,” “and” and “the” are understood to be singular or plural.

[0038] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about.

[0039] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

[0040] All references cited herein are hereby incorporated by reference in their entirety.

[0041] Production of Azelaic Acid

[0042] In certain embodiments, the subject invention provides a method for improving the solubility of AzA, wherein the method comprises utilizing a sophorolipid (SUP) as a starting material in the production of AzA (as opposed to oleic acid). Through oxidation, the SUP is converted into AzA and a short chain, truncated (C9) SUP (tSUP). FIG. 1.

[0043] Advantageously, the tSUP can bring the AzA into solution at higher concentrations and lower pH than when AzA is produced using oleic acid and methods that are known in the art. Improved solubility can improve the function of the AzA in a formulation as intended, as well as improve the overall appearance and clarity of the solution containing the AzA. For example, with improved solubilization, the AzA appears clear or transparent in solution, rather than white or cloudy.

[0044] Furthermore, the tSUP can provide additional benefits in an end product formulation. For example, the tSUP can have similar utility to traditional Cl 8 linear SUP, but providing improved water solubility, improved bioavailability for other active components, improved solvency for other active components and improved biodegradability.

[0045] In certain embodiments, the methods of the subject invention comprise converting a SLP molecule into a linear tSLP and AzA via oxidation. The linear SLP starting material can be prepared by a fermentation process known in the art in the presence of select fatty acids and carbohydrate feedstocks. As used herein “fermentation” refers to growth of cells under controlled conditions. The growth could be aerobic or anaerobic. The fermentation processes known in the art include, but are not limited to, solid-state fermentation, submerged fermentation, or modifications, hybrids and / or combinations thereof.

[0046] In some embodiments, the SLP produced by microorganisms of interest may be retained in the microorganisms or secreted into their growth medium. The SLP content of the growth medium can be, for example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80 %, or 90% by weight.

[0047] The growth medium may contain compounds that stabilize the activity of the SLP. The SLP can be purified, or the SLP can be used in crude form, meaning they are not separated from the fermentation broth in which they were produced. In some embodiments, the crude form contains residual cellular matter, including cell wall components and other metabolites produced during fermentation.

[0048] In certain embodiments, the SLP is isolated and / or purified from the growth medium resulting from fermentation of a biosurfactant-producing microorganism. Isolation and purification can be easily achieved using standard methods or techniques described in the literature. The SLP can be further concentrated, if desired.

[0049] Two-Step Oxidation

[0050] A) In preferred embodiments, the SLP starting material for the production of AzA is a C18 linear SLP derived from oleic acid. The linear SLP is first subjected to primary oxidation in a vessel. A cosolvent is added to the linear SLP at a ratio of 1: 1, 1:2 or 1:3, wherein the cosolvent is, e.g., isopropanol, ethanol, methanol, acetone or acetonitrile.

[0051] In certain preferred embodiments, the ratio of cosolvent to linear SLP is 1:2. In other preferred embodiments, the cosolvent is acetone.

[0052] The mixture of linear SLP and cosolvent is ozonated using a gas sparger for 12 hours to 28 days, or 1 day to 14 days, with agitation. Preferably, the ozone generator is supplied with compressed air, although oxygen gas can also be used. The ozone flow rate should be between 0.5 to 10 L / min, or 1 to 5 L / min.

[0053] B) After primary oxidation is determined to be complete (using, e.g., LCMS), the method comprises subjecting the ozonated linear SLP and co-solvent mixture to secondary oxidation. During secondary oxidation, the ozone generator is stopped, but air flow continues into the vessel.

[0054] A secondary oxidant is added to the vessel, wherein the secondary oxidant is, e.g., hydrogen peroxide, ozone gas, oxygen gas or oxone salt. In certain preferred embodiments, the secondary oxidant is ozone gas.

[0055] C) The two-step oxidation reaction is next “cleaned up.” If the secondary oxidant of B) is oxone salts, two product streams can be created: a blended material and a separate tSLP and SLP -derived AzA.

