FORMULATIONS AND METHODS FOR SKIN REGENERATION

FIELD OF THE DISCLOSURE

The present disclosure is directed to formulations and their methods of use in improving skin regeneration, including skin barrier integrity, skin barrier repair, and skin moisture, and methods of treating musculoskeletal diseases or conditions, including for example, osteoarthritis, or symptoms thereof.

BACKGROUND

Skin diseases and conditions can cause significant physical discomfort and emotional distress for patients afflicted with such diseases and conditions. Common skin diseases and conditions include atopic dermatitis, psoriasis, and ichthyosis. Xerosis cutis (dry skin) is a very common dermatological disorder that afflicts a significant subset of the population. Osteoarthritis is the most common arthropathy and leading cause of pain and disability in the Western world. Skin diseases, conditions, and disorders that afflict patients frequently vary with age and are impacted by lifestyle and geographical area. Despite the high prevalence of skin diseases, conditions, and disorders in the population and the adverse effects of same on people's quality-of-life and the economic burden associated therewith, there are few effective treatments available. Effective formulations for addressing cosmetic skin care objectives and treatments for diseases, conditions, and disorders of the skin and musculoskeletal system represent unmet needs.

SUMMARY

Cosmetic and pharmaceutical formulations for skin care and treatments for diseases, conditions, and disorders of the skin and musculoskeletal system are described herein. Such formulations include those for skin regeneration that strengthen and enhance skin epithelial cell barrier repair, function, and integrity, thereby reducing moisture loss from the skin and restoring skin's structural integrity, intercellular adhesion, and biochemistry and/or promoting maintenance of these features, as well as for treating musculoskeletal system diseases or disorders, for example, osteoarthritis, and its symptoms. Other features and advantages of the amino acid formulations and methods of use will be apparent from the detailed description, and from the embodiments.

Described herein are topical formulations comprising, consisting essentially of, or consisting of, as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount comprising, consisting essentially of, or consisting of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), where each amino acid present comprises a ratio to each of the other amino acids of 1:100 or greater (e.g., 2:100; 4:100; 6:100; 8:100; 10:100; 12:100; 14:100; 16:100; 18:100; 20:100; 22:100; 24:100; 26:100; 28:100; 30:100; 32:100; 34:100; 36:100; 38:100; 40:100; 42:100; 44:100; 46:100; 48:100; 50:100; 52:100; 54:100; 56:100; 58:100; 60:100; 62:100; 64:100; 66:100; 68:100; 70:100; 72:100; 74:100; 76:100; 78:100; 80:100; 82:100; 84:100; 86:100; 88:100; 90:100; 92:100; 94:100; 96:100; 98:100; 100:100; 110:100; 120:100; 130:100; 140:100:150:100; 160:100; 170:100; 180:100; 190:100; 200:100); or 100:1 or less (e.g., 99:1; 97:1; 95:1; 93:1; 91:1; 89:1; 87:1; 85:1; 83:1; 81:1; 79:1; 77:1; 75:1; 73:1; 71:1; 69:1; 67:1; 65:1; 63:1; 61:1; 59:1; 57:1; 55:1; 53:1; 51:1; 49:1; 47:1; 45:1; 43:1; 41:1; 39:1; 37:1; 35:1; 33:1; 31:1; 29:1; 27:1; 25:1; 23:1; 21:1; 19:1; 17:1; 15:1; 13:1; 11:1; 9:1; 7:1; 5:1; 3:1; 1:1; 0.9:1; 0.7:1; 0.5:1; 0.3:1; 0.1:1); where the therapeutically effective amount improves skin regeneration (e.g., skin barrier integrity, function, repair; improves skin moisture; treats or ameliorates musculoskeletal diseases or disorders, e.g., osteoarthritis, or its symptoms thereof, or any combination thereof), when tested by a method detecting expression of a skin regeneration marker gene (e.g., skin barrier marker gene (e.g., EGF, FGF2, PDGF, PPARD, FLG, MKI67, CPT2, TGM4)) and determining that the marker gene is modulated by a 1-7 average fold change over an amino acid concentration in a range of 0.4 mM-10 mM of formulation-treated skin cells as compared to untreated skin cells; and optionally, a dermatologically-acceptable carrier; where the formulation improves skin regeneration (e.g., skin barrier integrity, function, or repair of the skin, improves skin moisture, or treats or ameliorates musculoskeletal diseases or disorders, e.g., osteoarthritis, or its symptoms thereof).

Another aspect of the disclosure provides a topical formulation, where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, or derivatives or salts thereof; where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, isoleucine, or derivatives or salts thereof; where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, isoleucine, valine, or derivatives or salts thereof; where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, isoleucine, arginine, valine, or derivatives or salts thereof; where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, isoleucine, arginine, valine, threonine, or derivatives or salts thereof; where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, isoleucine, arginine, valine, threonine, tryptophan, or derivatives or salts thereof; or where the therapeutically effective amount of free amino acids or a peptide combination of amino acids comprise glutamine, glycine, alanine, serine, isoleucine, arginine, valine, threonine, or derivatives or salts thereof, Such formulations described here can have in some aspects, a ratio where the glutamine to glycine ratio is 192:100; where the glutamine to alanine ratio is 164:100; where the glutamine to serine ratio is 139:100; where the glutamine to isoleucine ratio is 111:100; where the glutamine to valine ratio is 124:100; where the glutamine to arginine ratio is 83:100; where the glutamine to threonine ratio is 122:100; or where the glutamine to tryptophan ratio is 71:100.

Other aspects of such formulations described here include those where EGF is upregulated by: a 1-7 average fold change of treated as compared to untreated skin cells; a 1-5 average fold change of treated as compared to untreated skin cells; a 1 or greater average fold change of treated as compared to untreated skin cells (e.g., 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5.1, 5.3, 5.5, 5.7, 5.9, 6.1, 6.3, 6.5, 6.7, 6.9, 7.1, 7.3, 7.5, 7.7, 7.9, 8.1, 8.3, 8.5, 8.7, 8.9, 9.1, 9.3, 9.5, 9.7, 9.9); a 10 or less average fold change of treated as compared to untreated skin cells (e.g., 9.8, 9.6, 9.4, 9.2, 9, 8.8, 8.6, 8.4, 8.2, 8, 7.8, 7.6, 7.4, 7.2, 7, 6.8, 6.6, 6.4, 6.2, 6, 5.8, 5.6, 5.4, 5.2, 5, 4.8, 4.6, 4.4, 4.2, 4, 3.8, 3.6, 3.4, 3.2, 3, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 1).

Some aspects provide such formulations, where each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or derivatives or salts thereof, is present at a concentration of 0.05 mM or greater; 20 mM or less; or ranging from 0.05 mM-20 mM.

Further embodiments are directed to a method for maintaining and/or improving skin regeneration of skin cells in a subject in need thereof, One aspect of the method comprises applying the formulation described here to the skin of the subject, thereby maintaining and/or improving skin regeneration of skin cells in the subject. A further aspect of the method provides such formulations that reduce the length, depth, and/or other dimension of lines and/or wrinkles in the skin; treat a skin condition selected from the group consisting of: atopic dermatitis, dermatitis, radiodermatitis, psoriasis, pruritus, eczema, a wound, an abrasion, and a burn, or combinations thereof.

Definitions

All terms used herein are intended to have their ordinary meaning in the art unless otherwise provided. All concentrations are in terms of percentage by weight of the specified component relative to the entire weight of the topical composition, unless otherwise defined.

As used herein, “a” or “an” shall mean one or more. As used herein when used in conjunction with the word “comprising,” the words “a” or “an” mean one or more than one. As used herein “another” means at least a second or more.

As used herein, all ranges of numeric values include the endpoints and all possible values disclosed between the disclosed values. The exact values of all half-integral numeric values are also contemplated as specifically disclosed and as limits for all subsets of the disclosed range. For example, a range of from 0.1% to 3% specifically discloses a percentage of 0.1%, 1%, 1.5%, 2.0%, 2.5%, and 3%. Additionally, a range of 0.1 to 3% includes subsets of the original range including from 0.5% to 2.5%, from 1% to 3%, from 0.1% to 2.5%, etc. It is understood that the sum of all weight % of individual components will not exceed 100%.

The terms “improving a cosmetic skin condition”, “improving a skin condition”, “improving skin regeneration”, or “treating a skin condition” include treating a cosmetic skin condition or therapeutically treating a skin condition and may involve at least one of the following benefits: improving moisture retention, reducing skin epithelial cell barrier permeability, thickening of skin (increasing the depth of the skin epithelial layer), restoring skin elasticity, preventing loss of skin elasticity, reducing skin cell flaking, improving skin texture to be smoother, reducing physical manifestations of lines or wrinkles (as determined by, e.g., visual appearance or touch perception) and any combination thereof.

The “skin” comprising skin cells, comprises three distinct layers: the stratum corneum (the outermost layer), the epidermis, and the dermis.

The term “barrier integrity of skin cells”, as used herein, refers to the ability of the epidermis and particularly the stratum corneum (SC) to function as a selective permeability barrier, which limits transcutaneous evaporative water loss and permits survival in desiccating external environments. The SC is a multilayered tissue composed of flattened, anucleate corneocytes, surrounded by multiple planar lamellae sheets, enriched in ceramides, cholesterol, and free fatty acids (FFA) that hold the entire matrix together. These components, together with tight junction proteins and antimicrobial peptides, contribute to the ability of the skin to act as a permeability barrier. Tight junction proteins contribute to the intercellular adhesive interface, which essentially serves as a seal between cells. Unregulated intercellular leakage from the outermost layer of the skin may, for example, result in loss of water from inner layers of the skin. Barrier integrity may be measured using a variety of assays, including: transepidermal water loss (TEWL) on the skin's surface (in situ): transepithelial/transendothelial electrical resistance (TEER) is also widely accepted as a quantitative technique to measure the integrity of tight junction dynamics in cell culture models of endothelial and epithelial monolayers; techniques that monitor diffusion of dyes (X-Gal or Lucifer Yellow, Toluidine Blue) through the upper epidermis are also used for skin explants and/or in situ skin; 2D or 3D keratinocyte cell cultures are used to assess epidermal differentiation processes that are directly related to the skin barrier permeability; upregulation of proteins that promote skin barrier function and/or downregulation of genes that impair skin barrier function; and immunohistochemistry, microscopical examination and/or other imaging techniques.

The term “improving”, when used with respect to improving “barrier integrity of skin cells”, refers to increasing the skin's permeability barrier, for example, by increasing the strength of adhesion between skin cells as reflected by an increased ability of the intercellular adhesive interface to prevent unregulated leakage between cells. Assays such as those described with respect to barrier integrity may be used to detect an increase in barrier integrity. The term “improving” may be used to describe an increase equal to or greater than 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% relative to a starting point measurement prior to contact with an amino acid formulation described herein. “Improving skin regeneration” refers to increasing dermal cells, including keratinocytes, skin cell production or speeding up skin cell regeneration and healing of skin, relative to damaged skin or compromised skin (e.g., abrasions, wounds, cuts, dry or cracked skin, eczema, psoriasis).

The term “maintaining”, when used with respect to maintaining “barrier integrity of skin cells”, refers to preserving the strength of the skin's permeability barrier, for example, by preserving the strength of adhesion between skin cells as reflected by the ability of the intercellular adhesive interface to prevent unregulated leakage between cells. With respect to maintaining “barrier integrity of skin cells”, measurements can be made to determine baseline strength of intercellular adhesion in, e.g., healthy skin so as to establish a level of intercellular adhesion that is the desired objective to maintain. Samples of the healthy skin can then be exposed to stressors, such as, for example, airborne pollution (e.g., cigarette smoke, noxious fumes, airborne irritants) or physical stressors (e.g., abrasion, harsh detergents, or chemicals) in the presence or absence of a formulation described herein. Amino acid formulations described herein that at least in part prevent a reduction in intercellular adhesion even when challenged with a stressor would be characterized as having the ability to maintain barrier integrity to the degree determined. In such a context, the term “maintaining” may be used to describe preservation of baseline intercellular strength at 100% pre-stressor levels, or equal to or greater than 99%, 98%, 97%. 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 2%, or 1% pre-stressor levels due to the presence of an amino acid formulation described herein which counteracts the negative impact of such a stressor.

The term “topical application,” as used herein, means to apply or spread the formulations described herein onto the surface of the epidermis tissue of a subject in need thereof.

The term “dermatologically-acceptable,” as used herein, means that the formulations or components thereof so described are suitable for use in contact with mammalian epidermal tissue without undue toxicity, incompatibility, instability, allergic response, and the like.

The term “therapeutically effective amount,” as used herein, refers to an amount of a compound or formulation sufficient to induce a positive benefit, an improvement in the appearance and/or texture of the skin, where in some embodiments the positive benefit of an amino acid or formulation treated skin describes an increase equal to or greater than 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% relative to a starting point measurement prior to contact with an amino acid formulation described herein or untreated skin; 1 fold or greater (e.g., 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41); 40 fold or less (e.g., 38, 36, 34, 32, 30, 28, 26, 24, 22, 20, 18, 16, 14, 12, 10, 8, 6, 4, 2, 0.8, 0.6); or 1-40 fold (e.g., 2-39; 3-38; 4-37; 5-36; 6-35; 7-34; 8-33; 9-32; 10-31; 11-30; 12-29; 13-28; 14-27; 15-26; 16-25; 17-24; 18-23; 19-22; 20-21) better than amino acid untreated skin. In accordance with the present disclosure, a therapeutically effective amount is an amount of a combination of free amino acids or a peptide combination thereof, either alone or in combination with other agents (e.g., plant extract, retinoids, additives), that regulates and/or improves the skin physically and/or visually; improves skin barrier integrity or repair and/or function; improves or maintains moisture; or any combination thereof.

The term “amelioration” or any grammatical variation thereof (e.g., ameliorate, ameliorating, and amelioration etc.), as used herein, includes, but is not limited to, delaying the onset, or reducing the severity of a disease or condition. Amelioration, as used herein, does not require the complete absence of symptoms.

The terms “effective amount” or “significant amount” as used herein, refers to an amount that is capable of treating or ameliorating a disease, condition, or disorder, or is otherwise capable of producing an intended therapeutic effect.

The term “functional” means a useful effect for human health, such as structural or functional regulation of nutrients, the immune system, inflammation, fluid balance, physiological action, or the like.

The term “carrier” or “vehicle” refers to a diluent, adjuvant, or excipient with which the compound or active ingredient, such as but not limited to, free amino acids or a peptide combination thereof (e.g., at least one of alanine, glutamine, glycine, and serine, or salts thereof, or any combination thereof), plant extract (e.g., Boswellia extract, which includes Boswellia serrata, Burseraceae family, olibanum, or frankincense) or plant extract combination (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); or retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof) described here, is formulated and administered. Non-limiting examples of a “carrier” or a “vehicle” include: water; oils (e.g., argan oil; jojoba oil; sweet almond oil; apricot oil; evening primrose oil; hemp seed oil; rosehip oil); solvents; lipids (e.g., liposomes, nanoemulsions, nanospheres, lipid nanoparticles, gold nanoparticles, niosomes), squalene; phyto-based nano-formulations; silicone (e.g., dimethicone); ceramides; a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications)), or any combinations thereof, Examples of suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” (23rd edition) by E. W. Martin, which is incorporated herein by reference.

The term “treatment” or any grammatical variation thereof (e.g., treat, treating, and treatment etc.), as used herein, includes but is not limited to, alleviating a symptom of a disease, condition, or disorder; and/or reducing, suppressing, inhibiting, lessening, or affecting the progression, severity, and/or scope of a disease, condition, or disorder.

The term “consisting essentially of,” as used herein, limits the scope of the ingredients and steps to the specified materials or steps and those that do not materially affect the basic and novel characteristic(s) of the present embodiments of the disclosure, i.e., formulations and methods for promoting at least one of skin integrity, function, texture, moisture, or appearance, or any combination thereof, or for skin regeneration, including skin barrier repair and strengthening, or for improving skin barrier integrity. For instance, by using “consisting essentially of,” the therapeutic formulation does not contain any unspecified ingredients including, but not limited to, free amino acids, di-, oligo-, or polypeptides or proteins; and mono-, di-, oligo-, polysaccharides, and carbohydrates that have a direct beneficial or adverse therapeutic effect on improving skin regeneration, including skin barrier integrity, repair, or function of skin cells. Also, by using the term “consisting essentially of,” the formulation may comprise substances that do not have therapeutic effects on improving barrier integrity of skin cells or improving musculoskeletal conditions, such as but not limited to osteoarthritis, or its symptoms; such ingredients include vehicles, carriers, excipients, adjuvants, fragrances, etc. that do not negatively affect barrier integrity of skin cells or treatment or amelioration of musculoskeletal conditions (e.g., osteoarthritis) or their symptoms.

The term “amino acid” encompasses all known amino acids comprising an amine (—NH₂) functional group, a carboxyl (—COOH) functional group, and/or a side chain (“R”) group specific to each amino acid. “Amino acids” encompasses the 21 amino acids encoded by the human genome (i.e., proteinogenic amino acids), amino acids encoded or produced by bacteria or single-celled organisms, and naturally derived amino acids. For the purposes of this disclosure, the conjugate acid form of amino acids with basic side chains (arginine, lysine, and histidine) or the conjugate base form of amino acids with acidic side chains (aspartic acid and glutamic acid) are essentially the same, unless otherwise noted. “Amino acids” also encompass derivatives and analogs thereof that retain substantially the same activity in terms of improving barrier integrity of skin cells in, for example, a TEER or TEWL assay as described above. The derivatives and analogs may be, for example, enantiomers, and include both the D- and L-forms of the amino acids. In some embodiments, at least one or more of the amino acids are in the D-form. In other embodiments, at least one or more of the amino acids are in the L-form. A “peptide combination” of amino acids (two or more amino acids; typically, 2-50 amino acids) described here can include one or more amino acids in the D-form, in the L-form, or a mixture of D-form and L-form linked by a peptide bond. An “amino acid combination” as used here can refer to free amino acids or a peptide combination of amino acids. The derivatives and analogs may be derivatives of “natural” or “non-natural” amino acids (e.g., β-amino acids, homo-amino acids, proline derivatives, pyruvic acid derivatives, 3-substituted alanine derivatives, glycine derivatives, ring-substituted cysteine derivatives, ring-substituted phenylalanine derivatives, linear core amino acids, and N-methyl amino acids), for example, selenocysteine, pyrrolysine, iodocysteine, norleucine, or norvaline. The derivatives and analogs may comprise a protecting group (α-amino group, α-carboxylic acid group, or suitable R group, wherein R contains a —NH₂, —OH, —SH, —COOH or other reactive functionality). Other amino acid derivatives include, but are not limited to, those that are synthesized by, for example, acylation, methylation, glycosylation, and/or halogenation of the amino acid. These include, for example, β-methyl amino acids, C-methyl amino acids, and N-methyl amino acids. The amino acids described herein may be present as free amino acids. The term “free amino acid” refers to an amino acid that is not part of a peptide or polypeptide (e.g., is not connected to another amino acid through a peptide bond). A free amino acid is free in solution (as opposed to being linked to at least one other amino acid via, for example, a dipeptide bond), but may be associated with a salt or other component in solution.

As used herein, the term “salt” refers to any and all salts or salt forms of compounds and encompasses pharmaceutically acceptable salts. For example, embodiments of the disclosure are directed to amino acids or salts thereof.

Exemplary salts for inclusion in a formulation described herein include sodium chloride, potassium chloride, calcium chloride, magnesium chloride, or tri-sodium citrate, sodium bicarbonate, sodium gluconate phosphate buffers using mono, di or tri-sodium phosphate or any combination thereof.

Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, cellulose, microcrystalline cellulose, kaolin, sodium chloride, and mixtures thereof.

Pharmaceutically acceptable excipients used in the manufacture of formulations, including cosmetic and pharmaceutical formulations, described herein include inert diluents, dispersing and/or granulating agents, surface active agents and/or an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine), disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, and perfuming agents may also be present in the composition.

Abbreviations used: Amino acids: Ala—alanine, Arg—arginine, Cys—cysteine, Gln—glutamine, Gly—glycine, His—histidine, Ile—isoleucine, Ser—serine, Thr—threonine, Tyr—tyrosine, Val—valine. Genes/products: CPT2—carnitine palmitoyltransferase 2; CASP8—caspase 8; EGF—epidermal growth factor; FGF—fibroblast growth factor; MKI67—marker of proliferation Ki-67; PDGF—platelet-derived growth factor; PPARs—peroxisome proliferator-activated receptors; SMAD—an acronym from the fusion of Caenorhabditis elegans Sma genes and the Drosophila Mad, Mothers against decapentaplegic; TGFB1—transforming grown factor beta 1; TGM—transglutaminase, TNFRSF10D-TNF Receptor Superfamily Member 10d; Other: FAA—free amino acids, NMF—natural moisturizing factor, NHEK—normal human epidermal keratinocytes, pPCR—quantitative polymerase chain reaction; TJ—tight junctions, SC—stratum corneum.

Any formulation or methods provided here can be combined with one or more of any of the other formulation and methods provided here.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 shows the effect of treatment of amino acid combinations on mRNA expression of barrier marker genes in differentiated normal human epidermal keratinocytes (NHEK) cells of 5 donors. Treatment time—4 h. All graphed data show mean±SE, *: p<0.05, **: p<0.01, ***: p<0.001 compared to untreated cells (CTR) in two-tailed t-test. Data from two independent experiments. Exemplary barrier marker genes examined as follows: Epidermal growth factor (EGF); fibroblast growth factor 2 (FGF2); platelet derived growth factor (PDGF); Carnitine Palmitoyltransferase 2 (CPT2); Transglutaminase 4 (TGM4); Peroxisome Proliferator Activated Receptor Delta (PPARD); TNF Receptor Superfamily Member 10d (TNFRSF10D).

FIG. 2 presents a Table listing combinations of amino acids. Amino acids in combos #1, 2, 4, 6 and 7 were used at a concentration of 4 mM for each amino acid. Amino acids in combos #3 and #4 were used at the lower indicated concentrations (in mM) due to toxic effects detected at concentrations 4 mM.

FIG. 3 presents a Table showing a transcript analysis of skin barrier integrity marker genes in primary keratinocytes. Differentiation Genes are designated with *, whereas Proliferation Genes are designated with +. EGF contributes to both differentiation and proliferation. Scoring system developed: # of points given to relative expression. (Average relative expression >1.5 (50% Increase); Untreated=1).

FIG. 4 presents a Table showing a transcript analysis of skin barrier integrity marker genes in primary keratinocytes. Differentiation Genes are designated with *, whereas Proliferation Genes are designated with +. EGF contributes to both differentiation and proliferation. Scoring system developed: # of points given to relative expression. (Average relative expression >1.5 (50% Increase); Untreated=1) (n=5).

FIG. 5 shows quantification of Western Blot analysis of involucrin (INV) expression at 24 h of treatment. Combo #1: Gln, Gly, Ala, Ser; Combo #2: Gln, Gly, Ala, Ser, Ile, Val; Combo #5: Cys, His, Leu, Asp. Quantitative analysis of involucrin expression determined based on results from two donor samples.

FIG. 6 shows quantification of Western Blot analysis of filaggrin (FLG) expression at 4 and 24 h of treatment. Combo #1: Gln, Gly, Ala, Ser; Combo #2: Gln, Gly, Ala, Ser, Ile, Val; Combo #3: Cys, His, Tyr; Combo #5: Cys, His, Leu, Asp.

FIGS. 7A-7D present graphs of EGF expression comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA) and Boswellia plant extract combination (BSW=BSXL) alone or in combination. FIG. 7A shows EGF expression of 4GAA alone (4 mM); BSW 1% alone; the combination of 4GAA 4 mM and BSW 1% with Donor O cells. FIG. 7B illustrates EGF expression of 4GAA alone (1 mM); BSW 0.25% alone; the combination of 4GAA 1 mM and BSW 0.25% with Donor O cells. FIG. 7C shows EGF expression of 4GAA alone (4 mM); BSW 0.5% alone; the combination of 4GAA 4 mM and BSW 0.5% with Donor G cells. FIG. 7D presents EGF expression of 4GAA alone (1 mM); BSW 0.25% alone; the combination of 4GAA 1 mM and BSW 0.25% with Donor G keratinocyte cells. *p value=0.05; **p value=0.01. “CTR” or “CTR untr” is untreated control. n=3

FIGS. 8A-8B illustrate the fold change of EGF mRNA in Donor G keratinocyte cells comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA) 1 mM or 4 mM and/or plant extract combination containing, e.g., Boswellia (BSXL) alone (0.1%, 0.25%, 0.5%) or in combination at different concentrations. The data are shown in graphical form (FIG. 8A) and tabular form (FIG. 8B). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 vs untreated control (ctr). n=3

FIGS. 9A-9B present the fold change of EGF mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA) and Boswellia plant extract combination (BSXL; Bosexil®) alone or in combination at different concentrations with Donor O keratinocyte cells. The data are shown in graphical form (FIG. 9A) and tabular form (FIG. 9B), **p<0.01, ***p≤0.001 versus untreated CTR control. n=2 or n=3

FIGS. 10A-10B present the fold change of EGF mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), 3 amino acid combination (Cys 1 mM, His 0.5 mM, Tyr 1 mM; 3BAA), 0.5% Boswellia plant extract combination (BSXL), and Piperonylic acid (100 μM; PA) alone or in combination with Donor G keratinocyte cells. The data are shown in graphical form (FIG. 10A) and tabular form (FIG. 10B), *p<0.05, **p≤0.01, ***p≤0.001 versus untreated CTR control. n=3

FIGS. 11A-11B demonstrate the fold change of EGF mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), 3 amino acid combination (Cys 1 mM, His 0.5 mM, Tyr 1 mM; 3BAA), Boswellia plant extract combination (BSXL), and Piperonylic acid (100 μM; PA) alone or in combination with Donor T keratinocyte cells. The data are shown in graphical form (FIG. 11A) and tabular form (FIG. 11B), *p<0.05, **p<0.01, ****p<0.001 versus untreated CTR control. n=3

FIGS. 12A-12B illustrate the fold change of EGF mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), Boswellia plant extract combination (BSXL), and Boswellic acid extract (BA) alone or in combination and in various concentrations or weight percentages (wt/wt %) with Donor O keratinocytes. The data are shown in graphical form (FIG. 12A) and tabular form (FIG. 12B), *p<0.05, **p≤0.01, ***p≤0.001 versus untreated CTR control. n=2 or n=3

FIGS. 13A-13B show the fold change of EGF mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), alternative 4 amino acid combination (Asp, Ile, Tyr, Val; 4OAA; 4 mM), and Boswellia plant extract combination (BSXL; 0.5% or 0.5 w/w % in combination), alone or in combination and in various concentrations or weight percentages (wt/wt %) with Donor A keratinocytes. The data are shown in graphical form (FIG. 13A) and tabular form (FIG. 13B), *** p≤0.001 versus untreated CTR control. n=3

FIGS. 14A-14C show the fold change of EGF mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and Boswellia plant extract combination (BSXL; 0.5 w/w %), alone or in combination with Donor A. Donor G, and Donor T keratinocytes. The data are shown in tabular form (FIG. 14A (Donor A (n=1), Donor G (n=3), and Donor T (n=2)); FIG. 14B (Donor A (n=3), Donor G (n=9), and Donor T (n=6)) and FIG. 14B data represented in graphical form (FIG. 14C). FIG. 14C shows CTR; 4GAA; BSXL; 4GAA+BSXL combination: Donor G (top 3). Donor T (center). Donor A (bottom 3), *p<0.05 versus untreated CTR control; #p<0.05 versus 4GAA and BSXL. Repeated measures analysis of variance (RMANOVA) with Tukey post-hoc was used for analysis.

FIGS. 15A-15B present the transepidermal water loss (TEWL) change after test material (Active (4GAA) in an oil/water (O/W) vehicle, left column; Placebo, right column) application for 10 days on 12 subjects who are then subjected to repeated tape strips (15 tape strips). FIG. 15A shows the TEWL change from baseline-Day 1 immediately after tape-stripping and Days 2, 3, and 5 (X-axis) are after two treatments per day of test materials. FIG. 15B demonstrates the relative TEWL increase after tape-stripping where all values are normalized to the baseline-Day 1 which represents TEWL immediately after tape-stripping. Active (left column); Placebo (right column). Mean+/−95% Confidence Interval (CI).

FIG. 16 presents skin barrier improvement measuring skin hydration (Y-axis) after test material application for 10 days on 25 subjects who were then subjected to repeated tape strips (25). Control represents untreated (top graphical line at Day 3; left column-inset); Placebo represents water/oil/water (W/O/W) vehicle without active (middle graphical line at Day 3; center column-inset); Amino Acid represents the 4 amino acid combination (Ala (4%). Gln (4%), Gly (4%). Ser (4%)) in the W/O/W vehicle (bottom graphical line at Day 3; right column-inset), *, p-value=0.05. Inset: Control (left), Placebo (center), Amino Acid (right).

FIGS. 17A-17C show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination, to control (CTR; untreated control without amino acids or retinol) with Donor E, Donor G. Donor O, Donor R, and Donor T human-derived epidermal keratinocytes (NHEK) with a 4 h treatment in amino acid (AA)-deprived medium. The data are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination. FIG. 17A shows the fold change of EGF mRNA (n=3). FIG. 17B shows the fold change of TGFB mRNA (n=3). FIG. 17C shows the fold change of COL1A1 mRNA (n=3).

FIGS. 18A-18C show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination with Donor E. Donor G. Donor O. Donor R. and Donor T human-derived epidermal keratinocytes (NHEK) to control (CTR; untreated control without amino acids or retinol) with a 4 h treatment in amino acid (AA)-deprived medium. The data of FIGS. 18A-18C are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination. FIG. 18A shows the fold change of HAS1 mRNA. FIG. 18B shows the fold change of HAS2 mRNA. FIG. 18C shows the fold change of HAS3 mRNA. FIGS. 18D-18F show the data in tabular form (Donor E (n=3). Donor G (n=3). Donor O (n=3). Donor R (n=3), and Donor T (n=3)) for fold change of HAS1 mRNA. HAS2 mRNA, and HAS3 mRNA, respectively, and in tabular form showing the fold change for each of the specified conditions under each respective graphical form.

FIGS. 19A-19D show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala. Gln. Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination to control (CTR; untreated control without amino acids or retinol) with Donor O human-derived epidermal keratinocytes (NHEK) with a 24 h treatment in complete medium. The data of FIGS. 19A-19D are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination. FIG. 19A shows the fold change of EGF mRNA. FIG. 19B shows the fold change of HAS1 mRNA. FIG. 19C shows the fold change of HAS2 mRNA. FIG. 19D shows the fold change of HAS3 mRNA. FIG. 21 shows the Donor O data in tabular form for fold change of EGF, HAS1, HAS2, and HAS3 mRNA under the specified conditions.

FIGS. 20A-20D show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination with Donor R human-derived epidermal keratinocytes (NHEK) to control (CTR; untreated control without amino acids or retinol) with a 24 h treatment in complete medium. The data of FIGS. 20A-20D are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination. FIG. 20A shows the fold change of EGF mRNA. FIG. 20B shows the fold change of HAS1 mRNA. FIG. 20C shows the fold change of HAS2 mRNA. FIG. 20D shows the fold change of HAS3 mRNA. FIG. 21 shows the Donor R data in tabular form for fold change of EGF mRNA, HAS1 mRNA, HAS2 mRNA, and HAS3 mRNA under the specified conditions.

FIG. 21 shows the Donor O and Donor R data in tabular form for fold change of EGF mRNA. HAS1 mRNA. HAS2 mRNA, and HAS3 mRNA under the specified conditions.

FIGS. 22A-22B show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 5 μM), alone or in combination (4GAA+ROL), with 3D skin equivalents (normal, human-derived epidermal keratinocytes (NHEK) cultured at air-liquid interface (ALI) on tissue culture inserts; EpiDerm™; MatTek Corp., Ashland, MA) to control (CTR; untreated control without amino acids or retinol) with two treatments over a 24 h period in complete medium. The data of FIGS. 22A-22B are shown in graphical form showing CTR; 4GAA (4 mM); ROL (5 μM); 4GAA (4 mM)+ROL (5 μM) combination and in tabular form showing the fold change for each of the specified conditions under each respective graphical form. FIG. 22A shows the fold change of HAS2 mRNA. FIG. 22B shows the fold change of HAS3 mRNA.

FIGS. 23A-23B show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination with fibroblasts (human dermal fibroblasts, adult (HDFa)) to control (CTR; untreated control without amino acids or retinol) with a 4 h treatment in EBS medium. The data of FIGS. 23A-23B are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination. FIG. 23A shows the fold change of collagen. COL1A1 mRNA in fibroblasts (n=3). FIG. 23B shows the fold change of elastin. ELN mRNA in fibroblasts (n=3).

FIGS. 24A-24C show the fold change of a specific gene mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination with fibroblasts (human dermal fibroblasts, adult (HDFa)) to control (CTR; untreated control without amino acids or retinol) with a 4 h treatment in complete medium. The data of FIGS. 24A-24C are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination and in tabular form showing the fold change for each of the specified conditions under each respective graphical form. FIG. 24A shows the fold change of HAS1 mRNA (n=3). FIG. 24B shows the fold change of HAS2 mRNA (n=3). FIG. 24C shows the fold change of HAS3 mRNA (n=3).

FIGS. 25A-25B show the fold change of HAS1 mRNA comparing the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), and retinol (ROL; 10 μM), alone or in combination with fibroblasts (human dermal fibroblasts, adult (HDFa)) to control (CTR; untreated control without amino acids or retinol) with a 4 h treatment in amino acid (AA)-deprived and complete medium (FIG. 25A) and with a 24 h treatment in complete medium (FIG. 25B). The data of FIGS. 25A-25B are shown in graphical form showing CTR; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination and in tabular form showing the fold change of HAS1 mRNA for each of the specified conditions under each respective graphical form. FIG. 25A shows the fold change of HAS1 mRNA (n=3) with a 4 h treatment in AA-derived medium and complete medium. FIG. 25B shows the fold change of HAS1 mRNA (n=3) with a 24 h treatment in complete medium.

FIG. 26 shows fibroblast data in tabular form for fold change of EGF mRNA, HAS1 mRNA, HAS2 mRNA, and HAS3 mRNA under the specified conditions: control (CTR; untreated control without amino acids or retinol; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination to control (CTR; complete medium with fibroblasts) with a 24 h treatment in complete medium.

FIGS. 27A-27E show Donor E. Donor G. Donor O. Donor R. and Donor T human-derived epidermal keratinocyte (NHEK) data (cycle threshold. CT=35) in tabular form for CT of endogenous expression of HAS2 mRNA under the specified conditions: control (CTR; untreated control without amino acids or retinol; 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination.

FIG. 28 shows the average effect of treatment of single amino acids and combinations of amino acids on mRNA expression of barrier marker genes in differentiated normal human epidermal keratinocytes (NHEK) cells of 2 donors. Treatment time-4 h. All graphed data show mean±SE compared to untreated cells (Control) in two-tailed t-test. Exemplary barrier marker genes examined as follows: Epidermal growth factor (EGF); fibroblast growth factor 2 (FGF2); platelet derived growth factor (PDGF); Peroxisome Proliferator Activated Receptor Delta (PPARD).

FIG. 29 shows the average fold change in EGF mRNA treated with the indicated amino acids from the average EGF mRNA Control (no treatment) across a range of amino acid concentrations from 0.4 mM to 10 mM.

DETAILED DESCRIPTION

Detailed embodiments of the present disclosure are disclosed herein; however, it is to be understood that the disclosed embodiments are merely illustrative of the disclosure that may be embodied in various forms. In addition, each of the examples given in connection with the various embodiments of the disclosure is intended to be illustrative, and not restrictive.

The skin acts as a physical barrier that separates an organism from its surrounding environment, but also serves as an interactive interface with that environment. The skin comprises three distinct layers: the stratum corneum, the epidermis, and the dermis. The stratum corneum (the outermost layer) comprises keratinized cells and provides a physical barrier with the external environment. Environmental stressors vary based, for example, on climate and/or lifestyle and may include xenobiotics, UV radiation, pollution (e.g., cigarette smoke or engine exhaust), and pathogens. In addition to acting as a shield that protects the lower layers of the skin from such environmental insults, the stratum corneum also reduces water loss from the skin. Skin keratinocytes are connected via tight junction (TJ) proteins, such as zona occludins (ZOs). TJ proteins play a significant role in barrier function, as well as contributing to keratinocyte proliferation and differentiation. Barrier function integrity serves as a major feature of the skin that prevents excessive water loss from the skin, thereby maintaining healthy skin.

Skin barrier dysregulation is implicated in a variety of common skin diseases like atopic dermatitis, psoriasis, and ichthyosis. Skin barrier dysregulation also contributes to xerosis cutis (dry skin), a common dermatological disorder, which is exacerbated by low humidity, repeated washing, and exposure to harsh chemicals. Such external stressors promote water loss from the skin. Dry skin may lead to or be accompanied by itching and/or skin irritation. Dry skin may be considered a symptom of decreasing barrier function of the skin. Dry skin is also a common feature of aging skin.

Optimal management of many skin diseases, conditions, or disorders includes appropriate skin care. Incorporation of a well-designed moisturizer/barrier repair formulation can help reduce disease-associated signs and symptoms, thereby ameliorating the disease. Effective moisturizer/barrier repair formulations for treating dry skin that enhance skin regeneration and strengthen skin barrier function are described herein. The formulations comprise specific amino acid combinations that confer cosmetic and/or therapeutic benefit in the context of treating a variety of cosmetic skin conditions and skin diseases, conditions, and disorders (for example, xerotic skin).

Various embodiments of the disclosure feature formulations and methods of improving skin regeneration, including skin barrier integrity, repair, and function. The disclosure is based, at least in part, on the discovery that one or more amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof, can modulate transcription of key regulators of differentiation and proliferation, such as but not limited to, EGF, PDGF, FGF2, and PPARD genes (or skin barrier markers), in normal primary human keratinocytes.

Without being bound by theory, delivering specific amino acids and combinations thereof to the skin, allows the skin's innate ability to form and maintain a proper barrier. The problem of how to maintain/restore skin barrier function is solved by developing amino acid formulations that target specific cellular pathways involved in skin barrier function and repair, thereby transforming amino acids from simple building blocks into small molecules capable of triggering signaling pathways that promote skin regeneration, including skin barrier integrity, repair, and function.

Results presented herein demonstrate that supplementation with particular amino acids or combinations thereof, either free amino acids or peptide combinations, can modulate transcription of key regulators of differentiation and proliferation, such as, but not limited to, EGF, PDGF, FGF2, and PPARD genes (or skin barrier markers), in normal primary human keratinocytes. Specific amino acids or combinations of amino acids (free amino acids or peptide combinations) that exhibit this functional ability were determined by analyzing their action on skin barrier markers. Notably, specific amino acids or combinations of certain amino acids, as free amino acids or peptide combinations of amino acids, exhibit a beneficial effect on skin regeneration including skin barrier function (such as alanine (Ala), arginine (Arg), glutamine (Gln), glycine (Gly), isoleucine (Ile), serine (Ser), threonine (Thr), tryptophan (Trp), and valine (Val), alone or in combination).

Described herein are formulations of specific combinations of amino acids for maintaining healthy skin, treating cosmetic skin conditions, and treating skin conditions, disorders, and diseases. In one aspect, described herein are formulations and methods for improving barrier integrity of the skin, promoting cellular proliferation, and/or development. As used herein, reference to “development” can include, for example, migration, maturation, and/or differentiation of skin cells. The disclosure also provides formulations and methods for treating and/or preventing a cosmetic skin condition or a skin condition associated with a disease or disorder of the skin. Such skin conditions include, without limitation, atopic dermatitis, psoriasis, conditions related to aging of the skin, pruritis, eczema, and/or a cosmetic condition. The disclosure also provides formulations and methods for treating wounds, abrasions, or burns.

Atopic Dermatitis (AD), for example, is the most common chronic inflammatory skin disease in people, affecting up to 15 million Americans (17% of children and 6% adults). Despite its high prevalence, effects on quality-of-life, and economic burden, there are still few effective treatments for AD and most have focused generally on inhibiting inflammation. AD is thought to develop in part as consequence of an acquired or genetic defect of the skin's barrier. The epidermis from AD subjects exhibit altered tight junctions (TJ), which was associated with reduced expression of selected TJ components (e.g., claudin). TJs seal the intercellular spaces between epithelial cells and the ‘tightness’ of this structure is dynamically regulated by endogenous or environmental factors. The regulation of the TJ seal is important for a variety of reasons, including appropriate trafficking patterns of ions, proteins, hydration, and even the penetration of immune cells.

The integrity of the tight junction dynamics between endothelial cells and epithelial cells, which regulates diffusion and maintains homeostasis of organs protected by physiological barriers, may be measured in-vitro using transepithelial/transendothelial electrical resistance (TEER). TEER can be used to identify agents that improve physiological barriers, and that therefore may have a positive impact on conditions associated with elevated levels of permeability of physiological barriers relative to a normal, healthy state. Conditions associated with elevated levels of permeability of physiological barriers include, e.g., atopic dermatitis and xerosis cutis. Results presented herein reveal that specific amino acid combinations improve/enhance barrier integrity of skin cells. Improvements in skin cell barrier integrity are thought to be mediated, at least in part, by tight junction dynamics and reflect reduced permeability of tight junctions. See, for example, FIGS. 1-6.

The subject may be a patient in which improving barrier integrity of the skin is needed. The patient may have this need due to, for example, exposure to chemical or physical irritants, age related predispositions, genetic predispositions, or skin inflammation. In one embodiment, the patient is asymptomatic. The subject can be any animal, including, for example, a human. In addition to humans, the animal may be, for example, mammals, such as rabbits, cattle, horses, sheep, pigs, goats, dogs, and cats.

