Modified sophorolipids for improved bioavailability of amino acids and methods of use - Patents.com

Abstract

The present invention provides modified sophorolipids modified with amino acids, for example to improve the bioavailability of the amino acids, which can efficiently deliver the amino acids to target sites and improve the efficacy of the amino acids and / or the sophorolipids. TIFF2025511856000017.tif92128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 328,883, filed April 8, 2022, which is incorporated by reference in its entirety. [Background technology]

[0002] 2. Background of the Invention Sophorolipids are classified as glycolipid biosurfactants because they are produced by fermentation of microorganisms such as Starmerella bombicola using carbohydrates and lipids as carbon sources. Sophorolipids are amphipathic molecules containing a hydrophilic carbohydrate group (sophorose) and a hydrophobic fatty acid. Sophorose is a disaccharide consisting of two glucose molecules linked via a β-1,2 bond. The sophorose in sophorolipids can be acetylated, particularly at the 6' and / or 6'' positions. The fatty acids in sophorolipids are β-glycosidically linked to the sophorose molecule through their hydroxyl groups, and their terminal carboxylic acid groups are either free (as shown in Figure 1) or internally esterified (i.e., lactone form), typically at the 4'' position. The fatty acids in sophorolipids generally have 16 or 18 carbon atoms and one unsaturated bond. However, they may consist of chain lengths from C12 to C22 with various degrees and positions of unsaturation. Thus, sophorolipids are often produced as a mixture of related molecules. The differences between related molecules arise primarily from their fatty acid structure (degree of unsaturation, chain length, position of unsaturation, and position of hydroxylation); whether they are produced in free or lactonic form, acetylation pattern; the presence of stereoisomers; and / or whether the β-glycosidic bond on the fatty acid is at the ω position (e.g., terminal) or the ω-1 position (subterminal).

[0003] Sophorolipids have many advantageous characteristics that make them superior to synthetic surfactants, such as biodegradability, low toxicity, high surface and interfacial activity, and stability over a wide range of temperatures, pressures and ionic strengths.

[0004] In recent years, chemical modifications of sophorolipids have been attempted with the aim of improving their properties. One example is poly(sophorolipid), a polymer of sophorolipids that has potential applications in biomaterials. Summary of the Invention

[0005] One aspect of the invention provides sophorolipids or "truncated" sophorolipids modified with amino acids. In some embodiments, the modified sophorolipids improve the bioavailability of the attached amino acid compared to the free amino acid. In other embodiments, the modified sophorolipids improve the efficacy of the sophorolipid and / or the amino acid.

[0006] According to a first set of embodiments, the present invention provides a modified sophorolipid of formula (I): TIFF2025511856000002.tif45128, or a salt thereof, R1 is an amino acid bound to the adjacent carbonyl moiety of the sophorolipid structure via an amide bond; Each R2 is independently H or -COCH3; R3 is a saturated or unsaturated aliphatic hydrocarbon chain having 5 to 15 carbons; R4 is H or -CH3.

[0007] According to a second set of aspects, the present invention provides a modified sophorolipid of formula (II): TIFF2025511856000003.tif43128, or a salt thereof, R1 is an amino acid bound to the adjacent carbonyl moiety of the sophorolipid structure via an amide bond; Each R2 is independently H or -COCH3; R4 is H or -CH3.

[0008] The present invention encompasses all of the compounds represented by general formulas (I) and (II), including their salts, their hydrates, their geometric and optical isomers, and their polymorphic forms.

[0009] Another aspect of the present invention is directed to a method for improving the bioavailability of an amino acid. The modified sophorolipid of the present invention can efficiently deliver and improve the efficacy of the bound amino acid by improving the bioavailability of the amino acid. In another embodiment, the modified sophorolipid also improves the efficacy of the sophorolipid and the amino acid.

[0010] Additionally, certain embodiments of the present invention provide methods of treating or preventing various diseases, disorders, and / or conditions by administering modified sophorolipids.

[0011] A further aspect of the present invention is directed to a method of improving the nutrition in a subject's diet by administering a modified sophorolipid that incorporates an amino acid that is or may be missing or insufficient in the subject's diet. [Brief description of the drawings]

[0012] [Figure 1] 1 shows an exemplary general structure of sophorolipids in their free acidic form, where each R is independently H or acetyl (-COCH3); Rc represents a carbon chain, typically C10-C20 in length, with varying degrees and positions of unsaturation. [Diagram 2] 1 shows exemplary structures of α-amino acids. [Diagram 3] 1 shows exemplary structures of β-amino acids. [Figure 4] 1 shows exemplary structures of γ-amino acids. [Diagram 5] 1 shows exemplary structures of δ-amino acids. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Detailed Description of the Invention modified sophorolipids In one aspect, the present invention provides modified sophorolipids of general formula (I), (II), or salts thereof, in which a sophorolipid or truncated sophorolipid is linked to an amino acid via an amide bond. By "truncated" it is meant that the carbon chain of the fatty acid is shorter than that normally present in the sophorolipid.

[0014] According to a first set of embodiments, the present invention provides a modified sophorolipid of formula (I): TIFF2025511856000004.tif43128, or a salt thereof, R1 is an amino acid bound to the adjacent carbonyl moiety of the sophorolipid structure via an amide bond; Each R2 is independently H or -COCH3; R3 is a saturated or unsaturated aliphatic hydrocarbon chain having 5 to 15 carbons; R4 is H or -CH3.

[0015] In some preferred embodiments, the modified sophorolipid of formula (I) or a salt thereof has R3 that is an eight carbon aliphatic hydrocarbon chain with one unsaturation in the form of a double bond, R4 that is methyl (-CH3), and R2 that are all H.

[0016] In a further embodiment, the modified sophorolipid of formula (I) or a salt thereof has the following structure: TIFF2025511856000005.tif39128, where R1 is an amino acid attached to the carbonyl moiety of the sophorolipid structure via an amide bond.

[0017] According to a second set of aspects, the present invention provides a modified sophorolipid of formula (II): TIFF2025511856000006.tif43128, or a salt thereof, R1 is an amino acid bound to the adjacent carbonyl moiety of the sophorolipid structure via an amide bond; Each R2 is independently H or -COCH3; R4 is H or -CH3.

[0018] In some preferred embodiments, the modified sophorolipid of formula (II) or a salt thereof has R4 that is methyl (-CH3) and R2 that are all H.

[0019] In a further embodiment, the modified sophorolipid of formula (II) or a salt thereof has the following structure: TIFF2025511856000007.tif41128, where R1 is an amino acid attached to the carbonyl moiety of the sophorolipid structure via an amide bond.

[0020] The present invention encompasses all compounds represented by general formula (I) and (II), including their salts, their hydrates, their geometric and optical isomers, and their polymorphic forms.Furthermore, those skilled in the art will recognize that the modified sophorolipids of formula (I) and (II) may exist in lactone form when the amino acid of the R1 group has a free acid, such as a carboxylic acid.That is, the free carboxylic acid may be internally esterified with one of the hydroxyl substituents of the sophorose group.

[0021] In some embodiments, the modified sophorolipid improves the bioavailability of the conjugated amino acid compared to the free amino acid, hi other embodiments, the modified sophorolipid improves the potency of the sophorolipid and / or the amino acid.