[0056] 1) If the product to be created is the blended material, the crude reaction is basified with the addition of barium hydroxide in a 2-molar ratio to the supplied oxone salt. Heat is used to ensure complete dissolution of the barium hydroxide. Precipitated barium sulfate is removed by filtration and / or centrifugation. The final product contains tSLP in equal molar parts to SLP -derived AzA, with the concentration being dependent upon the starting material at this stage, where the concentration of tSLP and the concentration of AzA being produced from SLP can be summed to equal the original concentration of SLP.

[0057] 2) If the product is the separate tSLP and SLP-derived AzA, the crude material is acidified to a pH between 2-3 using sulfuric acid. The precipitated AzA is recovered by filtration, and contains some tSLP, which can be removed, e.g., by recrystallization in hot water.

[0058] The crude material is then basified with barium hydroxide using 2 molar equivalents per oxone salt and added sulfuric acid. Heat can be used to ensure complete dissolution of the barium hydroxide. Precipitated barium sulfate is removed by filtration and / or centrifugation. The final product contains the tSLP and small parts (0 - 5 wt %) SLP-derived AzA.

[0059] 3) If the secondary oxidant is not oxone salts, no further processing is required for a blended product. AzA can be isolated from the crude secondary oxidation product as described above in paragraph C2, by acidifying the crude reaction to a pH between 2 - 3 using sulfuric acid and collecting the material via filtration.

[0060] Further, the tSLP can be isolated as described above in C2; however, barium hydroxide is not needed. Instead, any hydroxide salt can be used. No sulfate precipitation will occur with this route.

[0061] Compositions

[0062] Other aspects of the subject invention are directed to improved formulations comprising an AzA and / or tSLP produced according to the subject invention, as well as methods of producing and / or formulating a pharmaceutical, cosmetic and / or personal care product by mixing the AzA and / or tSLP with one or more active and / or inert ingredients that are typical for such compositions (e.g., carriers, pH modifiers, emollients, moisturizers, antimicrobials, and the like). Advantageously, in addition to improved solubility of the AzA, the compositions produced according to the subject invention can exhibit improved bioavailability, solubility and / or dermal penetrability of other active compounds.

[0063] Acceptable carriers may be included in the composition, depending on the form of the composition and / or mode of administration. Pharmaceutically or cosmetically acceptable carries include but are not limited to, pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while com starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid, or talc. If desired or suitable, a coating material may also be used such as glyceryl monostearate or glyceryl distearate, for example, to delay absorption in the gastrointestinal tract if appropriate and the pharmaceutical composition is in the form of a solid form.

[0064] In certain embodiments, the composition according to the present invention further comprises microbial growth by-products. Particularly, embodiments of the present invention provide a topical therapeutic composition for promoting the healing of skin conditions or an ingestible composition for improving gut health, wherein the composition comprises therapeutically effective amounts of one or more biochemicals produced by the cultivation of microorganisms.

[0065] In certain embodiments, the microbial growth by-products are amphiphilic molecules, enzymes and / or proteins. In one embodiment, the microbial growth by-products have antimicrobial and / or anti-biofdm properties. In one embodiment, the microbial growth by-products have skin rejuvenating properties, meaning they help make skin appear younger, smoother, and brighter.

[0066] In certain embodiments, the composition further comprises one or more amphiphilic molecules, wherein the amphiphilic molecules are biosurfactants selected from, for example, low molecular weight glycolipids (e.g., sophorolipids, rhamnolipids, mannosylerythritol lipids and trehalose lipids), lipopeptides (e.g., surfactin, iturin, fengycin, athrofactin and lichenysin), cellobiose lipids, flavolipids, phospholipids (e.g., cardiolipins), and high molecular weight polymers such as lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein- fatty acid complexes. The amphiphilic molecules may be a modified form, derivative, fraction, isoform, isomer, or subtype of a biosurfactant, including forms that are biologically or synthetically modified. In one embodiment, the one or more biosurfactants are present in the composition in critical micelle concentration (CMC).