As indicated above, skin barrier malfunction is implicated in common skin diseases like atopic dermatitis, psoriasis, and ichthyosis and a very common dermatological disorder, xerosis cutis (dry skin). Dry skin is a condition experienced by most people at some point in their lives. Seasonal xerosis is common during the cold, dry winter months, and the frequency of xerosis increases with age. Optimal management of many skin diseases includes appropriate skin care. Incorporation of a well-designed moisturizer/barrier repair formulation into a skin care regimen could contribute to improvement or mitigation of condition- or disease-associated symptoms.

An effective moisturizer/barrier repair formulation for dry xerotic skin that enhances skin's own ability to regenerate, restore and strengthen its own barrier function is desired, wherein the formulation uses an amino acid-based technology. These formulations offer new treatment options for dry scaly skin that benefit a large number of people suffering from same and at risk for developing more serious conditions that arise from unattended dry skin.

Physiologically, amino acids and their derivatives are important elements of the skin, as they are components of the Natural Moisturizing Factor (NMF) in the outermost layer of the skin, the stratum corneum (SC). Free amino acids (FAA) are produced by the hydrolysis of the keratinocyte protein filaggrin, corneodesmosomes, and SC keratins during normal physiological processes of epidermal maturation and terminal differentiation. NMF contributes to maintenance of adequate skin hydration. Some amino acids can effectively bind water and may, therefore, play a role in the regulation of skin hydration. Thus, specific amino acid supplementation to the skin is proposed herein to ensure proper levels of NMF and protein synthesis or production of certain significant amino acid metabolites [pyrrolidone carboxylic acid (PCA) and trans-urocanic acid (t-UCA)] that condition, nourish, and/or replenish the proteins in normal and xerotic skin, as well other skin conditions where the epidermal barrier is impaired.

Without being bound by theory, by delivering specific amino acid combinations to the skin, the skin's innate ability to form a proper barrier might be “fertilized”. The problem of how to maintain/restore skin barrier function is solved by developing amino acid formulations that target specific cellular pathways involved in skin barrier function and repair, thereby transforming amino acids from simple building blocks into small molecules capable of triggering signaling pathways that promote skin barrier function.

Results presented herein demonstrate that supplementation with particular amino acid combinations can modulate transcription of key regulators of differentiation and proliferation, such as, e.g., EGF and PDGF genes, in normal primary human keratinocytes. Specific combinations of amino acids that exhibit this functional ability was determined by analyzing their action on skin barrier markers. Notably, specific combinations of certain amino acids exhibit a beneficial effect on skin barrier function (such as Ser, Ala, Gly and Gln), while combinations of other amino acids (such as Cys, His and Tyr) have no effect or even negative effects on skin barrier function. Additional screening assays to identify the most effective combination/s of amino acids for improving barrier integrity of the skin/skin barrier function are performed, tested, and validated in preclinical studies using different skin models and different modes of administration (e.g., systemic and/or topical).

Specific aims include defining efficacious formulations of amino acids that promote skin barrier function (see FIGS. 1-6) and further evaluating and developing such amino acid formulations using 2D monolayer cultures (human differentiated keratinocytes) and 3D models (human epidermal skin equivalents). The effect of media supplementation with several amino acid combinations (e.g., the four core amino acid combination of Ser, Ala, Gly and Gln described above with and without Ile, Trp and/or Arg) on mRNA and protein levels of a panel of skin barrier markers that was identified based on reiterative analyses are determined. See, e.g., EXAMPLE 1.

Briefly, genes and proteins related to epidermal differentiation, proliferation and lipid regulation in skin were iteratively evaluated and weighted with respect to their structural/functional impact on skin barrier integrity. Determination of the panel of markers utilized herein that reflect barrier integrity of the skin was determined by a series of multivariate, iterative analyses. The complexity of the task is underscored by the voluminous array of potential genes that have been implicated in, e.g., barrier function, proliferation, differentiation, and inflammation, each of which biological processes could contribute to some degree in barrier integrity of the skin. Indeed, hundreds of genes have been implicated in these processes. The starting point for determining a suitable panel of markers that reflect barrier integrity of the skin, therefore, offers an indeterminant number of different potential skin barrier marker panels. Choosing a panel from this myriad of options required experimentation using multiple sample sources to avoid sample bias, reiterative analyses studying different endpoints, weighted significance of different genes in the skin barrier marker panel, and careful analysis of all the variables in these analytic approaches. The panel of skin barrier markers chosen to reflect barrier integrity of the skin is the result of the above analytical approach and thus, was determined following a mathematical, biological approach involving experimental results, statistical analyses thereof, and algorithmic analyses of all inputs collected.

Once the skin barrier marker panel was identified, a scoring system by which to evaluate different amino acid combinations under consideration was developed. The scoring system was developed to address the numerous variables arising from the experimental results. Such variables included, for example, degree of transcriptional upregulation, the number of different genes up- or down-regulated in response to an amino acid combination examined, and variation in readout observed among different sample sources. See, e.g., FIGS. 3 and 4.

Specific aims also include evaluating the effect of topical administration of the most promising amino acid formulations on epithelial barrier markers in in vitro and ex vivo models of normal and compromised skin. Briefly, amino acid formulations identified based on their superior ability to improve barrier function are used for topical treatment of normal and compromised epidermal skin equivalents and normal skin histocultures (skin explants). To generate an inflammatory skin phenotype, immature epidermal skin equivalents are pretreated with T-helper cell (Th-2)-derived cytokines in combination with TNFα and skin explants are delipidated with a tape-strip approach. The effect of treatment on the barrier function and integrity of tight junction (TJ) dynamics are investigated through a combination of gene expression, biochemical, morphological, and functional analyses (e.g., transepithelial/transendothelial electrical resistance (TEER) technique). Sec, e.g., EXAMPLES 4 and 5.

Specific aims include defining efficacious formulations of amino acids that promote skin barrier function and further evaluating and developing such amino acid formulations using 2D monolayer cultures (human differentiated keratinocytes) and 3D models (human epidermal skin equivalents). The effect of media supplementation with individual or several amino acid combinations (e.g., amino acids: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, alone or in combination, either as free amino acids or peptide combinations) on mRNA and protein levels of a panel of skin barrier markers that were identified based on reiterative analyses are determined. See, e.g., EXAMPLE 6.

Once the skin barrier marker panel was identified, a scoring system by which to evaluate different amino acids or combinations thereof under consideration was developed. The scoring system was developed to address the numerous variables arising from the experimental results. Such variables included, for example, degree of transcriptional upregulation, the number of different genes up- or down-regulated in response to an amino acid combination examined, and variation in readout observed among different sample sources. The average score of the different samples as compared to control on regulation of different skin barrier markers is shown, where a score of: 0-4 demonstrates a 1-fold to 2-fold change; 4-8 demonstrates a 2-fold to 3-fold change; and 8-13 demonstrates a 3-fold and greater fold change. Sec, e.g., FIG. 28. Additionally, the average fold change in skin regeneration marker, such as EGF, mRNA treated with the formulations described here from the average EGF mRNA control (no treatment) across a range of amino acid concentrations (i.e., 0.4 mM to 10 mM) comprises 1 average fold change or greater; 7 average fold change or less; or an average fold change of 1 to 7. See, e.g., FIG. 29.

Specific aims also include evaluating the effect of topical administration or parenteral administration (e.g., injection that is subcutaneous, intramuscular, or intravenous) of the most promising amino acid formulations on epithelial barrier markers in in vitro and ex vivo models of normal and compromised skin. Briefly, amino acid formulations identified based on their superior ability to improve barrier function are used for topical or parenteral treatment of normal and compromised epidermal skin equivalents and normal skin histocultures (skin explants). To generate an inflammatory skin phenotype, immature epidermal skin equivalents are pretreated with T-helper cell (Th-2)-derived cytokines in combination with TNFα and skin explants are delipidated with a tape-strip approach. The effect of treatment on the barrier function and integrity of tight junction (TJ) dynamics are investigated through a combination of gene expression, biochemical, morphological, and functional analyses (e.g., transepithelial/transendothelial electrical resistance (TEER) technique).

The identification of effective amino acid formulations and use thereof in skin regeneration, including skin barrier repair or treatments of skin diseases, skin conditions, or osteoarthritis, or symptoms thereof, therefore, provides a solution to the problem of how to maintain and restore healthy skin as people age and under conditions of duress and/or disease or how to treat osteoarthritis. Formulations identified herein, therefore, satisfy a long-sought need for scientifically validated barrier repair or treatments of skin diseases, skin conditions, or osteoarthritis, or symptoms thereof that address the underlying biological causes of skin conditions (e.g., xerosis cutis) or osteoarthritis pain/discomfort, rather than just the symptoms thereof, Proof-of-concept studies are conducted on human subjects using the most promising amino acid-based formulations.

The identification of effective amino acid formulations and use thereof in barrier repair formulations/moisturizers, therefore, provides a solution to the problem of how to maintain and restore healthy skin as people age and under conditions of duress and/or disease. Formulations identified herein, therefore, satisfy a long-sought need for scientifically validated barrier repair formulations/moisturizers that address the underlying biological causes of skin conditions (e.g., xerosis cutis), rather than just the symptoms thereof, Proof-of-concept studies are conducted on human subjects using the most promising amino acid-based cosmetic formulations.

Formulations of the Disclosure

Various embodiments of the disclosure provide a formulation for improving skin regeneration or treating a musculoskeletal disease or condition, comprising: as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or derivatives or salts thereof, or combinations thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1. For example, if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine). Such formulations improve skin regeneration, including skin barrier repair or integrity, or treats osteoarthritis, when tested by a method detecting expression of a skin regeneration or skin barrier marker gene (e.g., CPT2, EGF, FGF2, FLG, MKI67, PDGF. PPARD, TGM4) and determining that the skin regeneration or skin barrier marker gene is modulated by a 1-7 average fold change (e.g., 1 average fold or greater (e.g., 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1; 11.3; 11.5; 11.7; 11.9; 12.1; 12.3; 12.5; 12.7; 12.9; 13.1; 13.3; 13.5; 13.7; 13.9; 14.1; 14.3; 14.5; 14.7; 14.9; 15.1; 15.3; 15.5; 15.7; 15.9; 16.1; 16.3; 16.5; 16.7; 16.9; 17.1; 17.3; 17.5; 17.7; 17.9; 18.1; 18.3; 18.5; 18.7; 18.9; 19.1; 19.3; 19.5; 19.7; 19.9; 20.1; 20.3; 20.5; 20.7; 20.9; 21.1; 21.3; 21.5; 21.7; 21.9; 22.1; 22.3; 22.5; 22.7; 22.9; 23.1; 23.3; 23.5; 23.7; 23.9; 23.1; 23.3; 23.5; 23.7; 23.9; 24.1; 24.3; 24.5; 24.7; 24.9; 25.1; 25.3; 25.5; 25.7; 25.9; 26.1; 26.3; 26.5; 26.7; 26.9; 27.1; 27.3; 27.5; 27.7; 27.9; 28.1; 28.3; 28.5; 28.7; 28.9; 29.1; 29.3; 29.5; 29.7; 29.9; 30.1; 30.3; 30.5; 30.7; 30.9; 31.1; 31.3; 31.5; 31.7; 31.9; 32.1; 32.3; 32.5; 32.7; 32.9; 33.1; 33.3; 33.5; 33.7; 33.9; 34.1; 34.3; 34.5; 34.7; 34.9; 35.1; 35.3; 35.5; 35.7; 35.9; 36.1; 36.3; 36.5; 36.7; 36.9; 37.1; 37.3; 37.5; 37.7; 37.9; 38.1; 38.3; 38.5; 38.7; 38.9; 39.1; 39.3; 39.5; 39.7; 39.9; 40.1; 40.3; 40.5; 40.7; 40.9; 41.1; 41.3; 41.5); 40 average fold or less (e.g., 39.8; 39.6; 39.4; 39.2; 39; 38.8; 38.6; 38.4; 38.2; 38; 37.8; 37.6; 37.4; 37.2; 37; 36.8; 36.6; 36.4; 36.2; 36; 35.8; 35.6; 35.4; 35.2; 35; 34.8; 34.6; 34.4; 34.2; 34; 33.8; 33.6; 33.4; 33.2; 33; 32.8; 32.6; 32.4; 32.2; 32; 31.8; 31.6; 31.4; 31.2; 31; 30.8; 30.6; 30.4; 30.2; 30; 29.8; 29.6; 29.4; 29.2; 29; 28.8; 28.6; 28.4; 28.2; 28; 27.8; 27.6; 27.4; 27.2; 27; 26.8; 26.6; 26.4; 26.2; 26; 25.8; 25.6; 25.4; 25.2; 25; 24.8; 24.6; 24.4; 24.2; 24; 23.8; 23.6; 23.4; 23.2; 23; 22.8; 22.6; 22.4; 22.2; 22; 21.8; 21.6; 21.4; 21.2; 21; 20.8; 20.6; 20.4; 20.2; 20; 19.8; 19.6; 19.4; 19.2; 19; 18.8; 18.6; 18.4; 18.2; 18; 17.8; 17.6; 17.4; 17.2; 17.2; 17; 16.8; 16.6; 16.4; 16.2; 16; 15.8; 15.6; 15.4; 15.2; 15; 14.8; 14.6; 14.4; 14.2; 14; 13.8; 13.6; 13.4; 13.2; 13; 12.8; 12.6; 12.4; 12.2; 12; 11.8; 11.6; 11.4; 11.2; 11; 10.8; 10.6; 10.4; 10.2; 10; 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6; 0.4); or 1 average fold-40 average fold (e.g., 1.1-39.9; 1.2-39.8; 1.3-39.7; 1.4-39.6; 1.5-39.5; 1.6-39.4; 1.7-39.3; 1.8-39.2; 1.9-39.1; 2-39; 2.1-38.9; 2.2-38.8; 2.3-38.7; 2.4-38.6; 2.5-38.5; 2.6-38.4; 2.7-38.3; 2.8-38.2; 2.9-38.1; 3-38; 3.1-37.9; 3.2-37.8; 3.3-37.7; 3.4-37.6; 3.5-37.5; 3.6-37.4; 3.7-37.3; 3.8-37.2; 3.9-37.1; 4-37; 4.1-36.9; 4.2-36.8; 4.3-36.7; 4.4-36.6; 4.5-36.5; 4.6-36.4; 4.7-36.3; 4.8-36.2; 4.9-36.1; 5-36; 5.1-35.9; 5.2-35.8; 5.3-35.7; 5.4-35.6; 5.5-35.5; 5.6-35.4; 5.7-35.3; 5.8-35.2; 5.9-35.1; 6-35; 6.1-34.9; 6.2-34.8; 6.3-34.7; 6.4-34.6; 6.5-34.5; 6.6-34.4; 6.7-34.3; 6.8-34.2; 6.9-34.1; 7-34; 7.1-33.9; 7.2-33.8; 7.3-33.7; 7.4-33.6; 7.5-33.5; 7.6-33.4; 7.7-33.3; 7.8-33.2; 7.9-33.1; 8-33; 8.1-32.9; 8.2-32.8; 8.3-32.7; 8.4-32.6; 8.5-32.5; 8.6-32.4; 8.7-32.3; 8.8-32.2; 8.9-32.1; 9-32; 9.1-31.9; 9.2-31.8; 9.3-31.7; 9.4-31.6; 9.5-31.5; 9.6-31.4; 9.7-31.3; 9.8-31.2; 9.9-31.1; 10-31; 10.1-30.9; 10.2-30.8; 10.3-30.7; 10.4-30.6; 10.5-30.5; 10.6-30.4; 10.7-30.3; 10.8-30.2; 10.9-30.1; 11 30; 11.1-29.9; 11.2-29.8; 11.3-29.7; 11.4-29.6; 11.5-29.5; 11.6-29.4; 11.7-29.3; 11.8-29.2; 11.9-29.1; 12-29; 12.1-28.9; 12.2-28.8; 12.3-28.7; 12.4-28.6; 12.5-28.5; 12.6-28.4; 12.7-28.3; 12.8-28.2; 12.9-28.1; 13-28; 13.1-27.9; 13.2-27.8; 13.3-27.7; 13.4-27.6; 13.5-27.5; 13.6-27.4; 13.7-27.3; 13.8-27.2; 13.9-27.1; 14-27; 14.1-26.9; 14.2-26.8; 14.3-26.7; 14.4-26.6; 14.5-26.5; 14.6-26.4; 14.7-26.3; 14.8-26.2; 14.9-26.1; 15-26; 15.1-25.9; 15.2-25.8; 15.3-25.7; 15.4-25.6; 15.5-25.5; 15.6-25.4; 15.7-25.3; 15.8-25.2; 15.9-25.1; 16-25; 16.1-24.9; 16.2-24.8; 16.3-24.7; 16.4-24.6; 16.5-24.5; 16.6-24.4; 16.7-24.3; 16.8-24.2; 16.9-24.1; 17-24; 17.1-23.9; 17.2-23.8; 17.3-23.7; 17.4-23.6; 17.5-23.5; 17.6-23.4; 17.7-23.3; 17.8-23.2; 17.9-23.1; 18-23; 18.1-22.9; 18.2-22.8; 18.3-22.7; 18.4-22.6; 18.5-22.5; 18.6-22.4; 18.7-22.3; 18.8-22.2; 18.9-22.1; 19-22; 19.1-21.9; 19.2-21.8; 19.3-21.7; 19.4-21.6; 19.5-21.5; 19.6-21.4; 19.7-21.3; 19.8-21.2; 19.9-21.1; 20-21; 20.1-20.9; 20.2-20.8; 20.3-20.7; 20.4-20.6)), of formulation-treated cells as compared to untreated cells; and optionally, a dermatologically or physiologically acceptable carrier; where the formulation improves barrier integrity of the skin or treats a musculoskeletal disease or condition, e.g., osteoarthritis, or its symptoms thereof. For example, the average fold change in EGF mRNA treated with the formulations containing the specified amino acids from the average EGF mRNA control (no treatment) over a range of amino acid concentrations from 0.4 mM to 10 mM comprises an average fold change of: 2.2 (Glutamine, Glycine); 1.9 (Glutamine, Glycine, Alanine, Serine); 4.4 (Glutamine, Glycine, Alanine, Serine, Isoleucine); 3.4 (Glutamine, Glycine, Alanine, Serine, Isoleucine; Valine); 3.1 (Glutamine, Glycine, Alanine, Serine, Isoleucine, Valine, Arginine); 1 (Glutamine, Glycine, Alanine, Serine, Isoleucine, Valine, Arginine, Threonine); 2.2 (Glutamine, Glycine, Alanine, Serine, Isoleucine, Valine, Arginine, Threonine, Tryptophan). In some embodiments, such formulations of the disclosure are formulated as topical formulations (e.g., transdermal, epicutaneous) or parenteral formulations (e.g., subcutaneous, intradermal, intra-articular, intrasynovial, periarticular, intramuscular, intravenous). In one embodiment, the formulation optionally comprises, for example, pharmaceutically acceptable carriers, adjuvants, other active agents, and additives (e.g., sugars, electrolytes, vitamins, minerals, etc.).

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: alanine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: glutamine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: glycine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: serine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: arginine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: isoleucine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount comprising, consisting essentially of, or consisting of: threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: alanine and glycine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: alanine and glutamine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: alanine and serine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: glutamine and serine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: alanine and arginine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: glutamine and isoleucine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: glutamine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: glutamine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of an amino acid combination (free amino acids or peptide combination) comprising, consisting essentially of, or consisting of: glutamine and valine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and arginine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and arginine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and arginine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and arginine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and arginine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and arginine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of arginine and threonine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of isoleucine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of isoleucine and arginine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of isoleucine and arginine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and arginine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and arginine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and serine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and arginine and threonine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and isoleucine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and isoleucine and arginine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and isoleucine and arginine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and arginine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and arginine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and arginine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and arginine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and arginine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and arginine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and arginine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and isoleucine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and arginine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and arginine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and isoleucine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and isoleucine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and isoleucine and arginine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and threonine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and arginine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and isoleucine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and arginine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and isoleucine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and valine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and threonine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and arginine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and arginine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and isoleucine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and isoleucine and threonine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and isoleucine and arginine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and arginine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and isoleucine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and arginine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and isoleucine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and valine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and arginine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and isoleucine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and valine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and arginine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and isoleucine and threonine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and isoleucine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and arginine and threonine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and arginine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and arginine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and arginine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and arginine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and arginine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and isoleucine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and arginine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and arginine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and isoleucine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and isoleucine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and isoleucine and arginine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and arginine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and isoleucine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and arginine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and isoleucine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and valine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and arginine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and isoleucine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and isoleucine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and isoleucine and arginine and threonine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and arginine and threonine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine and arginine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and arginine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and arginine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine and arginine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and arginine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and arginine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and arginine and threonine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and threonine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and arginine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of isoleucine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and arginine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and arginine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and isoleucine and arginine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and arginine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and arginine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and arginine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and arginine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and arginine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and arginine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and isoleucine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and isoleucine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and arginine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and isoleucine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and isoleucine and arginine and

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and isoleucine and arginine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and arginine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and isoleucine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and arginine and threonine.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and isoleucine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and isoleucine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and isoleucine and arginine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and valine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and valine and arginine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and arginine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and isoleucine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and isoleucine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and arginine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and isoleucine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and isoleucine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and isoleucine and arginine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and arginine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and isoleucine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and isoleucine and arginine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and valine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and valine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and threonine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and arginine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and isoleucine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and isoleucine and arginine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and valine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and valine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and serine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and serine and arginine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and serine and valine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and isoleucine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and arginine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and arginine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and arginine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and arginine and threonine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and arginine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and arginine and threonine.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and isoleucine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and isoleucine and arginine.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and arginine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and isoleucine and threonine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and isoleucine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and isoleucine and arginine and threonine.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and arginine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and isoleucine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and isoleucine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and arginine and threonine.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and isoleucine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and isoleucine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and isoleucine and arginine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and valine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and valine and arginine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and isoleucine and arginine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine and arginine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and arginine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and isoleucine and arginine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and arginine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine and threonine.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine and arginine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and isoleucine and arginine and threonine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and arginine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and arginine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and arginine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of valine and isoleucine and arginine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and valine and isoleucine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and isoleucine and arginine and threonine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and arginine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and serine and valine and isoleucine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and isoleucine and threonine and tryptophan.

In various implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and valine and isoleucine and arginine and threonine.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and arginine and threonine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and isoleucine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and valine and isoleucine and threonine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and isoleucine and arginine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and arginine and threonine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and isoleucine and arginine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and valine and isoleucine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and valine and isoleucine and arginine.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and glycine and isoleucine and arginine and tryptophan.

In some embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and valine and isoleucine and arginine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and isoleucine and arginine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and arginine and threonine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and isoleucine and arginine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and valine and isoleucine and arginine and threonine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and isoleucine and arginine and tryptophan.

In various embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and valine and isoleucine and arginine.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and valine and isoleucine and arginine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and arginine and threonine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine and arginine and tryptophan.

In certain embodiments, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of serine and valine and isoleucine and arginine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and valine and isoleucine and arginine and threonine and tryptophan.

In some aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and isoleucine and arginine and threonine and tryptophan.

In various aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glutamine and serine and valine and isoleucine and threonine and tryptophan.

In some implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and serine and valine and isoleucine and arginine and threonine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glutamine and glycine and serine and valine and isoleucine and arginine and tryptophan.

In certain implementations, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of glycine and serine and valine and isoleucine and arginine and threonine and tryptophan.

In certain aspects, the formulation described here comprises, consists essentially of, or consists of a therapeutically effective amount of alanine and glycine and serine and valine and isoleucine and arginine and threonine and tryptophan.

Accordingly, in certain embodiments, the present disclosure provides methods for treating and/or preventing a skin condition in a subject in need thereof, the method comprising topically administering to the subject any of the formulations described herein. In certain aspects, the skin condition is dry skin, dermatitis, atopic dermatitis, radiodermatitis, eczema, ichthyosis, pruritus (itch), psoriasis, aging of skin, a condition related to the aging of skin (e.g., wrinkles, loss of suppleness, increase is roughness), burns, or wounds. In further aspects, the skin condition is a cosmetic skin condition. The formulation of the present disclosure can also be administered at a surgical site, including at a site of minimally invasive surgery, to improve healing and the surgical outcome.

In certain aspects, the disclosure provides methods and uses of the described formulation for improving skin barrier integrity and skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration); and optionally, a dermatologically acceptable carrier, where the formulation improves barrier integrity of skin cells. In a various embodiments, the formulation is a topical or a parenteral skin formulation that improves skin barrier integrity, when tested by a method detecting expression of a barrier marker gene (e.g., EGF, PDGF, FGF2, PPARD, CPT2, FLG, MKI67, TGM4) and determining that the barrier marker gene is modulated by a 1-7 average fold change of formulation-treated skin cells as compared to amino acid-untreated skin cells; and optionally, a dermatologically acceptable carrier; where the formulation improves barrier integrity of the skin. The average fold increase of the skin regeneration marker gene expression over an amino acid concentration of 0.4 mM to 10 mM is; 1 average fold or greater (e.g., 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1; 11.3; 11.5; 11.7; 11.9; 12.1; 12.3; 12.5; 12.7; 12.9; 13.1; 13.3; 13.5; 13.7; 13.9; 14.1; 14.3; 14.5; 14.7; 14.9; 15.1; 15.3; 15.5; 15.7; 15.9; 16.1; 16.3; 16.5; 16.7; 16.9; 17.1; 17.3; 17.5; 17.7; 17.9; 18.1; 18.3; 18.5; 18.7; 18.9; 19.1; 19.3; 19.5; 19.7; 19.9; 20.1; 20.3; 20.5; 20.7; 20.9; 21.1; 21.3; 21.5; 21.7; 21.9; 22.1; 22.3; 22.5; 22.7; 22.9; 23.1; 23.3; 23.5; 23.7; 23.9; 23.1; 23.3; 23.5; 23.7; 23.9; 24.1; 24.3; 24.5; 24.7; 24.9; 25.1; 25.3; 25.5; 25.7; 25.9; 26.1; 26.3; 26.5; 26.7; 26.9; 27.1; 27.3; 27.5; 27.7; 27.9; 28.1; 28.3; 28.5; 28.7; 28.9; 29.1; 29.3; 29.5; 29.7; 29.9; 30.1; 30.3; 30.5; 30.7; 30.9; 31.1; 31.3; 31.5; 31.7; 31.9; 32.1; 32.3; 32.5; 32.7; 32.9; 33.1; 33.3; 33.5; 33.7; 33.9; 34.1; 34.3; 34.5; 34.7; 34.9; 35.1; 35.3; 35.5; 35.7; 35.9; 36.1; 36.3; 36.5; 36.7; 36.9; 37.1; 37.3; 37.5; 37.7; 37.9; 38.1; 38.3; 38.5; 38.7; 38.9; 39.1; 39.3; 39.5; 39.7; 39.9; 40.1; 40.3; 40.5; 40.7; 40.9; 41.1; 41.3; 41.5); 40 average fold or less (e.g., 39.8; 39.6; 39.4; 39.2; 39; 38.8; 38.6; 38.4; 38.2; 38; 37.8; 37.6; 37.4; 37.2; 37; 36.8; 36.6; 36.4; 36.2; 36; 35.8; 35.6; 35.4; 35.2; 35; 34.8; 34.6; 34.4; 34.2; 34; 33.8; 33.6; 33.4; 33.2; 33; 32.8; 32.6; 32.4; 32.2; 32; 31.8; 31.6; 31.4; 31.2; 31; 30.8; 30.6; 30.4; 30.2; 30; 29.8; 29.6; 29.4; 29.2; 29; 28.8; 28.6; 28.4; 28.2; 28; 27.8; 27.6; 27.4; 27.2; 27; 26.8; 26.6; 26.4; 26.2; 26; 25.8; 25.6; 25.4; 25.2; 25; 24.8; 24.6; 24.4; 24.2; 24; 23.8; 23.6; 23.4; 23.2; 23; 22.8; 22.6; 22.4; 22.2; 22; 21.8; 21.6; 21.4; 21.2; 21; 20.8; 20.6; 20.4; 20.2; 20; 19.8; 19.6; 19.4; 19.2; 19; 18.8; 18.6; 18.4; 18.2; 18; 17.8; 17.6; 17.4; 17.2; 17.2; 17; 16.8; 16.6; 16.4; 16.2; 16; 15.8; 15.6; 15.4; 15.2; 15; 14.8; 14.6; 14.4; 14.2; 14; 13.8; 13.6; 13.4; 13.2; 13; 12.8; 12.6; 12.4; 12.2; 12; 11.8; 11.6; 11.4; 11.2; 11; 10.8; 10.6; 10.4; 10.2; 10; 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6; 0.4); or 1 average fold-40 average fold (e.g., 1.1-39.9; 1.2-39.8; 1.3-39.7; 1.4-39.6; 1.5-39.5; 1.6-39.4; 1.7-39.3; 1.8-39.2; 1.9-39.1; 2-39; 2.1-38.9; 2.2-38.8; 2.3-38.7; 2.4-38.6; 2.5-38.5; 2.6-38.4; 2.7-38.3; 2.8-38.2; 2.9-38.1; 3-38; 3.1-37.9; 3.2-37.8; 3.3-37.7; 3.4-37.6; 3.5-37.5; 3.6-37.4; 3.7-37.3; 3.8-37.2; 3.9-37.1; 4-37; 4.1-36.9; 4.2-36.8; 4.3-36.7; 4.4-36.6; 4.5-36.5; 4.6-36.4; 4.7-36.3; 4.8-36.2; 4.9-36.1; 5-36; 5.1-35.9; 5.2-35.8; 5.3-35.7; 5.4-35.6; 5.5-35.5; 5.6-35.4; 5.7-35.3; 5.8-35.2; 5.9-35.1; 6-35; 6.1-34.9; 6.2-34.8; 6.3-34.7; 6.4-34.6; 6.5-34.5; 6.6-34.4; 6.7-34.3; 6.8-34.2; 6.9-34.1; 7-34; 7.1-33.9; 7.2-33.8; 7.3-33.7; 7.4-33.6; 7.5-33.5; 7.6-33.4; 7.7-33.3; 7.8-33.2; 7.9-33.1; 8-33; 8.1-32.9; 8.2-32.8; 8.3-32.7; 8.4-32.6; 8.5-32.5; 8.6-32.4; 8.7-32.3; 8.8-32.2; 8.9-32.1; 9-32; 9.1-31.9; 9.2-31.8; 9.3-31.7; 9.4-31.6; 9.5-31.5; 9.6-31.4; 9.7-31.3; 9.8-31.2; 9.9-31.1; 10-31; 10.1-30.9; 10.2-30.8; 10.3-30.7; 10.4-30.6; 10.5-30.5; 10.6-30.4; 10.7-30.3; 10.8-30.2; 10.9-30.1; 11-30; 11.1-29.9; 11.2-29.8; 11.3-29.7; 11.4-29.6; 11.5-29.5; 11.6-29.4; 11.7-29.3; 11.8-29.2; 11.9-29.1; 12-29; 12.1-28.9; 12.2-28.8; 12.3-28.7; 12.4-28.6; 12.5-28.5; 12.6-28.4; 12.7-28.3; 12.8-28.2; 12.9-28.1; 13-28; 13.1-27.9; 13.2-27.8; 13.3-27.7; 13.4-27.6; 13.5-27.5; 13.6-27.4; 13.7-27.3; 13.8-27.2; 13.9-27.1; 14-27; 14.1-26.9; 14.2-26.8; 14.3-26.7; 14.4-26.6; 14.5-26.5; 14.6-26.4; 14.7-26.3; 14.8-26.2; 14.9-26.1; 15-26; 15.1-25.9; 15.2-25.8; 15.3-25.7; 15.4-25.6; 15.5-25.5; 15.6-25.4; 15.7-25.3; 15.8-25.2; 15.9-25.1; 16-25; 16.1-24.9; 16.2-24.8; 16.3-24.7; 16.4-24.6; 16.5-24.5; 16.6-24.4; 16.7-24.3; 16.8-24.2; 16.9-24.1; 17-24; 17.1-23.9; 17.2-23.8; 17.3-23.7; 17.4-23.6; 17.5-23.5; 17.6-23.4; 17.7-23.3; 17.8-23.2; 17.9-23.1; 18-23; 18.1-22.9; 18.2-22.8; 18.3-22.7; 18.4-22.6; 18.5-22.5; 18.6-22.4; 18.7-22.3; 18.8-22.2; 18.9-22.1; 19-22; 19.1-21.9; 19.2-21.8; 19.3-21.7; 19.4-21.6; 19.5-21.5; 19.6-21.4; 19.7-21.3; 19.8-21.2; 19.9-21.1; 20-21; 20.1-20.9; 20.2-20.8; 20.3-20.7; 20.4-20.6). Improving barrier integrity and function also improves skin moisture

In some embodiments of the formulations described herein, where, if present, amino acids of alanine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of glutamine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of glycine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of serine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of arginine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of isoleucine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of threonine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of tryptophan are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of valine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); or any combination thereof.

Or, in certain embodiments, even if these amino acids are present in the formulation, they are not present in an amount that would affect barrier integrity of the skin. By “negligible” it is meant that the specific amino acid present has no effect on barrier integrity of the skin. By “negligible” it is meant that the specific amino acid present has no effect on a disease or condition that is related to barrier integrity of the skin, e.g., wound healing, treating skin conditions (e.g., atopic dermatitis, psoriasis, or condition related to the aging of skin) in a subject in need thereof.

The formulations described here comprise a therapeutically effective amount of an amino acid or combination of amino acids (free amino acids or peptide combination), where the amino acid comprises a concentration of a concentration ranging from 0.1 mM or greater (e.g., 0.3, 0.5, 0.7; 0.9; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6, 1; 6.3; 6.5; 6, 7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5); of 10 mM or less (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0, 6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02); or ranging from 0.1 mM to 10 mM (e.g., 0.2 mM to 9.9 mM; 0.3 mM to 9.8 mM; 0.4 mM to 9.7 mM; 0.5 mM to 9.6 mM; 0.6 mM to 9.5 mM; 0.7 mM to 9.4 mM; 0.8 mM to 9.3 mM; 0.9 mM to 9.2 mM; 1 mM to 9.1 mM; 1.1 mM to 9 mM; 1.2 mM to 8.9 mM; 1.3 mM to 8.8 mM; 1.4 mM to 8.7 mM; 1.5 mM to 8.6 mM; 1.6 mM to 8.5 mM; 1.7 mM to 8.4 mM; 1.8 mM to 8.3 mM; 1.9 mM to 8.2 mM; 2 mM to 8.1 mM; 2.1 mM to 8 mM; 2.2 mM to 7.9 mM; 2.3 mM to 7.8 mM; 2.4 mM to 7.7 mM; 2.5 mM to 7.6 mM; 2.6 mM to 7.5 mM; 2.7 mM to 7.4 mM; 2.8 mM to 7.3 mM; 2.9 mM to 7.2 mM; 3 mM to 7.1 mM; 3.1 mM to 7 mM; 3.2 mM to 6.9 mM; 3.3 mM to 6.8 mM; 3.4 mM to 6.7 mM; 3.5 mM to 6.6 mM; 3.6 mM to 6.5 mM; 3.7 mM to 6.4 mM; 3.8 mM to 6.3 mM; 3.9 mM to 6.2 mM; 4 mM to 6.1 mM; 4.1 mM to 6 mM; 4.2 mM to 5.9 mM; 4.3 mM to 5.8 mM; 4.4 mM to 5.7 mM; 4.5 mM to 5.6 mM; 4.6 mM to 5.5 mM; 4.7 mM to 5.4 mM; 4.8 mM to 5.3 mM; 4.9 mM to 5.2 mM; 5 mM to 5.1 mM), For example, 4 mM of each amino acid comprises: 0.0356% alanine; 0.0584% glutamine; 0.03% glycine; and 0.042% serine. In some aspects, each of the amino acids (4% or 4 mM) in the formulations described here are about 100-fold greater than the concentrations that have been used in tissue culture (TC) experiments.

In some embodiments described here, the disclosed formulation or the topical formulation are provided, wherein the therapeutically effective amount of each amino acid, or each amino acid of a combination of free amino acids or a peptide combination thereof, is present at a weight percent to the formulation (wt/wt %) of: 0.001 wt % or greater (e.g., 0.002; 0.003; 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9; 1); 1 wt % or less (e.g., 0.95; 0.85; 0.75; 0.65; 0.55; 0.45; 0.35; 0.25; 0.15; 0.095; 0.085; 0.075; 0.065; 0.055; 0.045; 0.035; 0.025; 0.015; 0.0095; 0.0085; 0.0075; 0.0065; 0.0055; 0.0045; 0.0035; 0.0025; 0.0015); or 0.001 wt % to 1 wt % (e.g., 0.0012 wt % to 0.99 wt %; 0.0014 wt % to 0.97 wt %; 0.0016 wt % to 0.95 wt %; 0.0018 wt % to 0.93 wt %; 0.002 wt % to 0.91 wt %; 0.0022 wt % to 0.89 wt %; 0.0024 wt % to 0.87 wt %; 0.0026 wt % to 0.85 wt %; 0.0028 wt % to 0.83 wt %; 0.003 wt % to 0.81 wt %; 0.0032 wt % to 0.79 wt %; 0.0034 wt % to 0.77 wt %; 0.0036 wt % to 0.75 wt %; 0.0038 wt % to 0.73 wt %; 0.004 wt % to 0.71 wt %; 0.0042 wt % to 0.69 wt %; 0.0044 wt % to 0.67 wt %; 0.0046 wt % to 0.65 wt %; 0.0048 wt % to 0.63 wt %; 0.005 wt % to 0.61 wt %; 0.0052 wt % to 0.59 wt %; 0.0054 wt % to 0.57 wt %; 0.0056 wt % to 0.55 wt %; 0.0058 wt % to 0.53 wt %; 0.006 wt % to 0.51 wt %; 0.0062 wt % to 0.49 wt %; 0.0064 wt % to 0.47 wt %; 0.0066 wt % to 0.45 wt %; 0.0068 wt % to 0.43 wt %; 0.007 wt % to 0.41 wt %; 0.0072 wt % to 0.39 wt %; 0.0074 wt % to 0.37 wt %; 0.0076 wt % to 0.35 wt %; 0.0078 wt % to 0.33 wt %; 0.008 wt % to 0.31 wt %; 0.0082 wt % to 0.29 wt %; 0.0084 wt % to 0.27 wt %; 0.0086 wt % to 0.25 wt %; 0.0088 wt % to 0.23 wt %; 0.009 wt % to 0.21 wt %; 0.0092 wt % to 0.19 wt %; 0.0094 wt % to 0.17 wt %; 0.0096 wt % to 0.15 wt %; 0.0098 wt % to 0.13 wt %; 0.01 wt % to 0.11 wt %; 0.012 wt % to 0.099 wt %; 0.014 wt % to 0.097 wt %; 0.016 wt % to 0.095 wt %; 0.018 wt % to 0.093 wt %; 0.02 wt % to 0.091 wt %; 0.022 wt % to 0.089 wt %; 0.024 wt % to 0.087 wt %; 0.026 wt % to 0.085 wt %; 0.028 wt % to 0.083 wt %; 0.03 wt % to 0.081 wt %; 0.032 wt % to 0.079 wt %; 0.034 wt % to 0.077 wt %; 0.036 wt % to 0.075 wt %; 0.038 wt % to 0.073 wt %; 0.04 wt % to 0.071 wt %; 0.042 wt % to 0.069 wt %; 0.044 wt % to 0.067 wt %; 0.046 wt % to 0.065 wt %; 0.048 wt % to 0.063 wt %; 0.05 wt % to 0.061 wt %; 0.052 wt % to 0.059 wt %; 0.054 wt % to 0.057 wt %).

In one embodiment, the total osmolarity of the formulation is from about 10 mOsm or greater (e.g., 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300); 280 mOsm or less (e.g., 275, 265, 255, 245, 235, 225, 215, 205, 195, 185, 175, 165, 155, 145, 135, 125, 115, 105, 95, 85, 75, 65, 55, 45, 35, 25, 15, 5); or 10 mOsm-280 mOsm (e.g., 20 mOsm-270 mOsm; 30 mOsm-260 mOsm; 40 mOsm-250 mOsm; 50 mOsm-240 mOsm; 60 mOsm-230 mOsm; 70 mOsm-220 mOsm; 80 mOsm-210 mOsm; 90 mOsm-200 mOsm; 100 mOsm-190 mOsm; 110 mOsm-180 mOsm; 120 mOsm-170 mOsm; 130 mOsm-160 mOsm; 140 mOsm-150 mOsm).

In some embodiments, the formulation has a pH from, for example, about 2.5 or greater (e.g., 3.5, 4.5, 5.5, 6.5, 7.5, 8.5, 9.5, 10.5); a pH of about 8.5 or less (e.g., 8, 7, 6, 5, 4, 3, 2, 1); or a pH ranging from 2.5-8.5 (e.g., 2.6-8.4; 2.7-8.3; 2.8-8.2; 2.9-8.1; 3-8; 3.1-7.9; 3.2-7.8; 3.3-7.7; 3.4-7.6; 3.5-7.5; 3.6-7.4; 3.7-7.3; 3.8-7.2; 3.9-7.1; 4-7; 4.1-6.9; 4.2-6.8; 4.3-6.7; 4.4-6.6; 4.5-6.5; 4.6-6.4; 4.7-6.3; 4.8-6.2; 4.9-6.1; 5-6; 5.1-5.9; 5.2-5.8; 5.3-5.7; 5.4-5.6).

In one embodiment, the formulation optionally comprises, for example, pharmaceutically acceptable carriers, adjuvants, other active agents, and additives (e.g., sugars, electrolytes, vitamins, minerals, etc.). Formulation may further comprise, for example, a prebiotic or probiotic substance.

In some embodiments of the formulation described here, the formulation does not comprise amino acids of cysteine, histidine, tyrosine, leucine, aspartic acid, taurate, taurine, or glutamate, or any combination thereof. In some embodiments of formulations described herein, wherein, if present, amino acids of cysteine are present at a concentration of equal to or less than 1 mM; if present, amino acids of histidine are present at a concentration of equal to or less than 0.5 mM; if present, amino acids of tyrosine are present at a concentration of equal to or less than 1 mM; if present, amino acids of leucine are present at a concentration of equal to or less than 4 mM; or if present, amino acids of aspartic acid are present at a concentration of equal to or less than 2 mM; or any combination thereof.