[0022] Sophorolipids Sophorolipids are generally obtained from microbial fermentation, using a fermentation process involving wild-type or genetically engineered yeast strains, using pure fatty acids, mixtures of fatty acids, pure fatty acid esters, mixtures of fatty acid esters, triglycerides as carbon sources along with carbohydrate sources such as corn syrup, dextrin and glucose. As shown in Figure 1, sophorolipids generally consist of sophorose, which may or may not be acetylated, particularly at the 6' and 6'' positions, and hydroxylated fatty acid tails with various degrees of unsaturation. Most commonly, the fatty acids consist of 16 or 18 carbon atoms and one unsaturated bond, although the fatty acid structure (e.g., degree of unsaturation, chain length) varies depending on the carbon source provided during production and the microorganism undergoing fermentation.

[0023] The production of sophorolipids using renewable substrates and different microbial species, as well as various cultivation parameters (cultivation time, stirring speed, pH value of the medium and added nutrients), allows obtaining compounds with distinct structural and physical properties, which allows the generation of a wide range of compounds that can exhibit different physical, chemical, biochemical and biophysical properties.

[0024] The hydrophobic fatty acid tails of sophorolipids are usually terminally hydroxylated and β-glycosidically linked to the sophorose molecule. The fatty acid carboxylic acid groups in sophorolipids are either free (acidic form as shown in Figure 1) or intramolecularly esterified (lactone form), typically at the 4'' position. The degree of sophorolipid lactonization and sophorose acetylation can be influenced by the carbon source provided during sophorolipid production. For example, sophorolipids derived from rapeseed, sunflower and palm oils, which are rich in C18:0 and C18:1 fatty acids, have been found to form with higher diacetylated lactone concentrations than sophorolipids produced from the corresponding fatty acid ester feedstocks.

[0025] Sophorolipids have environmental compatibility, high biodegradability, low toxicity, high selectivity, and specific activity in a wide range of temperature, pH and salinity conditions. Suggested benefits of sophorolipids include, but are not limited to, killing pathogens in the skin, regulating the skin's immune system, killing melanocytes to allow replacement cell growth, reducing oxidative stress, improving keratinocyte and fibroblast proliferation and function, for example, improving skin penetration of both the biosurfactant and one or more other active ingredients in the composition. See, for example, U.S. Patent Application Publication No. 2020 / 0155444.

[0026] fermentation In some embodiments, the sophorolipids modified with the amino acids of the present invention are produced by fermentation processes known in the art in the presence of selected fatty acids and carbohydrate feedstocks. In other embodiments, the modified sophorolipids are first produced by fermentation processes known in the art and then chemically modified using known synthetic techniques to attach the desired amino acids. As used herein, "fermentation" refers to the growth of cells under controlled conditions. Growth can be aerobic or anaerobic. Fermentation processes known in the art include, but are not limited to, solid-state fermentation, liquid-state fermentation, or modifications, hybrids and / or combinations thereof.

[0027] The modified sophorolipids of formula (I) or (II), or modified according to the present invention, may be derived via a fermentation process from a recombinant organism or by a strain that naturally produces sophorolipids. Non-limiting examples of sophorolipid producing organisms include Candida bombicola, Candida apicola, Candida bogoriensis, Yarrowia lipolytica, Starmerella bombicola, Starmerella clade, Rhodotorula bogoriensis, Wickerhamiella domericqiae, and Wickerhamomyces anomalus. Several recombinant sophorolipid-producing microorganisms have been reported that allow control of sophorolipid structure. As a non-limiting example, certain recombinant S. bombicola strains may be utilized to produce sophorolipids that are either fully lactonic or fully acidic (see Roelants et. al., Towards the Industrialization of New Biosurfactants: Biotechnological Opportunities for the Lactone Esterase Gene from Starmerella Bombicola, 113 Biotechnology and Bioengineering 3, 550-559 (2015)). As an additional example, recombinant Candida bombicola strains with an acetyltransferase gene knocked out may be used to produce sophorolipids without acetylation (see, for example, WO2012 / 080116).

[0028] The subject microorganisms may be cultivated on-site on a small or large scale and may be utilized while still mixed with their medium. The modified sophorolipids of the present invention may be produced in large quantities wherever needed. The growth vessel used to grow the sophorolipid producing organisms may be any fermenter or culture reactor for industrial use.

[0029] In one embodiment, a single type of microorganism is grown in the reactor system. In alternative embodiments, multiple microorganisms may be grown in a single reactor system that can be grown together without adverse effects on the growth or the resulting product. For example, two or more different types of microorganisms may be grown simultaneously in a single reactor. In some embodiments, more than one type of microorganism is grown symbiotically in the reactor.

[0030] In one embodiment, the vessel may have or be connected to functional controllers / sensors for measuring key factors in the culture process, such as pH, oxygen, pressure, temperature, agitator shaft power, humidity, viscosity and / or microbial density and / or metabolite concentration.

[0031] The culture may be supplemented with a carbon source. The carbon source is typically a carbohydrate such as glucose, sucrose, lactose, fructose, trehalose, mannose, mannitol, and / or maltose; an organic acid such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid, malonic acid, and / or pyruvic acid; an alcohol such as ethanol, isopropyl, propanol, butanol, pentanol, hexanol, isobutanol, and / or glycerol; an oil such as soybean oil, rice bran oil, canola oil, olive oil, corn oil, sesame oil, and / or linseed oil, and the like. These carbon sources may be used alone or in combination of two or more.

[0032] In some embodiments, the carbon source utilized in the fermentation process has already been chemically modified to bear a desired functional group. For example, the fatty acid may already be conjugated to a selected amino acid prior to being introduced into the reaction vessel containing the sophorolipid producing microorganism. In other embodiments, the fatty acid utilized in the fermentation process has not yet been conjugated to an amino acid.

[0033] The culture may be supplemented with a nitrogen source. The nitrogen source may be, for example, potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium phosphate, ammonia, urea, and / or ammonium chloride. These nitrogen sources may be used independently or in combination of two or more.

[0034] The growth medium in the culture may also be supplied with oxygen. One embodiment is to slowly move air to remove oxygen-poor air and introduce oxygen-laden air. In the case of liquid-phase fermentation, the oxygen-laden air may be ambient air that is replenished daily through a mechanism that includes an impeller for mechanically stirring the liquid and an air sparger for providing gas bubbles to the liquid to dissolve oxygen in the liquid.

[0035] In one embodiment, the microorganisms may be grown on a solid or semi-solid substrate, such as, for example, corn, wheat, soybeans, chickpeas, beans, oatmeal, pasta, rice, and / or flours or meals of these or other similar substances.

[0036] In one embodiment, the medium contains growth factors and micronutrients for the microorganism. This is particularly preferred when culturing a microorganism that is not capable of producing all the vitamins required. Inorganic nutrients, including trace elements such as iron, zinc, copper, manganese, molybdenum and / or cobalt, may also be included in the medium. In addition, sources of vitamins, essential amino acids, and microelements may be included, for example in the form of powders or meals, 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, may also be included. Such amino acids include amino acids that are not intended to be part of the modified sophorolipid of formula (I) or (II).

[0037] In one embodiment, inorganic salts may be included. Usable inorganic salts may be potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, magnesium chloride, ferrous sulfate, ferrous chloride, manganese sulfate, manganese chloride, zinc sulfate, lead chloride, copper sulfate, calcium chloride, calcium carbonate, sodium chloride, and / or sodium carbonate. These inorganic salts may be used alone or in combination of two or more.