[0067] In one embodiment, the composition comprises a combination of SLP and mannosylerythritol lipids (MEL). The SLP can be present in the formulation at, e.g., 0.1 to 500 ppm, or 1 to 250 ppm, or 5 to 100 ppm. The MEL can be present in the formulation at, e.g., 0.001 ppm to 100 ppm, or 0.1 to 50 ppm, or 0.5 to 25 ppm, or 1 to 5 ppm.

[0068] The composition according to the present invention may come in various physical forms. In some embodiments, the composition is in the form of solids including tablets, filled capsules, powder and pellet forms. In another embodiment, the pharmaceutical composition may be in the powder form, in which the pharmaceutically accepted carrier is a finely divided solid that is in a mixture with the finely divided active ingredient. In a further embodiment, the pharmaceutical composition according to the present invention is a sustained release system such as semipermeable matrices of solid hydrophobic polymers containing the tSLP of the present invention. In another embodiment, the pharmaceutical composition is in a liquid form such as aqueous or non-aqueous solutions, suspensions, emulsions, elixirs, and capsules filled with the same.

[0069] Any appropriate mode of administration can be utilized to administer the compositions according to the present invention. Such mode includes, but is not limited to, oral, rectal, bronchial, nasal, topical, buccal, sub-lingual, transdermal, vaginal, intramuscular, intraperitoneal, intravenous, intra-arterial, subcutaneous, intracerebral, intraocular administration or in a form suitable for administration by inhalation or insufflation, including powders and liquid aerosol administration, or intraparenteral infusion. Administration may be also by way of other carriers or vehicles such as patches, micelles, liposomes, vesicles, implants (e.g., microimplants), synthetic polymers, and the like. Factors that determine the mode of administration of a particular composition include the nature of the therapeutic agent cargo that is being delivered to the subject.

[0070] Orally administered compositions according to the invention are any preparations or compositions suitable for consumption, for nutrition, for oral hygiene and / or for pleasure, and are products intended to be introduced into the human or animal oral cavity, to remain there for a certain period of time and then to either be swallowed (e.g. , food ready for consumption) or to be removed from the oral cavity again (e.g., chewing gums or products of oral hygiene or medical mouth washes). These products include all substances or products intended to be ingested by humans or animals in a processed, semi-processed or unprocessed state. This also includes substances that are added to orally consumable products (particularly food and pharmaceutical products) during their production, treatment or processing and intended to be introduced into the human or animal oral cavity. In certain embodiments, the composition can further comprise components that are, for example, sources of energy, nutrients and / or other health-promoting supplements, flavorings, preservatives, pH adjusters, sweeteners and / or dyes. The composition may also be carbonated. In one embodiment, the composition is formulated as a dehydrated powder or concentrate that can be reconstituted into a drinkable fluid by the addition of water. In one embodiment, the composition is formulated as a blended smoothie or milkshake.

[0071] Orally administered compositions can also include substances intended to be swallowed by humans or animals and then digested in an unmodified, prepared, or processed state; the orally administered compositions according to the invention therefore also include casings, coatings or other encapsulations that are intended also to be swallowed together with the product or for which swallowing is to be anticipated.

[0072] In other embodiments, the compositions according to the present invention are formulated for direct administration into the GI tract. For example, the composition can be formulated for administration to the proximal lower GI by hand or colonoscopy, to the distal lower GI tract via enema or rectal tubes, and the upper GI tract via nasogastric tubes, duodenal tubes, and endoscopy / gastroscopy.

[0073] In certain embodiments, the present invention provides topical skin care compositions. In some embodiments, the topical composition can further comprise a dermatologically acceptable carrier, and one or more active or inactive cosmetic ingredients, such as, e.g. , vitamins, moisturizers, dyes, fragrances, sunscreens, exfoliants, essential oils, botanical extracts, and so on.