In certain aspects, the formulation for use in methods of treating and/or preventing skin conditions (e.g., Xerosis cutis, atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, a wound, burn, or a condition relating to aging of the skin), improve skin barrier integrity or function, or improve skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))) comprises: as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount comprising, consisting essentially of, or consisting of one or more of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine; and optionally, a dermatologically acceptable or pharmaceutically acceptable carrier; where the therapeutically effective amount improves barrier integrity of skin cells; where the formulation improves barrier integrity of the skin. Some aspects of the disclosure are directed to such formulations for treating and/or preventing Xerosis cutis, atopic dermatitis, dermatitis, radiodermatits, psoriasis, pruritus, eczema, a wound, burn, or a condition relating to aging of the skin, that are administered topically or by injection (e.g., intradermal, subcutaneous, intramuscular).

In some embodiments, the formulation is a solution. The formulation can be administered with other therapeutic agents.

Additional embodiments described here provides the formulation or the topical formulation disclosed here, where the therapeutically effective amount of the free amino acid or combination of free amino acids, or peptide combination of amino acids, comprises, consists essentially of, or consists of a therapeutically effective amount of one or more amino acids selected from: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, for skin barrier integrity (repair and/or strengthening), or for improving skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))). In some embodiments, the formulation described here comprising a therapeutically effective amount of amino acids comprising, consisting essentially of, or consisting of, if present, alanine (4 mM) of 0.0356%; glutamine (4 mM) of 0.0584%; glycine (4 mM) of 0.03%; or serine (4 mM) of 0.042%).

Other embodiments described here provide the formulation or the topical formulation disclosed here, wherein the therapeutically effective amount of free amino acids, a combination of free amino acids or a peptide combination of amino acids comprises (or consists essentially of or consists of a therapeutically effective amount of), if present, one or more of the amino acids selected from: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, wherein each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1.

In some embodiments, the formulation is administered systemically or locally. In some embodiments, the formulation is used for treating a disease or condition that is related to barrier integrity of the skin, e.g., xerotic skin, wound healing, treating skin conditions (e.g., is atopic dermatitis, dermatitis, psoriasis, aging of skin, a condition related to the aging of skin, pruritus, or eczema), and/or improving barrier integrity of the skin. The therapeutic formulation can be administered topically or parenterally. In certain embodiments, the formulation is therapeutic or cosmetic.

In certain embodiments, the formulations may comprise natural amino acids or derivatives thereof that retain substantially the same, or better, activity in terms of improving barrier integrity of skin. In certain embodiments, the formulations may comprise natural amino acids or derivatives thereof that retain substantially the same, or better, activity in terms of treating xerosis cutis, wound healing, and/or treating skin conditions (e.g., atopic dermatitis, psoriasis, or condition related to the aging of skin) and/or improving skin barrier integrity and/or function in a subject in need thereof. The derivatives may be, for example, enantiomers, and include both the D- and L-forms of the amino acids. The derivatives may be, for example, iodotyrosine, or norvaline. Other amino acid derivatives include, but are not limited to, those that are synthesized by, for example, acylation, methylation, and/or halogenation of the amino acid. These include, for example, β-methyl amino acids, C-methyl amino acids, and N-methyl amino acids. Additional amino acid derivatives include, for example, norleucine, ornithine, penicillamine, pyroglutamine derivatives, or other derivatives of the amino acid or combination of amino acids (free amino acids or peptide combination) described here, or any combination of any of the foregoing.

In some embodiments, the formulation and methods described herein are useful for cosmetic applications where, for example, rejuvenation of the various layers of the skin and/or the underlying tissues is desired. This rejuvenation can be aided by, for example, the improving barrier integrity of the skin.

In this embodiment, the methods of the present disclosure generally include the step of topically applying the formulations to the skin (e.g., epidermis) of the patient needing such treatment, wherein a therapeutically effective amount of such formulation is applied. In one embodiment, the formulation is applied to the face. Some embodiments provide for formulations for application to the body.

Advantageously, the present invention provides formulations and methods that combat the aging of skin, wherein combating the aging of skin can include, for example, treating the appearance of wrinkles, fine lines, and other forms of undesirable skin texture. By presenting the formulation to the dermal and/or epidermal layer(s) of the skin, the form, strength, as well as function of the skin is enhanced. In certain embodiments, the formulation and methods described herein are useful for beauty applications where, for example, rejuvenation of the various layers of the skin and/or the underlying tissues is desired.

In some embodiments, the formulations of the present disclosure comprise agents, in addition to the amino acids, that are useful in delaying, minimizing, or eliminating skin aging, wrinkling, and/or other histological changes typically associated with the intrinsic conditions (such as aging, menopause, etc.) and extrinsic conditions (such as environmental pollution (e.g., cigarette smoke), wind, heat, sunlight, radiation, low humidity, harsh surfactants, etc.).

The present disclosure is useful for therapeutically and/or prophylactically improving visible and/or tactile characteristics in skin. For example, in one embodiment, the length, depth, and/or other dimension of lines and/or wrinkles are decreased.

Formulations and Methods for Treating a Cosmetic Skin Condition or Other Skin Conditions

Dry, sensitive, xerotic skin [xerosis cutis (also referred to as xeroderma, dry skin, asteatosis)] is the most common dermatological disorder impacting approximately 50% of the world's population. The severity of this disorder increases with age. It affects patients' quality of life and, due to the impaired skin barrier, is a risk factor for the development of atopic or allergic dermatitis and other skin diseases.

Dry skin is associated with the development of cutaneous fissures and pressure-related ulcers that could be avoided through adequate skin care. Many skin care products used for the management of dry skin mimic the various components of the skin barrier. They are designed to incorporate lipophilic (lipid-replenishing, film-forming) and hydrophilic (remoisturizing) ingredients into the skin. Such products can improve the hydration of the SC, thereby dampening inflammation, but they do not address the underlying biochemical abnormalities in dry skin. Moreover, many skin care products sold on the market provide no scientifically proven benefits.

A need, therefore, exists for effective topical skin care for xerosis cutis that not only mimics the skin barrier components but enhances the skin's own ability to maintain and/or repair the strength of its barrier. A protective barrier so formed restores, at least in part, the structural and functional properties of healthy skin. Topical skin care products that enhance the skin's ability to maintain and/or repair barrier integrity, therefore, address the underlying causes of xerosis cutis, rather than just its symptoms. Formulations directed to addressing this objective were developed.

Xerotic aged skin, for example, has decreased levels of profilaggrin and filaggrin—major sources of free amino acids (FAA) in SC—so decreased levels of these proteins lead, in turn, to a decreased pool of available FAA. Results presented herein show that specific amino acid combinations described herein supplement the age-depleted FAA pool with targeted specificity. Results presented herein further demonstrate that specific combinations of amino acids synergize to confer benefit to structural/functional properties of skin barrier integrity and surprisingly, that certain amino acids can even be detrimental to skin barrier integrity.

Data presented herein also demonstrate that treatment with specific amino acid combinations upregulates a panel of skin barrier marker genes in normal human keratinocytes (e.g., EGF, PDGF, CPT2, TGM4) that play important roles in skin development and homeostasis.

The mechanism(s) whereby amino acids strengthen barrier integrity of the skin, with focused attention on their effect on TJs, and/or improve skin regeneration, including skin barrier repair, are described here. To the best of their knowledge, these mechanisms have not been previously investigated in the skin with scientific rigor.

As emphasized above, amino acids do not have equal beneficial effects on skin barrier integrity. Indeed, some amino acids, alone or in combination, are contraindicated with respect to skin barrier integrity. Therefore, a targeted approach is needed to identify the most effective amino acids for promoting skin barrier health. A new scientifically proven amino acid-based technology targeting compromised epithelial skin is proposed herein to provide a better solution for people who suffer from dry and sensitive skin.

Accordingly, in certain embodiments, the present disclosure provides methods for treating and/or preventing a skin condition in a subject in need thereof, the method comprising administering to the subject a formulation described herein. In certain aspects, the skin condition is dry skin, atopic dermatitis, dermatitis, radiodermatitis, ichthyosis, psoriasis, aging of skin, a condition related to the aging of skin (e.g., wrinkles, loss of suppleness, increase is roughness), pruritus, or eczema. In further aspects, the skin condition is a cosmetic skin condition.

In certain aspects, the formulation for treating and/or preventing a cosmetic skin condition or treating and/or preventing a skin condition comprises: (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1; and optionally, a physiologically acceptable, pharmaceutically acceptable, or dermatologically acceptable carrier; wherein the formulation improves skin regeneration, including skin barrier integrity or repair of the skin. For example, if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine). In a particular embodiment, the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine; and at least one additional amino acid of isoleucine, arginine, threonine, or tryptophan, or any combination thereof. In a more particular embodiment, the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine; and at least one additional amino acid of arginine, or threonine, or any combination thereof. In more particular embodiments, the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and arginine; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, and serine; and at least one additional amino acid of isoleucine, arginine, threonine, or tryptophan, or any combination thereof; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine; and at least one additional amino acid of arginine, or threonine, or tryptophan, or any combination thereof; the free amino acids or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and arginine; the free amino or peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, threonine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, threonine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine; and at least one additional amino acid of isoleucine, threonine, or tryptophan, or any combination thereof; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine; and at least one additional amino acid of isoleucine, arginine, or tryptophan, or any combination thereof; or the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; and at least one additional amino acid of isoleucine, arginine, or threonine, or any combination thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)). Some aspects of the disclosure are directed to such formulations for treating and/or preventing a skin condition that are administered topically or by injection (e.g., intradermal, subcutaneous, intramuscular).

In some embodiments of formulations described herein, wherein, if present, amino acids of cysteine are present at a concentration of equal to or less than 1 mM; if present, amino acids of histidine are present at a concentration of equal to or less than 0.5 mM; if present, amino acids of tyrosine are present at a concentration of equal to or less than 1 mM; if present, amino acids of leucine are present at a concentration of equal to or less than 4 mM; or if present, amino acids of aspartic acid are present at a concentration of equal to or less than 2 mM; or any combination thereof. In some embodiments of the formulation, the formulation does not comprise amino acids of cysteine, histidine, tyrosine, leucine, aspartic acid, taurate, taurine, or glutamate, or any combination thereof.

In some embodiments of formulations described herein, wherein, if present, amino acids of alanine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of glutamine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of glycine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); if present, amino acids of serine are present at a concentration of equal to or less than 4% (e.g., 3.8, 3.6, 3.4, 3.2, 2.8, 2.6, 2.4, 2.2, 2, 1.8, 1.6, 1.4, 1.2, 0.8, 0.6, 0.4, 0.2, 0.18, 0.16, 0.14, 0.12, 0.08, 0.06, 0.058, 0.056, 0.054, 0.052, 0.05, 0.048, 0.046, 0.044, 0.042, 0.04, 0.038, 0.036, 0.034, 0.032, 0.03, 0.02, 0.018, 0.016, 0.014, 0.012); 0.01% or greater (e.g., 0.011, 0.013, 0.015, 0.017, 0.019, 0.021, 0.023, 0.025, 0.027, 0.029, 0.03, 0.031, 0.033, 0.035, 0.037, 0.039, 0.041, 0.043, 0.045, 0.047, 0.049, 0.05, 0.051, 0.053, 0.055, 0.057, 0.059, 0.07, 0.09, 0.1, 0.11, 0.13, 0.15, 0.17, 0.19, 0.3, 0.5, 0.7, 0.9, 1, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, 2.3, 2.5, 2.7, 2.9, 3, 3.1, 3.3, 3.5, 3.7, 3.9, 4.1, 4.3, 4.5, 4.7, 4.9, 5); or any combination thereof.

Amino acids, if present in a formulation described herein, may be present in, for example, at a concentration of 0.5 mM, about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM or about 10 mM. For example, 4 mM of each amino acid comprises: 0.0356% alanine; 0.0584% glutamine; 0.03% glycine; and 0.042% serine. In some aspects, each of the amino acids (4% or 4 mM) in the formulations described here are about 100-fold greater than the concentrations that have been used in tissue culture (TC) experiments.

In one embodiment, the total osmolarity of the formulation is from about 10 mosm to about 280 mosm 50 mosm to about 280 mosm, 100 mosm to about 280 mosm, or about 150 to about 260 mosm.

In some embodiments, the formulation has a pH from, for example, about 2.5 to about 8.5. In some embodiments, the formulation has a pH from about 2.5 to about 6.5, about 2.5 to about 6.0, about 3.0 to about 6.0, about 3.5 to about 6.0, about 3.9 to about 6.0, about 4.2 to about 6.0, about 3.5 to about 5.5, about 3.9 to about 5.0, or about 4.2 to about 4.6. In some embodiments, the pH is about 4.0 to about 6.0 or about 4.5 to about 5.7.

In some embodiments, the formulation is administered systemically or locally. In some embodiments, the formulation is used for treating a disease or condition that is related to barrier integrity of the skin, e.g., xerotic skin, wound healing, treating skin conditions (e.g., is atopic dermatitis, dermatitis, psoriasis, aging of skin, a condition related to the aging of skin, pruritus, or eczema), and/or improving barrier integrity of the skin. The therapeutic formulation can be administered via an enteral route or parenterally or topically or by inhalation. In certain embodiments, the formulation is therapeutic, cosmetic, or nutritional.

In some embodiments, the formulation is a solution. The formulation can be administered with other therapeutic agents.

Formulations and Uses Thereof for Treating and/or Preventing Xerosis Cutis, Atopic Dermatitis, Dermatitis, Radiodermatitis, Psoriasis, Pruritus, Eczema, a Wound, Burn, or a Condition Relating to Aging of Skin

In certain embodiments, the present disclosure provides methods for treating and/or preventing Xerosis cutis, atopic dermatitis, dermatitis, radiodermatits, psoriasis, pruritus, eczema, a wound, burn, or a condition relating to aging of the skin, the method comprising administering to the subject a formulation described herein.

In certain aspects, the formulation for use in treating and/or preventing Xerosis cutis, atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, a wound, an abrasion, a burn, or a condition relating to aging of the skin comprises: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount comprising (consisting essentially of, consisting of) a combination of glutamine, glycine, alanine, and serine, or salts or derivatives thereof; and optionally, a therapeutically effective amount of at least one additional free amino acid or a peptide combination of amino acids of valine, isoleucine, arginine, threonine, or tryptophan, or salts or derivatives thereof, or any combination thereof; wherein the therapeutically effective amount of the combination of glutamine, glycine, alanine, and serine and the therapeutically effective amount of the at least one additional free amino acid or peptide combination of amino acids improves skin regeneration, including skin barrier integrity or repair of skin cells; and optionally, a dermatologically-, pharmaceutically-, or physiologically-acceptable carrier; wherein the formulation improves skin regeneration, including barrier integrity or repair of the skin. In a particular embodiment, the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine; and at least one additional amino acid of isoleucine, arginine, threonine, or tryptophan, or any combination thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)). In a more particular embodiment, the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine; and at least one additional amino acid of arginine, or threonine, or any combination thereof. In more particular embodiments, the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and arginine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, and serine; and at least one additional amino acid of isoleucine, arginine, threonine, or tryptophan, or any combination thereof; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine; and at least one additional amino acid of arginine, or threonine, or tryptophan, or any combination thereof; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and arginine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, threonine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, threonine, and tryptophan; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine; and at least one additional amino acid of isoleucine, threonine, or tryptophan, or any combination thereof; the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine; and at least one additional amino acid of isoleucine, arginine, or tryptophan, or any combination thereof; or the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; and at least one additional amino acid of isoleucine, arginine, or threonine, or any combination thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)). Additional embodiments of the disclosure, provide for formulations comprising as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount comprising, consisting essentially of, or consisting of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1. Some aspects of the disclosure are directed to such formulations for treating and/or preventing Xerosis cutis, atopic dermatitis, dermatitis, radiodermatits, psoriasis, pruritus, eczema, a wound, burn, or a condition relating to aging of the skin, that are administered topically or by injection (e.g., intradermal, subcutaneous, intramuscular).

Amino acids, if present in a formulation described herein, may be present in, for example, at a concentration of about 0.5 mM, about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM or about 10 mM.

In one embodiment, the total osmolarity of the formulation is from about 10 mosm to about 280 mosm 50 mosm to about 280 mosm, 100 mosm to about 280 mosm, or about 150 to about 260 mosm.

In some embodiments, the formulation is administered systemically or locally. In some embodiments, the formulation is used for treating a disease or condition that is related to barrier integrity of the skin, e.g., xerotic skin, wound healing, treating skin conditions (e.g., is atopic dermatitis, dermatitis, psoriasis, aging of skin, a condition related to the aging of skin, pruritus, or eczema), and/or improving barrier integrity of the skin. The therapeutic formulation can be administered via an enteral route or parenterally or topically or by inhalation, or any combination thereof. In certain embodiments, the formulation is therapeutic, cosmetic, or nutritional.

In some embodiments, the formulation is a solution. The formulation can be administered with other therapeutic agents.

In some embodiments, the formulation is administered systemically or locally. The therapeutic formulation can be administered via an enteral route or parenterally or topically or by inhalation. In certain embodiments, the formulation is therapeutic, cosmetic, or nutritional.

In certain embodiments, the formulations may comprise natural amino acids or derivatives thereof that retain substantially the same, or better, activity in terms of improving barrier integrity of skin. In certain embodiments, the formulations may comprise natural amino acids or derivatives thereof that retain substantially the same, or better, activity in terms of treating xerosis cutis, wound healing, and/or treating skin conditions (e.g., atopic dermatitis, psoriasis, or condition related to the aging of skin) in a subject in need thereof. The derivatives may be, for example, enantiomers, and include both the D- and L-forms of the amino acids. The derivatives may be, for example, iodotyrosine, or norvaline. Other amino acid derivatives include, for example, norleucine, ornithine, penicillamine, pyroglutamine derivatives, or other derivatives of alanine, asparagine, aspartic acid, cysteine, glutamic acid, glycine, isoleucine, leucine, lysine, methionine, proline, phenylalanine, serine, threonine, tryptophan, or valine. In certain embodiments, the amino acid derivatives are derivatives of at least one of glutamine, glycine, alanine, serine, valine, isoleucine, arginine, threonine, or tryptophan, or any combination thereof, Other amino acid derivatives include, but are not limited to, those that are synthesized by, for example, acylation, methylation, and/or halogenation of the amino acid. These include, for example, β-methyl amino acids, C-methyl amino acids, and N-methyl amino acids.

In certain embodiments, the formulation also comprises additives (e.g., nutrients, electrolytes, vitamins, minerals, etc.). In certain embodiments, the formulation comprises iron or zinc. In certain embodiments, the therapeutic formulation comprises one or more electrolytes selected from, for example, Na⁺; K⁺; HCO₃—; CO₃²⁻; Ca²⁺; Mg²⁺; Fe₂; Cl⁻; phosphate ions, such as H₂PO₄⁻, HPO₄²⁻, and PO₄³—; zinc; iodine; copper; iron; selenium; chromium; and molybdenum. In an alternative embodiment, the formulation does not contain HCO₃⁻ or CO₃²⁻. In another alternative embodiment, the formulation comprises HCO₃⁻ or CO₃²⁻ at a total concentration of less than 5 mg/l, or concentrations lower than 5 mg/l. In certain embodiments, the formulation does not contain electrolytes. For example, in certain embodiments, the formulation does not include one or more, or any, of Na⁺; K⁺; HCO₃⁻; CO₃²⁻; Ca²⁺; Mg²⁺; Fe₂; Cl⁻; phosphate ions, such as H₂PO₄⁻, HPO₄²⁻, and PO₄³⁻; zinc; iodine; copper; iron; selenium; chromium; and molybdenum.

In one embodiment, phosphate ions, such as H₂PO₄—, HPO₄²⁻, and PO₄³⁻, are used to buffer the formulation described herein. In one embodiment, the therapeutic formulation uses HCO₃⁻ or CO₃²⁻ as a buffer. In another embodiment, the therapeutic formulation does not use HCO₃⁻ or CO₃²⁻ as buffer.

The present invention is useful for therapeutically and/or prophylactically improving visible and/or tactile characteristics in skin. For example, in one embodiment, the length, depth, and/or other dimension of lines and/or wrinkles are decreased.

In one embodiment, the formulation applied to the skin or other tissue can further comprise collagen, hyaluronic acid (HA), and/or ceramides. In one embodiment, the HA is cross-linked HA. The formulation can further comprise components and/or additives such as, but not limited to, dermatologically-acceptable vehicles, carriers, or additives described here, such as but not limited to collagen, hyaluronic acid, and ceramides,

In another aspect, the present disclosure provides methods of treating and/or preventing a skin condition (e.g., atopic dermatitis, radiodermatitis, psoriasis, or condition related to the aging of skin) in a subject in need thereof, the method comprising administering to the subject a formulation described herein. In certain embodiments, the skin condition is atopic dermatitis, radiodermatitis, psoriasis, the aging of skin, or a condition related to the aging of skin. In some embodiments, the skin condition is pruritus (itch), psoriasis, eczema, burns, or dermatitis. In certain embodiments, the skin condition is psoriasis. In certain embodiments, the skin condition is pruritis.

The formulation of the present disclosure can also be administered at a surgical site, including at a site of minimally invasive surgery, to improve healing and the surgical outcome.

Formulations and Uses Thereof for Skin Regeneration, Including Skin Barrier Repair and Strengthening

In additional aspects, the therapeutically effective amount of a free amino acid, a combination of free amino acids, or a peptide combination thereof of the formulations described here, improves skin barrier repair and strengthening. The improvement of skin barrier repair includes, and is not limited to, skin barrier correction, skin barrier strengthening, skin barrier preservation, skin barrier integrity, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocytes, endothelial cells, and fibroblasts; facilitation of dermal regeneration, or combinations thereof. Further aspects are directed to the modulation of a skin barrier marker (e.g., epidermal growth factor (EGF); platelet-derived growth factor (PDGF), fibroblast growth factor 2 (FGF2), peroxisome proliferator activated receptor delta (PPARD); carnitine palmitoyltransferase 2 (CPT2); filaggrin (FLG); marker of proliferation Ki-67 (MKI67); transglutaminase 4 (TGM4)) using a therapeutically effective amount of the amino acid combination or the formulation comprising (or consisting essentially of or consisting of) the amino acid combination described here, where the amino acid combination comprises, consists essentially of, or consists of a therapeutically effective amount of as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts or derivatives thereof, where if in combination, each amino acid can be in a ratio in relation to another amino acid in a ratio of, for example, 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the ratio of glutamine at 4 mM to other amino acids comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)); where if present, each amino acid comprises: a concentration of 0.01% or greater, 4% or less, or 0.01% to 4%; and/or a concentration of 0.1 mM or greater, 10 mM or less, or 0.1 mM to 10 mM; where if present, each amino acid comprises a weight percent to the formulation of: 0.001 wt % or greater; 5 wt % or less; or 0.001 wt % to 5 wt %; or any combinations of features thereof; and where the formulation can optionally comprise a dermatologically acceptable vehicle and/or additive, where when tested by a method detecting expression of a barrier marker gene such as but not limited to EGF and determining that the barrier marker gene is increased by a 1.5-40 average fold increase of amino acid-treated as compared to amino acid-untreated skin. The use of any of the formulations described here and its methods of improving skin barrier repair includes but is not limited to, correcting the skin barrier, strengthening the skin barrier, preserving the skin barrier, increasing skin barrier integrity, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocytes, endothelial cells, and fibroblasts; facilitating dermal regeneration, and any combinations thereof.

In some embodiments, a formulation or a topical formulation of the disclosure comprises (or consists essentially of or consists of) therapeutically effective amounts of as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1; a therapeutically effective amount of a plant extract, wherein the plant extract comprises (or consists essentially of or consists of): a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) or a plant extract combination (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica) comprising (or consisting essentially of, or consisting of): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically acceptable vehicle and/or additive. For example, a formulation comprising as free amino acids (including free amino acid combinations), or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): at least one of: alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; a therapeutically effective amount of a plant extract, wherein the plant extract comprises (or consists essentially of or consists of): a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) or a plant extract combination (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica) comprising (or consisting essentially of, or consisting of): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically acceptable vehicle and/or additive. Additional aspects of the formulation or the topical formulation comprising, consisting essentially of, or consisting of, a therapeutically effective amount of an amino acid or an amino acid combination (of free amino acids or a peptide combination of amino acids), where the therapeutically effective amount of the amino acids or the amino acid combination of the formulation comprise, consist essentially of, or consist of: alanine; glutamine; glycine; serine; alanine and glutamine; glycine and serine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; alanine, glutamine, and glycine; glutamine, glycine, and serine; alanine, glutamine, and serine; alanine, glycine, and serine; or alanine, glutamine, glycine, and serine, or salts thereof. Further aspects of the formulation described here provide a therapeutically effective amount of the amino acids or the amino acid combination and/or a therapeutically effective amount of the plant extract (e.g., Boswellia extract) sufficient to improve skin regeneration, including skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration) as indicated by an average fold increase of a skin regeneration, including skin barrier marker gene, EGR, expression of: 1.5 or greater (e.g., 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6, 1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1; 11.3; 11.5; 11.7; 11.9; 12.1; 12.3; 12.5; 12.7; 12.9; 13.1; 13.3; 13.5; 13.7; 13.9; 14.1; 14.3; 14.5; 14.7; 14.9; 15.1; 15.3; 15.5; 15.7; 15.9; 16.1; 16.3; 16.5; 16.7; 16.9; 17.1; 17.3; 17.5; 17.7; 17.9; 18.1; 18.3; 18.5; 18.7; 18.9; 19.1; 19.3; 19.5; 19.7; 19.9; 20.1; 20.3; 20.5; 20.7; 20.9; 21.1; 21.3; 21.5; 21.7; 21.9; 22.1; 22.3; 22.5; 22.7; 22.9; 23.1; 23.3; 23.5; 23.7; 23.9; 23.1; 23.3; 23.5; 23.7; 23.9; 24.1; 24.3; 24.5; 24.7; 24.9; 25.1; 25.3; 25.5; 25.7; 25.9; 26.1; 26.3; 26.5; 26.7; 26.9; 27.1; 27.3; 27.5; 27.7; 27.9; 28.1; 28.3; 28.5; 28.7; 28.9; 29.1; 29.3; 29.5; 29.7; 29.9; 30.1; 30.3; 30.5; 30.7; 30.9; 31.1; 31.3; 31.5; 31.7; 31.9; 32.1; 32.3; 32.5; 32.7; 32.9; 33.1; 33.3; 33.5; 33.7; 33.9; 34.1; 34.3; 34.5; 34.7; 34.9; 35.1; 35.3; 35.5; 35.7; 35.9; 36.1; 36.3; 36.5; 36.7; 36.9; 37.1; 37.3; 37.5; 37.7; 37.9; 38.1; 38.3; 38.5; 38.7; 38.9; 39.1; 39.3; 39.5; 39.7; 39.9; 40.1; 40.3; 40.5; 40.7; 40.9; 41.1; 41.3; 41.5); 40 or less (e.g., 39.8; 39.6; 39.4; 39.2; 39; 38.8; 38.6; 38.4; 38.2; 38; 37.8; 37.6; 37.4; 37.2; 37; 36.8; 36.6; 36.4; 36.2; 36; 35.8; 35.6; 35.4; 35.2; 35; 34.8; 34.6; 34.4; 34.2; 34; 33.8; 33.6; 33.4; 33.2; 33; 32.8; 32.6; 32.4; 32.2; 32; 31.8; 31.6; 31.4; 31.2; 31; 30.8; 30.6; 30.4; 30.2; 30; 29.8; 29.6; 29.4; 29.2; 29; 28.8; 28.6; 28.4; 28.2; 28; 27.8; 27.6; 27.4; 27.2; 27; 26.8; 26.6; 26.4; 26.2; 26; 25.8; 25.6; 25.4; 25.2; 25; 24.8; 24.6; 24.4; 24.2; 24; 23.8; 23.6; 23.4; 23.2; 23; 22.8; 22.6; 22.4; 22.2; 22; 21.8; 21.6; 21.4; 21.2; 21; 20.8; 20.6; 20.4; 20.2; 20; 19.8; 19.6; 19.4; 19.2; 19; 18.8; 18.6; 18.4; 18.2; 18; 17.8; 17.6; 17.4; 17.2; 17.2; 17; 16.8; 16.6; 16.4; 16.2; 16; 15.8; 15.6; 15.4; 15.2; 15; 14.8; 14.6; 14.4; 14.2; 14; 13.8; 13.6; 13.4; 13.2; 13; 12.8; 12.6; 12.4; 12.2; 12; 11.8; 11.6; 11.4; 11.2; 11; 10.8; 10.6; 10.4; 10.2; 10; 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6); or 1.5-40 (e.g., 1.6-39.9; 1.7-39.8; 1.8-39.7; 1.9-39.6; 2-39.5; 2.1-39.4; 2.2-39.3; 2.3-39.2; 2.4-39.1; 2.5-39; 2.6-38.9; 2.7-38.8; 2.8-38.7; 2.9-38.6; 3-38.5; 3.1-38.4; 3.2-38.3; 3.3-38.2; 3.4-38.1; 3.5-38; 3.6-37.9; 3.7-37.8; 3.8-37.7; 3.9-37.6; 4-37.5; 4.1-37.4; 4.2-37.3; 4.3-37.2; 4.4-37.1; 4.5-37; 4.6-36.9; 4.7-36.8; 4.8-36.7; 4.9-36.6; 5-36.5; 5.1-36.4; 5.2-36.3; 5.3-36.2; 5.4-36.1; 5.5-36; 5.6-35.9; 5.7-35.8; 5.8-35.7; 5.9-35.6; 6-35.5; 6.1-35.4; 6.2-35.3; 6.3-35.2; 6.4-35.1; 6.5-35; 6.6-34.9; 6.7-34.8; 6.8-34.7; 6.9-34.6; 7-34.5; 7.1-34.4; 7.2-34.3; 7.3-34.2; 7.4-34.1; 7.5-34; 7.6-33.9; 7.7-33.8; 7.8-33.7; 7.9-33.6; 8-33.5; 8.1-33.4; 8.2-33.3; 8.3-33.2; 8.4-33.1; 8.5-33; 8.6-32.9; 8.7-32.8; 8.8-32.7; 8.9-32.6; 9-32.5; 9.1-32.4; 9.2-32.3; 9.3-32.2; 9.4-32.1; 9.5-32; 9.6-31.9; 9.7-31.8; 9.8-31.7; 9.9-31.6; 10-31.5; 10.1-31.4; 10.2-31.3; 10.3-31.2; 10.4-31.1; 10.5-31; 10.6-30.9; 10.7-30.8; 10.8-30.7; 10.9-30.6; 11-30.5; 11.1-30.4; 11.2-30.3; 11.3-30.2; 11.4 30.1; 11.5-30; 11.6-29.9; 11.7-29.8; 11.8-29.7; 11.9-29.6; 12-29.5; 12.1-29.4; 12.2-29.3; 12.3-29.2; 12.4-29.1; 12.5-29; 12.6-28.9; 12.7-28.8; 12.8-28.7; 12.9-28.6; 13-28.5; 13.1-28.4; 13.2-28.3; 13.3-28.2; 13.4-28.1; 13.5-28; 13.6-27.9; 13.7-27.8; 13.8-27.7; 13.9-27.6; 14-27.5; 14.1-27.4; 14.2-27.3; 14.3-27.2; 14.4-27.1; 14.5-27; 14.6-26.9; 14.7-26.8; 14.8-26.7; 14.9-26.6; 15-26.5; 15.1-26.4; 15.2-26.3; 15.3-26.2; 15.4-26.1; 15.5-26; 15.6-25.9; 15.7-25.8; 15.8-25.7; 15.9-25.6; 16-25.5; 16.1-25.4; 16.2-25.3; 16.3-25.2; 16.4-25.1; 16.5 25; 16.6-24.9; 16.7-24.8; 16.8-24.7; 16.9-24.6; 17-24.5; 17.1-24.4; 17.2-24.3; 17.3-24.2; 17.4-24.1; 17.5-24; 17.6-23.9; 17.7-23.8; 17.8-23.7; 17.9-23.6; 18-23.5; 18.1-23.4; 18.2-23.3; 18.3-23.2; 18.4-23.1; 18.5-23; 18.6-22.9; 18.7-22.8; 18.8-22.7; 18.9-22.6; 19-22.5; 19.1-22.4; 19.2-22.3; 19.3-22.2; 19.4-22.1; 19.5-22; 19.6-21.9; 19.7-21.8; 19.8-21.7; 19.9 21.6; 20-21.5; 20.1-21.4; 20.2-21.3; 20.3-21.2; 20.4-21.1; 20.5-21; 20.6-20.9; 20.7-20.8). See, e.g., FIGS. 8-14.

Other embodiments provide a formulation or a topical formulation, comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): alanine, glutamine, glycine, and serine, or salts thereof; a therapeutically effective amount of a plant extract (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) or a plant extract combination (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising (or consisting essentially of, or consisting of): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically acceptable vehicle and/or additive.

In additional embodiments, a formulation or a topical formulation for administration to skin of a subject, comprises (or consists essentially of or consists of): a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising (or consisting essentially of or consisting of): alanine, glutamine, glycine, and serine, or salts thereof; a therapeutically effective amount of a plant extract (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) or a plant extract combination comprising (consisting essentially of or consisting of) (e.g., BOSEXIL™ or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine, soy lecithin); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically acceptable vehicle and/or additive, wherein the therapeutically effective amount of the combination of free amino acids or a peptide combination thereof and the therapeutically effective amount of the plant extract or plant extract combination improves skin regeneration, including skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration), wherein the formulation or the topical formulation improves skin regeneration, including skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration) when tested by a method detecting expression of a barrier marker gene. EGF, and determining that the barrier marker gene is increased by a 1.5-40 average fold increase of amino acid treated as compared to amino acid untreated skin (see, e.g., FIGS. 8-14). Without being bound by theory, the formulation or the topical formulation comprising the free amino acids and/or peptide combination of amino acids described here increase barrier marker gene expression, e.g., EGF, as well as, increase the synthesis of such barrier marker genes, e.g., EGF, which is a precursor to downstream responses for proliferation, which can be specific for EGF and diseases or conditions, such as but not limited to those in need of improved skin regeneration, including skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration); ichthyosis, psoriasis, dermatitis, atopic dermatitis, radiodermatitis; and the like. Impaired skin barriers can be caused by genetics, aging, environmental factors (e.g., sun radiation (e.g., ultraviolet radiation, visible light, infra-red radiation); air pollution; chemical pollutants (e.g., detergents, cosmetics, skin products containing alcohol, artificial fragrances); smoking); overuse of acids or abrasives; health factors (e.g., poor nutrition; poor sleep; stress; dehydration); and the like.

In additional embodiments, the formulation or the topical formulation described here comprising the free amino acids or the peptide combination of amino acids, a therapeutically effective amount comprising (consisting essentially of, consisting of) (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1, in a ratio where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the ratio of glutamine at 4 mM to other amino acids comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine); and the therapeutically effective amount of the plant extract (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) or plant extract combination (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising (or consisting essentially of, or consisting of): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); or a retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof); optionally a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically-, pharmaceutically-, or physiologically-acceptable carrier and/or additive, where the plant extract or the plant extract combination, or the retinoid is present at a weight percentage (w/w %) of the formulation of: 0.01 w/w % or greater (e.g., 0.011; 0.013; 0.015; 0.017; 0.019; 0.021; 0.023; 0.025; 0.027; 0.029; 0.031; 0.033; 0.035; 0.037; 0.039; 0.041; 0.043; 0.045; 0.047; 0.049; 0.051; 0.053; 0.055; 0.057; 0.059; 0.061; 0.063; 0.065; 0.067; 0.069; 0.071; 0.073; 0.075; 0.077; 0.079; 0.081; 0.083; 0.085; 0.087; 0.089; 0.091; 0.093; 0.095; 0.097; 0.099; 0.1; 0.15; 0.3; 0.35; 0.5; 0.55; 0.7; 0.75; 0.9; 0.95; 1, 1; 1.3; 1.5; 1.7; 1.9; 2, 1; 2.3; 2.5; 2.7; 2.9; 3, 1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3); 5 w/w % or less (e.g., 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.85; 0.8; 0.65; 0.6; 0.45; 0.4; 0.25; 0.2; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.008; 0.006; 0.004); or 0.01 w/w %-5 w/w % (e.g., 0.011 w/w %-4.9 w/w %; 0.012 w/w %-4.8 w/w %; 0.013 w/w %-4.7 w/w %; 0.014 w/w %-4.6 w/w %; 0.015 w/w %-4.5 w/w %; 0.016 w/w %-4.4 w/w %; 0.017 w/w %-4.3 w/w %; 0.018 w/w %-4.2 w/w %; 0.019 w/w %-4.1 w/w %; 0.02 w/w %-4 w/w %; 0.021 w/w %-3.9 w/w %; 0.022 w/w %-3.8 w/w %; 0.023 w/w %-3.7 w/w %; 0.024 w/w %-3.6 w/w %; 0.025 w/w %-3.5 w/w %; 0.026 w/w %-3.4 w/w %; 0.027 w/w %-3.3 w/w %; 0.028 w/w %-3.2 w/w %; 0.029 w/w %-3.1 w/w %; 0.03 w/w %-3 w/w %; 0.031 w/w %-2.9 w/w %; 0.032 w/w %-2.8 w/w %; 0.033 w/w %-2.7 w/w %; 0.034 w/w %-2.6 w/w %; 0.035 w/w %-2.5 w/w %; 0.036 w/w %-2.4 w/w %; 0.037 w/w %-2.3 w/w %; 0.038 w/w %-2.2 w/w %; 0.039 w/w %-2.1 w/w %; 0.04 w/w %-2 w/w %; 0.041 w/w %-1.9 w/w %; 0.042 w/w %-1.8 w/w %; 0.043 w/w %-1.7 w/w %; 0.044 w/w %-1.6 w/w %; 0.045 w/w %-1.5 w/w %; 0.046 w/w %-1.4 w/w %; 0.047 w/w %-1.3 w/w %; 0.048 w/w %-1.2 w/w %; 0.049 w/w %-1.1 w/w %; 0.05 w/w %-1 w/w %; 0.051 w/w %-0.99 w/w %; 0.052 w/w %-0.98 w/w %; 0.053 w/w %-0.97 w/w %; 0.054 w/w %-0.96 w/w %; 0.055 w/w %-0.95 w/w %; 0.056 w/w %-0.94 w/w %; 0.057 w/w %-0.93 w/w %; 0.058 w/w %-0.92 w/w %; 0.059 w/w %-0.91 w/w %; 0.06 w/w %-0.9 w/w %; 0.061 w/w %-0.89 w/w %; 0.062 w/w %-0.88 w/w %; 0.063 w/w %-0.87 w/w %; 0.064 w/w %-0.86 w/w %; 0.065 w/w %-0.85 w/w %; 0.066 w/w %-0.84 w/w %; 0.067 w/w %-0.83 w/w %; 0.068 w/w %-0.82 w/w %; 0.069 w/w %-0.81 w/w %; 0.07 w/w %-0.8 w/w %; 0.071 w/w %-0.79 w/w %; 0.072 w/w %-0.78 w/w %; 0.073 w/w %-0.77 w/w %; 0.074 w/w %-0.76 w/w %; 0.075 w/w %-0.75 w/w %; 0.076 w/w %-0.74 w/w %; 0.077 w/w %-0.73 w/w %; 0.078 w/w %-0.72 w/w %; 0.079 w/w %-0.71 w/w %; 0.08 w/w %-0.7 w/w %; 0.081 w/w %-0.69 w/w %; 0.082 w/w %-0.68 w/w %; 0.083 w/w %-0.67 w/w %; 0.084 w/w %-0.66 w/w %; 0.085 w/w %-0.65 w/w %; 0.086 w/w %-0.64 w/w %; 0.087 w/w %-0.63 w/w %; 0.088 w/w %-0.62 w/w %; 0.089 w/w %-0.61 w/w %; 0.09 w/w %-0.6 w/w %; 0.091 w/w %-0.59 w/w %; 0.092 w/w %-0.58 w/w %; 0.093 w/w %-0.57 w/w %; 0.094 w/w %-0.56 w/w %; 0.095 w/w %-0.55 w/w %; 0.096 w/w %-0.54 w/w %; 0.097 w/w %-0.53 w/w %; 0.098 w/w %-0.52 w/w %; 0.099 w/w %-0.51 w/w %; 0.1 w/w %-0.5 w/w %; 0.101 w/w %-0.49 w/w %; 0.102 w/w %-0.48 w/w %; 0.103 w/w %-0.47 w/w %; 0.104 w/w %-0.46 w/w %; 0.105 w/w %-0.45 w/w %; 0.106 w/w %-0.44 w/w %; 0.107 w/w %-0.43 w/w %; 0.108 w/w %-0.42 w/w %; 0.109 w/w %-0.41 w/w %; 0.110 w/w %-0.4 w/w %; 0.111 w/w %-0.39 w/w %; 0.112 w/w %-0.38 w/w %; 0.113 w/w %-0.37 w/w %; 0.114 w/w %-0.36 w/w %; 0.115 w/w %-0.35 w/w %; 0.116 w/w %-0.34 w/w %; 0.117 w/w %-0.33 w/w %; 0.118 w/w %-0.32 w/w %; 0.119 w/w %-0.31 w/w %; 0.120 w/w %-0.3 w/w %; 0.121 w/w %-0.29 w/w %; 0.122 w/w %-0.28 w/w %; 0.123 w/w %-0.27 w/w %; 0.124 w/w %-0.26 w/w %; 0.125 w/w %-0.25 w/w %; 0.126 w/w %-0.24 w/w %; 0.127 w/w %-0.23 w/w %; 0.128 w/w %-0.22 w/w %; 0.129 w/w %-0.21 w/w %; 0.130 w/w %-0.2 w/w %; 0.0131 w/w %-0.19 w/w %; 0.132 w/w %-0.18 w/w %; 0.133 w/w %-0.17 w/w %; 0.134 w/w %-0.16 w/w %; 0.135 w/w %-0.15 w/w %; 0.136 w/w %-0.14 w/w %; 0.137 w/w %-0.139 w/w %). In some aspects, the plant extract comprises, consists essentially of, or consists of a Boswellia extract, which includes Boswellia serrata, Burseraceae family, olibanum, or frankincense. Other aspects are directed to the plant extract combination comprising, consisting essentially of, or consists of: Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications). In additional aspects, the plant extract combination comprises, consists essentially of, or consists of: Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, and silica.