[0038] In some embodiments, for example, when the microorganisms used to inoculate the substrate are in the form of spores (e.g., bacterial spores), a germination promoter may be added to the substrate. Examples of germination promoters according to the present invention include, but are not limited to, L-alanine, manganese, L-valine, and L-asparagine, or any other known germination promoter.

[0039] In some embodiments, additional acid and / or antibacterial agents may be added to the liquid medium before and / or during the culture process. Antibacterial agents or antibiotics are used to protect the culture from contamination. In addition, if gas is generated during culture, antifoam agents may also be added to prevent foam formation and / or accumulation.

[0040] The pH of the mixture should be suitable for the target microorganism. Buffers and pH adjusters such as carbonates and phosphates may be used to stabilize the pH near the preferred value. If metal ions are present in high concentrations, it may be necessary to use chelating agents in the liquid medium.

[0041] The methods and apparatus for culturing microorganisms and producing microbial by-products may be carried out in batch, semi-continuous, or continuous processes.

[0042] In one embodiment, the microorganism is cultured at about 5° C. to about 100° C., preferably about 15° C. to about 60° C., and more preferably about 25° C. to about 50° C. In a further embodiment, the culture may be continuously performed at a constant temperature. In another embodiment, the culture may be subjected to temperature changes.

[0043] In one embodiment, the equipment used in the method and culture process is sterilized. The culture equipment, such as the reactor / container, is separate from but may be connected to a sterilization unit, e.g., an autoclave. The culture equipment may also have a sterilization unit that sterilizes in situ before inoculation begins. Air may be sterilized by methods known in the art. For example, ambient air may pass through at least one filter before being introduced into the container. In another embodiment, the medium may be pasteurized or may not be heated at all, where unwanted bacterial growth may be controlled by using low water activity and low pH.

[0044] In one embodiment, all of the microbial culture composition is removed upon completion of the culture (e.g., upon achieving a desired cell density or density of a specified metabolite).In this batch procedure, an entirely new batch is started upon harvest of the first batch.

[0045] 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 microbial-free medium or may contain cells, spores, hyphae, conidia, or other microbial propagules. In this way, a semi-continuous system is created.

[0046] In some embodiments, the modified sophorolipid or modified sophorolipid of formula (I) or (II) is produced by the target microorganisms and is retained within the microorganisms or secreted into their growth medium. The modified sophorolipid or modified sophorolipid can be, for example, at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% w / v of the total composition removed.

[0047] The growth medium may contain compounds that stabilize the activity of the sophorolipids. The modified sophorolipids or the sophorolipids to be modified may be purified, or such sophorolipids may be used in crude form, meaning that they have not been separated from the fermentation broth in which they were produced.

[0048] In certain embodiments, the modified sophorolipid or the sophorolipid to be modified is isolated and / or purified from the growth medium resulting from the fermentation of the biosurfactant-producing microorganism. Isolation and purification can be readily accomplished using known methods or techniques described in the literature. If desired, the sophorolipid can be further concentrated.

[0049] As used herein, the terms "isolated" or "purified" when used in reference to a biological or natural substance, such as a glycolipid, mean that the substance is substantially free of other compounds with which it is naturally associated, such as cellular material, i.e., the substance does not exist in nature without these other compounds and / or has different or distinctive properties compared to those found in the native material.

[0050] 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 any appropriate standard method, for example, by column chromatography, thin layer chromatography, or high performance liquid chromatography (HPLC) analysis.

[0051] Chemical modification method In some embodiments, the modified sophorolipids of the present invention are obtained by chemically modifying sophorolipids produced by sophorolipid-producing microorganisms. The chemically modified sophorolipids may be obtained by the fermentation methods described above and / or any other methods known in the art, and then isolated or purified using known methods or techniques. Any suitable techniques and chemical reactions known in the art may be used to modify the obtained sophorolipids. For example, amino acids may be attached to the sophorolipid structure via various reactions, including but not limited to amidation, reductive amination, and esterification. In a preferred embodiment, amino acids may be attached to the sophorolipid structure via amidation, including but not limited to amide coupling reactions.

[0052] Chemical modifications may also be carried out, for example to alter the degree of unsaturation of the fatty acid chains. Commonly known hydrogenation or dehydrogenation reactions, or addition or elimination reactions, with or without a catalyst, may be used.

[0053] amino acid The modified sophorolipids of the present invention incorporate within their structure an amino acid that is linked to the fatty acid chain of the sophorolipid structure via an amide bond. In a preferred embodiment, the amino acid is released from the sophorolipid structure upon digestion of the sophorolipid structure to impart its therapeutic, pharmaceutical, nutritional, or other benefits. By covalently linking an amino acid to a sophorolipid or truncated sophorolipid via an amide bond (i.e., formulas (I) and (II), respectively), the amino acid can be delivered, released, and absorbed by the subject's body more efficiently than if the amino acid alone were administered to the subject.

[0054] As used herein, an amino acid has at least one amino group and at least one acid, e.g., carboxyl group, in its backbone, with different side chains for each amino acid. The amino group in the backbone can form an amide bond with the carbonyl moiety of the sophorolipid structure of formulas (I) and (II). The side chain can be a variety of substituents, including but not limited to hydrogen (e.g., in the case of glycine), organic, semi-organic or inorganic, and rings cyclized on the amine (e.g., in the case of proline). If a suitable amino group is present in the side chain, it can also form an amide bond with the carbonyl moiety of the sophorolipid structure of formulas (I) and (II). The amino group and acid (e.g., carboxyl group) of the backbone can be attached to the same carbon (α-carbon) or different carbons.

[0055] Furthermore, it is within the spirit and scope of the present invention that amino acids contain acidic groups other than carboxyl groups in their backbones.For example, sulfonic acid or thiocarboxylic acid (i.e., one of the oxygen atoms is replaced by a sulfur atom) may be present in the amino acid structure instead of carboxyl group.A non-limiting example of such an amino acid is taurine (also called 2-aminoethanesulfonic acid), which contains sulfonic acid and an amino group attached to the β-carbon.

[0056] The amino acid used herein may be an L-amino acid or a D-amino acid. It may be a natural amino acid (such as an essential amino acid, a non-essential amino acid, or a conditionally essential amino acid) or a synthetic / modified amino acid that does not occur naturally. The present invention encompasses both the amino acid that is incorporated into proteins and the amino acid that is not incorporated into proteins. The naturally occurring amino acid may be a purified product or may be artificially synthesized.

[0057] In aqueous solution, amino acids, particularly the common naturally occurring forms of amino acids, exist as zwitterions, where both the amino group and the acid (e.g., carboxyl group) are zwitterions, e.g., -NH3 + or -NH2 + , - and -CO2 - Depending on the pH of the environment, certain side chains may also exist in a charged state.

[0058] In some embodiments, the amino acids of the present invention are alpha (α) amino acids. Exemplary structures are shown in Figure 2, which contain an amino group and a carboxyl group attached to the same carbon adjacent to the carboxyl group, designated as the α-carbon. Also attached to the α-carbon are substituents X and Y, which may be the same or different, at least one of which is typically H. The amino group may be a substituted, primary amine, where Q is H or where Q is a substituent. Non-limiting examples of α-amino acids include glycine, methionine, lysine, and glutamine.