[0074] The dermatologically acceptable carrier may include, for example, water; saline; physiological saline; ointments; creams; oil-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 octal 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 and isohexadecane; silicone oils such as cyclomethicone, dimethicone, dimethicone cross-polymer, polysiloxanes and their derivatives, preferably organo-modified derivatives including PDMS, dimethicone copolyol, dimethiconols, and amodimethiconols; hydrocarbon oils such as mineral oil, petrolatum, isoeicosane and polyolefins, e.g., (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 waxes; or any combinations or mixtures of the foregoing. Aqueous carriers may include one or more solvents miscible with water, including lower alcohols, such as ethanol, isopropanol, and the like. The carrier may comprise from about 1% to about 99% by weight of the composition, from 10% to about 85%, from 25% to 75%, or from 50% to about 65%.

[0075] 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 sulfates and sodium / ammonium laureth sulfates), amphoterics (e.g., amphoacetates and amphopropionates), sulfosuccinates, alkyl polyglucosides, betaines (e.g., cocamidopropyl betaine), sultaines, sacrosinates, isethionates, taurates, ethoxylated sorbitan esters, alkanolamides and amino acid-based surfactants.

[0076] Viscosity modifiers can also be added to the compositions, including, for example, cocamide DEA, oleamide DEA, sodium chloride, cellulosic polymers, polyacrylates, ethoxylated esters, alcohol, glycols, xylene sulfonates, polysorbate 20, alkanolamides, and cellulose derivatives (e.g., hydroxypropyl methylcellulose and hydroxyethyl cellulose).

[0077] Polymers can also be added, including, for example, xanthan gum, guar gum, polyquatemium-10, PEG-120 methyl glucose dioleate, PEG-150 distearate, PEG-150 polyglyceryl- 2 tristearate and PEG- 150 pentaerythrityl tetrastearate

[0078] In one embodiment, the composition may include additional skin actives, including but not limited to, keratolytic agents, desquamating agents, keratinocyte proliferation enhancers, collagenase inhibitors, elastase inhibitors, depigmenting agents, anti-inflammatory agents, steroids, anti -acne agents, antioxidants (e.g., ascorbic acid), advanced glycation end-product (AGE) inhibitors, exfoliating agents (e.g., glycolic acid, 3,6,9-trioxaundecanedioic acid, etc.), estrogen synthetase stimulating compounds (e.g., caffeine and derivatives), antibiotics (e.g., bacitracin, neomycin, vancomycin), compounds capable of inhibiting 5 alpha-reductase activity (e.g., linolenic acid, linoleic acid, finasteride, and mixtures thereof), barrier function enhancing agents (e.g., ceramides, glycerides, cholesterol and its esters, alpha-hydroxy and omega-hydroxy fatty acids and esters thereof), retinoids, sunscreen, to name a few.

[0079] The composition may optionally comprise additives, adjuvants, or other components of topical compositions known to those skilled in the art including, but not limited to: skin penetration enhancers; emollients (e.g., isopropyl myristate, petrolatum, volatile or non-volatile silicones oils, such as methicone and dimethicone, ester oils, mineral oils, and fatty acid esters); emulsifiers (e.g., glyceryl stearate, Candelilla / jojoba / rice bran polyglyceryl-3 esters); humectants (e.g., colloidal oatmeal, glycerin, hexylene glycol, caprylyl glycol); skin plumpers (e.g., palmitoyl oligopeptide, collagen, collagen and / or glycosaminoglycan (GAG) enhancing agents); anti-inflammatory agents (e.g., Aloe vera, bioflavonoids, diclofenac, salicylic acid); chelating agents (e.g., EDTA or a salt thereof, such as disodium EDTA); vitamins (e.g., retinyl palmitate, tocopherol and ascorbic acid); vitamin derivatives (e.g., ascorbyl monopalmitate, tocopheryl acetate, Vitamin E palmitate); thickeners (e.g., hydroxyalkyl cellulose, carboxymethylcellulose, carbomers, and vegetable gums, such as xanthan gum); gelling agents (e.g., ester-terminated polyester amides); structuring agents; proteins (e.g., lactoferrin); immune modulators (e.g., corticosteroids and non-steroidal immune modulators). Further examples include, but are not limited to: stabilizers; fragrances; fdm formers; insect repellents; skin cooling compounds; skin protectants; lubricants; preservatives; pearls; chromalites; micas; conditioners; anti-allergenics; pH adjusters; and antimicrobials.