Further embodiments provide the formulation or the topical formulation described here, where the therapeutically effective amount of the combination of free amino acids or a peptide combination thereof and the therapeutically effective amount of the plant extract or plant extract combination inhibit 5-lipoxygenase (an inflammation-inducing enzyme); decreases mobilization of calcium ions; decreases activation of MAP kinases. Without wishing to be bound by theory, the plant extract or plant extract combination comprising Boswellia extract comprises anti-inflammatory properties, which taper down the inflammation induced by, for example, 5-lipoxygenasc. See, e.g., D. Pocckel and O. Werz (e.g., Curr Med Chem. 13(28):3359-3369, 2006; Singh et al. Phytomedicine. 15(6-7):400-407, 2008.

In some of the embodiments, the formulation or the topical formulation comprises, consists essentially of, or consists of: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of) alanine, glutamine, glycine, and serine; a therapeutically effective amount of a plant extract, wherein the plant extract comprises (or consists essentially of or consists of): a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); and optionally, a dermatologically acceptable vehicle and/or additive, such as but not limited to: lecithin, microcrystalline cellulose, and silica.

In some embodiments, the formulation or the topical formulation of the disclosure, comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): at least one of: alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine); a therapeutically effective amount of a plant extract or a plant extract combination; a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically acceptable vehicle and/or additive. Non-limiting examples of at least one phospholipid useful in the formulation of the disclosure include any phospholipids comprising two fatty acids per phosphate backbone, where the phospholipid can include: phosphatidic acid (phosphatidate) (PA); phosphatidylethanolamine (cephalin) (PE); phosphatidylcholine (lecithin) (PC); phosphatidylserine (PS); phosphatidylinositol (PI); phosphatidylinositol phosphate (PIP); phosphatidylinositol bisphosphate (PIP2); and phosphatidylinositol trisphosphate (PIP3); ceramides comprising phosphate group; ceramide phosphorylcholine (Sphingomyelin) (SPH); ceramide phosphorylethanolamine (Sphingomelin) (Cer-PE); phospholipids comprising fatty acids selected from the group consisting of: saturated fatty acids (e.g., butyric acid, lauric acid, myristic acid, palmitic acid, stearic acid); unsaturated fatty acids (e.g., myristoleic acid (cis-9-tetradecenoic acid); sapienic acid (cis-6-hexadecenoic acid); palmitoleic acid (cis-9-hexadecenoic acid); oleic acid (cis-9-octadecenoic acid); petroselinic acid (cis-Octadec-6-enoic acid); cis-vaccenic acid (cis-11-octadecenoic acid); vaccenic acid (trans-11-octadecenoic acid); elaidic acid (trans-9-octadecenoic acid (trans-oleic acid)); linoleic acid; linolenic acid; paullinic acid (cis-13-eicosenoic acid); gadoleic acid (cis-9-icosenoic acid); gondoic acid (cis-11-eicosenoic acid); erucic acid (cis-15-docosenoic acid); brassidic acid (trans-15-docosenoic acid); nervonic acid (cis-15-tetracosenoic acid); arachidonic acid; and combinations thereof, Other embodiments provide the formulation or the topical formulation comprising, consisting essentially of, or consisting of: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of) (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1; and (i) a therapeutically effective amount of a plant extract or plant extract combination comprising (or consisting essentially of or consisting of) a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or in combination with at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications), or (ii) a therapeutically effective amount of a retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof), where the amino acids are in a ratio of: 1:100 or greater (e.g., 2:100; 4:100; 6:100; 8:100; 10:100; 12:100; 14:100; 16:100; 18:100; 20:100; 22:100; 24:100; 26:100; 28:100; 30:100; 32:100; 34:100; 36:100; 38:100; 40:100; 42:100; 44:100; 46:100; 48:100; 50:100; 52:100; 54:100; 56:100; 58:100; 60:100; 62:100; 64:100; 66:100; 68:100; 70:100; 72:100; 74:100; 76:100; 78:100; 80:100; 82:100; 84:100; 86:100; 88:100; 90:100; 92:100; 94:100; 96:100; 98:100; 100:100; 110:100; 120:100; 130:100; 140:100:150:100; 160:100; 170:100; 180:100; 190:100; 200:100); 100:1 or less (e.g., 99:1; 97:1; 95:1; 93:1; 91:1; 89:1; 87:1; 85:1; 83:1; 81:1; 79:1; 77:1; 75:1; 73:1; 71:1; 69:1; 67:1; 65:1; 63:1; 61:1; 59:1; 57:1; 55:1; 53:1; 51:1; 49:1; 47:1; 45:1; 43:1; 41:1; 39:1; 37:1; 35:1; 33:1; 31:1; 29:1; 27:1; 25:1; 23:1; 21:1; 19:1; 17:1; 15:1; 13:1; 11:1; 9:1; 7:1; 5:1; 3:1; 1:1; 0.9:1; 0.7:1; 0.5:1; 0.3:1; 0.1:1; 99:100; 97:100; 95:100; 93:100; 91:100; 89:100; 87:100; 85:100; 83:100; 81:100; 79:100; 77:100; 75:100; 73:100; 71:100; 69:100; 67:100; 65:100; 63:100; 61:100; 59:100; 57:100; 55:100; 53:100; 51:100; 49:100; 47:100; 45:100; 43:100; 41:100; 39:100; 37:100; 35:100; 33:100; 31:100; 29:100; 27:100; 25:100; 23:100; 21:100; 19:100; 17:100; 15:100; 13:100; 11:100; 9:100; 7:100; 5:100; 3:100; 1:100); 1:100 to 100:1 (e.g., 1:99 to 99:1; 1:98 to 98:1; 1:97 to 97:1; 1:96 to 96:1; 1:95 to 95:1; 1:94 to 94:1; 1:93 to 93:1; 1:92 to 92:1; 1:91 to 91:1; 1:90 to 90:1; 1:89 to 89:1; 1:88 to 88:1; 1:87 to 87:1; 1:86 to 86:1; 1:85 to 85:1; 1:84 to 84:1; 1:83 to 83:1; 1:82 to 82:1; 1:81 to 81:1; 1:80 to 80:1; 1:79 to 79:1; 1:78 to 78:1; 1:77 to 77:1; 1:76 to 76:1; 1:75 to 75:1; 1:74 to 74:1; 1:73 to 73:1; 1:72 to 72:1; 1:71 to 71:1; 1:70 to 70:1; 1:69 to 69:1; 1:68 to 68:1; 1:67 to 67:1; 1:66 to 66:1; 1:65 to 65:1; 1:64 to 64:1; 1:63 to 63:1; 1:62 to 62:1; 1:61 to 61:1; 1:60 to 60:1; 1:59 to 59:1; 1:58 to 58:1; 1:57 to 57:1; 1:56 to 56:1; 1:55 to 55:1; 1:54 to 54:1; 1:53 to 53:1; 1:52 to 52:1; 1:51 to 51:1; 1:50 to 50:1; 1:49 to 49:1; 1:48 to 48:1; 1:47 to 47:1; 1:46 to 46:1; 1:45 to 45:1; 1:44 to 44:1; 1:43 to 43:1; 1:42 to 42:1; 1:41 to 41:1; 1:40 to 40:1; 1:39 to 39:1; 1:38 to 38:1; 1:37 to 37:1; 1:36 to 36:1; 1:35 to 35:1; 1:34 to 34:1; 1:33 to 33:1; 1:32 to 32:1; 1:31 to 31:1; 1:30 to 30:1; 1:29 to 29:1; 1:28 to 28:1; 1:27 to 27:1; 1:26 to 26:1; 1:25 to 25:1; 1:24 to 24:1; 1:23 to 23:1; 1:22 to 22:1; 1:21 to 21:1; 1:20 to 20:1; 1:19 to 19:1; 1:18 to 18:1; 1:17 to 17:1; 1:16 to 16:1; 1:15 to 15:1; 1:14 to 14:1; 1:13 to 13:1; 1:12 to 12:1; 1:11 to 11:1; 1:10 to 10:1; 1:9 to 9:1; 1:8 to 8:1; 1:7 to 7:1; 1:6 to 6:1; 1:5 to 5:1; 1:4 to 4:1; 1:3 to 3:1; 1:2 to 2:1), and where the free amino acids or the peptide combination of amino acids and (i) the plant extract or the plant extract combination, or (ii) the retinoid, are in a ratio of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1.

In additional embodiments, the formulation or the topical formulation described here can be prepared by mixing and/or stirring (concurrently, simultaneously, sequentially): (a) a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising, consisting essentially of at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine); (b) a therapeutically effective amount of a plant extract or a plant extract combination comprising, consisting essentially of, or consisting of: a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); and optionally, (c) a dermatologically acceptable vehicle and/or additive. The mixture of (a), (b), and optionally (c) can be heated (either together or separately) to a temperature sufficient to completely mix, homogenize, or emulsify (a), (b), and optionally (c), where the temperature is 20° C. or greater (e.g., 22; 24; 26; 28; 30; 32; 34; 36; 38; 40; 42; 44; 46; 48; 50; 52; 54; 56; 58; 60; 62; 64; 66; 68; 70; 72; 74; 76; 78; 80; 82; 84; 86; 88; 90; 92; 94; 96; 98; 100; 102; 104; 106; 108; 110); 100° C. or less (e.g., 99; 97; 95; 93; 91; 89; 87; 85; 83; 81; 79; 77; 75; 73; 71; 69; 67; 65; 63; 61; 59; 57; 55; 53; 51; 49; 47; 45; 43; 41; 39; 37; 35; 33; 31; 29; 27; 25; 23; 21; 19; 17); or 20° C.-100° C. (e.g., 21° C.-99° C.; 22° C.-98° C.; 23° C.-97° C.; 24° C.-96° C.; 25° C.- 95° C.; 26° C.-94° C.; 27° C.-93° C.; 28° C.-92° C.; 29° C.-91° C.; 30° C.-90° C.; 31° C.-89° C.; 32° C.-88° C.; 33° C.-87° C.; 34° C.-86° C.; 35° C.-85° C.; 36° C.-84° C.; 37° C.-83° C.; 38° C.-82° C.; 39° C.-81° C.; 40° C.-80° C.; 41° C.-79° C.; 42° C.-78° C.; 43° C.-77° C.; 44° C.-76° C.; 45° C.-75° C.; 46° C.-74° C.; 47° C.-73° C.; 48° C.-72° C.; 49° C.-71° C.; 50° C.-70° C.; 51° C.-69° C.; 52° C.-68° C.; 53° C.-67° C.; 54° C.-66° C.; 55° C.-65° C.; 56° C.-64° C.; 57° C.-63° C.; 58° C.-62° C.; 59° C.-61° C., for a time sufficient to completely mix, homogenize, or emulsify (e.g., seconds to minutes). In some aspects, the plant extract combination of the formulation or the topical formulation described here, further comprises (or consists essentially of or consists of): a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications). See, e.g., EXAMPLE 7.

Formulations and Uses Thereof for Improving Skin Moisture

Aging skin exhibits wrinkles, elastosis, abnormal formation and/or distribution of pigmentation, loss of skin firmness, and a decrease in moisture. Hyaluronic acid (HA; also known as hyaluronan or hyaluronate, used interchangeably here), a major constituent of the extracellular matrix (ECM), is a glycosaminoglycan (GAG) known to be an important molecule in matrix assembly, cell proliferation, cell migration, tissue development, and retaining skin moisture because of its water-binding capacity. HA is a polymer of disaccharides (D-glucuronic acid; N-acetyl-D-glucosamine) linked with alternating β-(1→4) and β-(1→3) glycosidic bonds, where the carboxylic groups make HA negatively charged and thereby enabling binding to water. HA levels in skin decline with age progression, contributing to a loss of moisture and/or firmness in aging skin. Hyaluronic acid synthase (HAS1, HAS2, HAS3) is the enzyme responsible for synthesizing HA at the plasma membrane, which is then released into the extracellular space. HAS1, HAS2, and HAS3 are three mammalian isoforms of HAS having different enzymatic properties and capable of synthesizing HA chains of varying lengths. Accordingly, the presence of HA is desirable for improving skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS)). HA induction associated with skin moisturization and anti-aging, where aging is associated with the collapse of the ECM and the loss of hydration. Without being bound by theory, upregulation of HA increases the hydration and the healthy structure of the ECM, thereby supporting the skin dermis and reducing recesses, wrinkles, or tissue collapse or herniation.

In various embodiments of the disclosure, a formulation, such as a topical formulation, described here comprises a therapeutically effective amount of a combination of: free amino acids or a peptide combination of amino acids comprising, consisting essentially of, or consisting of: (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)); and a retinoid (e.g., a retinoid, a retinoid derivative, a retinoid precursor, or combinations thereof), where the retinoid includes and is not limited to: retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof; and optionally a dermatologically acceptable vehicle and/or additive. Some aspects are directed to the formulation or the topical formulation of the disclosure comprising, as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids that comprises, consists essentially of, or consists of at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, or any combination thereof; and a therapeutically effective amount of a retinol or vitamin A; or combinations thereof; and optionally a dermatologically acceptable vehicle and/or additive. Further aspects of the formulation described here provide a therapeutically effective amount of the free amino acids or the peptide combination of amino acids and a therapeutically effective amount of the retinoid, a retinoid derivative, a retinoid precursor, or combinations thereof (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof); and optionally, a dermatologically acceptable vehicle and/or additive, sufficient to improve skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))) as indicated by, for example, a fold increase or fold change of a skin moisture marker or an HA marker gene, e.g., HAS1, HAS2, HAS3, or EGF expression of: 1.4 or greater (e.g., 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3, 1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6, 1; 6.3; 6.5; 6.7; 6.9; 7, 1; 7.3; 7.5; 7.7; 7.9; 8, 1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1; 11.3; 11.5; 11.7; 11.9; 12.1; 12.3; 12.5; 12.7; 12.9; 13.1; 13.3; 13.5; 13.7; 13.9; 14.1; 14.3; 14.5; 14.7; 14.9; 15.1; 15.3; 15.5; 15.7; 15.9; 16.1; 16.3; 16.5; 16.7; 16.9; 17.1; 17.3; 17.5; 17.7; 17.9; 18.1; 18.3; 18.5; 18.7; 18.9; 19.1; 19.3; 19.5; 19.7; 19.9; 20.1; 20.3; 20.5; 20.7; 20.9; 21.1; 21.3; 21.5; 21.7; 21.9; 22.1; 22.3; 22.5; 22.7; 22.9; 23.1; 23.3; 23.5; 23.7; 23.9; 23.1; 23.3; 23.5; 23.7; 23.9; 24.1; 24.3; 24.5; 24.7; 24.9; 25.1; 25.3; 25.5; 25.7; 25.9; 26.1; 26.3; 26.5; 26.7; 26.9; 27.1; 27.3; 27.5; 27.7; 27.9; 28.1; 28.3; 28.5; 28.7; 28.9; 29.1; 29.3; 29.5; 29.7; 29.9; 30.1; 30.3; 30.5; 30.7; 30.9; 31.1; 31.3; 31.5; 31.7; 31.9; 32.1; 32.3; 32.5; 32.7; 32.9; 33.1; 33.3; 33.5; 33.7; 33.9; 34.1; 34.3; 34.5; 34.7; 34.9; 35.1; 35.3; 35.5; 35.7; 35.9; 36.1; 36.3; 36.5; 36.7; 36.9; 37.1; 37.3; 37.5; 37.7; 37.9; 38.1; 38.3; 38.5; 38.7; 38.9; 39.1; 39.3; 39.5; 39.7; 39.9; 40.1; 40.3; 40.5; 40.7; 40.9; 41.1; 41.3; 41.5; 55; 70; 90; 105; 120; 170; 195; 205; 255; 270; 295; 305; 320; 355; 370; 405; 420; 455); 450 or less (e.g., 425; 405; 400; 375; 350; 325; 300; 290; 275; 250; 225; 200; 190; 175; 150; 125; 100; 95; 75; 50; 39.8; 39.6; 39.4; 39.2; 39; 38.8; 38.6; 38.4; 38.2; 38; 37.8; 37.6; 37.4; 37.2; 37; 36.8; 36.6; 36.4; 36.2; 36; 35.8; 35.6; 35.4; 35.2; 35; 34.8; 34.6; 34.4; 34.2; 34; 33.8; 33.6; 33.4; 33.2; 33; 32.8; 32.6; 32.4; 32.2; 32; 31.8; 31.6; 31.4; 31.2; 31; 30.8; 30.6; 30.4; 30.2; 30; 29.8; 29.6; 29.4; 29.2; 29; 28.8; 28.6; 28.4; 28.2; 28; 27.8; 27.6; 27.4; 27.2; 27; 26.8; 26.6; 26.4; 26.2; 26; 25.8; 25.6; 25.4; 25.2; 25; 24.8; 24.6; 24.4; 24.2; 24; 23.8; 23.6; 23.4; 23.2; 23; 22.8; 22.6; 22.4; 22.2; 22; 21.8; 21.6; 21.4; 21.2; 21; 20.8; 20.6; 20.4; 20.2; 20; 19.8; 19.6; 19.4; 19.2; 19; 18.8; 18.6; 18.4; 18.2; 18; 17.8; 17.6; 17.4; 17.2; 17.2; 17; 16.8; 16.6; 16.4; 16.2; 16; 15.8; 15.6; 15.4; 15.2; 15; 14.8; 14.6; 14.4; 14.2; 14; 13.8; 13.6; 13.4; 13.2; 13; 12.8; 12.6; 12.4; 12.2; 12; 11.8; 11.6; 11.4; 11.2; 11; 10.8; 10.6; 10.4; 10.2; 10; 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6); or 1.4-450 (e.g., 1.5-440; 1.6 445; 1.7-430; 1.8-420; 1.9-410; 2-400; 2.1-390; 2.2-380; 2.3-370; 2.4-360; 2.5-350; 2.6-340; 2.7-330; 2.8-320; 2.9-310; 3-300; 3.1-290; 3.2-280; 3.3-270; 3.4-260; 3.5-250; 3.6-240; 3.7-230; 3.8-220; 3.9-210; 4-200; 4.1-190; 4.2-180; 4.3-170; 4.4-160; 4.5 150; 4.6-140; 4.7-130; 4.8-120; 4.9-110; 5-100; 5.1-90; 5.2-80; 5.3-70; 5.4-60; 5.5-50; 5.6-40; 5.7-35.8; 5.8-35.7; 5.9-35.6; 6-35.5; 6.1-35.4; 6.2-35.3; 6.3-35.2; 6.4-35.1; 6.5-35; 6.6-34.9; 6.7-34.8; 6.8-34.7; 6.9-34.6; 7-34.5; 7.1-34.4; 7.2-34.3; 7.3-34.2; 7.4-34.1; 7.5-34; 7.6-33.9; 7.7-33.8; 7.8-33.7; 7.9-33.6; 8-33.5; 8.1-33.4; 8.2-33.3; 8.3-33.2; 8.4-33.1; 8.5-33; 8.6-32.9; 8.7-32.8; 8.8-32.7; 8.9-32.6; 9-32.5; 9.1-32.4; 9.2-32.3; 9.3-32.2; 9.4-32.1; 9.5-32; 9.6-31.9; 9.7-31.8; 9.8-31.7; 9.9-31.6; 10-31.5; 10.1-31.4; 10.2-31.3; 10.3-31.2; 10.4-31.1; 10.5-31; 10.6-30.9; 10.7-30.8; 10.8-30.7; 10.9-30.6; 11-30.5; 11.1-30.4; 11.2-30.3; 11.3-30.2; 11.4-30.1; 11.5-30; 11.6-29.9; 11.7-29.8; 11.8-29.7; 11.9-29.6; 12-29.5; 12.1-29.4; 12.2-29.3; 12.3-29.2; 12.4-29.1; 12.5-29; 12.6-28.9; 12.7-28.8; 12.8-28.7; 12.9-28.6; 13-28.5; 13.1-28.4; 13.2-28.3; 13.3-28.2; 13.4-28.1; 13.5-28; 13.6-27.9; 13.7-27.8; 13.8-27.7; 13.9-27.6; 14-27.5; 14.1-27.4; 14.2-27.3; 14.3-27.2; 14.4-27.1; 14.5-27; 14.6-26.9; 14.7-26.8; 14.8-26.7; 14.9-26.6; 15-26.5; 15.1-26.4; 15.2-26.3; 15.3-26.2; 15.4-26.1; 15.5-26; 15.6-25.9; 15.7-25.8; 15.8-25.7; 15.9-25.6; 16-25.5; 16.1-25.4; 16.2-25.3; 16.3-25.2; 16.4-25.1; 16.5-25; 16.6-24.9; 16.7-24.8; 16.8-24.7; 16.9-24.6; 17-24.5; 17.1-24.4; 17.2-24.3; 17.3-24.2; 17.4-24.1; 17.5-24; 17.6-23.9; 17.7-23.8; 17.8-23.7; 17.9-23.6; 18-23.5; 18.1-23.4; 18.2-23.3; 18.3-23.2; 18.4-23.1; 18.5-23; 18.6-22.9; 18.7-22.8; 18.8-22.7; 18.9-22.6; 19-22.5; 19.1-22.4; 19.2-22.3; 19.3-22.2; 19.4-22.1; 19.5-22; 19.6-21.9; 19.7-21.8; 19.8-21.7; 19.9-21.6; 20-21.5; 20.1-21.4; 20.2-21.3; 20.3-21.2; 20.4 21.1; 20.5-21; 20.6-20.9; 20.7-20.8). See, e.g., FIGS. 17-22, 24-27 (in part).

Further embodiments described here provide the formulation or the topical formulation of the disclosure comprising: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising, consisting essentially of, or consisting of at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and (i) a therapeutically effective amount of a plant extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense; or combinations thereof) or a plant extract combination (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications)) for skin regeneration, including skin barrier repair and strengthening; or (ii) a therapeutically effective amount of a retinoid, a retinoid derivative, or a retinoid precursor (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof) for improving skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))), wherein each of the free amino acids or the peptide combination of amino acids (at least one of: alanine, glutamine, glycine, and serine, or salts thereof) is present at a concentration: of 0.1 mM or greater (e.g., 0.3, 0.5, 0.7; 0.9; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5); of 10 mM or less (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0, 6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02); or ranging from 0.1 mM to 10 mM (e.g., 0.2 mM to 9.9 mM; 0.3 mM to 9.8 mM; 0.4 mM to 9.7 mM; 0.5 mM to 9.6 mM; 0.6 mM to 9.5 mM; 0.7 mM to 9.4 mM; 0.8 mM to 9.3 mM; 0.9 mM to 9.2 mM; 1 mM to 9.1 mM; 1.1 mM to 9 mM; 1.2 mM to 8.9 mM; 1.3 mM to 8.8 mM; 1.4 mM to 8.7 mM; 1.5 mM to 8.6 mM; 1.6 mM to 8.5 mM; 1.7 mM to 8.4 mM; 1.8 mM to 8.3 mM; 1.9 mM to 8.2 mM; 2 mM to 8.1 mM; 2.1 mM to 8 mM; 2.2 mM to 7.9 mM; 2.3 mM to 7.8 mM; 2.4 mM to 7.7 mM; 2.5 mM to 7.6 mM; 2.6 mM to 7.5 mM; 2.7 mM to 7.4 mM; 2.8 mM to 7.3 mM; 2.9 mM to 7.2 mM; 3 mM to 7.1 mM; 3.1 mM to 7 mM; 3.2 mM to 6.9 mM; 3.3 mM to 6.8 mM; 3.4 mM to 6.7 mM; 3.5 mM to 6.6 mM; 3.6 mM to 6.5 mM; 3.7 mM to 6.4 mM; 3.8 mM to 6.3 mM; 3.9 mM to 6.2 mM; 4 mM to 6.1 mM; 4.1 mM to 6 mM; 4.2 mM to 5.9 mM; 4.3 mM to 5.8 mM; 4.4 mM to 5.7 mM; 4.5 mM to 5.6 mM; 4.6 mM to 5.5 mM; 4.7 mM to 5.4 mM; 4.8 mM to 5.3 mM; 4.9 mM to 5.2 mM; 5 mM to 5.1 mM).

In some embodiments described here, the disclosed formulation or the topical formulation are provided, wherein the therapeutically effective amount of free amino acids or a peptide combination thereof, wherein each of the free amino acids or the peptide combination thereof (at least one of: alanine, glutamine, glycine, and serine, or salts thereof), is present at a weight percent to the formulation (wt/wt %) of: 0.001 wt % or greater (e.g., 0.002; 0.003; 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9; 1); 1 wt % or less (e.g., 0.95; 0.85; 0.75; 0.65; 0.55; 0.45; 0.35; 0.25; 0.15; 0.095; 0.085; 0.075; 0.065; 0.055; 0.045; 0.035; 0.025; 0.015; 0.0095; 0.0085; 0.0075; 0.0065; 0.0055; 0.0045; 0.0035; 0.0025; 0.0015); or 0.001 wt % to 1 wt % (e.g., 0.0012 wt % to 0.99 wt %; 0.0014 wt % to 0.97 wt %; 0.0016 wt % to 0.95 wt %; 0.0018 wt % to 0.93 wt %; 0.002 wt % to 0.91 wt %; 0.0022 wt % to 0.89 wt %; 0.0024 wt % to 0.87 wt %; 0.0026 wt % to 0.85 wt %; 0.0028 wt % to 0.83 wt %; 0.003 wt % to 0.81 wt %; 0.0032 wt % to 0.79 wt %; 0.0034 wt % to 0.77 wt %; 0.0036 wt % to 0.75 wt %; 0.0038 wt % to 0.73 wt %; 0.004 wt % to 0.71 wt %; 0.0042 wt % to 0.69 wt %; 0.0044 wt % to 0.67 wt %; 0.0046 wt % to 0.65 wt %; 0.0048 wt % to 0.63 wt %; 0.005 wt % to 0.61 wt %; 0.0052 wt % to 0.59 wt %; 0.0054 wt % to 0.57 wt %; 0.0056 wt % to 0.55 wt %; 0.0058 wt % to 0.53 wt %; 0.006 wt % to 0.51 wt %; 0.0062 wt % to 0.49 wt %; 0.0064 wt % to 0.47 wt %; 0.0066 wt % to 0.45 wt %; 0.0068 wt % to 0.43 wt %; 0.007 wt % to 0.41 wt %; 0.0072 wt % to 0.39 wt %; 0.0074 wt % to 0.37 wt %; 0.0076 wt % to 0.35 wt %; 0.0078 wt % to 0.33 wt %; 0.008 wt % to 0.31 wt %; 0.0082 wt % to 0.29 wt %; 0.0084 wt % to 0.27 wt %; 0.0086 wt % to 0.25 wt %; 0.0088 wt % to 0.23 wt %; 0.009 wt % to 0.21 wt %; 0.0092 wt % to 0.19 wt %; 0.0094 wt % to 0.17 wt %; 0.0096 wt % to 0.15 wt %; 0.0098 wt % to 0.13 wt %; 0.01 wt % to 0.11 wt %; 0.012 wt % to 0.099 wt %; 0.014 wt % to 0.097 wt %; 0.016 wt % to 0.095 wt %; 0.018 wt % to 0.093 wt %; 0.02 wt % to 0.091 wt %; 0.022 wt % to 0.089 wt %; 0.024 wt % to 0.087 wt %; 0.026 wt % to 0.085 wt %; 0.028 wt % to 0.083 wt %; 0.03 wt % to 0.081 wt %; 0.032 wt % to 0.079 wt %; 0.034 wt % to 0.077 wt %; 0.036 wt % to 0.075 wt %; 0.038 wt % to 0.073 wt %; 0.04 wt % to 0.071 wt %; 0.042 wt % to 0.069 wt %; 0.044 wt % to 0.067 wt %; 0.046 wt % to 0.065 wt %; 0.048 wt % to 0.063 wt %; 0.05 wt % to 0.061 wt %; 0.052 wt % to 0.059 wt %; 0.054 wt % to 0.057 wt %).