[0059] In another embodiment, the amino acid of the present invention is a beta (β)-amino acid. Exemplary structures containing an amino group attached to the β-carbon (the second carbon from the carboxyl group) are shown in Figure 3. X, Y, W, and Z are optional substituents that may be the same or different, at least one of which is typically H. The amino group may be a substituted, primary amine where Q is H or where Q is a substituent. Non-limiting examples of β-amino acids include β-alanine and taurine.

[0060] In yet another embodiment, the amino acid of the present invention is a gamma (γ)-amino acid. Exemplary structures containing an amino group attached to the γ-carbon (the third carbon from the carboxyl group) are shown in Figure 4. X, Y, W, Z, A, and G are optional substituents that may be the same or different, at least one of which is typically H. The amino group may be a substituted, primary amine where Q is H or where Q is a substituent. Non-limiting examples of γ-amino acids include vigabatrin (4-amino-5-hexenoic acid) and gamma-aminobutyric acid (GABA).

[0061] In further embodiments, the amino acids of the present invention are delta (δ)-amino acids. Exemplary structures containing an amino group attached to the δ-carbon (the fourth carbon from the carboxyl group) are shown in Figure 5. X, Y, W, Z, A, G, J, and L are optional substituents that may be the same or different, at least one of which is typically H. The amino group may be a substituted, primary amine where Q is H or where Q is a substituent. Non-limiting examples of δ-amino acids include 5-aminopentanoic acid and 5-amino-2-oxopentanoic acid.

[0062] Those skilled in the art will appreciate that the general structure of an amino acid may be further modified without departing from the spirit and scope of the invention, for example, the backbone carbon chain between the amino group and the acid may be longer than that of the δ-amino acids described above, or the acid may be esterified.

[0063] In some embodiments, the amino acid incorporated into the modified sophorolipids of the present invention is methionine (Met), lysine (Lys), or glutamine (Gln). In specific embodiments, these modified sophorolipids function to reduce factors that contribute to stress in a subject, such as reactive oxygen species (ROS) and cortisol.

[0064] Methionine (Met) is an essential amino acid and an important precursor of S-adenosylmethionine, the cellular methyl donor. Methionine residues in proteins readily react with a variety of reactive oxygen species (ROS) to scavenge the reactive species. After reaction, methionine forms methionine sulfoxide, which can be reduced back to methionine within the cell. Thus, methionine residues may function as catalytic antioxidants that protect both the proteins in which they reside and other macromolecules, including the cells themselves. Methionine is further believed to provide health benefits, including aiding in the absorption of nutrients such as selenium and zinc, promoting the excretion of heavy metals, preventing excess fat accumulation in the liver, and lowering cholesterol levels by increasing lecithin production in the liver. Methionine has also been shown to be effective in treating Tylenol (acetaminophen) poisoning.

[0065] Lysine (Lys) is an essential amino acid that has been shown to act as a partial antagonist of the serotonin receptor 4 (5-HT4) to reduce blood levels of cortisol (the so-called "stress hormone") and stress-induced brain-gut responses. In laboratory animals, prolonged insufficient dietary lysine increases stress-induced anxiety. Lysine supplementation has been shown to reduce anxiety in humans with low dietary L-lysine intake. In one study conducted in otherwise healthy adults, oral treatment with a combination of L-lysine and L-arginine reduced both predisposing and stress-induced anxiety states. Furthermore, the combination reduced basal levels of salivary cortisol and chromogranin A (a salivary marker of the sympathetic adrenal system) in male subjects. See, e.g., Smigra, M. et al., Oral Treatment with L-lysine and L-arginine Reduces Anxiety and Basal Cortisol Levels in Healthy Humans, Biomed Res. 28(2), 85-90 (2007). Thus, lysine supplementation has been suggested to be an effective reducer of anxiety and stress. Lysine supplementation has also been linked to a variety of other health benefits, including suppression of cold sores, calcium absorption and retention, and wound healing.

[0066] Glutamine (Gln) is a conditionally essential amino acid. This means that glutamine can be synthesized by the body under normal physiological conditions, but under pathological conditions, there is a relative lack of glutamine due to increased demand from the body. Glutamine has anti-inflammatory and antioxidant properties, which makes it a useful supplement for athletes and other subjects who perform intense physical activity in high-stress environments. Insufficient glutamine levels lead to disrupted energy metabolism, immunosuppression, and increased oxidative stress.

[0067] In another embodiment, the amino acid incorporated into the modified sophorolipid of the invention is taurine. Taurine is a conditionally essential amino acid known to have antioxidant properties and performs various physiological functions in the body. Non-exhaustive examples of the functions of taurine are listed below. Modified sophorolipids incorporating taurine can be used to treat various diseases and / or maintain general well-being.

[0068] First, various studies suggest that taurine supplementation may delay the initiation and progression of atherosclerosis and / or exert preventive effects against other cardiovascular disorders and diseases. For example, a reduction in cholesterol levels and inflammation has been observed in patients with heart failure. Ahmadian, M., Taurine Supplementation Has Anti-atherogenic and Anti-inflammatory Effects Before and After Incremental Exercise in Heart Failure, Ther. Adv. Cardiovasc. Dis. 11(7): 185-194 (2017). Furthermore, taurine supplementation provided significant improvements in antihypertensive effects, as well as endothelium-dependent and endothelium-independent vasodilation in prehypertension. Sun, Q. et al., Taurine Supplementation Lowers Blood Pressure and Improves Vascular Function in Prehypertension, Hypertension 67, 541-549 (2016). Taurine's ability to lower blood pressure, combined with its anti-inflammatory effects, makes it an effective preventative agent against coronary heart disease.

[0069] Taurine is also present in high concentrations in mammalian skeletal muscle. Taurine is thought to regulate muscle metabolism and gene expression. Alterations in intracellular taurine concentrations in skeletal muscle have been linked to various pathophysiological conditions, such as disuse muscle atrophy, muscular dystrophies, and / or aging. Taurine supplementation has therapeutic potential to restore skeletal muscle function and performance. Furthermore, taurine treatment may be beneficial in reducing muscle fiber membrane hyperexcitability in myotonia-related syndromes. De Luca, A., Taurine: the appeal of a safe amino acid for skeletal muscle disorders, J. Transl. Med. 13, 243 (2015). Taurine supplementation may also improve exercise performance, e.g., time performance in long-distance running.

[0070] Similarly, taurine has been shown to induce the expression of connective tissue growth factor (CTGF) in osteoblasts, suggesting a role for taurine in connective tissue strength.

[0071] Further benefits of taurine supplementation are related to the central and peripheral nervous systems. Taurine is necessary for the development and function of the central and peripheral nervous systems, where it exerts osmoregulatory, neuromodulatory and anti-apoptotic effects. Deficiency of taurine can lead to some neurological deficits and sensory loss. Animal studies have shown that taurine helps to alleviate symptoms of neurotoxicity and neurological disorders in rodents. It has also been suggested that taurine may improve or protect brain function, especially learning and memory.

[0072] Taurine is also sometimes used as a treatment for eye diseases such as glaucoma and diabetic retinopathy, and for diabetes to reduce blood sugar levels and insulin resistance.