[0080] The composition may be formulated as a suspension, emulsion, hydrogel, multiphase solution, vesicular dispersion or in any other known form of topical skin composition.

[0081] In certain embodiments, the topical composition may be formulated so that it can be applied, for example, via pen, tube, bottle, brush, stick, sponge, cotton swab, towelette (wipe), sprayer, dropper, hand, or finger.

[0082] The topical composition may be formulated in a variety of product forms, such as, for example, a lotion, cream, serum, spray, aerosol, liquid cake, ointment, essence, gel, paste, patch, pencil, powder, towelette, soap or other cleanser, shampoo, conditioner, stick, foam, mousse, elixir or concentrate.

[0083] In one embodiment, the composition can be provided in the form of a facial mask. The mask may be a peel-off mask or wash-off mask comprising the composition of the present invention.

[0084] In other embodiments, the composition of the present invention can be provided in a wound dressing, bandage, or a patch that may be applied, attached, or coupled to one or more layers of the skin or tissue of the subject. For example, the composition may be applied to a wound dressing, bandage, or a patch, which can then be placed over the area of skin being treated.

[0085] A wound dressing, bandage, and a patch can be made, for example, from any material that is dermatologically acceptable and suitable for placing on a wound or the skin. In exemplary embodiments, the A wound dressing, bandage, and a patch may be made from a woven or nonwoven fabric of synthetic or non-synthetic fibers, or any combination thereof. The dressing may also include a support, such as a polymer foam, a natural or man-made sponge, a gel or a membrane that may absorb or have disposed thereon, a composition. By way of example, the support can be a film, a natural or synthetic polymer, or a rigid or malleable material (e.g., gauze). The wound dressing, bandage, and a patch may be absorbent and can be, for example, wetted with a composition of the present invention before applying the gauze to a wound, scar or other sites.

[0086] In one embodiment, the wound dressing, bandage, or a patch may be impregnated with a composition of the subject invention and, optionally, dried. This allows the impregnated material to be stored for later use, or to avoid excessively dampening of the skin. In other embodiments, the pharmaceutical composition may be formulated for parenteral administration (e.g. , by injection, for example bolus injection or continuous infusion). In addition, the composition may be presented in unit dose form in ampoules, pre-fdled syringes, and small volume infusion or in multi-dose containers with or without an added preservative. The composition may be in forms of suspensions, solutions, or emulsions in oily or aqueous carriers. The composition may further contain formulation agents such as suspending, stabilizing and / or dispersing agents. In a further embodiment, the active ingredient of the composition according to the invention may be in a powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution for constitution with a suitable carrier, e.g., sterile, pyrogen-free water, before use.

[0087] In yet other embodiments, the pharmaceutical composition may be formulated for direct administration to the nasal cavity by conventional means, for example with a dropper, pipette, or spray. The compositions may be provided in single or multi-dose form. Administration to the respiratory tract may also be achieved by means of an aerosol formulation in which the active ingredient is provided in a pressurized pack with a suitable propellant such as a chlorofluorocarbon (CFC) for example dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. The aerosol may conveniently also contain a surfactant such as lecithin.