Other embodiments provide the formulation or the topical formulation described here, wherein the amino acid, alanine, is present at a weight percent to the formulation (wt/wt %) of: 0.0035 wt % or greater (e.g., 0.00352; 0.00354; 0.00356; 0.00358; 0.0036; 0.00362; 0.00364; 0.00366; 0.00368; 0.0037; 0.00372; 0.00374; 0.00376; 0.00378; 0.0038; 0.00382; 0.00384; 0.00386; 0.00388; 0.004; 0.0042; 0.0044; 0.0046; 0.0048; 0.005; 0.0052; 0.0054; 0.0056; 0.0058; 0.006; 0.0062; 0.0064; 0.0066; 0.0068; 0.007; 0.0072; 0.0074; 0.0076; 0.0078; 0.008; 0.0082; 0.0084; 0.0086; 0.0088; 0.009; 0.0092; 0.0094; 0.0096; 0.0098; 0.01; 0.012; 0.014; 0.016; 0.018; 0.02; 0.022; 0.024; 0.026; 0.028; 0.03; 0.032; 0.034; 0.036; 0.038; 0.04; 0.042; 0.044; 0.046; 0.048; 0.05; 0.052; 0.054; 0.056; 0.058; 0.06; 0.062; 0.064; 0.066; 0.068; 0.07; 0.072; 0.074; 0.076; 0.078; 0.08; 0.082; 0.084; 0.086; 0.088; 0.09; 0.092; 0.094; 0.096; 0.098; 0.1; 0.102; 0.104; 0.106; 0.108; 0.11; 0.112; 0.114; 0.116; 0.118; 0.12; 0.122; 0.124; 0.126; 0.128; 0.13; 0.132; 0.134; 0.136; 0.138; 0.14; 0.142; 0.144; 0.146; 0.148; 0.15; 0.152; 0.154; 0.156; 0.158; 0.16; 0.162; 0.164; 0.166; 0.168; 0.17; 0.172; 0.174; 0.176; 0.178; 0.18; 0.182; 0.184; 0.186; 0.188; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.202; 0.204; 0.206; 0.208; 0.21; 0.212; 0.214; 0.216; 0.218; 0.22; 0.222; 0.224; 0.226; 0.228; 0.23; 0.232; 0.234; 0.236; 0.238; 0.24; 0.242; 0.244; 0.246; 0.248; 0.25; 0.252; 0.254; 0.256; 0.258; 0.26; 0.262; 0.264; 0.266; 0.268; 0.27; 0.272; 0.274; 0.276; 0.278; 0.28; 0.282; 0.284; 0.286; 0.288; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.202; 0.204; 0.206; 0.208; 0.21; 0.212; 0.214; 0.216; 0.218; 0.22; 0.222; 0.224; 0.226; 0.228; 0.23; 0.232; 0.234; 0.236; 0.238; 0.24; 0.242; 0.244; 0.246; 0.248; 0.25; 0.252; 0.254; 0.256; 0.258; 0.26; 0.262; 0.264; 0.266; 0.268; 0.27; 0.272; 0.274; 0.276; 0.278; 0.28; 0.282; 0.284; 0.286; 0.288; 0.3; 0.302; 0.304; 0.306; 0.308; 0.31; 0.312; 0.314; 0.316; 0.318; 0.32; 0.322; 0.324; 0.326; 0.328; 0.33; 0.332; 0.334; 0.336; 0.338; 0.34; 0.342; 0.344; 0.346; 0.348; 0.35; 0.352; 0.354; 0.356; 0.358; 0.36; 0.362; 0.364; 0.366; 0.368; 0.37; 0.372; 0.374; 0.376; 0.378; 0.38; 0.382; 0.384; 0.386; 0.388; 0.39; 0.392; 0.394; 0.396; 0.398; 0.4; 0.402; 0.404; 0.406; 0.408; 0.41; 0.412; 0.414; 0.416; 0.418; 0.42; 0.422; 0.424; 0.426; 0.428; 0.43; 0.432; 0.434; 0.436; 0.438; 0.44; 0.442; 0.444; 0.446; 0.448; 0.45; 0.452; 0.454; 0.456; 0.458; 0.46; 0.462; 0.464; 0.466; 0.468; 0.47; 0.472; 0.474; 0.476; 0.478; 0.48; 0.482; 0.484; 0.486; 0.488; 0.5); 0.4 wt % or less (e.g., 0.399; 0.397; 0.395; 0.393; 0.391; 0.389; 0.387; 0.385; 0.383; 0.381; 0.379; 0.377; 0.375; 0.373; 0.371; 0.369; 0.367; 0.365; 0.363; 0.361; 0.359; 0.357; 0.355; 0.353; 0.351; 0.349; 0.347; 0.345; 0.343; 0.341; 0.339; 0.337; 0.335; 0.333; 0.331; 0.329; 0.327; 0.325; 0.323; 0.321; 0.319; 0.317; 0.315; 0.313; 0.311; 0.309; 0.307; 0.305; 0.303; 0.301; 0.299; 0.297; 0.295; 0.293; 0.291; 0.289; 0.287; 0.285; 0.283; 0.281; 0.279; 0.277; 0.275; 0.273; 0.271; 0.269; 0.267; 0.265; 0.263; 0.261; 0.259; 0.257; 0.255; 0.253; 0.251; 0.249; 0.247; 0.245; 0.243; 0.241; 0.239; 0.237; 0.235; 0.233; 0.231; 0.229; 0.227; 0.225; 0.223; 0.221; 0.219; 0.217; 0.215; 0.213; 0.211; 0.209; 0.207; 0.205; 0.203; 0.201; 0.199; 0.197; 0.195; 0.193; 0.191; 0.189; 0.187; 0.185; 0.183; 0.181; 0.179; 0.177; 0.175; 0.173; 0.171; 0.169; 0.167; 0.165; 0.163; 0.161; 0.159; 0.157; 0.155; 0.153; 0.151; 0.149; 0.147; 0.145; 0.143; 0.141; 0.139; 0.137; 0.135; 0.133; 0.131; 0.129; 0.127; 0.125; 0.123; 0.121; 0.119; 0.117; 0.115; 0.113; 0.111; 0.109; 0.107; 0.105; 0.103; 0.101; 0.099; 0.097; 0.095; 0.093; 0.091; 0.089; 0.087; 0.085; 0.083; 0.081; 0.079; 0.077; 0.075; 0.073; 0.071; 0.069; 0.067; 0.065; 0.063; 0.061; 0.059; 0.057; 0.055; 0.053; 0.051; 0.049; 0.047; 0.045; 0.043; 0.041; 0.039; 0.037; 0.035; 0.033; 0.031; 0.029; 0.027; 0.025; 0.023; 0.021; 0.019; 0.017; 0.015; 0.013; 0.011; 0.009; 0.007; 0.005; 0.003; 0.002); or 0.0035 wt % to 0.36 wt % (e.g., 0.00351 wt % to 0.359 wt %; 0.00352 wt % to 0.358 wt %; 0.00353 wt % to 0.357 wt %; 0.00354 wt % to 0.356 wt %; 0.00355 wt % to 0.355 wt %; 0.00356 wt % to 0.354 wt %; 0.00357 wt % to 0.353 wt %; 0.00358 wt % to 0.352 wt %; 0.00359 wt % to 0.351 wt %; 0.0036 wt % to 0.35 wt %; 0.00361 wt % to 0.349 wt %; 0.00362 wt % to 0.348 wt %; 0.00363 wt % to 0.347 wt %; 0.00364 wt % to 0.346 wt %; 0.00365 wt % to 0.345 wt %; 0.00366 wt % to 0.344 wt %; 0.00367 wt % to 0.343 wt %; 0.00368 wt % to 0.342 wt %; 0.00369 wt % to 0.341 wt %; 0.0037 wt % to 0.34 wt %; 0.00371 wt % to 0.339 wt %; 0.00372 wt % to 0.338 wt %; 0.00373 wt % to 0.337 wt %; 0.00374 wt % to 0.336 wt %; 0.00375 wt % to 0.335 wt %; 0.00376 wt % to 0.334 wt %; 0.00377 wt % to 0.333 wt %; 0.00378 wt % to 0.332 wt %; 0.00379 wt % to 0.331 wt %; 0.0038 wt % to 0.33 wt %; 0.00381 wt % to 0.329 wt %; 0.00382 wt % to 0.328 wt %; 0.00383 wt % to 0.327 wt %; 0.00384 wt % to 0.326 wt %; 0.00385 wt % to 0.325 wt %; 0.00386 wt % to 0.324 wt %; 0.00387 wt % to 0.323 wt %; 0.00388 wt % to 0.322 wt %; 0.00389 wt % to 0.321 wt %; 0.0039 wt % to 0.32 wt %; 0.00391 wt % to 0.319 wt %; 0.00392 wt % to 0.318 wt %; 0.00393 wt % to 0.317 wt %; 0.00394 wt % to 0.316 wt %; 0.00395 wt % to 0.315 wt %; 0.00396 wt % to 0.314 wt %; 0.00397 wt % to 0.313 wt %; 0.00398 wt % to 0.312 wt %; 0.00399 wt % to 0.311 wt %; 0.004 wt % to 0.31 wt %; 0.00401 wt % to 0.309 wt %; 0.00402 wt % to 0.308 wt %; 0.00403 wt % to 0.307 wt %; 0.00404 wt % to 0.306 wt %; 0.00405 wt % to 0.305 wt %; 0.00406 wt % to 0.304 wt %; 0.00407 wt % to 0.303 wt %; 0.00408 wt % to 0.302 wt %; 0.00409 wt % to 0.301 wt %; 0.0041 wt % to 0.3 wt %; 0.00411 wt % to 0.299 wt %; 0.00412 wt % to 0.298 wt %; 0.00413 wt % to 0.297 wt %; 0.00414 wt % to 0.296 wt %; 0.00415 wt % to 0.295 wt %; 0.00416 wt % to 0.294 wt %; 0.00417 wt % to 0.293 wt %; 0.00418 wt % to 0.292 wt %; 0.00419 wt % to 0.291 wt %; 0.0042 wt % to 0.29 wt %; 0.00421 wt % to 0.289 wt %; 0.00422 wt % to 0.288 wt %; 0.00423 wt % to 0.287 wt %; 0.00424 wt % to 0.286 wt %; 0.00425 wt % to 0.285 wt %; 0.00426 wt % to 0.284 wt %; 0.00427 wt % to 0.283 wt %; 0.00428 wt % to 0.282 wt %; 0.00429 wt % to 0.281 wt %; 0.0043 wt % to 0.28 wt %; 0.00431 wt % to 0.279 wt %; 0.00432 wt % to 0.278 wt %; 0.00433 wt % to 0.277 wt %; 0.00434 wt % to 0.276 wt %; 0.00435 wt % to 0.275 wt %; 0.00436 wt % to 0.274 wt %; 0.00437 wt % to 0.273 wt %; 0.00438 wt % to 0.272 wt %; 0.00439 wt % to 0.271 wt %; 0.0044 wt % to 0.27 wt %; 0.00441 wt % to 0.269 wt %; 0.00442 wt % to 0.268 wt %; 0.00443 wt % to 0.267 wt %; 0.00444 wt % to 0.266 wt %; 0.00445 wt % to 0.265 wt %; 0.00446 wt % to 0.264 wt %; 0.00447 wt % to 0.263 wt %; 0.00448 wt % to 0.262 wt %; 0.00449 wt % to 0.261 wt %; 0.0045 wt % to 0.26 wt %; 0.00451 wt % to 0.259 wt %; 0.00452 wt % to 0.258 wt %; 0.00453 wt % to 0.257 wt %; 0.00454 wt % to 0.256 wt %; 0.00455 wt % to 0.255 wt %; 0.00456 wt % to 0.254 wt %; 0.00457 wt % to 0.253 wt %; 0.00458 wt % to 0.252 wt %; 0.00459 wt % to 0.251 wt %; 0.0046 wt % to 0.25 wt %; 0.00461 wt % to 0.249 wt %; 0.00462 wt % to 0.248 wt %; 0.00463 wt % to 0.247 wt %; 0.00464 wt % to 0.246 wt %; 0.00465 wt % to 0.245 wt %; 0.00466 wt % to 0.244 wt %; 0.00467 wt % to 0.243 wt %; 0.00468 wt % to 0.242 wt %; 0.00469 wt % to 0.241 wt %; 0.0047 wt % to 0.24 wt %; 0.00471 wt % to 0.239 wt %; 0.00472 wt % to 0.238 wt %; 0.00473 wt % to 0.237 wt %; 0.00474 wt % to 0.236 wt %; 0.00475 wt % to 0.235 wt %; 0.00476 wt % to 0.234 wt %; 0.00477 wt % to 0.233 wt %; 0.00478 wt % to 0.232 wt %; 0.00479 wt % to 0.231 wt %; 0.0048 wt % to 0.23 wt %; 0.00481 wt % to 0.229 wt %; 0.00482 wt % to 0.228 wt %; 0.00483 wt % to 0.227 wt %; 0.00484 wt % to 0.226 wt %; 0.00485 wt % to 0.225 wt %; 0.00486 wt % to 0.224 wt %; 0.00487 wt % to 0.223 wt %; 0.00488 wt % to 0.222 wt %; 0.00489 wt % to 0.221 wt %; 0.0049 wt % to 0.22 wt %; 0.00491 wt % to 0.219 wt %; 0.00492 wt % to 0.218 wt %; 0.00493 wt % to 0.217 wt %; 0.00494 wt % to 0.216 wt %; 0.00495 wt % to 0.215 wt %; 0.00496 wt % to 0.214 wt %; 0.00497 wt % to 0.213 wt %; 0.00498 wt % to 0.212 wt %; 0.00499 wt % to 0.211 wt %; 0.005 wt % to 0.21 wt %; 0.00501 wt % to 0.209 wt %; 0.00502 wt % to 0.208 wt %; 0.00503 wt % to 0.207 wt %; 0.00504 wt % to 0.206 wt %; 0.00505 wt % to 0.205 wt %; 0.00506 wt % to 0.204 wt %; 0.00507 wt % to 0.203 wt %; 0.00508 wt % to 0.202 wt %; 0.00509 wt % to 0.201 wt %; 0.0051 wt % to 0.2 wt %; 0.00511 wt % to 0.199 wt %; 0.00512 wt % to 0.198 wt %; 0.00513 wt % to 0.197 wt %; 0.00514 wt % to 0.196 wt %; 0.00515 wt % to 0.195 wt %; 0.00516 wt % to 0.194 wt %; 0.00517 wt % to 0.193 wt %; 0.00518 wt % to 0.192 wt %; 0.00519 wt % to 0.191 wt %; 0.0052 wt % to 0.19 wt %; 0.00521 wt % to 0.189 wt %; 0.00522 wt % to 0.188 wt %; 0.00523 wt % to 0.187 wt %; 0.00524 wt % to 0.186 wt %; 0.00525 wt % to 0.185 wt %; 0.00526 wt % to 0.184 wt %; 0.00527 wt % to 0.183 wt %; 0.00528 wt % to 0.182 wt %; 0.00529 wt % to 0.181 wt %; 0.0053 wt % to 0.18 wt %; 0.00531 wt % to 0.179 wt %; 0.00532 wt % to 0.178 wt %; 0.00533 wt % to 0.177 wt %; 0.00534 wt % to 0.176 wt %; 0.00535 wt % to 0.175 wt %; 0.00536 wt % to 0.174 wt %; 0.00537 wt % to 0.173 wt %; 0.00538 wt % to 0.172 wt %; 0.00539 wt % to 0.171 wt %; 0.0054 wt % to 0.17 wt %; 0.00541 wt % to 0.169 wt %; 0.00542 wt % to 0.168 wt %; 0.00543 wt % to 0.167 wt %; 0.00544 wt % to 0.166 wt %; 0.00545 wt % to 0.165 wt %; 0.00546 wt % to 0.164 wt %; 0.00547 wt % to 0.163 wt %; 0.00548 wt % to 0.162 wt %; 0.00549 wt % to 0.161 wt %; 0.0055 wt % to 0.16 wt %; 0.00551 wt % to 0.159 wt %; 0.00552 wt % to 0.158 wt %; 0.00553 wt % to 0.157 wt %; 0.00554 wt % to 0.156 wt %; 0.00555 wt % to 0.155 wt %; 0.00556 wt % to 0.154 wt %; 0.00557 wt % to 0.153 wt %; 0.00558 wt % to 0.152 wt %; 0.00559 wt % to 0.151 wt %; 0.0056 wt % to 0.15 wt %; 0.00561 wt % to 0.149 wt %; 0.00562 wt % to 0.148 wt %; 0.00563 wt % to 0.147 wt %; 0.00564 wt % to 0.146 wt %; 0.00565 wt % to 0.145 wt %; 0.00566 wt % to 0.144 wt %; 0.00567 wt % to 0.143 wt %; 0.00568 wt % to 0.142 wt %; 0.00569 wt % to 0.141 wt %; 0.0057 wt % to 0.14 wt %; 0.00571 wt % to 0.139 wt %; 0.00572 wt % to 0.138 wt %; 0.00573 wt % to 0.137 wt %; 0.00574 wt % to 0.136 wt %; 0.00575 wt % to 0.135 wt %; 0.00576 wt % to 0.134 wt %; 0.00577 wt % to 0.133 wt %; 0.00578 wt % to 0.132 wt %; 0.00579 wt % to 0.131 wt %; 0.0058 wt % to 0.13 wt %; 0.00581 wt % to 0.129 wt %; 0.00582 wt % to 0.128 wt %; 0.00583 wt % to 0.127 wt %; 0.00584 wt % to 0.126 wt %; 0.00585 wt % to 0.125 wt %; 0.00586 wt % to 0.124 wt %; 0.00587 wt % to 0.123 wt %; 0.00588 wt % to 0.122 wt %; 0.00589 wt % to 0.121 wt %; 0.0059 wt % to 0.12 wt %; 0.00591 wt % to 0.119 wt %; 0.00592 wt % to 0.118 wt %; 0.00593 wt % to 0.117 wt %; 0.00594 wt % to 0.116 wt %; 0.00595 wt % to 0.115 wt %; 0.00596 wt % to 0.114 wt %; 0.00597 wt % to 0.113 wt %; 0.00598 wt % to 0.112 wt %; 0.00599 wt % to 0.111 wt %; 0.0056 wt % to 0.11 wt %; 0.00561 wt % to 0.109 wt %; 0.00562 wt % to 0.108 wt %; 0.00563 wt % to 0.107 wt %; 0.00564 wt % to 0.106 wt %; 0.00565 wt % to 0.105 wt %; 0.00566 wt % to 0.104 wt %; 0.00567 wt % to 0.103 wt %; 0.00568 wt % to 0.102 wt %; 0.00569 wt % to 0.101 wt %; 0.0057 wt % to 0.1 wt %; 0.00571 wt % to 0.099 wt %; 0.00572 wt % to 0.098 wt %; 0.00573 wt % to 0.097 wt %; 0.00574 wt % to 0.096 wt %; 0.00575 wt % to 0.095 wt %; 0.00576 wt % to 0.094 wt %; 0.00577 wt % to 0.093 wt %; 0.00578 wt % to 0.092 wt %; 0.00579 wt % to 0.091 wt %; 0.0058 wt % to 0.09 wt %; 0.00581 wt % to 0.089 wt %; 0.00582 wt % to 0.088 wt %; 0.00583 wt % to 0.087 wt %; 0.00584 wt % to 0.086 wt %; 0.00585 wt % to 0.085 wt %; 0.00586 wt % to 0.084 wt %; 0.00587 wt % to 0.083 wt %; 0.00588 wt % to 0.082 wt %; 0.00589 wt % to 0.081 wt %; 0.0059 wt % to 0.08 wt %; 0.00591 wt % to 0.079 wt %; 0.00592 wt % to 0.078 wt %; 0.00593 wt % to 0.077 wt %; 0.00594 wt % to 0.076 wt %; 0.00595 wt % to 0.075 wt %; 0.00596 wt % to 0.074 wt %; 0.00597 wt % to 0.073 wt %; 0.00598 wt % to 0.072 wt %; 0.00599 wt % to 0.071 wt %; 0.006 wt % to 0.07 wt %; 0.00601 wt % to 0.069 wt %; 0.00602 wt % to 0.068 wt %; 0.00603 wt % to 0.067 wt %; 0.00604 wt % to 0.066 wt %; 0.00605 wt % to 0.065 wt %; 0.00606 wt % to 0.064 wt %; 0.00607 wt % to 0.063 wt %; 0.00608 wt % to 0.062 wt %; 0.00609 wt % to 0.061 wt %; 0.0061 wt % to 0.06 wt %; 0.00611 wt % to 0.059 wt %; 0.00612 wt % to 0.058 wt %; 0.00613 wt % to 0.057 wt %; 0.00614 wt % to 0.056 wt %; 0.00615 wt % to 0.055 wt %; 0.00616 wt % to 0.054 wt %; 0.00617 wt % to 0.053 wt %; 0.00618 wt % to 0.052 wt %; 0.00619 wt % to 0.051 wt %; 0.0062 wt % to 0.05 wt %; 0.00621 wt % to 0.049 wt %; 0.00622 wt % to 0.048 wt %; 0.00623 wt % to 0.047 wt %; 0.00624 wt % to 0.046 wt %; 0.00625 wt % to 0.045 wt %; 0.00626 wt % to 0.044 wt %; 0.00627 wt % to 0.043 wt %; 0.00628 wt % to 0.042 wt %; 0.00629 wt % to 0.041 wt %; 0.0063 wt % to 0.04 wt %; 0.00631 wt % to 0.0399 wt %; 0.00632 wt % to 0.0398 wt %; 0.00633 wt % to 0.0397 wt %; 0.00534 wt % to 0.0396 wt %; 0.00535 wt % to 0.0395 wt %; 0.00536 wt % to 0.0394 wt %; 0.00537 wt % to 0.0393 wt %; 0.00538 wt % to 0.0392 wt %; 0.00539 wt % to 0.0391 wt %; 0.0054 wt % to 0.039 wt %; 0.00541 wt % to 0.0389 wt %; 0.00542 wt % to 0.0388 wt %; 0.00543 wt % to 0.0387 wt %; 0.00544 wt % to 0.0386 wt %; 0.00545 wt % to 0.0385 wt %; 0.00546 wt % to 0.0384 wt %; 0.00547 wt % to 0.0383 wt %; 0.00548 wt % to 0.0382 wt %; 0.00549 wt % to 0.0381 wt %; 0.0055 wt % to 0.038 wt %; 0.00551 wt % to 0.0379 wt %; 0.00552 wt % to 0.0378 wt %; 0.00553 wt % to 0.0377 wt %; 0.00554 wt % to 0.0376 wt %; 0.00555 wt % to 0.0375 wt %; 0.00556 wt % to 0.0374 wt %; 0.00557 wt % to 0.0373 wt %; 0.00558 wt % to 0.0372 wt %; 0.00559 wt % to 0.0371 wt %; 0.0056 wt % to 0.037 wt %; 0.00561 wt % to 0.0369 wt %; 0.00562 wt % to 0.0368 wt %; 0.00563 wt % to 0.0367 wt %; 0.00564 wt % to 0.0366 wt %; 0.00565 wt % to 0.0365 wt %; 0.00566 wt % to 0.0364 wt %; 0.00567 wt % to 0.0363 wt %; 0.00568 wt % to 0.0362 wt %; 0.00569 wt % to 0.0361 wt %; 0.0057 wt % to 0.036 wt %; 0.00571 wt % to 0.0359 wt %; 0.00572 wt % to 0.0358 wt %; 0.00573 wt % to 0.0357 wt %; 0.00574 wt % to 0.0356 wt %; 0.00575 wt % to 0.0355 wt %; 0.00576 wt % to 0.0354 wt %; 0.00577 wt % to 0.0353 wt %; 0.00578 wt % to 0.0352 wt %; 0.00579 wt % to 0.0351 wt %; 0.0058 wt % to 0.035 wt %; 0.00581 wt % to 0.0349 wt %; 0.00582 wt % to 0.0348 wt %; 0.00583 wt % to 0.0347 wt %; 0.00584 wt % to 0.0346 wt %; 0.00585 wt % to 0.0345 wt %; 0.00586 wt % to 0.0344 wt %; 0.00587 wt % to 0.0343 wt %; 0.00588 wt % to 0.0342 wt %; 0.00589 wt % to 0.0341 wt %; 0.0059 wt % to 0.034 wt %; 0.00591 wt % to 0.0339 wt %; 0.00592 wt % to 0.0338 wt %; 0.00593 wt % to 0.0337 wt %; 0.00594 wt % to 0.0336 wt %; 0.00595 wt % to 0.0335 wt %; 0.00596 wt % to 0.0334 wt %; 0.00597 wt % to 0.0333 wt %; 0.00598 wt % to 0.0332 wt %; 0.00599 wt % to 0.0331 wt %; 0.0056 wt % to 0.033 wt %; 0.00561 wt % to 0.0329 wt %; 0.00562 wt % to 0.0328 wt %; 0.00563 wt % to 0.0327 wt %; 0.00564 wt % to 0.0326 wt %; 0.00565 wt % to 0.0325 wt %; 0.00566 wt % to 0.0324 wt %; 0.00567 wt % to 0.0323 wt %; 0.00568 wt % to 0.0322 wt %; 0.00569 wt % to 0.0321 wt %; 0.0057 wt % to 0.032 wt %; 0.00571 wt % to 0.0319 wt %; 0.00572 wt % to 0.0318 wt %; 0.00573 wt % to 0.0317 wt %; 0.00574 wt % to 0.0316 wt %; 0.00575 wt % to 0.0315 wt %; 0.00576 wt % to 0.0314 wt %; 0.00577 wt % to 0.0313 wt %; 0.00578 wt % to 0.0312 wt %; 0.00579 wt % to 0.0311 wt %; 0.0058 wt % to 0.031 wt %; 0.00581 wt % to 0.0309 wt %; 0.00582 wt % to 0.0308 wt %; 0.00583 wt % to 0.0307 wt %; 0.00584 wt % to 0.0306 wt %; 0.00585 wt % to 0.0305 wt %; 0.00586 wt % to 0.0304 wt %; 0.00587 wt % to 0.0303 wt %; 0.00588 wt % to 0.0302 wt %; 0.00589 wt % to 0.0301 wt %; 0.0059 wt % to 0.03 wt %; 0.00591 wt % to 0.0299 wt %; 0.00592 wt % to 0.0298 wt %; 0.00593 wt % to 0.0297 wt %; 0.00594 wt % to 0.0296 wt %; 0.00595 wt % to 0.0295 wt %; 0.00596 wt % to 0.0294 wt %; 0.00597 wt % to 0.0293 wt %; 0.00598 wt % to 0.0292 wt %; 0.00599 wt % to 0.0291 wt %; 0.006 wt % to 0.029 wt %; 0.00601 wt % to 0.0289 wt %; 0.00602 wt % to 0.0288 wt %; 0.00603 wt % to 0.0287 wt %; 0.00604 wt % to 0.0286 wt %; 0.00605 wt % to 0.0285 wt %; 0.00606 wt % to 0.0284 wt %; 0.00607 wt % to 0.0283 wt %; 0.00608 wt % to 0.0282 wt %; 0.00609 wt % to 0.0281 wt %; 0.0061 wt % to 0.028 wt %; 0.00611 wt % to 0.0279 wt %; 0.00612 wt % to 0.0278 wt %; 0.00613 wt % to 0.0277 wt %; 0.00614 wt % to 0.0276 wt %; 0.00615 wt % to 0.0275 wt %; 0.00616 wt % to 0.0274 wt %; 0.00617 wt % to 0.0273 wt %; 0.00618 wt % to 0.0272 wt %; 0.00619 wt % to 0.0271 wt %; 0.0062 wt % to 0.027 wt %; 0.00621 wt % to 0.0269 wt %; 0.00622 wt % to 0.0268 wt %; 0.00623 wt % to 0.0267 wt %; 0.00624 wt % to 0.0266 wt %; 0.00625 wt % to 0.0265 wt %; 0.00626 wt % to 0.0264 wt %; 0.00627 wt % to 0.0263 wt %; 0.00628 wt % to 0.0262 wt %; 0.00629 wt % to 0.0261 wt %; 0.0063 wt % to 0.026 wt %; 0.00631 wt % to 0.0259 wt %).

In additional embodiments, the formulation or the topical formulation described here comprises the amino acid, glutamine, present at a weight percent (wt %) to the formulation of: 0.00584 wt % or greater (e.g., 0.0059; 0.006; 0.007; 0.008; 0.009; 0.01; 0.011; 0.012; 0.013; 0.014; 0.015; 0.016; 0.017; 0.018; 0.019; 0.02; 0.021; 0.022; 0.023; 0.024; 0.025; 0.026; 0.027; 0.028; 0.029; 0.03; 0.031; 0.032; 0.033; 0.034; 0.035; 0.036; 0.037; 0.038; 0.039; 0.04; 0.041; 0.042; 0.043; 0.044; 0.045; 0.046; 0.047; 0.048; 0.049; 0.05; 0.051; 0.052; 0.053; 0.054; 0.055; 0.056; 0.057; 0.058; 0.059; 0.06; 0.061; 0.062; 0.063; 0.064; 0.065; 0.066; 0.067; 0.068; 0.069; 0.07; 0.071; 0.072; 0.073; 0.074; 0.075; 0.076; 0.077; 0.078; 0.079; 0.08; 0.081; 0.082; 0.083; 0.084; 0.085; 0.086; 0.087; 0.088; 0.089; 0.09; 0.091; 0.092; 0.093; 0.094; 0.095; 0.096; 0.097; 0.098; 0.099; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.2; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.3; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.4; 0.41; 0.42; 0.43; 0.44; 0.45; 0.46; 0.47; 0.48; 0.49; 0.5; 0.51; 0.52; 0.53; 0.54; 0.55; 0.56; 0.57; 0.58; 0.59; 0.6); 0.584 wt % or less (e.g., 0.583; 0.582; 0.581; 0.58; 0.579; 0.578; 0.577; 0.576; 0.575; 0.574; 0.573; 0.572; 0.571; 0.57; 0.569; 0.568; 0.567; 0.566; 0.565; 0.564; 0.563; 0.562; 0.561; 0.56; 0.559; 0.558; 0.557; 0.556; 0.554; 0.553; 0.552; 0.551; 0.55; 0.549; 0.548; 0.547; 0.546; 0.545; 0.544; 0.543; 0.542; 0.541; 0.54; 0.539; 0.538; 0.537; 0.536; 0.535; 0.534; 0.533; 0.532; 0.531; 0.53; 0.529; 0.528; 0.527; 0.526; 0.525; 0.524; 0.523; 0.522; 0.521; 0.52; 0.519; 0.518; 0.517; 0.516; 0.515; 0.514; 0.513; 0.512; 0.511; 0.51; 0.509; 0.508; 0.507; 0.506; 0.505; 0.504; 0.503; 0.502; 0.501; 0.5; 0.499; 0.498; 0.497; 0.496; 0.495; 0.494; 0.493; 0.492; 0.491; 0.49; 0.489; 0.488; 0.487; 0.486; 0.485; 0.484; 0.483; 0.482; 0.481; 0.48; 0.479; 0.478; 0.477; 0.476; 0.475; 0.474; 0.473; 0.472; 0.471; 0.47; 0.469; 0.468; 0.467; 0.466; 0.465; 0.464; 0.463; 0.462; 0.461; 0.46; 0.459; 0.458; 0.457; 0.456; 0.455; 0.454; 0.453; 0.452; 0.451; 0.45; 0.449; 0.448; 0.447; 0.446; 0.445; 0.444; 0.443; 0.442; 0.441; 0.44; 0.439; 0.438; 0.437; 0.436; 0.435; 0.434; 0.433; 0.432; 0.431; 0.43; 0.429; 0.428; 0.427; 0.426; 0.425; 0.424; 0.423; 0.422; 0.421; 0.42; 0.419; 0.418; 0.417; 0.416; 0.415; 0.414; 0.413; 0.412; 0.411; 0.41; 0.409; 0.408; 0.407; 0.406; 0.405; 0.404; 0.403; 0.402; 0.401; 0.4; 0.3; 0.2; 0.1; 0.09; 0.08; 0.07; 0.06; 0.05; 0.04; 0.03; 0.02; 0.01; 0.009; 0.008; 0.007; 0.006; 0.005; 0.004; 0.003; 0.002; 0.001); or 0.00584 wt % to 0.584 wt % (e.g., 0.00585 wt % to 0.582 wt %; 0.0059 wt % to 0.58 wt %; 0.0065 wt % to 0.578 wt %; 0.007 wt % to 0.576 wt %; 0.0075 wt % to 0.574 wt %; 0.008 wt % to 0.572 wt %; 0.0085 wt % to 0.57 wt %; 0.009 wt % to 0.568 wt %; 0.0095 wt % to 0.566 wt %; 0.01 wt % to 0.564 wt %; 0.015 wt % to 0.562 wt %; 0.02 wt % to 0.56 wt %; 0.025 wt % to 0.558 wt %; 0.03 wt % to 0.556 wt %; 0.035 wt % to 0.554 wt %; 0.04 wt % to 0.552 wt %; 0.045 wt % to 0.55 wt %; 0.05 wt % to 0.548 wt %; 0.055 wt % to 0.546 wt %; 0.06 wt % to 0.544 wt %; 0.065 wt % to 0.542 wt %; 0.07 wt % to 0.54 wt %; 0.075 wt % to 0.538 wt %; 0.08 wt % to 0.536 wt %; 0.085 wt % to 0.534 wt %; 0.09 wt % to 0.532 wt %; 0.095 wt % to 0.53 wt %; 0.1 wt % to 0.528 wt %; 0.15 wt % to 0.526 wt %; 0.2 wt % to 0.524 wt %; 0.25 wt % to 0.522 wt %; 0.3 wt % to 0.52 wt %; 0.35 wt % to 0.518 wt %; 0.4 wt % to 0.516 wt %; 0.45 wt % to 0.514 wt %; 0.5 wt % to 0.512 wt %).

Further embodiments described here provide the formulation or the topical formulation described here, wherein the amino acid, glycine, is present at a weight percent (wt %) to the formulation of: 0.003 wt % or greater (e.g., 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.2; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.3; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.4); 0.3 wt % or less (e.g., 0.298; 0.296; 0.294; 0.292; 0.29; 0.288; 0.286; 0.284; 0.282; 0.28; 0.278; 0.276; 0.274; 0.272; 0.27; 0.268; 0.266; 0.264; 0.262; 0.26; 0.258; 0.256; 0.254; 0.252; 0.25; 0.248; 0.246; 0.244; 0.242; 0.24; 0.238; 0.236; 0.234; 0.232; 0.23; 0.228; 0.226; 0.224; 0.222; 0.22; 0.218; 0.216; 0.214; 0.212; 0.21; 0.208; 0.206; 0.204; 0.202; 0.2; 0.198; 0.196; 0.194; 0.192; 0.19; 0.188; 0.186; 0.184; 0.182; 0.18; 0.178; 0.176; 0.174; 0.172; 0.17; 0.168; 0.166; 0.164; 0.162; 0.16; 0.158; 0.156; 0.154; 0.152; 0.15; 0.148; 0.146; 0.144; 0.142; 0.14; 0.138; 0.136; 0.134; 0.132; 0.13; 0.128; 0.126; 0.124; 0.122; 0.12; 0.118; 0.116; 0.114; 0.112; 0.11; 0.108; 0.106; 0.104; 0.102; 0.1; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.008; 0.006; 0.004; 0.002); or 0.003 wt % to 0.3 wt % (e.g., 0.0035 wt % to 0.29 wt %; 0.004 wt % to 0.27 wt %; 0.0045 wt % to 0.25 wt %; 0.005 wt % to 0.23 wt %; 0.0055 wt % to 0.21 wt %; 0.006 wt % to 0.19 wt %; 0.0065 wt % to 0.17 wt %; 0.007 wt % to 0.15 wt %; 0.0075 wt % to 0.13 wt %; 0.008 wt % to 0.11 wt %; 0.0085 wt % to 0.099 wt %; 0.009 wt % to 0.097 wt %; 0.0095 wt % to 0.095 wt %; 0.01 wt % to 0.093 wt %; 0.015 wt % to 0.091 wt %; 0.02 wt % to 0.089 wt %; 0.025 wt % to 0.087 wt %; 0.03 wt % to 0.085 wt %; 0.035 wt % to 0.083 wt %; 0.04 wt % to 0.081 wt %; 0.045 wt % to 0.079 wt %; 0.05 wt % to 0.077 wt %; 0.055 wt % to 0.075 wt %; 0.06 wt % to 0.073 wt %; 0.065 wt % to 0.071 wt %).

In some embodiments, the formulation or the topical formulation of the disclosure comprises the amino acid, serine, present at a weight percent (wt %) to the formulation of: 0.0042 wt % or greater (e.g., 0.0044; 0.0046; 0.0048; 0.005; 0.0052; 0.0054; 0.0056; 0.0058; 0.006; 0.0062; 0.0064; 0.0066; 0.0068; 0.007; 0.0072; 0.0074; 0.0076; 0.0078; 0.008; 0.0082; 0.0084; 0.0086; 0.0088; 0.009; 0.0092; 0.0094; 0.0096; 0.0098; 0.01; 0.0102; 0.0104; 0.0106; 0.0108; 0.011; 0.0112; 0.0114; 0.0116; 0.0118; 0.012; 0.0122; 0.0124; 0.0126; 0.0128; 0.013; 0.0132; 0.0134; 0.0136; 0.0138; 0.014; 0.0142; 0.0144; 0.0146; 0.0148; 0.015; 0.0152; 0.0154; 0.0156; 0.0158; 0.016; 0.0162; 0.0164; 0.0166; 0.0168; 0.017; 0.0172; 0.0174; 0.0176; 0.0178; 0.018; 0.0182; 0.0184; 0.0186; 0.0188; 0.019; 0.0192; 0.0194; 0.0196; 0.0198; 0.02; 0.0202; 0.0204; 0.0206; 0.208; 0.021; 0.0212; 0.0214; 0.0216; 0.0218; 0.022; 0.0222; 0.0224; 0.0226; 0.0228; 0.023; 0.0232; 0.0234; 0.0236; 0.0238; 0.024; 0.0242; 0.0244; 0.0246; 0.0248; 0.025; 0.0252; 0.0254; 0.0256; 0.0258; 0.026; 0.0262; 0.0264; 0.0266; 0.0268; 0.027; 0.0272; 0.0274; 0.0276; 0.0278; 0.028; 0.0282; 0.0284; 0.0286; 0.0288; 0.029; 0.0292; 0.0294; 0.0296; 0.0298; 0.03; 0.0302; 0.0304; 0.0306; 0.0308; 0.031; 0.0312; 0.0314; 0.0316; 0.0318; 0.032; 0.0322; 0.0324; 0.0326; 0.0328; 0.033; 0.0332; 0.0334; 0.0336; 0.0338; 0.034; 0.0342; 0.0344; 0.0346; 0.0348; 0.035; 0.0352; 0.0354; 0.0356; 0.0358; 0.036; 0.0362; 0.0364; 0.0366; 0.0368; 0.037; 0.0372; 0.0374; 0.0376; 0.0378; 0.038; 0.0382; 0.0384; 0.0386; 0.0388; 0.039; 0.0392; 0.0394; 0.0396; 0.0398; 0.04; 0.0402; 0.0404; 0.0406; 0.0408; 0.041; 0.0412; 0.0414; 0.0416; 0.0418; 0.042; 0.0422; 0.0424; 0.0426; 0.0428; 0.043; 0.0432; 0.0434; 0.0436; 0.0438; 0.044; 0.0442; 0.0444; 0.0446; 0.0448; 0.045; 0.0452; 0.0454; 0.0456; 0.0458; 0.046; 0.0462; 0.0464; 0.0466; 0.0468; 0.047; 0.0472; 0.0474; 0.0476; 0.0478; 0.048; 0.0482; 0.0484; 0.0486; 0.0488; 0.049; 0.0492; 0.0494; 0.0496; 0.0498; 0.05; 0.0502; 0.0504; 0.0506; 0.0508; 0.051; 0.0512; 0.0514; 0.0516; 0.0518; 0.052; 0.0522; 0.0524; 0.0526; 0.0528; 0.053; 0.0532; 0.0534; 0.0536; 0.0538; 0.054; 0.0542; 0.0544; 0.0546; 0.0548; 0.055; 0.0552; 0.0554; 0.0556; 0.0558; 0.056; 0.0562; 0.0564; 0.0566; 0.0568; 0.057; 0.0572; 0.0574; 0.0576; 0.0578; 0.058; 0.0582; 0.0584; 0.0586; 0.0588; 0.059; 0.0592; 0.0594; 0.0596; 0.0598; 0.06; 0.0602; 0.0604; 0.0606; 0.0608; 0.061; 0.0612; 0.0614; 0.0616; 0.0618; 0.062; 0.0622; 0.0624; 0.0626; 0.0628; 0.063; 0.0632; 0.0634; 0.0636; 0.0638; 0.064; 0.0642; 0.0644; 0.0646; 0.0648; 0.065; 0.0652; 0.0654; 0.0656; 0.0658; 0.066; 0.0662; 0.0664; 0.0666; 0.0668; 0.067; 0.0672; 0.0674; 0.0676; 0.0678; 0.068; 0.0682; 0.0684; 0.0686; 0.0688; 0.069; 0.0692; 0.0694; 0.0696; 0.0698; 0.07; 0.0702; 0.0704; 0.0706; 0.0708; 0.071; 0.0712; 0.0714; 0.0716; 0.0718; 0.072; 0.0722; 0.0724; 0.0726; 0.0728; 0.073; 0.0732; 0.0734; 0.0736; 0.0738; 0.074; 0.0742; 0.0744; 0.0746; 0.0748; 0.075; 0.0752; 0.0754; 0.0756; 0.0758; 0.076; 0.0762; 0.0764; 0.0766; 0.0768; 0.077; 0.0772; 0.0774; 0.0776; 0.0778; 0.078; 0.0782; 0.0784; 0.0786; 0.0788; 0.079; 0.0792; 0.0794; 0.0796; 0.0798; 0.08; 0.0802; 0.0804; 0.0806; 0.0808; 0.081; 0.0812; 0.0814; 0.0816; 0.0818; 0.082; 0.0822; 0.0824; 0.0826; 0.0828; 0.083; 0.0832; 0.0834; 0.0836; 0.0838; 0.084; 0.0842; 0.0844; 0.0846; 0.0848; 0.085; 0.0852; 0.0854; 0.0856; 0.0858; 0.086; 0.0862; 0.0864; 0.0866; 0.0868; 0.087; 0.0872; 0.0874; 0.0876; 0.0878; 0.088; 0.0882; 0.0884; 0.0886; 0.0888; 0.089; 0.0892; 0.0894; 0.0896; 0.0898; 0.09; 0.0902; 0.0904; 0.0906; 0.0908; 0.091; 0.0912; 0.0914; 0.0916; 0.0918; 0.092; 0.0922; 0.0924; 0.0926; 0.0928; 0.093; 0.0932; 0.0934; 0.0936; 0.0938; 0.094; 0.0942; 0.0944; 0.0946; 0.0948; 0.095; 0.0952; 0.0954; 0.0956; 0.0958; 0.096; 0.0962; 0.0964; 0.0966; 0.0968; 0.097; 0.0972; 0.0974; 0.0976; 0.0978; 0.098; 0.0982; 0.0984; 0.0986; 0.0988; 0.099; 0.0992; 0.0994; 0.0996; 0.0998; 0.1; 0.102; 0.104; 0.106; 0.108; 0.11; 0.112; 0.114; 0.116; 0.118; 0.12; 0.122; 0.124; 0.126; 0.128; 0.13; 0.132; 0.134; 0.136; 0.138; 0.14; 0.142; 0.144; 0.146; 0.148; 0.15; 0.152; 0.154; 0.156; 0.158; 0.16; 0.162; 0.164; 0.166; 0.168; 0.17; 0.172; 0.174; 0.176; 0.178; 0.18; 0.182; 0.184; 0.186; 0.188; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.22; 0.24; 0.26; 0.28; 0.3; 0.32; 0.34; 0.36; 0.38; 0.4; 0.42; 0.44; 0.46; 0.48; 0.5); 0.42 wt % or less (e.g., 0.418; 0.416; 0.414; 0.412; 0.41; 0.408; 0.406; 0.404; 0.402; 0.4; 0.398; 0.396; 0.394; 0.392; 0.39; 0.388; 0.386; 0.384; 0.382; 0.38; 0.378; 0.376; 0.374; 0.372; 0.37; 0.368; 0.366; 0.364; 0.362; 0.36; 0.358; 0.356; 0.354; 0.352; 0.35; 0.348; 0.346; 0.344; 0.342; 0.34; 0.338; 0.336; 0.334; 0.332; 0.33; 0.328; 0.326; 0.324; 0.322; 0.32; 0.318; 0.316; 0.314; 0.312; 0.31; 0.308; 0.306; 0.304; 0.302; 0.3; 0.298; 0.296; 0.294; 0.292; 0.294; 0.292; 0.29; 0.288; 0.286; 0.284; 0.282; 0.28; 0.278; 0.276; 0.274; 0.272; 0.27; 0.268; 0.266; 0.264; 0.262; 0.26; 0.258; 0.256; 0.254; 0.252; 0.25; 0.248; 0.246; 0.244; 0.242; 0.24; 0.238; 0.236; 0.234; 0.232; 0.23; 0.228; 0.226; 0.224; 0.222; 0.22; 0.218; 0.216; 0.214; 0.212; 0.21; 0.208; 0.206; 0.204; 0.202; 0.2; 0.198; 0.196; 0.194; 0.192; 0.19; 0.188; 0.186; 0.184; 0.182; 0.18; 0.178; 0.176; 0.174; 0.172; 0.17; 0.168; 0.166; 0.164; 0.162; 0.16; 0.158; 0.156; 0.154; 0.152; 0.15; 0.148; 0.146; 0.144; 0.142; 0.14; 0.138; 0.136; 0.134; 0.132; 0.13; 0.128; 0.126; 0.124; 0.122; 0.12; 0.118; 0.116; 0.114; 0.112; 0.11; 0.108; 0.106; 0.104; 0.102; 0.1; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.0088; 0.0086; 0.0084; 0.0082; 0.008; 0.0078; 0.0076; 0.0074; 0.0072; 0.007; 0.0068; 0.0066; 0.0064; 0.0062; 0.006; 0.0058; 0.0056; 0.0054; 0.0052; 0.005; 0.0048; 0.0046; 0.0044; 0.0042; 0.004); or 0.0042 wt % to 0.42 wt % (e.g., 0.0043 wt % to 0.415 wt %; 0.0045 wt % to 0.41 wt %; 0.0047 wt % to 0.405 wt %; 0.0049 wt % to 0.4 wt %; 0.0051 wt % to 0.395 wt %; 0.0053 wt % to 0.39 wt %; 0.0055 wt % to 0.385 wt %; 0.0057 wt % to 0.38 wt %; 0.0059 wt % to 0.375 wt %; 0.0061 wt % to 0.37 wt %; 0.0063 wt % to 0.365 wt %; 0.0065 wt % to 0.36 wt %; 0.0067 wt % to 0.355 wt %; 0.0069 wt % to 0.35 wt %; 0.0061 wt % to 0.345 wt %; 0.0063 wt % to 0.34 wt %; 0.0065 wt % to 0.335 wt %; 0.0067 wt % to 0.33 wt %; 0.0069 wt % to 0.325 wt %; 0.0071 wt % to 0.32 wt %; 0.0073 wt % to 0.315 wt %; 0.0075 wt % to 0.31 wt %; 0.0077 wt % to 0.305 wt %; 0.0079 wt % to 0.3 wt %; 0.0081 wt % to 0.295 wt %; 0.0083 wt % to 0.29 wt %; 0.0085 wt % to 0.285 wt %; 0.0087 wt % to 0.28 wt %; 0.0089 wt % to 0.275 wt %; 0.0091 wt % to 0.27 wt %; 0.0093 wt % to 0.265 wt %; 0.0095 wt % to 0.26 wt %; 0.0097 wt % to 0.255 wt %; 0.0099 wt % to 0.35 wt %; 0.0101 wt % to 0.345 wt %; 0.0103 wt % to 0.34 wt %; 0.0105 wt % to 0.335 wt %; 0.0107 wt % to 0.33 wt %; 0.0109 wt % to 0.325 wt %; 0.0111 wt % to 0.32 wt %; 0.0113 wt % to 0.315 wt %; 0.0115 wt % to 0.31 wt %; 0.0117 wt % to 0.305 wt %; 0.0119 wt % to 0.3 wt %; 0.0121 wt % to 0.295 wt %; 0.0123 wt % to 0.29 wt %; 0.0125 wt % to 0.285 wt %; 0.0127 wt % to 0.28 wt %; 0.0129 wt % to 0.275 wt %; 0.0131 wt % to 0.27 wt %; 0.0133 wt % to 0.265 wt %; 0.0135 wt % to 0.26 wt %; 0.0137 wt % to 0.255 wt %; 0.0139 wt % to 0.25 wt %; 0.0141 wt % to 0.245 wt %; 0.0143 wt % to 0.24 wt %; 0.0145 wt % to 0.235 wt %; 0.0147 wt % to 0.23 wt %; 0.0149 wt % to 0.225 wt %; 0.0151 wt % to 0.22 wt %; 0.0153 wt % to 0.215 wt %; 0.0155 wt % to 0.21 wt %; 0.0157 wt % to 0.205 wt %; 0.0159 wt % to 0.2 wt %; 0.0161 wt % to 0.195 wt %; 0.0163 wt % to 0.19 wt %; 0.0165 wt % to 0.185 wt %; 0.0167 wt % to 0.18 wt %; 0.0169 wt % to 0.175 wt %; 0.0171 wt % to 0.17 wt %; 0.0173 wt % to 0.165 wt %; 0.0175 wt % to 0.16 wt %; 0.0177 wt % to 0.155 wt %; 0.0179 wt % to 0.15 wt %; 0.0181 wt % to 0.145 wt %; 0.0183 wt % to 0.14 wt %; 0.0185 wt % to 0.135 wt %; 0.0187 wt % to 0.13 wt %; 0.0189 wt % to 0.125 wt %; 0.0191 wt % to 0.12 wt %; 0.0193 wt % to 0.115 wt %; 0.0195 wt % to 0.11 wt %; 0.0197 wt % to 0.105 wt %; 0.0199 wt % to 0.1 wt %; 0.0201 wt % to 0.095 wt %; 0.0203 wt % to 0.09 wt %; 0.0205 wt % to 0.085 wt %; 0.0207 wt % to 0.08 wt %; 0.0209 wt % to 0.075 wt %; 0.0211 wt % to 0.07 wt %; 0.0213 wt % to 0.065 wt %; 0.0215 wt % to 0.06 wt %; 0.0217 wt % to 0.055 wt %; 0.0219 wt % to 0.05 wt %; 0.0221 wt % to 0.045 wt %; 0.0223 wt % to 0.04 wt %; 0.0225 wt % to 0.035 wt %; 0.0227 wt % to 0.03 wt %; 0.0229 wt % to 0.025 wt %).

Additional embodiments described here provides the formulation or the topical formulation disclosed here, where the therapeutically effective amount of the amino acid combination comprises, consists essentially of, or consists of: alanine (4 mM) of 0.0356%; glutamine (4 mM) of 0.0584%; glycine (4 mM) of 0.03%; and serine (4 mM) of 0.042%. In some embodiments, the formulation or the topical formulation disclosed here comprising the therapeutically effective amount of the free amino acid or a peptide combination thereof, where the free amino acid or the peptide combination comprises, consists essentially of, or consists of: alanine (4 mM) of 0.0356%; glutamine (4 mM) of 0.0584%; glycine (4 mM) of 0.03%; and serine (4 mM) of 0.042%, and further comprises, the therapeutically effective amount of the plant extract or plant extract combination comprising (or consisting essentially of or consisting of) a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or the plant extract combination is in combination with at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications) described here, for skin regeneration, including skin barrier repair and strengthening, or the therapeutically effective amount of a retinoid or at least one retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof) for improving skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))).