[0073] Other non-limiting examples of amino acids that may be used in the present invention include alanine, β-alanine, arginine, asparagine, aspartic acid, cysteine, homocysteine, glutamic acid, glycine, histidine, homotaurine, isoleucine, leucine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, theanine, D-2-amino-3-guanidinopropionic acid, GABA, citrulline, tranexamic acid, aminocaproic acid, 4-amino-5-hexenoic acid, 4-oxaproline, 4-thioproline, 2-azaproline, 4-hydroxyproline, 1,5 -disubstituted tetrazoles, 2-aminoisobutyric acid, sarcosine, 1-aminocyclopentane-1-carboxylic acid, beta alanine, 2-amino-cyclopentanecarboxylic acid (beta-proline), 5-hydroxylysine, hydroxylysine-5-sulfate, hydroxylysine-5-nitrate, hydroxylysine-5-phosphate, serine-3-sulfate, threonine-3-sulfate, serine-3-nitrate, threonine-3-nitrate, serine-3-phosphate, threonine-3-phosphate, 2-hydroxyalkanoic acids, 5-aminopentanoic acid, and 5-amino-2-oxopentanoic acid.

[0074] Administration of modified sophorolipids according to the present invention provides a wide range of benefits depending on the amino acids incorporated therein. Various amino acids regulate glucose and lipid metabolism and energy balance, increase mitochondrial biogenesis, and maintain immune homeostasis. Dietary amino acids are also the main fuel for the small intestinal mucosa and are important substrates for the synthesis of intestinal proteins, nitric oxide, polyamines, and other biologically important products. Research supports that amino acids, including glutamine, glutamate, arginine, glycine, lysine, threonine, and / or sulfur-containing amino acids, are useful in treating gastrointestinal-related diseases. For example, glutamine intake has been shown to improve gastrointestinal health by supporting gastrointestinal microflora, gastrointestinal mucosal wall integrity, and by modulating inflammatory responses.

[0075] Pharmaceutically or cosmetically acceptable salts In some embodiments, the modified sophorolipids of the present invention are provided in the form of pharma- ceutically or cosmetically acceptable salts. In a specific embodiment, the acid group of the amino acid incorporated in the modified sophorolipid may be deprotonated to form a salt. The side chain of the amino acid may also form the corresponding salt if it contains a suitable functional group. The salt may be formed by procedures well known and described in the art.

[0076] Examples of pharma- ceutically acceptable cations for salts include, but are not limited to, aluminum, arginine, benzathine, calcium, chloroprocaine, choline, diethanolamine, ethanolamine, ethylenediamine, lysine, magnesium, histidine, lithium, meglumine, potassium, procaine, sodium, triethanolamine, and zinc. Preferred salts include the lithium (Li + ) salt, sodium (Na + ) salts, and potassium (K + ) salt.

[0077] Depending on the chemical characteristics of the amino acids in the modified sophorolipid, acid addition salts may be formed. Examples of pharma- ceutically acceptable addition salts include acetate from acetic acid, aconitate from aconitic acid, ascorbate from ascorbic acid, benzenesulfonate from benzenesulfonic acid, benzoate from benzoic acid, cinnamate from cinnamic acid, citrate from citric acid, embonate from embonic acid, enanthate from enanthic acid, formate from formic acid, fumarate from fumaric acid, glutamate from glutamic acid, glycolate from glycolic acid, hydrochloride from hydrochloric acid, hydrobromide from hydrobromic acid, lactate from lactic acid, maleate from maleic acid, maloic acid, and malonic acid. non-toxic inorganic and organic acid addition salts include, but are not limited to, malonate from carboxylic acid, mandelate from mandelic acid, methanesulfonate from methanesulfonic acid, naphthalene-2-sulfonate from naphthalene-2-sulfonic acid, nitrate from nitric acid, perchlorate from perchloric acid, phosphate from phosphoric acid, phthalate from phthalic acid, salicylate from salicylic acid, sorbate from sorbic acid, stearate from stearic acid, succinate from succinic acid, sulfate from sulfuric acid, tartrate from tartaric acid, and toluene-p-sulfonate from p-toluenesulfonic acid.

[0078] Other salts, which may not be considered pharma- ceutically acceptable, may be useful in the preparation of salts useful as intermediates in obtaining the modified sophorolipids of the present invention and their pharma- ceutically acceptable salts.

[0079] In another embodiment, the modified sophorolipids of the present invention are used in accordance with the present invention in their individual acid forms.

[0080] The modified sophorolipids of the present invention may be provided in unsolvated or solvated forms with a pharma- ceutically acceptable solvent such as water, ethanol, etc. Solvated forms may also include hydrated forms such as a monohydrate, dihydrate, hemihydrate, trihydrate, tetrahydrate, etc.

[0081] composition Another aspect of the invention is directed to a composition comprising the modified sophorolipids of the invention administered to a subject. Such compositions may be cosmetics, pharmaceuticals, foods, dietary supplements, etc. In some embodiments, the modified sophorolipids of the invention function as an active ingredient in the composition.

[0082] The term "subject" or "patient" as used herein refers to organisms including mammals such as primates. Mammalian species that may benefit from the disclosed therapeutic methods include, but are not limited to, apes, chimpanzees, orangutans, humans, and monkeys; domesticated animals such as dogs and cats; farm animals such as horses, cows, pigs, sheep, goats, and chickens; and other animals such as mice, rats, guinea pigs, and hamsters.

[0083] The particular mode of administration or dosing regimen designed to deliver an effective amount of modified sophorolipid is appropriately determined taking into account factors such as the nature of the amino acid, the age, sex, and details of the subject, the associated condition, disease, and pathology, and whether the purpose is prophylactic. As used herein, the terms "effective amount" and "effective dose" are used to refer to an amount of something (e.g., a compound, composition, time) that can cause a desired outcome (e.g., reducing cortisol levels, lowering blood pressure). Administration of an effective amount of modified sophorolipid may be achieved by administering modified sophorolipid once a day or multiple doses a day for days to months or even years. When modified sophorolipid is administered with another therapeutic agent, the effective amount of modified sophorolipid may include the same range of amino acids as are typical for the use of that amino acid as a monotherapy, or the amount may be less than a typical monotherapy amount, especially when the combination therapy produces a synergistic effect.

[0084] The compositions of the invention may include isolated and / or purified forms of modified sophorolipids as active ingredients. In another embodiment, the modified sophorolipids in the compositions of the invention are provided together with the fermentation broth in which they were produced. Such a "crude form" may contain, for example, at least 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% w / v of the modified sophorolipids in the broth.

[0085] The composition may further comprise additional active or inactive ingredients appropriate to the mode of administration and intended purpose, provided that such additions do not adversely affect the function of the modified sophorolipid.

[0086] In yet another embodiment, acceptable carriers may be included in the compositions of the present invention depending on the form of the composition and / or the mode of administration. Pharmaceutically or cosmetically acceptable carriers include, but are not limited to, pharma-ceutically acceptable excipients such as inert diluents, disintegrants, binders, lubricants, sweeteners, flavorings, colorants and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while cornstarch and alginic acid are suitable disintegrants. Binders may include starch and gelatin, while lubricants, if present, will generally be magnesium stearate, stearic acid or talc. If desired or suitable, coating materials such as glyceryl monostearate or glyceryl distearate may also be used if appropriate and the pharmaceutical composition is in solid form, for example, to delay absorption in the digestive tract.