Claims

CLAIMSWe claim:

1. A method for improving the solubility of azelaic acid (AzA), the method comprising subjecting a linear sophorolipid to two-step oxidation, wherein the two step-oxidation yields a truncated (C9) SLP (tSLP) and AzA, and wherein the presence of the tSLP brings the AzA into solution at higher AzA concentrations and lower pH than when AzA is produced using oxidation of oleic acid.

2. The method of claim 1, wherein the AzA is soluble a pH less than 8 in the presence of the tSLP.

3. The method of claim 1, wherein step one of the two-step oxidation comprises primary oxidation, said primary oxidation comprising adding the linear SLP to a vessel, adding a cosolvent to the vessel, and ozonating the linear SLP and cosolvent in the vessel for 12 hours to 28 days with agitation.

4. The method of claim 3, wherein the cosolvent is added to the linear SLP at a ratio of 1: 1, 1:2 or 1:3.

5. The method of claim 3, wherein the cosolvent is isopropanol, ethanol, methanol, acetone or acetonitrile.

6. The method of claim 3, wherein the ozone is generated using an ozone generator supplied with compressed air or oxygen gas, and wherein the ozone generator provides ozone at a flow rate of between 0.5 to 10 L / min.

7. The method of claim 3, wherein step two of the two-step oxidation comprises subjecting the ozonated linear SLP and cosolvent mixture to secondary oxidation, wherein secondary oxidation comprises stopping ozonation but allowing air flow to continue into the vessel, and adding a secondary oxidant to the vessel to produce a secondary oxidation crude product.

8. The method of claim 7, wherein the secondary oxidant is hydrogen peroxide, ozone gas, oxygen gas or oxone salt.

9. The method of claim 7, wherein the secondary oxidant is oxone salts, and wherein secondary oxidation is used to produce either a blended material or a separate material, said blended material comprising a blend of tSLP and SLP-derived AzA and said separate material comprising tSLP and SLP-derived AzA that are separated from one another.

10. The method of claim 9, comprising basifying the blended material by adding barium hydroxide in a 2-molar ratio to the supplied oxone salt, adding heat to ensure complete dissolution of the barium hydroxide, thereby producing precipitated barium sulfate, and removing the precipitated barium sulfate by fdtration and / or centrifugation, wherein tSLP and AzA are produced in equal molar parts.

11. The method of claim 9, comprising acidifying the separate material to a pH of 2-3 by adding sulfuric acid to the separate material, wherein AzA precipitates from the separate material, and recovering the precipitated AzA from the remaining separate material by filtration.

12. The method of claim 11, further comprising basifying the remaining separate material by adding barium hydroxide in a 2-molar ratio to the supplied oxone salt and sulfuric acid added, adding heat to ensure complete dissolution of the barium hydroxide, thereby producing precipitated barium sulfate, and removing the precipitated barium sulfate by filtration and / or centrifugation, wherein tSLP is produced, said tSLP comprising 0-5 wt% SLP-derived AzA.

13. The method of claim 8, wherein the secondary oxidant is not oxone salts, further comprising acidifying the secondary oxidation crude product to a pH of 2-3 by adding sulfuric acid, wherein AzA precipitates from the crude product, recovering the precipitated AzA from the remaining crude product by filtration, and isolating tSLP from the crude product by adding a hydroxide salt in a 2- molar ratio to the supplied secondary oxidant, and adding heat to ensure complete dissolution of the hydroxide salt.

14. A composition comprising AzA and a tSLP produced according any of claims 1-13 and one or more substances selected from carriers, 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, clarifying agents, non-biological surfactants, antioxidants, thickeners, softeners, sunscreens, moisturizers, colorants, and fragrances, antibiotics, anesthetics, keratolytic agents, desquamating agents, keratinocyte proliferation enhancers, collagenase inhibitors, elastase inhibitors, depigmentingagents, anti-inflammatory agents, steroids, anti-acne agents, and advanced glycation end-product (AGE) inhibitors, wherein the composition has a pH of 2-12.

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