Other embodiments described here provide the formulation or the topical formulation disclosed here, wherein the therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids comprises (or consists essentially of or consists of): Alanine (Ala), Glutamine (Gln), Glycine (Gly), and Serine (Ser), wherein each amino acid comprises a ratio to each of the other amino acids of: 1:100 [0.1 to 10] or greater (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):1 (Gln):100 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):100 (Gln):1 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):100 (Gln):100 (Gly):1 (Ser) [0.1 to 10]; 1 (Ala):90 (Gln):90 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):1 (Gln):90 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):90 (Gln):1 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):90 (Gln):90 (Gly):1 (Ser) [0.1 to 9]; 1 (Ala):80 (Gln):80 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):1 (Gln):80 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):80 (Gln):1 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):80 (Gln):80 (Gly):1 (Ser) [0.1 to 8]; 1 (Ala):70 (Gln):70 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):1 (Gln):70 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):70 (Gln):1 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):70 (Gln):70 (Gly):1 (Ser) [0.1 to 7]; 1 (Ala):60 (Gln):60 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):1 (Gln):60 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):60 (Gln):1 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):60 (Gln):60 (Gly):1 (Ser) [0.1 to 6]; 1 (Ala):50 (Gln):50 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):1 (Gln):50 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):50 (Gln):1 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):50 (Gln):50 (Gly):1 (Ser) [0.1 to 5]; 1 (Ala):40 (Gln):40 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):1 (Gln):40 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):40 (Gln):1 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):40 (Gln):40 (Gly):1 (Ser) [0.1 to 4]; 1 (Ala):30 (Gln):30 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):1 (Gln):30 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):30 (Gln):1 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):30 (Gln):30 (Gly):1 (Ser) [0.1 to 3]; 1 (Ala):20 (Gln):20 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):1 (Gln):20 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):20 (Gln):1 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):20 (Gln):20 (Gly):1 (Ser) [0.1 to 2]; 1 (Ala):10 (Gln):10 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):1 (Gln):10 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):10 (Gln):1 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):10 (Gln):10 (Gly):1 (Ser) [0.1 to 1]; 1 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):1 (Gln):5 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):5 (Gln):1 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):5 (Gln):5 (Gly):1 (Ser) [0.1 to 0.5]; 2 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):2 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):2 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):5 (Gly):2 (Ser) [0.1 to 0.25]; 1 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.1 to 0.1]; 2 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):2 (Gln):1 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):1 (Gln):2 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):2 (Ser) [0.2 to 0.1]; 3 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):3 (Gln):1 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):1 (Gln):3 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):3 (Ser) [0.3 to 0.1]; 4 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):4 (Gln):1 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):1 (Gln):4 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):4 (Ser) [0.4 to 0.1]; 1 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.5 to 0.1]; 1 (Ala):5 (Gln):1 (Gly):1 (Ser) [0.5 to 0.1]; 1 (Ala):1 (Gln):5 (Gly):1 (Ser) [0.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):5 (Ser) [0.5 to 0.1]; 6 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):6 (Gln):1 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):1 (Gln):6 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):6 (Ser) [0.6 to 0.1]; 7 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):7 (Gln):1 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):1 (Gln):7 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):7 (Ser) [0.7 to 0.1]; 8 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):8 (Gln):1 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):1 (Gln):8 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):8 (Ser) [0.8 to 0.1]; 9 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):9 (Gln):1 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):1 (Gln):9 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):9 (Ser) [0.9 to 0.1]; 10 (Ala):1 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):10 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):10 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):10 (Ser) [1 to 0.1]; 20 (Ala):1 (Gln):1 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):20 (Gln):1 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):1 (Gln):20 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):20 (Ser) [2 to 0.1]; 30 (Ala):1 (Gln):1 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):30 (Gln):1 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):1 (Gln):30 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):30 (Ser) [3 to 0.1]; 40 (Ala):1 (Gln):1 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):40 (Gln):1 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):1 (Gln):40 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):40 (Ser) [4 to 0.1]; 50 (Ala):1 (Gln):1 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):50 (Gln):1 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):1 (Gln):50 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):50 (Ser) [5 to 0.1]; 60 (Ala):1 (Gln):1 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):60 (Gln):1 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):1 (Gln):60 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):60 (Ser) [6 to 0.1]; 70 (Ala):1 (Gln):1 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):70 (Gln):1 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):1 (Gln):70 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):70 (Ser) [7 to 0.1]; 80 (Ala):1 (Gln):1 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):80 (Gln):1 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):1 (Gln):80 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):80 (Ser) [8 to 0.1]; 90 (Ala):1 (Gln):1 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):90 (Gln):1 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):1 (Gln):90 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):90 (Ser) [9 to 0.1]; 100 (Ala):1 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):100 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):1 (Gln):100 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):100 (Ser) [10 to 0.1]):100:1 [10 to 0.1] or less (e.g., 95 (Ala):1 (Gln):1 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):95 (Gln):1 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):1 (Gln):95 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):95 (Ser) [9.5 to 0.1]; 85 (Ala):1 (Gln):1 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):85 (Gln):1 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):1 (Gln):85 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):85 (Ser) [8.5 to 0.1]; 75 (Ala):1 (Gln):1 (Gly):1 (Ser) [7.5 to 0.1]; 1 (Ala):75 (Gln):1 (Gly):1 (Ser) [7.5 to 0.1]; 1 (Ala):1 (Gln):75 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):55 (Ser) [5.5 to 0.1]; 65 (Ala):1 (Gln):1 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):65 (Gln):1 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):1 (Gln):65 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):65 (Ser) [6.5 to 0.1]; 55 (Ala):1 (Gln):1 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):55 (Gln):1 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):55 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):55 (Ser) [5.5 to 0.1]; 45 (Ala):1 (Gln):1 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):45 (Gln):1 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):1 (Gln):45 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):45 (Ser) [4.5 to 0.1]; 35 (Ala):1 (Gln):1 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):35 (Gln):1 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):1 (Gln):35 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):35 (Ser) [3.5 to 0.1]; 25 (Ala):1 (Gln):1 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):25 (Gln):1 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):1 (Gln):25 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):25 (Ser) [0.1 to 0.5]; 15 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):15 (Gln):1 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):1 (Gln):15 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):15 (Ser) [1.5 to 0.1]; 14 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):14 (Gln):1 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):1 (Gln):14 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):14 (Ser) [1.4 to 0.1]; 13 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):13 (Gln):1 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):1 (Gln):13 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):13 (Ser) [1.3 to 0.1]; 12 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):12 (Gln):1 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):1 (Gln):12 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):12 (Ser) [1.2 to 0.1]; 11 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):11 (Gln):1 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):1 (Gln):11 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):11 (Ser) [1.1 to 0.1]; 10 (Ala):1 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):10 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):10 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):10 (Ser) [1 to 0.1]; 19 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):19 (Gln):2 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):2 (Gln):19 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):19 (Ser) [0.95 to 0.1]; 17 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):17 (Gln):2 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):2 (Gln):17 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):17 (Ser) [0.85 to 0.1]; 15 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):15 (Gln):2 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):2 (Gln):15 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):15 (Ser) [0.75 to 0.1]; 13 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):13 (Gln):2 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):2 (Gln):13 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):13 (Ser) [0.65 to 0.1]; 11 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):11 (Gln):2 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):2 (Gln):11 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):11 (Ser) [0.55 to 0.1]; 9 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):9 (Gln):2 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):2 (Gln):9 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):9 (Ser) [0.45 to 0.1]; 7 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):7 (Gln):2 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):2 (Gln):7 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):7 (Ser) [0.35 to 0.1]; 5 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):5 (Gln):2 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):2 (Gln):5 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):5 (Ser) [0.25 to 0.1]; 3 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):3 (Gln):2 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):2 (Gln):3 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):3 (Ser) [0.15 to 0.1]; 2 (Ala):3 (Gln):3 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):2 (Gln):3 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):3 (Gln):2 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):3 (Gln):3 (Gly):2 (Ser) [0.1 to 0.15]; 2 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):2 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):2 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):5 (Gly):2 (Ser) [0.1 to 0.25]; 2 (Ala):7 (Gln):7 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):2 (Gln):7 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):7 (Gln):2 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):7 (Gln):7 (Gly):2 (Ser) [0.1 to 0.35]; 2 (Ala):9 (Gln):9 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):2 (Gln):9 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):9 (Gln):2 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):9 (Gln):9 (Gly):2 (Ser) [0.1 to 0.45]; 2 (Ala):11 (Gln):11 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):2 (Gln):11 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):11 (Gln):2 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):11 (Gln):11 (Gly):2 (Ser) [0.1 to 0.55]; 2 (Ala):13 (Gln):13 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):2 (Gln):13 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):13 (Gln):2 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):13 (Gln):13 (Gly):2 (Ser) [0.1 to 0.65]; 2 (Ala):15 (Gln):15 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):2 (Gln):15 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):15 (Gln):2 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):15 (Gln):15 (Gly):2 (Ser) [0.1 to 0.75]; 2 (Ala):17 (Gln):17 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):2 (Gln):17 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):17 (Gln):2 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):17 (Gln):17 (Gly):2 (Ser) [0.1 to 0.85]; 2 (Ala):19 (Gln):19 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):2 (Gln):19 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):19 (Gln):2 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):19 (Gln):19 (Gly):2 (Ser) [0.1 to 0.95]); or from 1:100 to 100:1 (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) [0.1 to 10] to 100 (Ala):1 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):1 (Gln):100 (Gly):100 (Ser) [0.1 to 10] to 1 (Ala):100 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):100 (Gln):1 (Gly):100 (Ser) [0.1 to 10] to 1 (Ala):1 (Gln):100 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):100 (Gln):100 (Gly):1 (Ser) [0.1 to 10] to 1 (Ala):1 (Gln):1 (Gly):100 (Ser) [10 to 0.1]). For example, if glutamine is present, the ratio of glutamine at 4 mM to other amino acids comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine).

In some embodiments, the formulation (or topical formulation) described here comprising: a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising, consisting essentially of, or consisting of at least one of alanine, glutamine, glycine, and serine, or salts thereof; and a therapeutically effective amount of a plant extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense; or combinations thereof) or a plant extract combination (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications)), or a therapeutically effective amount of a retinoid or at least one retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof).

Formulations and Uses Thereof for Treating Osteoarthritis

In some embodiments, a formulation for and methods of treating or ameliorating musculoskeletal disorders, such as but not limited to osteoarthritis, or ameliorating the symptoms of osteoarthritis is provided. Such a formulation comprises (or consists essentially of or consists of): as free amino acids or a peptide combination of amino acids, a therapeutically effective amount comprising (consisting essentially of, consisting of): (a) one or more amino acids of alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or salts or derivatives thereof, or combinations thereof; (b) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof, or combinations thereof; (c) alanine, glutamine, glycine, and serine, or derivatives or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or derivatives or salts thereof, or combinations thereof; or (d) at least one of alanine, glutamine, glycine, and serine, or derivatives or salts thereof, or combinations thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)). Other aspects of such embodiments are directed to such formulations for treating or ameliorating the symptoms of musculoskeletal diseases or conditions, such as osteoarthritis, where the formulations comprise (or consist essentially of or consist of): as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of: (a) at least one of: alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, or combinations thereof; and (b) a therapeutically effective amount of a retinoid (e.g., a retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof). Additional aspects of such formulations include those formulations optionally comprising a pharmaceutically acceptable vehicle and/or additive. Such formulations for treating or ameliorating musculoskeletal diseases or conditions, such as osteoarthritis, or the symptoms thereof osteoarthritis (e.g., pain (in the joints, hands, hip, knees, back, neck); joint stiffness, crackles, swelling, tenderness; bony outgrowths in fingers or toes; deformities or limping) as described here can be in the form of a parenteral formulation, for example, an injectable formulation. In additional aspects, use of such formulations for treating or ameliorating the symptoms of musculoskeletal diseases or disorders, e.g., osteoarthritis, is also provided, where the formulation is injected into an affected area of a subject suffering from osteoarthritis. For example, osteoarthritis presents in joints, such as joints of the hands, hips, knees, ankles, feet, among other places. The formulations for treating or ameliorating the symptoms of osteoarthritis comprise, consist essentially of, or consist of a therapeutically effective amount of: as free amino acids or peptide combinations of amino acids ranging from; 0.05 mM or greater (e.g., 0.07, 0.09, 0.11, 0.13, 0.15, 0.17, 0.19, 0.21, 0.23, 0.25, 0.27, 0.29, 0.31, 0.33, 0.35, 0.37, 0.39, 0.41, 0.43, 0.45, 0.47, 0.49, 0.51, 0.53, 0.55, 0.57, 0.59, 0.61, 0.63, 0.65, 0.67, 0.69, 0.71, 0.73, 0.75, 0.77, 0.79, 0.81, 0.83, 0.85, 0.87, 0.89, 0.91, 0.93, 0.95, 0.97, 0.99, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25); 20 mM or less (e.g., 18, 16, 14, 12, 10, 8, 6, 4, 2, 0.98, 0.96, 0.94, 0.92, 0.9, 0.88, 0.86, 0.84, 0.82, 0.8, 0.78, 0.76, 0.74, 0.72, 0.7, 0.68, 0.66, 0.64, 0.62, 0.6, 0.58, 0.56, 0.54, 0.52, 0.5, 0.48, 0.46, 0.44, 0.42, 0.4, 0.38, 0.36, 0.34, 0.32, 0.3, 0.28, 0.26, 0.24, 0.22, 0.2, 0.18, 0.16, 0.14, 0.12, 0.1, 0.08, 0.06, 0.04, 0.02); or 0.05 mM to 20 mM (e.g., 0.06-19; 0.07-18; 0.08-17; 0.09-16; 0.1-15; 0.11-14; 0.12-13; 0.13-12; 0.14-11; 0.15-10; 0.16-9; 0.17-8; 0.18-7; 0.19-6; 0.2-5; 0.21-4; 0.22-3; 0.23-2; 0.24-1; 0.25-0.99; 0.26-0.98; 0.27-0.97; 0.28-0.96; 0.29-0.95; 0.3-0.94; 0.31-0.93; 0.32-0.92; 0.33-0.91; 0.34-0.9; 0.35-0.89; 0.36-0.88; 0.37-0.87; 0.38-0.86; 0.39-0.85; 0.4-0.84; 0.41-0.83; 0.42-0.82; 0.43-0.81; 0.44-0.8; 0.45-0.79; 0.46-0.78; 0.47-0.77; 0.48-0.76; 0.49-0.75; 0.5-0.74; 0.51-0.73; 0.52-0.72; 0.53-0.71; 0.54-0.7; 0.55-0.69; 0.56-0.68; 0.57-0.67; 0.58-0.66; 0.59-0.65; 0.6-0.64; 0.61-0.63). In some aspects, where the formulation comprises a retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof), the retinoid comprises (or consists essentially of or consists of) 0.01% or greater by weight of the formulation (e.g., 0.03; 0.05; 0.07; 0.09; 0.11; 0.13; 0.15; 0.17; 0.19; 0.21; 0.23; 0.25; 0.27; 0.29; 0.31; 0.33; 0.35; 0.37; 0.39; 0.41; 0.43; 0.45; 0.47; 0.49; 0.51; 0.53; 0.55; 0.57; 0.59; 0.61; 0.63; 0.65; 0.67; 0.69; 0.71; 0.73; 0.75; 0.77; 0.79; 0.81; 0.83; 0.85; 0.87; 0.89; 0.91; 0.93; 0.95; 0.97; 0.99; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3, 1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1); 10% or less by weight of the formulation (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02; 0.008; 0.006; 0.004; 0.002); or 0.01%-10% by weight of the formulation (e.g., 0.02%-9.9%; 0.03%-9.8%; 0.04%-9.7%; 0.05%-9.6%; 0.06%-9.5%; 0.07%-9.4%; 0.08%-9.3%; 0.09%-9.2%; 0.1%-9.1%; 0.2%-9%; 0.3%-8.9%; 0.4%-8.8%; 0.5%-8.7%; 0.6%-8.6%; 0.7%-8.5%; 0.8%-8.4%; 0.9%-8.3%; 1%-8.2%; 1.1%-8.1%; 1.2%-8%; 1.3%-7.9%; 1.4%-7.8%; 1.5%-7.7%; 1.6%-7.6%; 1.7%-7.5%; 1.8%-7.4%; 1.9%-7.3%; 2%-7.2%; 2.1%-7.1%; 2.2%-7%; 2.3%-6.9%; 2.4%-6.8%; 2.5%-6.7%; 2.6%-6.6%; 2.7%-6.5%; 2.8%-6.4%; 2.9%-6.3%; 3%-6.2%; 3.1%-6.1%; 3.2%-6%; 3.3%-5.9%; 3.4%-5.8%; 3.5%-5.7%; 3.6%-5.6%; 3.7%-5.5%; 3.8%-5.4%; 3.9%-5.3%; 4%-5.2%; 4.1%-5.1%; 4.2%-5%; 4.3%-4.9%; 4.4%-4.8%; 4.5%-4.7%).

In some embodiments, provided here is a method of treating a subject suffering from osteoarthritis, where the method comprises (or consists essentially of or consists of): administering at a joint or around a joint of a subject suffering from a muculoskeletal disease or condition, including for example, osteoarthritis (e.g., joints of the hands, hips, knees, ankles, or feet) by injecting in the subject a therapeutically effective amount of a formulation comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination, (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts or derivatives thereof, where the therapeutically effective amount of any one of (a)-(d) is alone, as free amino acids or peptide combinations of amino acids, in a formulation of the disclosure or is in combination with a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof. For example, at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof, Some aspects are directed to such formulations that further optionally comprise a pharmaceutically acceptable vehicle and/or additive. In one aspect, the pharmaceutically acceptable vehicle and/or additive is sterile. Such methods of treating the subject suffering from osteoarthritis or symptoms thereof by administering these injectable formulations comprising a therapeutically effective amount of amino acids described here cither alone or in combination with a therapeutically effective amount of a retinoid described here, result in one or more of: a decrease in pain, an increase in hyaluronic acid synthesis, an increase in proliferation of elastin and/or collagen, or combinations thereof, as compared to a control or an untreated subject (a healthy subject or a subject who does not suffer from osteoarthritis). In some embodiments, such formulations described here, such as but not limited to a therapeutically effective amount of a formulation comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (a) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine; and a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof; and optionally, a pharmaceutically acceptable vehicle and/or additive, increase hyaluronic acid synthesis, thereby increasing hyaluronic acid lubrication and hydration, and increasing proliferation of elastin and/or collagen in a subject treated with such formulations as compared to a control or an untreated subject. Administration of such injectable formulations are on a daily, weekly, or monthly basis. Some aspects provide for a single injection or multiple injections (e.g., 2, 3, 4, 5, 6) on a daily, weekly, or monthly basis. In additional aspects, the therapeutically effective amounts of amino acids (free amino acids or peptide combinations thereof) either alone or in combination with a retinoid as described here, can be administered simultaneously or sequentially, and/or together or separately. Further aspects provide for a single- or multi-unit dosage form of the injectable formulations described here.

Exemplary Vehicles and Additives

Other embodiments directed to any of the formulations described here, optionally further comprise, consist essentially of, or consist of a dermatologically acceptable vehicle and/or additive. Non-limiting examples of vehicles and additives useful in any of the formulations (topical or parenteral/injectable formulations) described here can be selected from the group consisting of: water, sodium phytate, xanthan gum, kaolin, glycerin, polyglycerides (e.g., oleic polyglycerides, linoleic polyglycerides, linolenic polyglycerides), triglycerides (e.g., caprylic triglycerides, capric triglycerides), sucrose (e.g., sucrose stearate, sucrose distearate), alcohol (e.g., behenyl alcohol, benzoic alcohol, benzyl alcohol, cetearyl alcohol, oleyl alcohol), cetearyl glucoside, sodium stearoyl lactylate, Cera alba (beeswax), Butyrospermum parkii (shea butter), Prunus amigdalus sulcis (sweet almond oil), Simmondsia chinenesis (jojoba oil) phenoxyethanol, lecithin, tocopherol, tocopherol acetate, ascorbyl palmitate, citric acid, potassium sorbate, lactic acid, UV protectant, hydrogenated olive oil decyl esters, decyl cocaote, astrocaryum, murumuru seed butter, Zanthoxylum bungeanum fruit extract, squalene, pentylene glycol, dehydroacetic acid, tamarinds indica seed polysaccharide, sodium benzoate, curcumin, Zanthoxylum bungeanume fruit extract, Passiflora edulis seed oil, cetearyl ethylhexanoate, dimethicone, isopropyl myristate, acrylates/C10-C30 alkyl acrylate crosspolymer, allantoin, sodium phytate, PEG-90 stearate/glyceryl stearate; menthol, sodium hydroxide, sodium benzoate, dehydroacetic acid, benzoic alcohol, hexyldecanol, cetaryl ethylhexanoate, dimethicone, cetearyl ethylhexanoate, PEG-90 stearate/glyceryl stearate, ascorbyl palmitate, citric acid, Zanthoxylum bungeanum fruit extract, allantoin, disodium EDTA, phenoxyethanol, dehydroacetic acid, allantoin, XILOGEL®, Vaccinium myrtillus fruit extract, Oryza sativa (e.g. rice bran oil), isoamyl laurate, polymethyl methacrylate, botanical extracts (e.g., chestnut extract; date palm extract; almond or almond kernel extract; pomegranate extract; pine bark extract; aloe vera extract; chamomile extract; calendula extract; coffeeberry extract), fragrance, nutrients, antioxidants, water, or combinations thereof. Additional formulation embodiments described here can further comprise at least one dermotologically acceptable additive, such as an inactive ingredient useful in, for example, the delivery of an active ingredient, preservation, the feel and/or the scent of the formulation, providing other skin benefits, and the like, or combinations thereof, such as but not limited to, a carrier, a diluent, an excipient, anti-aging ingredients (e.g., collagen stimulators), antioxidants (e.g., butylated hydroxytoluene (BHT), ascorbic acid, sodium ascorbate, ascorbyl palmitate, beta-carotene, glycolic acid, hyaluronic acid), antibacterial agents, antifungal agents, antihistamines, anti-inflammatory agents, anti-irritants, antimycobacterial agents, antiseptics, analgesics, a colorant, an emollient (dimethicone oils, ester oils, or hydrocarbon oils), a natural or synthetic fragrance, humectants (e.g., polyols, including propylene glycol, glycerin), beta-hydroxy acids (e.g., salicylic acid), depigmenting agents (e.g., kojic acid, thiodipropionic acid (TDPA), niacinamide), an emulsifying agent, a film former, a filler—that creates, for example, bulk, texture, or lubrication—(e.g., silica, talc, zinc stearate, mica, kaolin, nylon powders), a gelling agent, a humectant, a mineral, a moisturizer, plant extracts or oils, a viscosity or rheology modifier (e.g., a thickener), skin protectants, skin penetration enhancers, stabilizers, a structuring agent, a sunscreen, a surfactant, vitamins (e.g., tocopherol, tocopheryl acetate), and the like, and combinations or mixtures thereof.

In certain embodiments, the formulation of the disclosure can also comprise additives (e.g., nutrients, electrolytes, vitamins, minerals, etc.). In certain embodiments, the formulation comprises iron or zinc. In certain embodiments, the therapeutic formulation comprises one or more electrolytes selected from, for example, Na⁺; K⁺; HCO₃—; CO₃²⁻; Ca²⁺; Mg²⁺; Fe₂; Cl⁻; phosphate ions, such as H₂PO₄⁻, HPO₄²⁻, and PO₄³—; zinc; iodine; copper; iron; selenium; chromium; and molybdenum.

In an alternative embodiment, the formulation does not contain HCO₃⁻ or CO₃²⁻. In another alternative embodiment, the formulation comprises HCO₃⁻ and CO₃²⁻ at a total concentration of less than 5 mg/l, or concentrations lower than 5 mg/l. In certain embodiments, the formulation does not contain electrolytes. For example, in certain embodiments, the formulation does not include one or more, or any, of Na⁺; K⁺; HCO₃⁻; CO₃²⁻; Ca²⁺; Mg²⁺; Fe₂; Cl⁻; phosphate ions, such as H₂PO₄⁻, HPO₄²⁻, and PO₄³⁻; zinc; iodine; copper; iron; selenium; chromium; and molybdenum.

In one embodiment, phosphate ions, such as H₂PO₄—, HPO₄²⁻, and PO₄³⁻, are used to buffer the formulation described herein. In one embodiment, the therapeutic formulation uses HCO; or CO₃²⁻ as a buffer. In another embodiment, the therapeutic formulation does not use HCO; or CO₃²⁻ as buffer.

A formulator will understand that vehicles, carriers, excipients, diluents, or combinations thereof, primarily serve to deliver a safe and stable formulation, and the formulation can contain additional inactive ingredients, where the formulation comprises, consists essentially of, or consists of: (a) as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount of a free amino acid(s), a combination of free amino acids, or a peptide combination of amino acids comprising, consisting essentially of, or consisting of: one or more of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine; (b) as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount of a free amino acid(s); a combination of free amino acids, or a peptide combination of amino acids comprising, consisting essentially of, or consisting of: alanine, glutamine, glycine, and serine; and optionally, a therapeutically effective amount of at least one additional free amino acid or peptide combination of amino acids of valine, isoleucine, arginine, threonine, or tryptophan, or any combination thereof; (c) as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids comprising, consisting essentially of, or consisting of: at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glutamine and serine; alanine and glycine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and a therapeutically effective amount of a plant extract or a plant extract combination, wherein the plant extract comprises (or consists essentially of or consists of): a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) or the plant extract combination (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica) comprises (or consists essentially of, or consists of): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); (d) as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids comprising, consisting essentially of, or consisting of: at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and a therapeutically effective amount of a retinoid, a retinoid derivative, or a retinoid precursor (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof); where the formulation of (a), (b), (c), or (d) optionally comprises, consists essentially of, or consists of, a dermatologically- or pharmaceutically-acceptable vehicle and/or additive.

In one embodiment, the formulation described here applied to the skin can further comprise a dermatologically- or pharmaceutically-acceptable vehicle and/or additive selected from: collagen, hyaluronic acid (HA), and/or ceramides. Various embodiments provide the formulation described here that further comprises dermatologically- or pharmaceutically-acceptable components and/or additives such as, but not limited to, dermatologically acceptable carriers, desquamation agents, anti-acne agents, anti-wrinkle agents/anti-atrophy agents, vitamin B3 compounds, retinoids, hydroxyl acids, anti-oxidants/radical scavengers, chelators, flavonoids, anti-inflammatory agents, anti-cellulite agents, topical anesthetics, tanning agents, skin lightening agents, skin soothing and skin healing agents, antimicrobial and antifungal agents, sunscreen agents, conditioning agents, structuring agents, thickening agent (including thickeners and gelling agents), a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); an emulsifier or a phospholipid, where in some embodiments, phospholipids are used as an emulsifier (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); formulation preparation and preservatives. In this regard, international PCT application publication, WO 2008/089408 is incorporated herein, by reference, in its entirety for teachings related to formulation preparation and additives.

Such carriers described here for use in any of the formulations of the disclosure can be liquids, such as water, saline, buffers, and the like. The therapeutic formulation can also comprise excipients, adjuvants, etc. that facilitate processing of the active compounds into preparations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen. In an embodiment of the disclosure, the formulation and all ingredients contained therein are sterile. Examples of suitable pharmaceutical carriers are described in “Remington's Pharmaceutical Sciences” by E. W. Martin. Such formulations contain a therapeutically effective amount of the therapeutic formulation, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should, in embodiments described here, suit the topical and/or parenteral mode of administration.

Use of the Formulation for Skin Barrier Repair and Strengthening to Improve Barrier Integrity

Some embodiments of the disclosure provide for a formulation or a topical formulation described here comprising: a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising, consisting essentially of, or consisting of at least one of alanine, glutamine, glycine, and serine, or salts thereof; and therapeutically effective amount of a plant extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense; or combinations thereof) or a plant extract combination (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications)). In additional aspects, the therapeutically effective amount of the combination of free amino acids or a peptide combination thereof and the therapeutically effective amount of the plant extract or the plant extract combination improves skin regeneration, including skin barrier repair. The improvement of skin regeneration, including skin barrier repair, includes, and is not limited to, skin barrier correction, skin barrier strengthening, skin barrier preservation, skin barrier integrity, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocytes, endothelial cells, and fibroblasts; facilitation of dermal regeneration, and combinations thereof. Further aspects are directed to the modulation of epidermal growth factor (EGF) using a therapeutically effective amount of the amino acid combination or the formulation comprising (or consisting essentially of or consisting of) the amino acid combination described here, where the amino acid combination comprises, consists essentially of, or consists of at least one of alanine, glutamine, glycine, and serine, or salts thereof, in a ratio of, for example, 1:1:1:1.

In additional embodiments, a method of improving skin regeneration, including skin barrier repair, of skin of a subject, comprises (or consists essentially of or consists of): administering a topical formulation, comprising (or consisting essentially of or consisting of) a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising (or consisting essentially of or consisting of) (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) a therapeutically effective amount of at least one of alanine, glutamine, glycine, and serine, or derivatives or salts thereof, or combinations thereof, where each amino acid present comprises a ratio to each of the other amino acids of: 1:100 or greater; 100:1 or less; or from 1:100 to 100:1 (e.g., if glutamine is present, the amino acids having a ratio to glutamine at 4 mM comprises: 25:13 (Glutamine:Glycine); 100:61 (Glutamine:Alanine); 25:18 (Glutamine:Serine); 5:6 (Glutamine:Arginine); 10:9 (Glutamine:Isoleucine); 50:41 (Glutamine:Threonine); 100:141 (Glutamine:Tryptophan); 100:81 (Glutamine:Valine)); and a therapeutically effective amount of a plant extract combination comprising (consisting essentially of or consisting of) (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or optionally in combination with at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and optionally, a dermatologically acceptable vehicle and/or additive, when tested by a method detecting expression of a barrier marker gene such as but not limited to EGF and determining that the barrier marker gene is increased by a 1.5-40 average fold increase of amino acid treated as compared to amino acid untreated skin (see, e.g., FIGS. 8-14). Improving skin regeneration, including skin barrier repair is measured or indicated by a 1.5-40 average fold increase of skin barrier marker gene expression, such as EGF, when tested by a method detecting expression the barrier marker gene expression when comparing skin treated with any of the formulations described here to untreated skin. The method of improving skin barrier repair includes but is not limited to, correcting the skin barrier, strengthening the skin barrier, preserving the skin barrier, increasing skin barrier integrity, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocytes, endothelial cells, and fibroblasts; facilitating dermal regeneration, and any combinations thereof.

Kits

Also encompassed by the disclosure are kits (e.g., pharmaceutical, therapeutic, cosmetic). The kits provided may comprise a formulation (e.g., cosmetic, therapeutic, or pharmaceutical formulation) described herein and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a vehicle or carrier, an excipient, or diluent (e.g., pharmaceutical excipient) for dilution or suspension of a formulation (e.g., pharmaceutical, therapeutic, or cosmetic formulation) or compound described herein. In some embodiments, the formulation or compound described herein provided in the first container and the second container are combined to form one unit dosage form.

Thus, in one aspect, provided are kits including at least one container (e.g., a first container) comprising any of the formulations described herein. In certain embodiments, the kits are useful for treating a disorder or condition (e.g., a skin or musculoskeletal disorder or condition) or symptoms thereof in a subject in need thereof. In certain embodiments, the kits are useful for preventing a disorder or condition (e.g., a skin disorder or condition (e.g., Xerosis cutis, atopic dermatitis, dermatitis, radiodermatitis, psoriasis, pruritus, eczema, a wound, an abrasion, a burn, or a condition relating to aging of skin; skin regeneration, including skin barrier repair and strengthening; or improving skin moisture); osteoarthritis) or symptom thereof in a subject in need thereof.

In certain embodiments, a kit described herein further includes instructions for using the formulation included in the kit. A kit described herein may also include information as required by a regulatory agency such as the U.S. Food and Drug Administration (FDA). In certain embodiments, the information included in the kits is prescribing information. In certain embodiments, the kits and instructions provide for treating and/or preventing a disorder or condition (e.g., a skin disorder or condition) in a subject in need thereof, A kit described here may include one or more additional pharmaceuticals or other agents described here as a separate formulation.

Methods of Administration

In various various embodiments of the disclosure, a formulation described herein is administered via administration to the skin of a subject in need thereof (administered via, e.g., topical, transdermal, intradermal, epicutaneous, and/or subcutaneous administration, or any combination thereof). Other embodiments provide a formulation described here, for example, a formulation for treating or ameliorating musculoskeletal diseases or disorders as described here, or where the formulation is administered parenterally (e.g., subcutaneous, intradermal, intra-articular, intrasynovial, intra-articular, intrasynovial, periarticular, intralesional, intramedullary, intrathecal, intramuscular, intravenous, topical, transdermal, intradermal, epicutaneous, subcutaneous), for treatment of diseases or disorders of the skin and/or musculoskeletal system, including joints, or their symptoms (e.g., rashes, plaques, scales, dry and/or cracked skin, flaky, irritated, tightness, itching, burning, soreness, hyperpigmentation; pain, stiffness, tenderness, swelling, inflexibility, grating sensation, bone spurs). Formulations described here can also be administered by any of a number of additional routes including, but not limited to, oral, intramedullary, intrathecal, intraventricular, intralesional, intraperitoneal, intranasal, sublingual, or rectal means. In some embodiments, a topical formulation or a parenteral formulation can be used to improve skin regeneration, including skin barrier integrity and repair and/or strengthening, treat and/or prevent a skin or musculoskeletal system, including joints, disease or condition (e.g., xerotic skin, wound healing, atopic dermatitis, dermatitis, psoriasis, aging of skin, a condition related to the aging of skin, pruritus, eczema, osteoarthritis), and/or symptoms thereof (e.g., damaged or compromised skin, dry, abraded, cracked, joint pain, stiffness, tenderness, swelling, inflexibility, grating sensation, bone spurs). Other embodiments can include administering such formulations as described here, including topical formulations or injectable formulations for treating a skin disease or condition or for treating a musculoskeletal disease or condition, respectively, or symptoms thereof, where administration occurs on a daily, multiple times per day, weekly, multiple times per week, or monthly basis. Some aspects provide for a single application (topically or injected, or by any other means described here) or multiple applications (e.g., 2, 3, 4, 5, 6) on a daily, weekly, or monthly basis. In additional aspects, the therapeutically effective amounts of an amino acid or combinations of amino acids (free amino acids or peptide combinations thereof), either alone or in combination with other active agents, inactive agents, or beneficial ingredients (e.g., Boswellia extract, retinoid) as described here, can be administered simultaneously or sequentially, and/or together or separately. Further aspects provide for a single- or multi-unit dosage form of the topical skin formulation or the parenteral formulations described here. The amount of topical formulation for administration comprises an amount sufficient to provide a layer of the formulation over the affected area.

In some aspects of the disclosure, the formulations comprising the therapeutically effective amount of amino acids comprising, consisting essentially of, or consisting of a free amino acid, a combination of free amino acids, or a peptide combination of amino acids (one or more of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives, or salst thereof), is administered simultaneously or sequentially and/or administered together or separately with another active agent or additive.

Other aspects provide for a single- or multi-unit dosage of the formulations described here. Additional aspects of the disclosure are directed to the formulations described here in a unit dosage form (e.g., single- or multi-unit dosage form).

In some embodiments, the formulation (e.g., topical or parenteral formulation) described here can be topically applied to skin of or injected in a subject in need of: (a) skin regeneration, including skin barrier repair, skin barrier strengthening, improving skin barrier integrity, (b) skin regeneration, including improving skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS)), or combinations thereof, or (c) treatments for osteoarthritis, where the formulation comprises, as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount of amino acids comprising, consisting essentially of, or consisting of at least one of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof, where the formulation comprises, as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising, consisting essentially of, or consisting of at least one of: alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and (i) a therapeutically effective amount of a plant extract (e.g., plant extract combination) comprising, consisting essentially of, or consisting of: a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or in combination of at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications), or alternatively, (ii) a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof; and the formulation optionally comprises, a dermatologically acceptable vehicle and/or additive, wherein the therapeutically effective amount of the amino acid concentration comprises (or consists essentially of or consists of) 0.1 mM or greater (e.g., 0.3; 0.5; 0.7; 0.9; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5); 10 mM or less (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02); or 0.1 mM-10 mM (e.g., 0.2 mM-9.9 mM; 0.3 mM-9.8 mM; 0.4 mM-9.7 mM; 0.5 mM-9.6 mM; 0.6 mM-9.5 mM; 0.7 mM-9.4 mM; 0.8 mM-9.3 mM; 0.9 mM-9.2 mM; 1 mM-9.1 mM); 1.1 mM-9 mM; 1.2 mM-8.9 mM; 1.3 mM-8.8 mM; 1.4 mM-8.7 mM; 1.5 mM-8.6 mM; 1.6 mM-8.5 mM; 1.7 mM-8.4 mM; 1.8 mM-8.3 mM; 1.9 mM-8.2 mM; 2 mM-8.1 mM; 2.1 mM-8 mM; 2.2 mM-7.9 mM; 2.3 mM-7.8 mM; 2.4 mM-7.7 mM; 2.5 mM-7.6 mM; 2.6 mM-7.5 mM; 2.7 mM-7.4 mM; 2.8 mM-7.3 mM; 2.9 mM-7.2 mM; 3 mM-7.1 mM; 3.1 mM-7 mM; 3.2 mM-6.9 mM; 3.3 mM-6.8 mM; 3.4 mM-6.7 mM; 3.5 mM-6.6 mM; 3.6 mM-6.5 mM; 3.7 mM-6.4 mM; 3.8 mM-6.3 mM; 3.9 mM-6.2 mM; 4 mM-6.1 mM; 4.1 mM-6 mM; 4.2 mM-5.9 mM; 4.3 mM-5.8 mM; 4.4 mM-5.7 mM; 4.5 mM-5.6 mM; 4.6 mM-5.5 mM; 4.7 mM-5.4 mM; 4.8 mM-5.3 mM; 4.9 mM-5.2 mM; 5 mM-5.1 mM), wherein the therapeutically effective amount of (i) the plant extract (or plant extract concentration) or (ii) the retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof) comprises (or consists essentially of or consists of): 0.01% or greater by weight of the formulation (e.g., 0.03; 0.05; 0.07; 0.09; 0.11; 0.13; 0.15; 0.17; 0.19; 0.21; 0.23; 0.25; 0.27; 0.29; 0.31; 0.33; 0.35; 0.37; 0.39; 0.41; 0.43; 0.45; 0.47; 0.49; 0.51; 0.53; 0.55; 0.57; 0.59; 0.61; 0.63; 0.65; 0.67; 0.69; 0.71; 0.73; 0.75; 0.77; 0.79; 0.81; 0.83; 0.85; 0.87; 0.89; 0.91; 0.93; 0.95; 0.97; 0.99; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3, 1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1); 10% or less (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02; 0.008; 0.006; 0.004; 0.002); or 0.01%-10% (e.g., 0.02%-9.9%; 0.03%-9.8%; 0.04%-9.7%; 0.05%-9.6%; 0.06%-9.5%; 0.07%-9.4%; 0.08%-9.3%; 0.09%-9.2%; 0.1%-9.1%; 0.2%-9%; 0.3%-8.9%; 0.4%-8.8%; 0.5%-8.7%; 0.6%-8.6%; 0.7%-8.5%; 0.8%-8.4%; 0.9%-8.3%; 1%-8.2%; 1.1%-8.1%; 1.2%-8%; 1.3%-7.9%; 1.4%-7.8%; 1.5%-7.7%; 1.6%-7.6%; 1.7%-7.5%; 1.8%-7.4%; 1.9%-7.3%; 2%-7.2%; 2.1%-7.1%; 2.2%-7%; 2.3%-6.9%; 2.4%-6.8%; 2.5%-6.7%; 2.6%-6.6%; 2.7%-6.5%; 2.8%-6.4%; 2.9%-6.3%; 3%-6.2%; 3.1%-6.1%; 3.2%-6%; 3.3%-5.9%; 3.4%-5.8%; 3.5%-5.7%; 3.6%-5.6%; 3.7%-5.5%; 3.8%-5.4%; 3.9%-5.3%; 4%-5.2%; 4.1%-5.1%; 4.2%-5%; 4.3%-4.9%; 4.4%-4.8%; 4.5%-4.7%); and the formulation optionally comprises, a dermatologically or physiologically acceptable vehicle and/or additive, in an amount sufficient to promote skin regeneration, including skin barrier repair; promote skin barrier strengthening; improve skin barrier integrity; improve skin moisture; treat osteoarthritis or its symptoms; treat a skin disease, a cosmetic skin condition, or other skin conditions; treat and/or prevent Xerosis cutis, atopic dermatitis, dermatitis, radiodermatitis, psoriasis, pruritus, eczema, a wound, burn, abrasion, or a condition relating to aging of skin; skin barrier repair and strengthening; or improving skin moisture are administered topically on the skin of a subject suffering from one of the skin conditions, in need of skin barrier repair and strengthening, or in need of improving skin moisture, at a site where the subject suffers from one of the skin conditions or is in need of skin barrier repair and strengthening or improving skin moisture.

In some aspects, methods of treating a subject suffering from osteoarthritis or the symptoms thereof, where the formulation of the disclosure is administered by injection at a joint or around a joint of the subject suffering from a musculoskeletal disease or condition, including osteoarthritis (e.g., joints of the hands, hips, knees, ankles, feet). Some aspects are directed to administration of the osteoarthritis injectable formulation described here, where the formulation is injected in the subject suffering from osteoarthritis and results in one or more of: a decrease in pain, an increase in hyaluronic acid synthesis, an increase in proliferation of elastin and/or collagen, or combinations thereof, as compared to an untreated subject.

Other embodiments are directed to a method comprising (or consisting essentially of or consisting of): administering a topical formulation comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); (c) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one additional free amino acid or peptide combination of amino acids of: arginine, isoleucine, threonine, tryptophan, valine; or salts thereof, or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, or any combinations thereof; where the therapeutically effective amount of any one of (a)-(d) is alone in any of the formulations described here or the therapeutically effective amount of any one of (a)-(d) is in combination with, in any of the formulations described here: (i) a therapeutically effective amount of a plant extract (e.g., plant extract combination) comprising, consisting essentially of, or consisting of: a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or in combination of at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); or (ii) a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof; and the formulation optionally comprises, a dermatologically acceptable, pharmaceutically acceptable, or physiologically acceptable vehicle and/or additive. In some aspects provided here, formulations of the disclosure can be used in a method for improving skin regeneration, including skin barrier repair (and strengthening) by topically administering a formulation of as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); (c) a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof, or combinations thereof; and a therapeutically effective amount of a plant extract (e.g., plant extract combination) comprising, consisting essentially of, or consisting of: a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or in combination of at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications)), where each of the amino acids are in a ratio to the other amino acids ranging from, for example, 1:100 or greater (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser):100 (Ala):1 (Gln):100 (Gly):100 (Ser):100 (Ala):100 (Gln):1 (Gly):100 (Ser):100 (Ala):100 (Gln):100 (Gly):1 (Ser)):100:1 or less (e.g., 95 (Ala):1 (Gln):1 (Gly):1 (Ser):1 (Ala):95 (Gln):1 (Gly):1 (Ser):1 (Ala):1 (Gln):95 (Gly):1 (Ser):1 (Ala):1 (Gln):1 (Gly):95 (Ser); or from 1:100 to 100:1 (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) to 100 (Ala):1 (Gln):1 (Gly):1 (Ser):100 (Ala):1 (Gln):100 (Gly):100 (Ser) to 1 (Ala):100 (Gln):1 (Gly):1 (Ser):100 (Ala):100 (Gln):1 (Gly):100 (Ser) to 1 (Ala):1 (Gln):100 (Gly):1 (Ser):100 (Ala):100 (Gln):100 (Gly):1 (Ser) to 1 (Ala):1 (Gln):1 (Gly):100 (Ser)), where, in some embodiments, the method detects an increase of a skin regeneration marker gene, e.g., skin barrier marker gene (e.g., EGF) by a 1.5-40 average fold change in formulation-treated skin as compared to untreated skin, thereby improving skin regeneration, including skin barrier repair. Other aspects of the disclosure are directed to formulations and their methods of improving skin moisture of a skin of a subject, where the method comprises (consists essentially of or consists of) administering a therapeutically effective amount of a topical formulation of as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof), where each of the amino acids are in a ratio to the other amino acids ranging from, for example, 1:100 or greater (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser):100 (Ala):1 (Gln):100 (Gly):100 (Ser):100 (Ala):100 (Gln):1 (Gly):100 (Ser):100 (Ala):100 (Gln):100 (Gly):1 (Ser)):100:1 or less (e.g., 95 (Ala):1 (Gln):1 (Gly):1 (Ser):1 (Ala):95 (Gln):1 (Gly):1 (Ser):1 (Ala):1 (Gln):95 (Gly):1 (Ser):1 (Ala):1 (Gln):1 (Gly):95 (Ser); or from 1:100 to 100:1 (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) to 100 (Ala):1 (Gln):1 (Gly):1 (Ser):100 (Ala):1 (Gln):100 (Gly):100 (Ser) to 1 (Ala):100 (Gln):1 (Gly):1 (Ser):100 (Ala):100 (Gln):1 (Gly):100 (Ser) to 1 (Ala):1 (Gln):100 (Gly):1 (Ser):100 (Ala):100 (Gln):100 (Gly):1 (Ser) to 1 (Ala):1 (Gln):1 (Gly):100 (Ser)), where, in some embodiments, the method detects an increase of a skin moisture marker gene or an hyaluronic acid (HA) marker gene (e.g., HAS1, HAS2, HAS3, or EGF) by a 1.4-450 fold change in formulation-treated skin as compared to untreated skin, thereby improving skin regeneration, including skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))). Further aspects provide for methods of treating a subject suffering from musculoskeletal diseases or conditions, or musculoskeletal manifestations (e.g., osteoarthritis, rheumatoid arthritis, psoriatic arthritis, gout, spondyloarthritis, osteoporosis, osteopenia, associated fragility fractures, traumatic fractures, sarcopenia, back and neck pain, fibromyalgia pain conditions, inflammatory diseases (e.g., connective tissue diseases and vasculitis, systemic lupus erythematosus, amputation resulting from disease or trauma)) or symptoms thereof, where the method comprises (or consists essentially of or consists of): administering at a site of the musculoskeletal disease or condition, for example, osteoarthritis in a subject suffering from osteoarthritis, such as a joint or surrounding areas of a subject suffering from osteoarthritis (e.g., joints of the hands, hips, or knees), by injecting in the subject a therapeutically effective amount of a formulation comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (a) a therapeutically effective amount of one or more amino acids as free amino acids or a peptide combination of amino acids selected from the group consisting of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof; (b) a therapeutically effective amount of a combination of amino acids, as free amino acids or a peptide combination of amino acids comprising (or consisting essentially of, or consisting of) glutamine and glycine, and at least one amino acid selected from the group consisting of: alanine, arginine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof (e.g., glutamine, glycine, alanine; glutamine, glycine, alanine, arginine; glutamine, glycine, alanine, arginine, isoleucine; glutamine, glycine, alanine, arginine, isoleucine, serine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan; glutamine, glycine, alanine, arginine, isoleucine, serine, threonine, tryptophan, valine); a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of): (c) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or (d), a therapeutically effective amount of: at least one of alanine, glutamine, glycine, serine, or salts thereof, or any combination thereof (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine; or salts thereof, or any combinations thereof); and a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or any combinations thereof); and each of the retinoid formulations of (a)-(d), optionally comprises a dermatologically acceptable, pharmaceutically acceptable, or physiologically acceptable vehicle and/or additive, thereby treating the subject suffering from a musculoskeletal disease or condition, such as osteoarthritis or its symptoms. One aspect is directed to methods of treating a subject suffering from musculoskeletal diseases or conditions, including osteoarthritis, or their symptoms thereof, where such injectable formulations described here comprise (or consist essentially of or consist of), as free amino acids or a peptide combination of amino acids, any of the aforementioned amino acids, or salts thereof, or combinations thereof; and a therapeutically effective amount of a retinoid (e.g., a retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof). In some aspects, such methods of treating a subject suffering from osteoarthritis or the symptoms thereof, where the formulation is administered by injection at a joint or around a joint of the subject suffering from osteoarthritis (e.g., joints of the hands, hips, knees, ankles, feet). Additional aspects are directed to the formulation in a unit dosage form (e.g., single- or multi-unit dosage form). Some aspects are directed to administration of the musculoskeletal disease or condition, for example osteoarthritis, injectable formulation described here, where the formulation is injected in the subject suffering from the musculoskeletal disease or condition (e.g., osteoarthritis) and results in one or more of: a decrease in pain, an increase in hyaluronic acid synthesis, an increase in proliferation of elastin and/or collagen, or combinations thereof, as compared to an untreated subject.