[0087] In certain embodiments, the compositions of the present invention further comprise by-products of microbial growth. In certain embodiments, the by-products of microbial growth are amphipathic molecules, enzymes and / or proteins. In one embodiment, the by-products of microbial growth have antibacterial and / or antibiofilm properties.

[0088] In certain embodiments, the composition further comprises one or more amphiphilic molecules, where 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, atrophactin and lichenicin), cellobiose lipids, flavolipids, phospholipids (e.g., cardiolipin), and high molecular weight polymers such as lipoproteins, lipopolysaccharide-protein complexes, and polysaccharide-protein-fatty acid complexes. The amphiphilic molecules may be 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 the critical micelle concentration (CMC).

[0089] The composition may be housed in a container of appropriate size, taking into account, for example, the intended use, the intended application method, the size of the fermentation vessel, and any mode of transportation from the microbial growth facility to the site of use. Thus, the container in which the composition is housed may be, for example, 0.1 gallons to 1,000 gallons or more. The composition may further be housed in smaller containers, such as bottles, for dispensing individual doses of the composition.

[0090] The compositions of the present invention may be in various physical forms. In some embodiments, the compositions are in solid form, including tablets, filled capsules, powders and pellet forms. In another embodiment, the pharmaceutical composition may be in powder form, where the pharma- ceutically acceptable carrier is a finely divided solid that is in admixture with the finely divided active ingredient, i.e., the modified sophorolipid. In a further embodiment, the pharmaceutical composition of the present invention is a sustained release system, such as a semipermeable matrix of solid hydrophobic polymers containing the modified sophorolipid of the present invention. In another embodiment, the pharmaceutical composition is in liquid form, such as an aqueous or non-aqueous solution, a suspension, an emulsion, an elixir, and capsules filled therewith.

[0091] Any suitable mode of administration may be utilized to administer compositions comprising modified sophorolipids according to the present invention. Such modes include, but are not limited to, oral, rectal, bronchial, nasal, topical, buccal, sublingual, transdermal, vaginal, intramuscular, intraperitoneal, intravenous, intraarterial, subcutaneous, intracerebral, intraocular administration, or forms suitable for administration by inhalation or insufflation, including powder and liquid aerosol administration, or parenteral injection. Administration may also be via other carriers or vehicles, such as patches, micelles, liposomes, vesicles, implants (e.g., microimplants), synthetic polymers, etc. Factors determining the mode of administration of a particular composition include the nature of the amino acid to be delivered to the subject. In some embodiments, the mode of administration is oral. In other embodiments, the mode of administration is rectal.

[0092] An orally administered composition according to the invention is any preparation or composition suitable for consumption, nutrition, oral hygiene and / or recreational purposes, intended to be introduced into the oral cavity of a human or animal, to remain there for a period of time and then to be swallowed (e.g. ready-to-eat foods) or removed again from the oral cavity (e.g. oral hygiene chewing gums or products, or medicinal mouthwashes). These products include any substance or product intended to be ingested by a human or animal in a processed, semi-processed or raw state. This also includes substances added to orally ingestible products (especially food and pharmaceutical products) during their manufacture, treatment or processing, which are intended to be introduced into the oral cavity of a human or animal.

[0093] In certain embodiments, the composition may further comprise ingredients such as energy sources, nutrients and / or other health-enhancing 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 drinkable liquid by adding water.In one embodiment, the composition is formulated as a blended smoothie or milkshake.

[0094] Orally administered compositions may also include substances that are intended to be swallowed by a human or animal and then digested, either unmodified, prepared, or processed; thus, orally administered compositions according to the present invention also include envelopes, coatings, or other encapsulating materials that are intended or anticipated to be swallowed along with the product.

[0095] In another embodiment, the compositions of the present invention are formulated for direct administration into the gastrointestinal tract. For example, the compositions may be formulated for administration to the proximal lower gastrointestinal tract by manual or colonoscopy, to the distal lower gastrointestinal tract via an enema or rectal tube, and to the upper gastrointestinal tract via a nasogastric tube, a duodenal tube, and endoscopy / gastroscopy.

[0096] In certain embodiments, the present invention provides topical skin care compositions comprising one or more modified sophorolipids according to the present invention. In some embodiments, the topical compositions may further comprise a dermatologically acceptable carrier and one or more active or inactive cosmetic ingredients, such as, for example, vitamins, moisturizers, dyes, fragrances, sunscreens, exfoliants, essential oils, botanical extracts, and the like.

[0097] Dermatologically acceptable carriers include, for example, water; saline; saline; ointments; creams; oil-in-water emulsions; water-in-oil emulsions; silicone-in-water emulsions; water-in-silicone emulsions; wax-in-water emulsions; water-in-oil 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; isooctane, isododecane and isohexadecane. which may include isoparaffins; silicone oils such as cyclomethicone, dimethicone, dimethicone crosspolymer, polysiloxanes and their derivatives, preferably organically modified derivatives including PDMS, dimethicone copolyol, dimethiconol, and amodimethiconol; 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 vegetable wax; or any combination or mixture of the foregoing. The aqueous carrier may include one or more solvents miscible with water, including lower alcohols such as ethanol, isopropanol, and the like. The carrier may comprise about 1% to about 99%, 10% to about 85%, 25% to 75%, or 50% to about 65% of the total composition by weight.

[0098] Non-biological surfactants may 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, amphopropionates), sulfosuccinates, alkyl polyglucosides, betaines (e.g., cocamidopropyl betaine), sultaines, sacrosinates, isethionates, taurates, ethoxylated sorbitan esters, alkanolamides, and amino acid surfactants.

[0099] Viscosity modifiers may also be added to the compositions, including, for example, cocamide DEA, oleamide DEA, sodium chloride, cellulose polymers, polyacrylates, ethoxylated esters, alcohols, glycols, xylene sulfonates, polysorbate 20, alkanolamides, and cellulose derivatives (e.g., hydroxypropyl methylcellulose and hydroxyethyl cellulose).

[0100] Polymers 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 may also be added.

[0101] In one embodiment, the composition may contain additional skin active agents, including, but not limited to, keratolytic agents, exfoliants, keratinocyte proliferation promoters, collagenase inhibitors, elastase inhibitors, depigmenting agents, anti-inflammatory agents, steroids, anti-acne agents, antioxidants (e.g., ascorbic acid), advanced glycation end products (AGE) inhibitors, exfoliants (e.g., glycolic acid, 3,6,9-trioxaundecanedioic acid, and the like), estrogen synthase stimulating compounds (e.g., caffeine and derivatives), compounds capable of inhibiting 5-alpha reductase activity (e.g., linoleic acid, finasteride, and mixtures thereof), barrier function promoters (e.g., ceramides, glycerides, cholesterol and its esters, alpha-hydroxy and omega-hydroxy fatty acids and their esters), retinoids, sunscreens, to name a few.

[0102] The composition may optionally include additives, adjuvants, or other components of topical compositions known to those of skill in the art, including, but not limited to, 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., 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 salts thereof such as disodium EDTA); vitamins (e.g., tocopherol and ascorbic acid); vitamin derivatives (e.g., ascorbyl monopalmitate, tocopheryl acetate, vitamin E palmitate); thickening agents (e.g., hydroxyalkylcellulose, carboxymethylcellulose, carbomer, and vegetable gums such as xanthan gum); gelling agents (e.g., ester-terminated polyesteramides); structuring agents; proteins (e.g., lactoferrin); immunomodulators (e.g., corticosteroids and nonsteroidal immunomodulators). Further examples include, but are not limited to, stabilizers; fragrances; film formers; insect repellents; skin cooling compounds; skin protectants; lubricants; preservatives; pearls; chromalite; mica; conditioners; antiallergic agents; pH adjusters; and antimicrobial agents.

[0103] The composition may be formulated as a suspension, emulsion, hydrogel, multiphase solution, vesicular dispersion, or any other known form of a topical skin composition.

[0104] In certain embodiments, a topical composition may be formulated so that it can be applied via, for example, a pen, tube, bottle, brush, stick, sponge, swab, towelette (wipe), sprayer, dropper, hand, or finger.

[0105] Topical compositions may be formulated into 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.

[0106] In one embodiment, the composition may be provided in the form of a facial mask. The mask may be a peel-off mask or a wash-off mask comprising the composition of the present invention.

[0107] In another embodiment, the compositions of the present invention may be provided in a wound dressing, bandage, or patch that may be applied, attached, or adhered to one or more layers of the skin or tissue of a subject. For example, the composition may be applied to a wound dressing, bandage, or patch, which may then be placed on the area of ​​skin to be treated.

[0108] Wound dressings, bandages, and patches can be made of any material that is, for example, dermatologically acceptable and suitable for placement on wounds or skin. In exemplary embodiments, wound dressings, bandages, and patches can be made of synthetic or non-synthetic woven or non-woven fabrics, or any combination thereof. Dressings can also include a support, such as a polymer foam, natural or artificial sponge, gel, or membrane, that absorbs or has the composition attached to it. By way of example, the support can be a film, a natural or synthetic polymer, or a rigid or malleable material (e.g., gauze). Wound dressings, bandages, and patches can be absorbent, for example, gauze can be moistened with the composition of the present invention before being applied to a wound, scar, or other site.

[0109] In one embodiment, a wound dressing, bandage, or patch is impregnated with a composition of the invention and optionally allowed to dry, allowing the impregnated material to be stored for later use and to avoid excessive moistening of the skin.

[0110] In another embodiment, the pharmaceutical composition may be formulated for parenteral administration (e.g., by injection, e.g., bolus injection or continuous infusion). Furthermore, the composition may be provided in unit dose form in ampoules, prefilled syringes, and small volume injections, or in multi-dose containers, with or without the addition of a preservative. The composition may be in the form of a suspension, solution, or emulsion in an oily or aqueous carrier. The composition may further contain formulating agents, such as suspending, stabilizing and / or dispersing agents. In a further embodiment, the active ingredient of the inventive composition may be in powder form, obtained by aseptic isolation of a sterile solid or by lyophilization from solution, for constitution with a suitable carrier, e.g., sterile pyrogen-free water, before use.

[0111] In yet another embodiment, the pharmaceutical composition may be prepared for direct administration to the nasal cavity by conventional means, for example, by dropper, pipette, or spray. The composition may be provided in single or multiple dose form. Administration to the respiratory tract may also be achieved by aerosol formulations in which the active ingredient is provided in a pressurized pack with a suitable propellant, such as chlorofluorocarbons (CFCs), for example, dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. The aerosol may conveniently also contain a surfactant, such as lecithin.

[0112] Methods for improving bioavailability, physiology, and nutrition One aspect of the present invention relates to a method for improving the bioavailability, preferably oral bioavailability, of an amino acid in a subject. The method comprises the steps of preparing a modified sophorolipid or a salt thereof of formula (I) and / or (II) as described herein, wherein R1 is an amino acid attached to the carbonyl moiety of the sophorolipid structure via an amide bond. In particular, the modified sophorolipid of the present invention can have one amino acid per molecule, where the amino acid is attached to the carbonyl moiety of the sophorolipid structure via an amide bond. A suitable amino acid has the chemical structure described herein, with at least one amino functional group capable of forming an amide bond with the carbonyl moiety at the defined attachment point of formula (I) and (II).

[0113] According to certain aspects of the invention, modified sophorolipids can improve and amplify the beneficial effects of the amino acids attached to them. Specifically, modified sophorolipids of formula (I) and (II) or their salts incorporating amino acids contribute to more efficient and better absorption of amino acids by target bodies when administered orally. Without being bound by theory, the bioavailability of amino acids may be improved, for example, by inhibiting P-glycoprotein and / or modulating other physical barrier mechanisms that, for example, reduce the penetration of amino acids into epithelial cells of a subject or through the blood-brain barrier. Additionally or alternatively, bioavailability may be improved by protecting the amino acids from degradation, either during storage or in vivo. Additionally or alternatively, bioavailability may be improved by improving the solubility or emulsification of amino acids by attachment to the sophorolipid structure, either in the composition prior to administration or in vivo after administration.

[0114] In another aspect, the modified sophorolipid enhances the efficacy of the sophorolipid or amino acid.

[0115] Depending on the nature and effect of the amino acids contained in the modified sophorolipids of the present invention, various diseases, illnesses, and / or conditions can be treated or prevented.

[0116] As used herein, the term "treatment" refers to eliminating, reducing, ameliorating, or reversing to any extent the extent, signs, or symptoms of a condition or disorder, and includes, but does not require, a complete cure of the condition or disorder. Treating may be curing, improving, or partially ameliorating a disease.

[0117] As used herein, "preventing" a condition or disorder refers to avoiding, delaying, forestalling, or minimizing the onset of a particular sign or symptom of a condition or disorder. Prevention can include, but does not require, absolute or complete prevention, meaning that there is still a possibility that a sign or symptom may develop later. Prevention can include reducing the severity of the onset of such a condition or disorder and / or inhibiting the condition or disorder from progressing to a more severe state.

[0118] One aspect of the present invention relates to a method of reducing a stress factor in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is methionine, lysine, and / or glutamine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. The stress factor may include cortisol concentration, ROS concentration, and / or biomarkers of oxidative stress known to those skilled in the art, such as those disclosed in Katerji M., Approaches and Methods to Measure Oxidative Stress in Clinical Samples: Research Applications in the Cancer Field, Oxid. Med. Cell. Longev. 2019 (2019).

[0119] Another aspect of the present invention relates to a method of treating or preventing a muscle disorder or disease in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. The muscle disorder or disease may be, for example, disuse muscle atrophy, muscular dystrophy and / or aging, or hyperexcitability of the muscle fiber membrane in myotonia-related syndromes.

[0120] In some embodiments, the present invention provides a method of increasing muscle vitality in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. Muscle vitality may be assessed by measuring the muscle mass, muscle strength, muscle power, and / or muscle endurance of the subject.

[0121] Yet another aspect of the present invention relates to a method for treating or preventing a cardiovascular disorder or disease in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. The cardiovascular disorder or disease may be, for example, arteriosclerosis, hyperhomocysteinemia, cardiomyopathy, and coronary heart disease.

[0122] In some embodiments, the present invention provides a method for improving the cardiovascular vitality of a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. Improvements may be observed, for example, in the subject's blood pressure, heart rate, and blood vessels.

[0123] A further aspect of the present invention relates to a method of treating or preventing hypertension in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond.

[0124] Another aspect of the present invention relates to a method of treating or preventing a central or peripheral nervous system disorder or disease in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. The central or peripheral nervous system disorder or disease may be, for example, memory loss and Alzheimer's disease.