In some aspects of the disclosure, the therapeutically effective amount of free amino acids or peptide combination of amino acids either alone or with a therapeutically effective amount of a plant extract or plant extract combination or with a therapeutically effective amount of a retinoid as described here, are administered simultaneously or sequentially and/or administered together or separately. Other aspects provide for a single- or multi-unit dosage of the formulations described here. The amount of parenteral formulation for administration comprises an amount sufficient to reduce or ameliorate the musculoskeletal disease or condition, or symptoms thereof. The amount of topical formulation for administration comprises an amount sufficient to provide a layer of the formulation over the affected area.

The present disclosure provides for therapeutic or pharmaceutical formulations comprising a therapeutically effective amount of the subject formulation and, optionally, one or more pharmaceutically acceptable carriers. The present disclosure provides for therapeutic, pharmaceutical, cosmetic, or nutritional formulations comprising a therapeutically effective amount of the subject formulation and, optionally, one or more pharmaceutically acceptable carriers. Such pharmaceutical carriers can be liquids, such as water.

In one embodiment, the administration of the formulation can be systemic. Oral, intravenous, intra-arterial, subcutaneous, intra-peritoneal, intra-muscular, intra-ventricular, intranasal, transmucosal, subcutaneous, topical, rectal, and other modes of administration are all contemplated, as are all combinations thereof. In a particular embodiment, a formulation described herein is administered via administration to the skin (administered via, e.g., topical, transdermal, and/or subcutaneous administration, or any combination thereof). Formulations described herein may be incorporated into gauze, pads (adhesive or non-adhesive), bandages, and/or dressings for longer term application directly to skin. In a particular embodiment, a formulation described herein is administered to the skin in combination with oral administration of a formulation described herein. In a particular embodiment thereof, oral administration of a formulation described herein is implemented before, during, or after administration of a formulation to the skin. As described herein, administration to the skin may be achieved via topical, transdermal, and/or subcutaneous administration.

In one embodiment, for injection, the active ingredient can be formulated in aqueous solutions, in one example in physiologically compatible buffers. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. For oral administration, the active ingredient can be combined with carriers suitable for inclusion into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like. Formulations can also be prepared for use in inhalation therapy. For administration by inhalation, the formulation can be delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant. The formulation can also be administered via inhalation or other route as a powder.

Therapeutically effective doses of the presently described formulation can be determined by one of skill in the art, with a goal of achieving a desired strength of intercellular adhesion reflecting a healthy barrier integrity of the skin. An increase in the expression of marker genes indicative of skin cell proliferation, skin cell differentiation, and/or intercellular adhesion can be assessed using assays described herein. Immunohistochemistry, behavioral assessments, and/or electrophysiological techniques can also be utilized to assess barrier integrity of the skin.

In some embodiments, the methods according to the present disclosure include administering the therapeutic formulation by sustained-release systems. Suitable examples of sustained-release systems include suitable polymeric materials (such as, semi-permeable polymer matrices in the form of shaped articles, for example films, or microcapsules), suitable hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, and sparingly soluble derivatives (such as, for example, a sparingly soluble salt). Sustained-release formulations can be administered orally, parenterally, intracistemally, intraperitoneally, topically (as by powders, ointments, gels, drops or transdermal patch), or as an oral or nasal spray. Sustained-release matrices include polylactides, copolymers of L-glutamic acid and gamma-ethyl-L-glutamate, poly(2-hydroxyethyl methacrylate), ethylene vinyl acetate, or poly-D-(−)-3-hydroxybutyric acid.

In one embodiment, implantable drug infusion devices may be used to provide patients with a constant and long-term dosage or infusion of a therapeutic formulation. Such device can be categorized as either active or passive.

In one embodiment, polymers can be used for ion-controlled release. Various degradable and nondegradable polymeric matrices for use in controlled drug delivery may be used. For example, the block copolymer, polaxamer 407, hydroxyapatite, and liposomes.

The pharmaceutical formulation of the present invention may be used either alone or in combination with one or more drugs to be effective for treating diseases. The formulations can also be formulated in combination with at least one other agent, such as stabilizing or buffer compounds, which can be administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, buffered saline, dextrose, and water. In addition to the critical components of formulations discussed herein, cells or influencing factors, the formulations can contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active compounds into preparations that can be used pharmaceutically. The formulation may be prepared as a single-dosage form using a pharmaceutically acceptable carrier or excipient or may be contained in a multiple-dosage container.

In one embodiment, the formulation may further contain other proliferation and/or differentiation inducing agents. Examples include fibroblast growth factor (FGF), epidermal growth factor (EGF), and retinoic acid.

The formulation may further contain other commonly used additives such as an anti-oxidant, a buffer, a bacteriostat, etc., and may be formulated into an injectable formulation such as aqueous solution, suspension, emulsion, etc. a pill, a capsule, a granule, a tablet, etc., by further adding a diluent, a dispersant, a surfactant, a binder, a lubricant, etc.

A food formulation of the present invention may be contained in a health functional food. The health functional food may be prepared according to a method commonly employed in the art, and commonly used raw materials and ingredients may be added when preparing the health functional food. When a formulation described herein is included in a health functional food, the formulation may be added alone or together with another health functional food or other food ingredient(s), according to commonly employed methods. The amount of the active ingredient may be determined appropriately depending on the purpose of use (e.g., prevention, health improvement, or therapeutic intervention). The food formulation may further comprise, for example, a prebiotic or probiotic substance.

The kind of food is not limited. Examples of the food to which the formulation can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, soup, beverage, tea, drink, alcoholic beverage, vitamin complex, etc.

Methods of Preparing the Formulations of the Disclosure

In additional embodiments, the formulation described here can be prepared by mixing and/or stirring (concurrently, simultaneously, sequentially): (a) the therapeutically effective amount of the free amino acids, the combination of free amino acids, or the peptide combination of amino acids thereof comprising, consisting essentially of at least one of alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine; and optionally (b) a dermatologically acceptable vehicle and/or additive. The mixture of (a) and optionally (b) can be heated (either together or separately) to a temperature sufficient to completely mix, homogenize, or emulsify (a), (b), and optionally (c), where the temperature is 20° C. or greater (e.g., 22; 24; 26; 28; 30; 32; 34; 36; 38; 40; 42; 44; 46; 48; 50; 52; 54; 56; 58; 60; 62; 64; 66; 68; 70; 72; 74; 76; 78; 80; 82; 84; 86; 88; 90; 92; 94; 96; 98; 100; 102; 104; 106; 108; 110); 100° C. or less (e.g., 99; 97; 95; 93; 91; 89; 87; 85; 83; 81; 79; 77; 75; 73; 71; 69; 67; 65; 63; 61; 59; 57; 55; 53; 51; 49; 47; 45; 43; 41; 39; 37; 35; 33; 31; 29; 27; 25; 23; 21; 19; 17); or 20° C. 100° C. (e.g., 21° C.-99° C.; 22° C.-98° C.; 23° C.-97° C.; 24° C.-96° C.; 25° C.-95° C.; 26° C.- 94° C.; 27° C.-93° C.; 28° C.-92° C.; 29° C.-91° C.; 30° C.-90° C.; 31° C.-89° C.; 32° C.-88° C.; 33° C.-87° C.; 34° C. 86° C.; 35° C.-85° C.; 36° C.-84° C.; 37° C.-83° C.; 38° C.-82° C.; 39° C.-81° C.; 40° C.-80° C.; 41° C.-79° C.; 42° C.-78° C.; 43° C.-77° C.; 44° C.-76° C.; 45° C.-75° C.; 46° C.-74° C.; 47° C.-73° C.; 48° C.-72° C.; 49° C.-71° C.; 50° C.-70° C.; 51° C.-69° C.; 52° C.-68° C.; 53° C.-67° C.; 54° C.-66° C.; 55° C.-65° C.; 56° C.-64° C.; 57° C.-63° C.; 58° C.-62° C.; 59° C.-61° C., for a time sufficient to completely mix, homogenize, or emulsify (e.g., seconds to minutes). In some aspects, the formulation further comprises (or consists essentially of or consists of a therapeutically effective amount of) other active agents beneficial for skin; and/or ingredients for preparing a desirable formulation (e.g., scent, texture, appearance, preservative, etc.): a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications).

In some embodiments, any of the skin formulations described herein may be incorporated into gauze, paper/pads (adhesive or non-adhesive), bandages, and/or dressings for longer term application directly to skin. As described herein, administration to the skin may be achieved via topical, transdermal, and/or subcutaneous administration.

In one embodiment, for injection, the therapeutically effective amount of a free amino acid, a combination of amino acids, or a peptide combination of amino acids (one or more amino acids of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof) described here or other active ingredient can be formulated in aqueous solutions, in one example in physiologically compatible buffers. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. For topical administration, skin formulations can be formulated into capsules, liquids, gels, syrups, slurries, suspensions and the like.

In some embodiments, the methods according to the present disclosure include administering the therapeutic formulation by sustained-release systems. Suitable examples of sustained-release systems include suitable polymeric materials (such as, semi-permeable polymer matrices in the form of shaped articles, for example films, or microcapsules), suitable hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, and sparingly soluble derivatives (such as, for example, a sparingly soluble salt). Sustained-release formulations can be administered parenterally or topically (as by creams, moisturizers, paper, powders, ointments, gels, drops, transdermal patch, and the like). Sustained-release matrices can include polylactides, copolymers of L-glutamic acid and gamma-ethyl-L-glutamate, poly(2-hydroxyethyl methacrylate), ethylene vinyl acetate, or poly-D-(−)-3-hydroxybutyric acid.

The formulations and pharmaceutical formulations of the present disclosure may be used either alone or in combination with one or more drugs to be effective for treating diseases, such as but not limited to skin diseases or conditions, musculoskeletal diseases or conditions, or those diseases or conditions related to skin or the musculoskeletal system. The formulations can also be formulated in combination with at least one other agent, such as stabilizing or buffer compounds, which can be administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, buffered saline, dextrose, and water. In addition to the critical components of formulations discussed herein, cells or influencing factors, the formulations can contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active compounds into preparations that can be used pharmaceutically. The formulation may be prepared as a single-dosage form using a pharmaceutically acceptable carrier or excipient or may be contained in a multiple-dosage container.

In one embodiment, the formulation may further contain other proliferation and/or differentiation inducing agents. Examples include, but are not limited to, fibroblast growth factor (FGF), epidermal growth factor (EGF), and retinoic acid.

EXAMPLES

The following examples illustrate specific aspects of the instant description. The examples and embodiments described herein are for illustrative purposes only and various modifications or changes in light thereof are suggested to persons skilled in the art and are included within the spirit and purview of this application. In addition, any elements or limitations of any embodiment thereof disclosed herein can be combined with any and/or all other elements or limitations (individually or in any combination) or any other embodiment thereof disclosed herein, and all such combinations are contemplated with the scope of the embodiments described here without limitation thereto. The examples should not be construed as limiting, as the example merely provides specific understanding and practice of the embodiments and its various aspects.

Example 1: Testing of Amino Acids

To test the effect of amino acid treatment on epithelial skin barrier, a cellular model of differentiated normal human epidermal keratinocytes (NHEK) was utilized. Commercially available NHEK (ATCC), derived from three diverse healthy donors, were used. Based on a multivariate analysis, three amino acids for the treatment: Ser, Gln and Gly were selected.

Microarray gene expression and RNA sequencing (RNA-seq) analyses were utilized to assess the transcriptome of a panel of differentiated NHEK, focusing on a particular set of marker genes whose upregulation or downregulation (depending on the marker gene) reflects a trend toward improved barrier function in the skin. Upregulation of marker genes that promote, for example, keratinocyte differentiation and/or proliferation and downregulation of marker genes that promote, for example, apoptosis/cell death and/or inflammation reflect a trend toward improved barrier function in the skin. In contrast, downregulation of marker genes that promote, for example, keratinocyte differentiation and/or proliferation and upregulation of marker genes that promote, for example, apoptosis/cell death and/or inflammation reflect a trend toward impaired barrier function in the skin.

Using gene arrays, comparative analyses of two datasets were performed in three diverse donors focusing on consistently differentially [filter criterion of fold change 1.5 (positive or negative) and p-value <0.05)] expressed genes following treatment with various amino acids and combinations thereof, qPCR was used to assess the mRNA expression results. The qPCR results confirmed the array results pertaining to differential expression, thereby validating the panel of genes.

The marker genes: EGF, FGF, CPT2, TGFB1, SMAD1, MKI67, PPARGC1B, CASP8 and TNFRSF10D, and PPARA and PPARD were also included in the panel of marker genes examined, in part because PPARGC1B, a transcription factor, encodes a coactivator of PPARα, PPARδ, and PPARγ. The significant upregulation of CPT2 (which is a primary target for PPARα) reinforced inclusion of PPARA and PPARD in the panel.

Each of the amino acids was also titrated to arrive at suitable concentrations for use in amino acid combinations. Accordingly, suitable (e.g., non-toxic, non-morphology changing, therapeutically effective) concentrations for each single amino acid were determined during the course of developing suitable amino acid combinations.

Comparative analysis of qPCR data from two donors, which included a scoring system (as described above), identified two sets of consistently most effective (Gly, Gln, Ser, Ala) and least effective (Asp, Thr, Cys, Pro, Tyr) amino acid combinations. See, e.g., FIG. 2.

Two combinations of the most effective amino acids were tested as an initial set of components for skin formulations in five different donors [combo #1 (Gln, Gly, Ala, Ser) and combo #2 (Gln, Gly, Ala, Ser, Ile, Val)] to evaluate the effect thereof on mRNA expression (transcripts) of ten genes: EGF. FGF2, PDGF, CPT2, TGM4, PPARD, TGFB1, FLG, MKI67, and TNERSF10D. For the comparison, a set of the least effective amino acids (combo #3) was used in qPCR analysis. See, e.g., FIG. 1.

The conclusion from the experiments was that the combination of the most effective amino acids (combos #1 and 2) significantly increased gene expression of six out of ten tested genes (FIG. 1), while combo #3 had no effect on transcripts of the majority of the genes, or even downregulated them as in the case of EGF. In contrast, mRNA expression of TNFRSF10D, which encodes a member of TNF receptor superfamily, was significantly downregulated by combo #1, suggesting an anti-inflammatory/anti-apoptotic functionality for combo #1. The most significant effect of combo #1 and #2 was detected on upregulation of the transcript of EGF. Collectively, these results demonstrate that particular combinations of specific amino acid combinations, but not all or random amino acid combinations, upregulate keratinocyte genes important for normal barrier function.

These results further show that careful testing of amino acid combinations, considering multivariate, interdependent data points, is required to obtain the best beneficial effect. It is noteworthy that each of combo #1 and combo #2 conferred statistically significant increases in the expression of many of the marker genes identified relative to controls in the context of healthy skin such as that provided by keratinocytes derived from three diverse healthy donors. Results determined in the context of injured/compromised skin may yield even more significant results demonstrating efficacy of combo #1 and combo #2 and other amino acid combinations described herein. Results presented herein underscore the demonstrated potency of combo #1 and combo #2 and are reasonably predictive of their potential for the enhancement of the skin barrier, especially in the context of injured/compromised skin.

Results presented herein also demonstrate upregulation of proteins so as to result in contributing positively to barrier integrity of the skin. Involucrin, for example, was upregulated to a statistically significant degree at 24 h post-treatment with either of combo #1 or combo #2 relative to control. Sec. e.g., FIG. 5. Filaggrin was also upregulated to a statistically significant degree at 24 h post-treatment with either of combo #1 or combo #2 relative to control. See, e.g., FIG. 6.

Specific combinations of amino acids in suitable ratios had a synergistic effect and an orchestrating ability to improve and/or restore the skin barrier integrity. Particular amino acid combinations trigger some cellular pathways that in totality provide a desired synergistic effect.

Example 2: Experimental Methods

Amino acids are mixed in previously established concentrations in water and added to amino acid-deprived culture medium.

Gene Expression:

mRNA is extracted, converted to cDNA, and analyzed by qPCR method using custom TaqMan array 384-well plates (Thermo Fisher Scientific) containing 34 of the above-mentioned genes. Four control housekeeping genes are included to normalize the mRNA expression: GAPDH, POLR2A, YWHAZ and PGK1. The array plate will contain 38 genes in total and 10 samples can be analyzed in one run of qPCR.

Protein Analysis:

Western Blot. Keratinocytes or epidermal skin equivalents are homogenized in lysis buffer, protein is extracted, and Western blots performed per standard protocols. The following commercial antibodies are used: filaggrin (#sc-66192, Santa Cruz Bio), loricrin (#ab183646), involucrin (#ab68), TGM2 (#ab2386) (all from Abcam). β-actin (#SAB3500350, Sigma).

Since Western Blot analysis is labor intensive and not a high throughput assay, for some proteins ELISA is used instead, utilizing already validated assays as previously published for protein ELISAs. ANGPTL4 and involucrin immunoassays are carried out using Milliplex Map Human Liver Protein Magnetic Bead Panel hANGPTL4-MAG and Milliplex Map Human Skin Magnetic Bead INVOL-MAG (EMD Millipore, Billerica, MA) on a Luminex xMAP platform.

Statistics:

The differences between control and treatment groups are evaluated by 1-way ANOVA. Statistically, significant variation is defined as ≥20% variation from the control for protein assay and positive or negative fold change threshold of 1.5 for gene expression with a p-value p<0.05.

Example 3: Experimental Design and Methods Defining Additional Efficacious Combinations of Amino Acids Promoting Skin Barrier Function

In addition to the amino acid combinations described above, several other combinations are tested that include, e.g., Trp and/or Arg to determine if inclusion of these amino acids confers additional dermatological benefits. These two amino acids exhibited positive effects in preliminary screens, making them attractive candidates for inclusion in the amino acid formulations described herein. At the outset, the following combinations of amino acids are tested: Ala, Gln, Gly, Ser (combo #1); Ala, Gln, Gly, Ser, Val, Ile (combo #2); Ala, Gln, Gly, Ser, Val; Ala, Gln, Gly, Ser, Ile; Ala, Gln, Gly, Ser, Ile, Val, Trp; Ala, Gln, Gly, Ser, Ile, Val, Arg; and Ala, Gln, Gly, Ser, Ile, Val, Trp, Arg.

The experiments are conducted using differentiated normal primary keratinocytes of five different donors as above and commercially available epidermal skin equivalents (3D model) (EPI-200, MatTech, Ashland, MA). Such 3D cultures approximate human epidermis and are commonly used in skin research. In order to ensure the appropriate experimental conditions, the experiments are conducted in amino acid deprived medium during the time of the treatment with specific amino acid combinations. Since amino acids constitute critical components of the formulations described herein, the presence of amino acids normally present in culture medium has the potential to obfuscate the results, hence leading to a choice of amino acid deprived medium as the control. The cells or equivalents are treated with a combination of amino acids for 4 or 24 h. Differentiation of keratinocytes is induced by culturing them in the presence of 1.2 mM CaCl₂) for 48 h prior the treatment in FBS-free medium (amino acid deprived medium).

First, an array of classical skin barrier related genes that includes a previously tested panel is examined. The following genes encoding key components of the barrier or barrier formation process are included in arrays: growth factors EGF, TGFB1, FGF2, PDGF, terminal differentiation markers such as filaggrin (FLG), loricrin (LOR), TGM4, CORIN, KRT6, KRT16, SFN, SPRR2H and the late cornified envelope genes (LCE1, LCE3, S100A8/A9); lipid metabolism regulators such as PPARA, PPARD, CPT2, PNPLA1, FASN, UGCG, HMGCR, PLA2G5, ELOVL; adhesive structures (CLDN4, CLND7, DSC1, DSC2, OCLN, TJP1 and TJP2); water channel AQP3 and skin barrier integrity genes: SPINK5, KLK5, KLK7. High throughput analysis of mRNA expression of the above genes is performed using custom TaqMan array plates (Thermo Fisher Scientific). The % of genes that are co-expressed after the treatment with different amino acid combinations will be determined. Expression levels of crosslinking proteins such as involucrin, loricrin. TGM2 and ANGPTL are assessed by Western Blot analysis or ELISA using standard protocols.

Methods are performed as described in EXAMPLE 2.

Example 4: Other Model Systems in which to Assess and Identify Efficacious Amino Acid Combinations

Alternative models of keratinocytes that have immature or impacted barrier are tested to assess the effect of amino acid formulations described herein. Such model systems include, for example: undifferentiated keratinocytes (cultured in low calcium concentrations (0.03 mM)). These cells have an immature epithelial barrier; differentiated keratinocytes switched to low Ca2+-containing medium (0.03 mM) for 24 h prior the treatment. It has been shown that when epithelial cells are depleted of Ca2+, the TJs open and thereby disrupt the permeability barrier. This process is reversible with Ca2+replenishment; and differentiated keratinocytes treated with Th2 cytokines for 24 hours: IL-4 (50 ng/ml) and IL-13 (50 ng/mL) that downregulate the expression of filaggrin.

Example 5: Experimental Design and Methods Evaluating Efficacy of Topical Administration of Amino Acid Combinations on Skin Barrier Repair in In Vitro and Ex Vivo Models of Normal and Compromised Skin

The next step in evaluating the efficacy of amino acid formulations described herein is tested via topical administration, which is achieved by direct application on the stratum corneum surface. Normal and immature epidermal skin equivalents (EPI-200 and EPI-20, MatTek Corporation, Ashland. MA) and normal and delipidated (tape-stripped) skin explants are used as models for normal and compromised skin. Full-thickness human skin obtained from normal human adults undergoing abdominoplasty surgeries is available commercially for research and obtained from Zen Bio (Durham, NC). In addition to assessing expression of key barrier markers described in, for example, EXAMPLE 1, TJ proteins will also be examined. TJ proteins are involved in normal barrier function that prevents transepidermal water loss (TEWL) in healthy skin. Accordingly, expression of TJ proteins are assessed by Western Blot analysis and immunohistochemistry. Transepithelial electrical resistance (TEER) measurements will also included as an indicator of barrier integrity and TJ dynamics. Expression of filaggrin, involucrin, and loricrin is also performed as in EXAMPLE 1.

Methods:
Ex Vivo Model:

Ex-vivo studies are performed using human skin explants. Subcutaneous fat is removed from abdominal skin (Zen Bio) and skin biopsies of 0.93 cm²are prepared under sterile conditions and placed in keratinocyte growth medium under a 5% CO₂humidified atmosphere. To artificially induce scaly delipidated skin, abdominal skin is first tape-stripped per standard protocol and then biopsies are prepared. After 24 h of adaptation, amino acid combinations are applied topically every 24 hours for 7 days in total. Skin explants are analyzed for gene and protein expression levels at 24, 48, 72 hours and at the end of the experiment.

Measurement of Transepithelial Electrical Resistance (TEER) in 3D Model.

For TEER, immature epidermal skin equivalents (EPI-20, MatTek Corporation, Ashland, MA) are transferred to medium containing 100 ng/ml IL-4, 100 ng/ml IL-13, 50 ng/ml IL-31 and 30 ng/ml TNFα (R&D Systems, Minneapolis, MN) as described previously. Amino acid combinations are applied topically every 24 hours after 4 hours of pretreatment with cytokines. TEER is measured at 0, 24, and 48 hours by using Millicell ERS-2 Epithelial Volt-Ohm Meter (Milipore). The percentage change in TEER between time 0 (100%) and time 24 hours is expressed as follows: (TEER24 hours/TEER0hours)×100. At the end of the experiment skin equivalents are used for mRNA and protein expression analysis.

Qpcr Analysis.

Expression of the array of 38 genes is performed as described in EXAMPLES 1 and 2 using custom TaqMan array plates.

Protein Analysis:
Western Blot:

Expression of filaggrin, involucrin, loricrin, ANGPTL4 and TGM-2 is performed as described in EXAMPLES 1 and 2.

Immunohistochemical Staining.

Skin explants topically treated with amino acid combinations are incubated for 7 days with treatment renewal every 24 hours and collected for staining at the end of the experiment. The tissues are fixed in a 10% buffered neutral formalin solution. Fixed tissues are embedded and 5-μm-thick sections prepared. Immunohistochemistry analysis is performed using standard protocols. Sections are deparaffinized in xylene, hydrated in ethanol, and washed in water. Nonspecific binding is blocked with the mouse IG blocking reagent. The following primary antibodies are used: claudin-1 (JAY.8), occludin (OC-3F10), ZO-1 (ZO1-1A12), all from Zymed, or isotype control as described. Tissue sections will then be incubated with appropriate biotinylated secondary antibody and antibody binding is visualized using a DAB Peroxidase Substrate Kit (Vector Laboratories). The staining is assessed through microscopic observation.

Example 6: Methods and Materials
Cell Culture and Treatments

Human primary keratinocytes were treated with a panel of amino acid(s), either single amino acids and/or amino acid combinations, in order to prepare experimental samples for RNA analysis.

Primary normal human adult keratinocytes (ATCC, Cat #PCS-200-011) were cultured in Dermal Cell Basal Medium (ATCC. Cat #PCS-200-030) with growth supplement (Keratinocyte Growth Kit, ATCC Cat #PCS-200-040). For the cell growth, the medium was changed three times per week. For the experiment, cells were seeded on 6-well plates with a density of 20,000 cells/well. When cells reached 100% confluency, CaCl₂) was added to the final concentration of 1.8 mM for 24 h, to induce differentiation. All treatments were performed in amino acid-deprived medium (Choline Chloride 0.0285 mM, D-Calcium Pantothenate 0.00838 mM, Folic Acid 0.009 mM, Niacinamide 0.03 mM, Pyridoxine hydrochloride 0.02 mM, Riboflavin 0.001 mM, Thiamine Hydrochloride 0.01 mM, Myo-inositol 0.04 mM, Calcium Chloride 1.8 mM, Ferric Nitrate 2.47E-4 mM, Magnesium Sulfate 0.8 mM. Potassium Chloride 5.3 mM, Sodium Bicarbonate 44 mM, Sodium Chloride 110.3 mM, Sodium Phosphate Monobasic 0.9 mM, D-Glucose 25 mM, Sodium Pyruvate 1 mM) in the presence of 1.8 mM CaCl2 for 4 h. The 4GAA combination contained Glycine, Glutamine, Serine, and Alanine and was used in a total concentration of 4 mM (1:1:1:1). The 3BAA combination contained Cysteine (1 mM), Histidine (0.5 mM), Tyrosine (1 mM), The plant extract combination containing Boswellia serrata resin extract, lecithin, microcrystalline cellulose, and silica was dissolved in the amino-acid-deprived medium, filtered, and used in concentrations 0.1 w/w %-1 w/w % of the formulation.

Real-Time RT-PCR Analysis:

Cellular RNA was isolated using a NucleoSpin RNA Plus kit (Macherey Nagel). cDNA synthesis was performed using High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific) with 1 μg of total RNA. RT product was amplified using EGF, MYC, GAPDH, YWHAZ TaqMan Gene Expression Assays (Thermo Fisher Scientific). Detection was performed using the StepOne Real-Time PCR System (Applied Biosystems). Differences in the levels of gene expression were determined by relative quantification using the delta Ct method.

Equipment, Materials, Reagents, and Supplies

Confluent differentiated primary human keratinocytes were used in 6-well plates. The following were used in experiments described here:

- Sterile RNase-free microcentrifuge (1.7 mL) and conical (50 mL and 15 mL) tubes.
- Pipette, Multi-Channel pipette, Pipette Controller (Aid), Serological Pipettes and Pipette Tips, Aspirating Pipettes.
- Sterile Isopropyl Alcohol (SIPA, 70%).
- Dulbecco's media.
- Amino Acids: Serine, Glycine, Glutamine, Histidine, Valine, Tyrosine, Threonine, Aspartic Acid, or others based on experimental needs.
- Vacuum line/Vacuum pump, and pump waste collection receptacles.
- Syringe (30 mL), Syringe Filter (25 mm PES; 0.22 Micron), Syringe Needles (18G).
- Weighing Supplies: Paper, Spatula, etc.
- Lab equipment: Biosafety cabinet (BSC), Water Bath (37° C.), 5% CO₂Incubator (37° C.) Refrigerator (4° C.), Analytical Balance, Heat/Stir Plate.

TABLE 1

Reagents/Supplies Storage Conditions

Room Temp
Refrigerator
CO₂incubator

Item
(~20° C.-25° C.)
(4° C.)
(37° C.)

AA9(s)
X

PBS

X

Primary Human

X

Keratinocytes

DMEM

X

AA solutions

X

Amino Acid Treatment Preparation

Treatments were prepared no more than one week prior to beginning the experiment. A water bath was pre-heated to 37° C. The DMEM was retrieved and aseptically transferred to the BSC. The needed volume of DMEM (2 ml/well) was pipetted into conical tubes (50 mL), which were pre-warmed in the heated water bath for 30 min. The other required materials: conical tubes (50 mL), syringes (30 mL), needles (18G), and micron syringe filters (25 mm; 0.22 micron) were aseptically transferred to the BSC. The amino acids required for treatment were prepared by labeling conical tubes (50 mL), either for single amino acids (4 mM, 20 mM), 4 amino acid combination (4 mM, 20 mM), or ENTERADE™ (see, TABLE 2).

The appropriate amount of each amino acid (See, TABLE 2) was weighed out before they were transferred to their respective conical tubes. The amino acids for the 4AA formula and “ENTERADE®” should be weighed out individually and then transferred into the designated tube and labeled together. All of the conical tubes (50 mL) containing the dry amino acids and pre-warmed DMEM were aseptically transferred to the BSC. DMEM was transferred into the tubes according to TABLE 2 using serological pipettes.

All of the tubes were vortexed until the mixture was free of debris and homogenous and then the homogenous mixtures were aseptically returned to the BSC.

Should a mixture not be fully in solution with just vortexing, a small stir bar was placed in the conical tube (50 mL), then the conical tube was placed in a beaker half-filled with water and used a heat/stir plate (50° C./800 rpm) for ˜10 min until the solution was homogenous.

In a separate rack, a new set of conical tubes (50 mL) was labeled for each amino acid or the amino acid combination with the AA name, concentration, and date.

A needle (18G) was attached to a syringe (10 mL) and the entire contents of one of the AA “Medium” tubes were drawn up. The needle was removed and a syringe filter (13 mm, 0.22 Micron) was attached to the syringe (10 mL). The syringe filter (25 mm, 0.22 Micron) was placed over the corresponding amino acid (AA) “Treatment” labeled tube, the mixture was pushed in the syringe through the filter into the tube, and capped immediately. This was repeated as needed, changing filters for every 100 mL filtered or in-between each amino acid sample.

The syringe filtration step was repeated for each AA or AA combination and the syringe, needle, and filter were replaced in between each AA or AA combination to avoid cross contamination.

TABLE 2

Amino Acids and Concentrations

1X Stock
Final

Weight
Volume
Concentration
Concentration

Amino Acid
(mg)
(mL)
(mM)
(mM)

Serine
16.816
40
4
4

Glycine
12.011
40
4
4

Histidine
23.383
40
4
4

Glutamine
24.862
40
4
4

Serine
84.80
40
20
20

Glycine
60.056
40
20
20

Histidine
116.916
40
20
20

Glutamine
124.128
40
20
20

4AA Serine
16.816
40
4
4

4AA Glycine
12.011
40
4
4

4AA Histidine
23.383
40
4
4

4AA Glutamine
24.862
40
4
4

4 AA Serine
84.08
40
20
20

4 AA Glycine
60.056
40
20
20

4AA Histidine
116.916
40
20
20

4AA Glutamine
124.128
40
20
20

ENTERADE ®
31.94
30
10
8

Serine

ENTERADE ®
28.5888
30
10
10

Valine

ENTERADE ®
6.52284
30
1.2
1.2

Tyrosine

ENTERADE ®
35.154
30
8
8

Threonine

ENTERADE ®
31.53
30
8
8

Aspartic Acid

Control
NA
15
NA
NA

Amino Acid Treatment

PBS and AA treatments were pre-warmed in the water bath (37° C.) for 30 minutes. The 6-well plates containing “Differentiated Keratinocytes” were aseptically transferred from the 5% CO₂Incubator (37° C.) into the BSC. The 6-well plate lids were labeled with Date, Initials, Passage #, Cell Family, and the Target treatment. The well imprints on the plate's lid were labeled with T #(Treatment Wells) or C#(PBS only Wells).

The medium from each well was aspirated and washed with 2.0 mL PBS.

The PBS from each well was aspirated and 2.0 mL of the AA treatments was added into the wells ensuring the treatment added corresponds to the plate's label. (Serine treatments only went into wells of the serine labeled plate).

The remaining control well was filled in each plate with 2 mL of DMEM and the 6-Well plates in a secondary container were transferred to the 5% CO₂incubator (37° C.) and incubated based on the experiment.

Once the incubation time was completed, the “Treated” cells were removed and transferred to the BSC for RNA/Protein extraction.

Example 7: Preparation of Formulation

An exemplary method of preparing the formulation or topical formulation disclosed here included the steps as follows:

- Dispersed the xanthan gum in glycerin.
- Mixed until homogenous and then added water.
- Mixed until homogenous.
- Heated Phase A ingredients of TABLE 3 to a temperature of 80° C.
- Separately heated Phase B ingredients of TABLE 3 to a temperature of 80° C.

With both phases at 80° C., Phase B was added to Phase A with vigorous mixing, then removed from heat, forming a batch of Phase A and Phase B.

When the batch temperature was below 40° C., Phase C ingredients of TABLE 3 were added to the batch.

Then q.s. batch to 100% with deionized water. Various amino acids tested here were combined with the formulation prepared here, including alanine, glutamine, glycine, and serine.

TABLE 3

Ingredients

INCI

% IN

PHASE
TRADENAME
NAME
COMPANY
FORMULA

A
Keltrol Cg-Sft
Xanthan Gum
Cp Kelco
0.600%

A
Glycerin
Glycerin
Independent
1.000%

Chemical

A-A
Deionized Water
Water

75.000%

B
Ips Deca-Dp
Polyglyceryl-
Ips Labs
1.750%

10 Dipalmitate

Ips Hexa-Ds
Polyglyceryl-6
Ips Labs
0.750%

Distearate

B
Stearyl Alcohol
Stearyl

5.000%

Alcohol

B
Sunflower Oil
Helianthus

12.000%

Annuus Seed

Oil

C
Lexgard Natural
Methylheptyl-
Inolex
2.000%

Mhg Mb
glycerin

C
Zeastat
Capryl-
Inolex
1.000%

hydroxamic

Acid (And)

Propanediol

Example 8: EGF Expression Comparison of Amino Acid Combination and Plant Extract Combination, Alone or Together

Keratinocytes from different donors were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino acid combination (4GAA; alanine, glutamine, glycine, serine) at 1 mM or 4 mM and/or Boswellia plant extract combination (BSW or BSXL) in 0.25%. 0.5%, or 1% (w/w %) concentrations (where the combinations are 4GAA 4 mM/1% BSW (FIG. 7A); 4GAA 1 mM/0.25% BSW (FIG. 7B); 4GAA 4 mM/0.5% BSW (FIG. 7C); 4GAA 1 mM/0.5% BSW (FIG. 7D)).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 7A-7D, *p<0.05, **p<0.01 vs untreated control (CTR or CTR untr).

Example 9: Comparison of Fold Change of EGF mRNA of Amino Acid Combination and Plant Extract Combination, Alone or Together, Donor G

Keratinocytes from Donor G were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino acid combination (4GAA; alanine, glutamine, glycine, serine) at 1 mM or 4 mM and/or plant extract combination (BSXL) in 0.1%. 0.25% or 0.5% w/w % concentrations (where the combinations are 4GAA 1 mM/0.25% BSXL and 4GAA 4 mM/0.5% BSXL).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 8A-8B, *p<0.05, **p<0.01, ***p<0.001. ****p<0.0001 vs untreated control (ctr).

Example 10: Comparison of Fold Change of EGF mRNA of Amino Acid Combination and Plant Extract Combination, Alone or Together, Donor O

Keratinocytes from Donor O were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino acid combination (4GAA; alanine, glutamine, glycine, serine) at 1 mM or 4 mM and/or plant extract combination (BSXL) in 0.25% or 1% w/w % concentrations (where the combinations are BSXL 0.25%/1 mM 4GAA and BSXL 1%/4 mM 4GAA).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 9A-9B, **p<0.01, ***p<0.001 vs untreated control (CTR untr).

Example 11: Comparison of Fold Change of Egf mRNA of Amino Acid Combination and Plant Extract Combination, Alone or Together

Keratinocytes from Donor G were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino combination (4GAA at 4 mM: alanine, glutamine, glycine, serine; 3BAA: cysteine (Cys), histidine (His), tyrosine (Tyr): Cys 1 mM. His 0.5 mM. Tyr 1 mM) and/or plant extract combination (BSXL) in 0.5% and/or piperonylic acid (100 μM; PA) (where the combinations are 4GAA 4 mM/0.5% BSXL; 3BAA 2.5 mM/0.5% BSXL; 4GAA 4 mM/100 μM PA; 3BAA 2.5 mM/100 μM PA).

The same experiment was performed with similar cells and different combinations of amino acids, as well as a compound (PA) that upregulates EGFR (EGF Receptor).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 10A-10B, *p<0.05, **p<0.01, ***p≤0.001 versus untreated control (CTR).

Example 12: Comparison of Fold Change of EGF mRNA of Amino Acid Combination and Plant Extract Combination, Alone or Together, Donor G

Keratinocytes from Donor G were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino combination (4GAA at 4 mM: alanine, glutamine, glycine, serine; 3BAA: cysteine (Cys), histidine (His), tyrosine (Tyr): Cys 1 mM. His 0.5 mM. Tyr 1 mM) and/or plant extract combination (BSXL) in 0.5% and/or piperonylic acid (200 μM; PA) (where the combinations are 4GAA 4 mM/0.5% BSXL; 3BAA 2.5 mM/0.5% BSXL; 4GAA 4 mM/200 μM PA; 3BAA 2.5 mM/200 μM PA).

The same experiment was performed with similar cells and different combinations of amino acids, as well as a compound (PA) that upregulates EGFR (EGF Receptor).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 11A-11B, *p<0.05, **p≤0.01, ***p≤0.001 versus untreated control (CTR).

Example 13: Comparison of Fold Change of EGF mRNA of Amino Acid Combination, Plant Extract, and Plant Extract Combination, Alone or Together, Donor O

Keratinocytes from Donor O were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino combination (4GAA; alanine, glutamine, glycine, serine) at 1 mM or 4 mM and/or plant extract combination (BSXL) in 0.25% and/or Boswellic acid (BA) in 2 μM. 10 μM. and 50 μM concentrations (where the combinations are 4GAA 1 mM/0.25% BSXL; 4GAA 4 mM/0.25% BSXL; 4GAA 1 mM/10 μM BA; 4GAA 1 mM/2 μM BA; 4GAA 1 mM/50 μM BA).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 12A-12B, *p<0.05, **p≤0.01, ***p≤0.001 vs untreated control (CTR).