[0125] In some embodiments, the present invention provides a method for improving central or peripheral nervous system function in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond. Improvement may be seen, for example, in the memory retention or motor performance of the subject.

[0126] A further aspect of the present invention relates to a method of treating or preventing a connective tissue disorder or disease in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II) or a salt thereof, wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond.

[0127] Another aspect of the present invention relates to a method of strengthening connective tissue in a subject by orally administering to the subject a modified sophorolipid of formula (I) and / or (II), wherein R1 is taurine attached to the carbonyl moiety of the sophorolipid structure via an amide bond.

[0128] A further aspect of the present invention is directed to a method of improving the nutrition in a subject's diet by administering a modified sophorolipid that incorporates an amino acid that is or may be missing or insufficient in the subject's diet.

[0129] In some embodiments, the various methods of the present invention involve the modified sophorolipid of formula (I) and / or (II) or salts thereof in a composition in solid form, including tablet, filled capsule, and pellet form. In another embodiment, the pharmaceutical composition may be in powder form, where the pharma- ceutically acceptable carrier is a finely divided active ingredient, i.e., a finely divided solid that is in admixture with the modified sophorolipid. In a further embodiment, the pharmaceutical composition of the present invention is a sustained release system, such as a semipermeable matrix of solid hydrophobic polymers containing the modified sophorolipid of the present invention. In another embodiment, the pharmaceutical composition is in liquid form, such as an aqueous or non-aqueous solution, suspension, emulsion, elixir, and capsules filled therewith.

[0130] The transitional word "comprising," which is synonymous with "including" or "containing," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. In contrast, the transitional phrase "consisting of" excludes any element, step, or material not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to those substances or steps specified and those that do not "significantly affect the basic and novel characteristics" of the claimed invention, such as the ability to improve the bioavailability of the substance. Use of the term "comprising" contemplates other embodiments that "consist" or "consist essentially of" the recited ingredients.

[0131] As used herein, the term "or" is understood to be inclusive unless specifically stated or obvious from the context. As used herein, the terms "a," "an," and "the" are understood to be singular or plural, unless specifically stated or obvious from the context.

[0132] Unless specifically stated or obvious from the context, as used herein, the term "about" is understood to mean within normal tolerances in the art, e.g., within 2 standard deviations of the mean. As a further example, "about" can 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.

[0133] The recitation of a list of chemical groups in any definition of a variable herein includes that definition of the variable as any single group or combination of the listed groups. The recitation of an embodiment of a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0134] Any composition or method provided herein can be combined with one or more of any of the other compositions and methods provided herein. EXAMPLES

[0135] The following are exemplary compounds of the present invention, which are presented for illustration and are not intended to limit the invention. The present invention encompasses all compounds represented by general formulas (I) and (II), including their salts, their hydrates, their geometric and optical isomers, and their polymorphic forms. Various modifications or changes within the spirit and scope of the present application will be suggested to those skilled in the art.

[0136] Exemplary compounds of formula (I): TIFF2025511856000008.tif43128In the formula, R2 is H, R3 is an 8 carbon aliphatic hydrocarbon chain having one unsaturation in the form of a double bond, R4 is methyl (-CH3), and R1 is as defined in Table 1.

[0137] Table 1: Compound numbers and corresponding R1 groups of formula (I) TIFF2025511856000009.tif124154

[0138] Exemplary compounds of formula (II): TIFF2025511856000010.tif43128 where R2 is H, R4 is methyl (-CH3), and R1 is as defined in Table 2.

[0139] Table 2: Compound numbers and corresponding R1 groups of formula (II) TIFF2025511856000011.tif125154

Claims

1. Compounds of formula (I): or a salt thereof, wherein R 1 is an amino acid linked to adjacent carbonyl moieties via amide bonds; R 2 are each independently H or -COCH 3 and R 3 is a saturated or unsaturated aliphatic hydrocarbon chain having 5 to 15 carbons; R 4 is H or -CH 3 That is, A compound or a salt thereof.

2. 2. The compound of claim 1, wherein the amino acid is taurine.

3. 2. The compound of claim 1, wherein the amino acid is methionine, lysine, or glutamine.

4. The sodium of the amino acid (Na + ) salt or potassium (K + 2. The compound of claim 1, which is a salt of

5. Compound of formula (II): or a salt thereof, wherein R 1 is an amino acid linked to adjacent carbonyl moieties via amide bonds; R 2 are each independently H or -COCH 3 and R 4 is H or -CH 3 That is, A compound or a salt thereof.

6. 6. The compound of claim 5, wherein the amino acid is taurine.

7. 6. The compound of claim 5, wherein the amino acid is methionine, lysine, or glutamine.

8. The sodium of the amino acid (Na + ) salt or potassium (K + 6. The compound of claim 5, which is a salt thereof.

9. A composition comprising a compound according to claim 1 in a pharma- ceutically or cosmetically acceptable carrier.

10. 10. The composition of claim 9, further comprising a compound of claim 5.

11. A composition comprising a compound according to claim 5 in a pharma- ceutically or cosmetically acceptable carrier.

12. 1. A method for improving oral bioavailability of an amino acid in a subject, comprising: Modified sophorolipids of formula (I) and / or (II): or a salt thereof to said subject. wherein R 1 is an amino acid linked to adjacent carbonyl moieties via an amide bond, R 2 are each independently H or -COCH 3 and R 3 is a saturated or unsaturated aliphatic hydrocarbon chain having 5 to 15 carbons; R 4 are each independently H or -CH 3 That is, method.

13. 13. The method of claim 12, wherein the amino acid is taurine, methionine, lysine, or glutamine.

14. 1. A method of treating or preventing a disease, disorder, or condition in a subject, comprising: Modified sophorolipids of formula (I) and / or (II): or a salt thereof to said subject. wherein R 1 are each independently an amino acid attached to an adjacent carbonyl moiety via an amide bond; R 2 are each independently H or -COCH 3 and R 3 is a saturated or unsaturated aliphatic hydrocarbon chain having 5 to 15 carbons; R 4 are each independently H or -CH 3 That is, method.

15. the disease, disorder, or condition is an increase in a stress factor; The amino acids are methionine, lysine, and / or glutamine; 15. The method of claim 14.

16. the disease, disorder, or condition is a muscle disorder or disease; The amino acid is taurine.

15. The method of claim 14.

17. the disease, disorder, or condition is a cardiovascular disorder or disease; The amino acid is taurine.

15. The method of claim 14.

18. the disease, disorder, or condition is hypertension; The amino acid is taurine.

15. The method of claim 14.

19. the disease, disorder, or condition is a disorder or condition of the central or peripheral nervous system; The amino acid is taurine.

15. The method of claim 14.

20. the disease, disorder, or condition is a disorder or disease of connective tissue; The amino acid is taurine.

15. The method of claim 14.

21. 1. A method of improving muscle vitality in a subject, comprising: Modified sophorolipids of formula (I) and / or (II): or a salt thereof to said subject in need of such improvement. wherein R 1 is an amino acid linked to adjacent carbonyl moieties via an amide bond, R 2 are each independently H or -COCH 3 and R 3 is a saturated or unsaturated aliphatic hydrocarbon chain having 5 to 15 carbons; R 4 are each independently H or -CH 3 and The amino acid is taurine. method.

Images