Example 14: Comparison of Fold Change of EGF mRNA of Amino Acid Combination and Plant Extract Combination, Alone or Together, Donor A

Keratinocytes from Donor A were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino combination (4GAA: alanine, glutamine, glycine, serine; 4OAA: aspartic acid, isoleucine, tyrosine, valine) at 4 mM and/or plant extract combination (BSXL) in 0.5% (where the combinations are 4GAA 4 mM/0.5% BSXL; 4OAA 4 mM/0.5% BSXL).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. FIGS. 13A-13B, ***p≤0.001 vs untreated control (CTR).

Example 15: Comparison of Fold Change of Egf mRNA of Amino Acid Combination and Plant Extract Combination, Alone or Together

Keratinocytes from Donor A (n=1). Donor G (n=3), and Donor T (n=2) were cultured in amino acid-free media for 4 hours after complete differentiation and, in parallel, were treated with an amino combination (4GAA: alanine, glutamine, glycine, serine) at 4 mM and/or plant extract combination (BSXL) in 0.5% (where the combination is 4GAA 4 mM/0.5% BSXL).

Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed with GAPDH as the housekeeping gene. See. Averages: FIGS. 14A-14C, *p≤0.05 vs untreated control (CTR).

Example 16: Testing Transepidermal Water Loss (TEWL) Change

In order to determine transepidermal water loss (TEWL) change, test materials (Active (4GAA), left column; Placebo, right column) were applied for 5 days on the skin of 12 subjects who were then subjected to 15 repeated tape strips. See. FIGS. 15A-15B. The active 4GAA amino acid combination contained alanine (4%), glutamine (4%), glycine (4%), serine (4%) at a total concentration of 12% in an oil/water (O/W) vehicle and placebo. The baseline. Day 1, was immediately after tape-stripping. Days 2, 3, and 5 are each after two treatments per day of test materials.

Example 17: Measurement of Skin Hydration

In order to determine skin barrier improvement, test materials (Untreated control; Placebo; Active Amino Acid (4GAA)) were applied for 5 days on the skin of 25 subjects who were then subjected to 15 repeated tape strips. See. FIG. 16. Control represents untreated (top graphical line at Day 3; left column-inset); Placebo represents water/oil/water (W/O/W) vehicle without active (middle graphical line at Day 3; center column-inset); Amino Acid represents the 4 amino acid combination (Ala (4%). Gln (4%). Gly (4%). Ser (4%) in the W/O/W vehicle (bottom graphical line at Day 3; right column-inset). *, p-value=<0.05. Inset: Control (left), Placebo (center), Amino Acid (right).

Corneometry was used to measure skin hydration of the superficial layers of the skin of the subjects (stratum corneum).

Example 18: Gene Expression Comparison of Amino Acid Combination and Retinoid, Alone or Together

Keratinocytes from different donors (e.g., human-derived epidermal keratinocytes (NHEK)), skin equivalents (e.g., 3D models (human epidermal skin equivalents)), or fibroblasts (e.g., human dermal fibroblasts, adult (HDFa)) were cultured in amino acid-deprived media or complete media for 4 hours or 24 hours after complete differentiation and, in parallel, were treated with an amino acid combination (4GAA; alanine, glutamine, glycine, serine) at 4 mM, 10 μM retinol (ROL), or the combination of 4 mM 4GAA and 10 μM ROL. Cells were harvested, RNA was extracted, and qPCR analysis was performed with a housekeeping gene, e.g., GAPDH.

FIGS. 17A-17C and 18A-18C show the graphical results of the fold change, where the average fold change is depicted by the horizontal bar for each condition and each of the donors (Donors E, G, O, R, T) is depicted by a colored circle. Donor keratinocytes were treated for 4 hours (4 h) in amino acid-deprived medium with the 4 amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), retinol (ROL; 10 μM), alone or in combination, or with control (CTR: amino acid-deprived medium with keratinocytes).

The 4GAA free amino acid combination upregulated EGF in donor keratinocytes (FIG. 17A), but no synergy was observed in TGFB or COL1A1, genes that are upregulated by retinol (FIGS. 17B-17C).

Retinol (ROL) upregulated HAS2 and HAS3 in donor keratinocytes (FIGS. 18B-18C, respectively) as did the 4GAA free amino acid combination alone. When both 4GAA and ROL were combined, synergy was observed in HAS2 and HAS3 gene expression (FIGS. 18B-18C, respectively; see also, FIGS. 18E-18F). In contrast, HAS1 expression was downregulated, and no synergy was observed. (FIGS. 18A, 18D).

Donor O and Donor R keratinocyte cells were treated for 24 h in complete medium with control (CTR; complete medium); the 4GAA amino acid combination (Ala, Gln, Gly, Ser; 4GAA; 4 mM), retinol (ROL; 10 μM), and the combination of 4GAA and ROL. FIGS. 19A-19D, 20A-20D, and 21 provide graphical and tabular forms of the data generated. Gene expression was tested under the indicated conditions, where FIG. 19A and FIG. 20A show the results of EGF mRNA gene expression in keratinocyte cells from Donors O and R, respectively; FIG. 19B and FIG. 20B show the results of HAS1 mRNA gene expression in keratinocyte cells from Donors O and R, respectively; FIG. 19C and FIG. 20C show the results of HAS2 mRNA gene expression in keratinocyte cells from Donors O and R, respectively; and FIG. 19D and FIG. 20D show the results of HAS3 mRNA gene expression in keratinocyte cells from Donors O and R, respectively. FIG. 21 presents the generated data for gene expression under the specified conditions from keratinocytes from Donors O and R. EGF mRNA gene expression was tested as a control. ROL upregulated HAS2 and HAS3 in the donor keratinocytes and 4GAA alone had the same effect. For the 4GAA and ROL combination, synergy was observed in HAS2 and HAS3 mRNA gene expression.

Donor keratinocytes (FIGS. 18A-18C) were compared to a 3D skin equivalent model (FIGS. 22A-22B) (EpiDerm™; MatTek Corp., Ashland, MA). Specifically, synergy for HAS2 and HAS3 with the combination of 4GAA+ROL was observed in both donor keratinocytes and in the 3D skin equivalents. Whereas there was no HAS1 gene expression in the 3D skin equivalents (data not shown) and no synergy observed in keratinocytes (scc. FIG. 18A).

Human dermal fibroblasts, adult (HDFa) were also used to test the various conditions (n=3)/condition: control (CTR: EBS medium); 4GAA (4 mM); ROL (10 μM); 4GAA (4 mM)+ROL (10 μM) combination, under treatment of 4 h. No synergy was observed in fibroblasts for the combination of 4GAA+ROL when testing for COL1A1 gene expression (n=3) (FIG. 23A), and no changes in COL1A1 expression level for collagen was observed. However, elastin expression (ELN) was upregulated by 4GAA and further upregulated with ROL, demonstrating synergy (FIG. 23B). There was very low expression of EGF in fibroblasts (cycle threshold. CT=35; data not provided).

FIGS. 24A-24C demonstrates the fold change of HAS1, HAS2, and HAS3 gene expression, respectively, in fibroblasts. A 4 h treatment of fibroblasts with 4GAA amino acid combination alone or in combination with ROL resulted in an upregulation of HAS1 and HAS2 gene expression (FIGS. 24A-24B, respectively), and most predominantly in HAS1 gene expression in fibroblasts with the 4GAA+ROL combination (FIG. 24A). In contrast, 4GAA alone or in combination with ROL resulted in downregulation of HAS3 expression in fibroblasts (FIG. 24C). When the fibroblast data of FIGS. 24A-24C were compared to the keratinocyte data of FIGS. 18A-18C. HAS2 and HAS3 were found to be upregulated by the combination of 4GAA+ROL in keratinocytes, while HAS1 was predominantly upregulated by the combination of 4GAA+ROL in fibroblasts.

A comparison of HAS1 gene expression with fibroblasts in amino acid-deprived media and in complete media was performed by treating the fibroblasts for 4 h with: control (CTR EBS media vs CTR complete media); 4GAA (4 mM); ROL (10 μM); and the combination of 4GAA+ROL, each in their respective media. Another comparison of HAS1 gene expression with fibroblasts in complete media and either a 4 h treatment (FIG. 25A) or a 24 h treatment (FIG. 25B) was also performed. Synergy was demonstrated with a treatment of the combined 4GAA+ROL in fibroblasts for HAS1 gene expression in both media (AA-deprived vs complete medium) and treatment times.

Fibroblasts in complete medium were treated for 24 h under specified conditions (CTR, complete media; 4GAA, 4 mM: ROL, 10 μM: or 4GAA+ROL) and gene expression of EGF, HAS1, HAS2, HAS3, and COL1A1 was determined. See, FIG. 26. The combination of 4GAA+ROL was compared in fibroblasts expressing HAS1 (i.e., 5.6; FIG. 26) and in keratinocytes expressing HAS2 and HAS3 (i.e., Donor O/Donor R; 404.6/290, 1.6/4.2, respectively; FIG. 21).

Quantitative polymerase chain reaction (qPCR) was performed to demonstrate the amount of a gene present in a sample. The cycle threshold (CT) value increases with a decreasing amount of template for amplification. FIGS. 27A-27E present the CT values in donor human-derived epidermal keratinocytes (NHEK) (Donor E (FIG. 27A), Donor G (FIG. 27B), Donor O (FIG. 27C), Donor R (FIG. 27D), and Donor T (FIG. 27E) for endogenously expressed HAS2 (CT=35), which is a very low number in that the gene that is present is in an almost undetectable quantity or amount. The treatment, however, increased this concentration. Endogenous expression of the gene in untreated cells is almost undetectable. The CT value for HAS1 in untreated keratinocytes was 30-31; for HAS3 in untreated was 26-27; for EGF in untreated was 29-32; and for GAPDH was 19-21.

Example 19: Testing of Amino Acids

Using gene arrays, comparative analyses of two datasets were performed in three diverse donors focusing on consistently differentially [filter criterion of fold change 1.5 (positive or negative) and p-value <0.05)] expressed genes following treatment with various amino acids and combinations thereof. qPCR was used to assess the mRNA expression results. The qPCR results confirmed the array results pertaining to differential expression, thereby validating the panel of genes.

The marker genes: epidermal growth factor (EGF); platelet-derived growth factor (PDGF), fibroblast growth factor 2 (FGF2), peroxisome proliferator activated receptor delta (PPARD).

Comparative analysis of qPCR data from two donors, which included a scoring system (as described above), identified amino acids and combinations of amino acids which upregulated skin barrier markers.

Example 20: Experimental Methods

Amino acid(s) are mixed in previously established concentrations in water and added to amino acid-deprived culture medium.

Gene Expression:

mRNA is extracted, converted to cDNA, and analyzed by qPCR method using custom TaqMan array 384-well plates (Thermo Fisher Scientific) containing the above-mentioned genes. Control housekeeping genes were included to normalize the mRNA expression: GAPDH and/or YWHAZ. The array plate contained the genes and samples were analyzed in by qPCR.

SPECIFIC EMBODIMENTS

Non-limiting specific embodiments are described below each of which is considered to be within the present disclosure.

- Embodiment 1. A formulation for administration to skin comprising:
  - as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination of glutamine, glycine, alanine, and serine, or salts thereof; and
  - optionally, a therapeutically effective amount of at least one additional free amino acid or a peptide combination of amino acids of valine, isoleucine, arginine, threonine, or tryptophan, or salts thereof, or any combination thereof;
  - wherein the therapeutically effective amount of the combination of glutamine, glycine, alanine, and serine, or salts thereof and the therapeutically effective amount of the at least one additional free amino acid or peptide combination of amino acids improves barrier integrity of skin cells, when tested by a method detecting expression of a barrier marker gene (e.g., CPT2, EGF, FGF2, FLG, MKI67, PDGF, PPARD, TGM4) and determining that the barrier marker gene is modulated by a 1-7 average fold change of amino acid treated as compared to amino acid untreated skin; and
- optionally, a dermatologically acceptable carrier;
- wherein the formulation improves barrier integrity of the skin.
- Embodiment 2. The formulation of embodiment 1, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine, or salts thereof; and at least one additional amino acid of isoleucine, arginine, threonine, or tryptophan, or salts thereof, or any combination thereof.
- Embodiment 3. The formulation according to embodiment 1 or embodiment 2, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine, or salts thereof; and at least one additional amino acid of arginine, or threonine, or salts thereof, or any combination thereof.
- Embodiment 4. The formulation according to embodiment 1 or embodiment 2, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine, or salts thereof.
- Embodiment 5. The formulation according to embodiment 1 or embodiment 2, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine, or salts thereof.
- Embodiment 6. The formulation according to any one of embodiments 1-3, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and arginine, or salts thereof.
- Embodiment 7. The formulation according to any one of embodiments 1-3, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and threonine, or salts thereof.
- Embodiment 8. The formulation of embodiment 1, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, and serine, or salts thereof; and at least one additional amino acid of isoleucine, arginine, threonine, or tryptophan, or salts thereof, or any combination thereof.
- Embodiment 9. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine, or salts thereof.
- Embodiment 10. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine, or salts thereof.
- Embodiment 11. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine, or salts thereof.
- Embodiment 12. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan, or salts thereof.
- Embodiment 13. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine, or salts thereof; and at least one additional amino acid of arginine, or threonine, or tryptophan, or salts thereof, or any combination thereof.
- Embodiment 14. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and arginine, or salts thereof.
- Embodiment 15. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and threonine, or salts thereof.
- Embodiment 16. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and tryptophan, or salts thereof.
- Embodiment 17. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine, or salts thereof.
- Embodiment 18. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and tryptophan, or salts thereof.
- Embodiment 19. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, threonine, and tryptophan, or salts thereof.
- Embodiment 20. The formulation according to embodiment 13, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, threonine, and tryptophan, or salts thereof.
- Embodiment 21. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine, or salts thereof; and at least one additional amino acid of isoleucine, threonine, or tryptophan, or any combination thereof.
- Embodiment 22. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine, or salts thereof; and at least one additional amino acid of isoleucine, arginine, or tryptophan, or salts thereof, or any combination thereof.
- Embodiment 23. The formulation according to embodiment 8, wherein the free amino acids or a peptide combination of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan, or salts thereof; and at least one additional amino acid of isoleucine, arginine, or threonine, or salts thereof, or any combination thereof.
- Embodiment 24. The formulation according to any one of embodiments 1-23, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration ranging from 0.05 mM to 20 mM.
- Embodiment 25. The formulation according to any one of embodiments 1-24, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration ranging from 0.05 mM to 10 mM.
- Embodiment 26. The formulation according to any one of embodiments 1-25, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration ranging from 0.05 mM to 5 mM.
- Embodiment 27. The formulation according to any one of embodiments 1-23, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration of ranging from 0.1 mM to 10 mM.
- Embodiment 28. The formulation according to any one of embodiments 1-23, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration of ranging from 0.1 mM to 5 mM.
- Embodiment 29. The formulation according to any one of embodiments 1-11, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration of ranging from 0.1 mM to equal to or less than 4 mM.
- Embodiment 30. The formulation according to any one of embodiments 1-23, wherein each of the free amino acids, or each of the amino acids in the peptide combination of amino acids, or salts thereof, is present at a concentration ranging from equal to or greater than 0.5 mM to equal to or less than 4 mM.
- Embodiment 31. The formulation according to any one of embodiments 1-30, wherein, if present, amino acids of cysteine are present at a concentration of equal to or less than 1 mM;
- if present, amino acids of histidine are present at a concentration of equal to or less than 0.5 mM;
- if present, amino acids of tyrosine are present at a concentration of equal to or less than 1 mM;
- if present, amino acids of leucine are present at a concentration of equal to or less than 4 mM; or
- if present, amino acids of aspartic acid are present at a concentration of equal to or less than 2 mM; or any combination thereof.
- Embodiment 32. The formulation according to any one of embodiments 1-31, wherein the formulation does not comprise amino acids of cysteine, histidine, tyrosine, leucine, aspartic acid, taurine, taurate, or glutamate, or any combination thereof.
- Embodiment 33. The formulation according to any one of embodiments 1-32, wherein the formulation is administered to the skin or administrable via transdermal, subcutaneous, or topical administration.
- Embodiment 34. A method for maintaining and/or improving barrier integrity of skin cells in a subject in need thereof, the method comprising administering a formulation of any one of embodiments 1-33 to the skin of the subject.
- Embodiment 35. The method according to embodiment 34, wherein the formulation is administered to improve a cosmetic skin condition.
- Embodiment 36. The method according to embodiment 35, wherein the cosmetic skin condition is associated with aging of the skin.
- Embodiment 37. The method according to any one of embodiments 34-36, wherein the formulation is administered to improve at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin.
- Embodiment 38. The method according to embodiment 37, wherein improving the at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin reduces the length, depth, and/or other dimension of lines and/or wrinkles in the skin.
- Embodiment 39. The method according to embodiment 34, wherein the subject in need thereof is afflicted with a skin condition.
- Embodiment 40. The method according to embodiment 39, wherein the skin condition comprises atopic dermatitis, dermatitis, radiodermatitis, psoriasis, pruritus, eczema, a wound, or a burn.
- Embodiment 41. The method according to any one of embodiments 34-40, wherein the subject is a human.
- Embodiment 42. A method for improving a cosmetic skin condition of a human, the method comprising administering the formulation according to any one of embodiments 1-33 to the human's skin in an amount effective to improve at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin.
- Embodiment 43. The method according to embodiment 42, wherein the cosmetic skin condition is associated with aging of the skin.
- Embodiment 44. The method according to embodiment 42 or embodiment 43, wherein improving the at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin reduces the length, depth, and/or other dimension of lines and/or wrinkles in the skin
- Embodiment 45. Use of a formulation according to any one of embodiments 1-33 for the treatment of a cosmetic skin condition.
- Embodiment 46. The use according to embodiment 45, wherein the cosmetic skin condition is associated with aging of the skin.
- Embodiment 47. The use according to embodiment 45 or embodiment 46, wherein the formulation is administered to improve at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin.
- Embodiment 48. The according to embodiment 47, wherein improving the at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin reduces the length, depth, and/or other dimension of lines and/or wrinkles in the skin.
- Embodiment 49. Use of a formulation according to any one of embodiments 1-33 for the treatment of a skin condition.
- Embodiment 50. The use according to embodiment 49, wherein the skin condition comprises atopic dermatitis, dermatitis, radiodermatitis, psoriasis, pruritus, eczema, a wound, or a burn.
- Embodiment 51. Use of a formulation according to any one of embodiments 1-33 in the preparation of a dermatological medicament for treating a cosmetic skin condition in a subject in need thereof.
- Embodiment 52. The dermatological medicament according to embodiment 51, wherein the cosmetic skin condition is associated with aging of the skin.
- Embodiment 53. The dermatological medicament according to embodiment 51 or embodiment 52, wherein the dermatological medicament is administrable to improve at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin.
- Embodiment 54. The dermatological medicament according to embodiment 53, wherein improving the at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin reduces the length, depth, and/or other dimension of lines and/or wrinkles in the skin.
- Embodiment 55. Use of a formulation according to any one of embodiments 1-33 in the preparation of a pharmaceutical medicament for treating a skin condition in a subject in need thereof,
- Embodiment 56. The pharmaceutical medicament according to embodiment 55, wherein the skin condition comprises atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, a wound, or a burn.
- Embodiment 57. A topical formulation, comprising (or consisting essentially of or consisting of):
  - as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of, or consisting of):
    - at least one of: alanine, glutamine, glycine, and serine, or salts thereof;
  - a therapeutically effective amount of a plant extract,
    - wherein the plant extract comprises (or consists essentially of or consists of): a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); and
  - optionally, a dermatologically acceptable vehicle and/or additive.
- Embodiment 58. A topical formulation, comprising (or consisting essentially of or consisting of):
  - as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of):
    - alanine, glutamine, glycine, and serine, or salts thereof;
  - a therapeutically effective amount of a plant extract combination (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising (or consisting essentially of, or consisting of):
    - Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense);
    - a texturing and/or a bulking agent (e.g., cellulose (microcrystalline));
    - a phospholipid (e.g., lecithin or phosphatidylcholine); and
    - an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and
  - optionally, a dermatologically acceptable vehicle and/or additive.
- Embodiment 59. A formulation for administration to skin comprising (or consisting essentially of or consisting of):
  - a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising (or consisting essentially of or consisting of):
    - alanine, glutamine, glycine, and serine, or salts thereof;
  - a therapeutically effective amount of a plant extract combination comprising (consisting essentially of or consisting of) (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica):
    - Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense);
    - a texturing and/or a bulking agent (e.g., cellulose (microcrystalline);
    - a phospholipid (e.g., lecithin or phosphatidylcholine); and
    - an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications); and
  - optionally, a dermatologically acceptable vehicle,
    - wherein the therapeutically effective amount of the combination of free amino acids or a peptide combination thereof and the therapeutically effective amount of the plant extract combination improves skin regeneration (e.g., skin barrier repair, skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration) when tested by a method detecting expression of a barrier marker gene (e.g., EGF) and determining that the barrier marker gene is increased by a 1.5-40 average fold increase of amino acid treated as compared to amino acid untreated skin,
- wherein the formulation improves skin regeneration (e.g., skin barrier repair, skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocyte, endothelial cells, and fibroblasts; facilitates dermal regeneration),
- wherein an improvement in skin regeneration, including skin barrier repair is indicated by an average fold increase of a skin regeneration, including skin barrier, marker gene (e.g., EGR) expression of: 1.5 or greater (e.g., 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7, 1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1; 11.3; 11.5; 11.7; 11.9; 12.1; 12.3; 12.5; 12.7; 12.9; 13.1; 13.3; 13.5; 13.7; 13.9; 14.1; 14.3; 14.5; 14.7; 14.9; 15.1; 15.3; 15.5; 15.7; 15.9; 16.1; 16.3; 16.5; 16.7; 16.9; 17.1; 17.3; 17.5; 17.7; 17.9; 18.1; 18.3; 18.5; 18.7; 18.9; 19.1; 19.3; 19.5; 19.7; 19.9; 20.1; 20.3; 20.5; 20.7; 20.9; 21.1; 21.3; 21.5; 21.7; 21.9; 22.1; 22.3; 22.5; 22.7; 22.9; 23.1; 23.3; 23.5; 23.7; 23.9; 23.1; 23.3; 23.5; 23.7; 23.9; 24.1; 24.3; 24.5; 24.7; 24.9; 25.1; 25.3; 25.5; 25.7; 25.9; 26.1; 26.3; 26.5; 26.7; 26.9; 27.1; 27.3; 27.5; 27.7; 27.9; 28.1; 28.3; 28.5; 28.7; 28.9; 29.1; 29.3; 29.5; 29.7; 29.9; 30.1; 30.3; 30.5; 30.7; 30.9; 31.1; 31.3; 31.5; 31.7; 31.9; 32.1; 32.3; 32.5; 32.7; 32.9; 33.1; 33.3; 33.5; 33.7; 33.9; 34.1; 34.3; 34.5; 34.7; 34.9; 35.1; 35.3; 35.5; 35.7; 35.9; 36.1; 36.3; 36.5; 36.7; 36.9; 37.1; 37.3; 37.5; 37.7; 37.9; 38.1; 38.3; 38.5; 38.7; 38.9; 39.1; 39.3; 39.5; 39.7; 39.9; 40.1; 40.3; 40.5; 40.7; 40.9; 41.1; 41.3; 41.5);
- 40 or less (e.g., 39.8; 39.6; 39.4; 39.2; 39; 38.8; 38.6; 38.4; 38.2; 38; 37.8; 37.6; 37.4; 37.2; 37; 36.8; 36.6; 36.4; 36.2; 36; 35.8; 35.6; 35.4; 35.2; 35; 34.8; 34.6; 34.4; 34.2; 34; 33.8; 33.6; 33.4; 33.2; 33; 32.8; 32.6; 32.4; 32.2; 32; 31.8; 31.6; 31.4; 31.2; 31; 30.8; 30.6; 30.4; 30.2; 30; 29.8; 29.6; 29.4; 29.2; 29; 28.8; 28.6; 28.4; 28.2; 28; 27.8; 27.6; 27.4; 27.2; 27; 26.8; 26.6; 26.4; 26.2; 26; 25.8; 25.6; 25.4; 25.2; 25; 24.8; 24.6; 24.4; 24.2; 24; 23.8; 23.6; 23.4; 23.2; 23; 22.8; 22.6; 22.4; 22.2; 22; 21.8; 21.6; 21.4; 21.2; 21; 20.8; 20.6; 20.4; 20.2; 20; 19.8; 19.6; 19.4; 19.2; 19; 18.8; 18.6; 18.4; 18.2; 18; 17.8; 17.6; 17.4; 17.2; 17.2; 17; 16.8; 16.6; 16.4; 16.2; 16; 15.8; 15.6; 15.4; 15.2; 15; 14.8; 14.6; 14.4; 14.2; 14; 13.8; 13.6; 13.4; 13.2; 13; 12.8; 12.6; 12.4; 12.2; 12; 11.8; 11.6; 11.4; 11.2; 11; 10.8; 10.6; 10.4; 10.2; 10; 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6); or 1.5-40 (e.g., 1.6-39.9; 1.7-39.8; 1.8-39.7; 1.9-39.6; 2-39.5; 2.1-39.4; 2.2-39.3; 2.3-39.2; 2.4-39.1; 2.5-39; 2.6-38.9; 2.7-38.8; 2.8-38.7; 2.9-38.6; 3-38.5; 3.1-38.4; 3.2-38.3; 3.3-38.2; 3.4-38.1; 3.5-38; 3.6-37.9; 3.7-37.8; 3.8-37.7; 3.9-37.6; 4-37.5; 4.1-37.4; 4.2-37.3; 4.3-37.2; 4.4-37.1; 4.5-37; 4.6-36.9; 4.7-36.8; 4.8-36.7; 4.9-36.6; 5-36.5; 5.1-36.4; 5.2-36.3; 5.3-36.2; 5.4-36.1; 5.5-36; 5.6-35.9; 5.7-35.8; 5.8-35.7; 5.9-35.6; 6-35.5; 6.1-35.4; 6.2-35.3; 6.3-35.2; 6.4-35.1; 6.5-35; 6.6-34.9; 6.7-34.8; 6.8-34.7; 6.9-34.6; 7-34.5; 7.1-34.4; 7.2-34.3; 7.3-34.2; 7.4-34.1; 7.5-34; 7.6-33.9; 7.7-33.8; 7.8-33.7; 7.9-33.6; 8-33.5; 8.1-33.4; 8.2-33.3; 8.3-33.2; 8.4-33.1; 8.5-33; 8.6-32.9; 8.7-32.8; 8.8-32.7; 8.9-32.6; 9-32.5; 9.1-32.4; 9.2-32.3; 9.3-32.2; 9.4-32.1; 9.5-32; 9.6-31.9; 9.7-31.8; 9.8-31.7; 9.9-31.6; 10-31.5; 10.1-31.4; 10.2-31.3; 10.3-31.2; 10.4-31.1; 10.5-31; 10.6-30.9; 10.7-30.8; 10.8-30.7; 10.9-30.6; 11-30.5; 11.1-30.4; 11.2-30.3; 11.3-30.2; 11.4-30.1; 11.5-30; 11.6-29.9; 11.7-29.8; 11.8-29.7; 11.9-29.6; 12-29.5; 12.1-29.4; 12.2-29.3; 12.3-29.2; 12.4 29.1; 12.5-29; 12.6-28.9; 12.7-28.8; 12.8-28.7; 12.9-28.6; 13-28.5; 13.1-28.4; 13.2-28.3; 13.3-28.2; 13.4-28.1; 13.5-28; 13.6-27.9; 13.7-27.8; 13.8-27.7; 13.9-27.6; 14-27.5; 14.1-27.4; 14.2-27.3; 14.3-27.2; 14.4-27.1; 14.5-27; 14.6-26.9; 14.7-26.8; 14.8-26.7; 14.9-26.6; 15-26.5; 15.1-26.4; 15.2-26.3; 15.3-26.2; 15.4-26.1; 15.5-26; 15.6-25.9; 15.7-25.8; 15.8-25.7; 15.9-25.6; 16-25.5; 16.1-25.4; 16.2-25.3; 16.3-25.2; 16.4-25.1; 16.5-25; 16.6-24.9; 16.7-24.8; 16.8-24.7; 16.9-24.6; 17-24.5; 17.1-24.4; 17.2-24.3; 17.3-24.2; 17.4-24.1; 17.5 24; 17.6-23.9; 17.7-23.8; 17.8-23.7; 17.9-23.6; 18-23.5; 18.1-23.4; 18.2-23.3; 18.3-23.2; 18.4-23.1; 18.5-23; 18.6-22.9; 18.7-22.8; 18.8-22.7; 18.9-22.6; 19-22.5; 19.1-22.4; 19.2-22.3; 19.3-22.2; 19.4-22.1; 19.5-22; 19.6-21.9; 19.7-21.8; 19.8-21.7; 19.9-21.6; 20-21.5; 20.1-21.4; 20.2-21.3; 20.3-21.2; 20.4-21.1; 20.5-21; 20.6-20.9; 20.7-20.8).
- Embodiment 60. The formulation according to any one of embodiments 57-59, wherein each of the amino acids is present at a concentration:
  - of 0.1 mM or greater (e.g., 0.3, 0.5, 0.7; 0.9; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5);
  - of 10 mM or less (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0, 6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02); or
  - ranging from 0.1 mM to 10 mM (e.g., 0.2 mM to 9.9 mM; 0.3 mM to 9.8 mM; 0.4 mM to 9.7 mM; 0.5 mM to 9.6 mM; 0.6 mM to 9.5 mM; 0.7 mM to 9.4 mM; 0.8 mM to 9.3 mM; 0.9 mM to 9.2 mM; 1 mM to 9.1 mM; 1.1 mM to 9 mM; 1.2 mM to 8.9 mM; 1.3 mM to 8.8 mM; 1.4 mM to 8.7 mM; 1.5 mM to 8.6 mM; 1.6 mM to 8.5 mM; 1.7 mM to 8.4 mM; 1.8 mM to 8.3 mM; 1.9 mM to 8.2 mM; 2 mM to 8.1 mM; 2.1 mM to 8 mM; 2.2 mM to 7.9 mM; 2.3 mM to 7.8 mM; 2.4 mM to 7.7 mM; 2.5 mM to 7.6 mM; 2.6 mM to 7.5 mM; 2.7 mM to 7.4 mM; 2.8 mM to 7.3 mM; 2.9 mM to 7.2 mM; 3 mM to 7.1 mM; 3.1 mM to 7 mM; 3.2 mM to 6.9 mM; 3.3 mM to 6.8 mM; 3.4 mM to 6.7 mM; 3.5 mM to 6.6 mM; 3.6 mM to 6.5 mM; 3.7 mM to 6.4 mM; 3.8 mM to 6.3 mM; 3.9 mM to 6.2 mM; 4 mM to 6.1 mM; 4.1 mM to 6 mM; 4.2 mM to 5.9 mM; 4.3 mM to 5.8 mM; 4.4 mM to 5.7 mM; 4.5 mM to 5.6 mM; 4.6 mM to 5.5 mM; 4.7 mM to 5.4 mM; 4.8 mM to 5.3 mM; 4.9 mM to 5.2 mM; 5 mM to 5.1 mM),
- Embodiment 61. The formulation according to one of embodiments 57-59, wherein each of the amino acids is present at a weight percent to the formulation (wt/wt %) of:
  - 0.001 wt % or greater (e.g., 0.002; 0.003; 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9; 1);
  - 1 wt % or less (e.g., 0.95; 0.85; 0.75; 0.65; 0.55; 0.45; 0.35; 0.25; 0.15; 0.095; 0.085; 0.075; 0.065; 0.055; 0.045; 0.035; 0.025; 0.015; 0.0095; 0.0085; 0.0075; 0.0065; 0.0055; 0.0045; 0.0035; 0.0025; 0.0015); or
  - 0.001 wt % to 1 wt % (e.g., 0.0012 wt % to 0.99 wt %; 0.0014 wt % to 0.97 wt %; 0.0016 wt % to 0.95 wt %; 0.0018 wt % to 0.93 wt %; 0.002 wt % to 0.91 wt %; 0.0022 wt % to 0.89 wt %; 0.0024 wt % to 0.87 wt %; 0.0026 wt % to 0.85 wt %; 0.0028 wt % to 0.83 wt %; 0.003 wt % to 0.81 wt %; 0.0032 wt % to 0.79 wt %; 0.0034 wt % to 0.77 wt %; 0.0036 wt % to 0.75 wt %; 0.0038 wt % to 0.73 wt %; 0.004 wt % to 0.71 wt %; 0.0042 wt % to 0.69 wt %; 0.0044 wt % to 0.67 wt %; 0.0046 wt % to 0.65 wt %; 0.0048 wt % to 0.63 wt %; 0.005 wt % to 0.61 wt %; 0.0052 wt % to 0.59 wt %; 0.0054 wt % to 0.57 wt %; 0.0056 wt % to 0.55 wt %; 0.0058 wt % to 0.53 wt %; 0.006 wt % to 0.51 wt %; 0.0062 wt % to 0.49 wt %; 0.0064 wt % to 0.47 wt %; 0.0066 wt % to 0.45 wt %; 0.0068 wt % to 0.43 wt %; 0.007 wt % to 0.41 wt %; 0.0072 wt % to 0.39 wt %; 0.0074 wt % to 0.37 wt %; 0.0076 wt % to 0.35 wt %; 0.0078 wt % to 0.33 wt %; 0.008 wt % to 0.31 wt %; 0.0082 wt % to 0.29 wt %; 0.0084 wt % to 0.27 wt %; 0.0086 wt % to 0.25 wt %; 0.0088 wt % to 0.23 wt %; 0.009 wt % to 0.21 wt %; 0.0092 wt % to 0.19 wt %; 0.0094 wt % to 0.17 wt %; 0.0096 wt % to 0.15 wt %; 0.0098 wt % to 0.13 wt %; 0.01 wt % to 0.11 wt %; 0.012 wt % to 0.099 wt %; 0.014 wt % to 0.097 wt %; 0.016 wt % to 0.095 wt %; 0.018 wt % to 0.093 wt %; 0.02 wt % to 0.091 wt %; 0.022 wt % to 0.089 wt %; 0.024 wt % to 0.087 wt %; 0.026 wt % to 0.085 wt %; 0.028 wt % to 0.083 wt %; 0.03 wt % to 0.081 wt %; 0.032 wt % to 0.079 wt %; 0.034 wt % to 0.077 wt %; 0.036 wt % to 0.075 wt %; 0.038 wt % to 0.073 wt %; 0.04 wt % to 0.071 wt %; 0.042 wt % to 0.069 wt %; 0.044 wt % to 0.067 wt %; 0.046 wt % to 0.065 wt %; 0.048 wt % to 0.063 wt %; 0.05 wt % to 0.061 wt %; 0.052 wt % to 0.059 wt %; 0.054 wt % to 0.057 wt %).
- Embodiment 62. The formulation according to any one of embodiments 57-59, wherein alanine is present at a weight percent to the formulation (wt/wt %) of:
  - 0.0035 wt % or greater (e.g., 0.00352; 0.00354; 0.00356; 0.00358; 0.0036; 0.00362; 0.00364; 0.00366; 0.00368; 0.0037; 0.00372; 0.00374; 0.00376; 0.00378; 0.0038; 0.00382; 0.00384; 0.00386; 0.00388; 0.004; 0.0042; 0.0044; 0.0046; 0.0048; 0.005; 0.0052; 0.0054; 0.0056; 0.0058; 0.006; 0.0062; 0.0064; 0.0066; 0.0068; 0.007; 0.0072; 0.0074; 0.0076; 0.0078; 0.008; 0.0082; 0.0084; 0.0086; 0.0088; 0.009; 0.0092; 0.0094; 0.0096; 0.0098; 0.01; 0.012; 0.014; 0.016; 0.018; 0.02; 0.022; 0.024; 0.026; 0.028; 0.03; 0.032; 0.034; 0.036; 0.038; 0.04; 0.042; 0.044; 0.046; 0.048; 0.05; 0.052; 0.054; 0.056; 0.058; 0.06; 0.062; 0.064; 0.066; 0.068; 0.07; 0.072; 0.074; 0.076; 0.078; 0.08; 0.082; 0.084; 0.086; 0.088; 0.09; 0.092; 0.094; 0.096; 0.098; 0.1; 0.102; 0.104; 0.106; 0.108; 0.11; 0.112; 0.114; 0.116; 0.118; 0.12; 0.122; 0.124; 0.126; 0.128; 0.13; 0.132; 0.134; 0.136; 0.138; 0.14; 0.142; 0.144; 0.146; 0.148; 0.15; 0.152; 0.154; 0.156; 0.158; 0.16; 0.162; 0.164; 0.166; 0.168; 0.17; 0.172; 0.174; 0.176; 0.178; 0.18; 0.182; 0.184; 0.186; 0.188; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.202; 0.204; 0.206; 0.208; 0.21; 0.212; 0.214; 0.216; 0.218; 0.22; 0.222; 0.224; 0.226; 0.228; 0.23; 0.232; 0.234; 0.236; 0.238; 0.24; 0.242; 0.244; 0.246; 0.248; 0.25; 0.252; 0.254; 0.256; 0.258; 0.26; 0.262; 0.264; 0.266; 0.268; 0.27; 0.272; 0.274; 0.276; 0.278; 0.28; 0.282; 0.284; 0.286; 0.288; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.202; 0.204; 0.206; 0.208; 0.21; 0.212; 0.214; 0.216; 0.218; 0.22; 0.222; 0.224; 0.226; 0.228; 0.23; 0.232; 0.234; 0.236; 0.238; 0.24; 0.242; 0.244; 0.246; 0.248; 0.25; 0.252; 0.254; 0.256; 0.258; 0.26; 0.262; 0.264; 0.266; 0.268; 0.27; 0.272; 0.274; 0.276; 0.278; 0.28; 0.282; 0.284; 0.286; 0.288; 0.3; 0.302; 0.304; 0.306; 0.308; 0.31; 0.312; 0.314; 0.316; 0.318; 0.32; 0.322; 0.324; 0.326; 0.328; 0.33; 0.332; 0.334; 0.336; 0.338; 0.34; 0.342; 0.344; 0.346; 0.348; 0.35; 0.352; 0.354; 0.356; 0.358; 0.36; 0.362; 0.364; 0.366; 0.368; 0.37; 0.372; 0.374; 0.376; 0.378; 0.38; 0.382; 0.384; 0.386; 0.388; 0.39; 0.392; 0.394; 0.396; 0.398; 0.4; 0.402; 0.404; 0.406; 0.408; 0.41; 0.412; 0.414; 0.416; 0.418; 0.42; 0.422; 0.424; 0.426; 0.428; 0.43; 0.432; 0.434; 0.436; 0.438; 0.44; 0.442; 0.444; 0.446; 0.448; 0.45; 0.452; 0.454; 0.456; 0.458; 0.46; 0.462; 0.464; 0.466; 0.468; 0.47; 0.472; 0.474; 0.476; 0.478; 0.48; 0.482; 0.484; 0.486; 0.488; 0.5);
  - 0.4 wt % or less (e.g., 0.399; 0.397; 0.395; 0.393; 0.391; 0.389; 0.387; 0.385; 0.383; 0.381; 0.379; 0.377; 0.375; 0.373; 0.371; 0.369; 0.367; 0.365; 0.363; 0.361; 0.359; 0.357; 0.355; 0.353; 0.351; 0.349; 0.347; 0.345; 0.343; 0.341; 0.339; 0.337; 0.335; 0.333; 0.331; 0.329; 0.327; 0.325; 0.323; 0.321; 0.319; 0.317; 0.315; 0.313; 0.311; 0.309; 0.307; 0.305; 0.303; 0.301; 0.299; 0.297; 0.295; 0.293; 0.291; 0.289; 0.287; 0.285; 0.283; 0.281; 0.279; 0.277; 0.275; 0.273; 0.271; 0.269; 0.267; 0.265; 0.263; 0.261; 0.259; 0.257; 0.255; 0.253; 0.251; 0.249; 0.247; 0.245; 0.243; 0.241; 0.239; 0.237; 0.235; 0.233; 0.231; 0.229; 0.227; 0.225; 0.223; 0.221; 0.219; 0.217; 0.215; 0.213; 0.211; 0.209; 0.207; 0.205; 0.203; 0.201; 0.199; 0.197; 0.195; 0.193; 0.191; 0.189; 0.187; 0.185; 0.183; 0.181; 0.179; 0.177; 0.175; 0.173; 0.171; 0.169; 0.167; 0.165; 0.163; 0.161; 0.159; 0.157; 0.155; 0.153; 0.151; 0.149; 0.147; 0.145; 0.143; 0.141; 0.139; 0.137; 0.135; 0.133; 0.131; 0.129; 0.127; 0.125; 0.123; 0.121; 0.119; 0.117; 0.115; 0.113; 0.111; 0.109; 0.107; 0.105; 0.103; 0.101; 0.099; 0.097; 0.095; 0.093; 0.091; 0.089; 0.087; 0.085; 0.083; 0.081; 0.079; 0.077; 0.075; 0.073; 0.071; 0.069; 0.067; 0.065; 0.063; 0.061; 0.059; 0.057; 0.055; 0.053; 0.051; 0.049; 0.047; 0.045; 0.043; 0.041; 0.039; 0.037; 0.035; 0.033; 0.031; 0.029; 0.027; 0.025; 0.023; 0.021; 0.019; 0.017; 0.015; 0.013; 0.011; 0.009; 0.007; 0.005; 0.003; 0.002); or
  - 0.0035 wt % to 0.36 wt % (e.g., 0.00351 wt % to 0.359 wt %; 0.00352 wt % to 0.358 wt %; 0.00353 wt % to 0.357 wt %; 0.00354 wt % to 0.356 wt %; 0.00355 wt % to 0.355 wt %; 0.00356 wt % to 0.354 wt %; 0.00357 wt % to 0.353 wt %; 0.00358 wt % to 0.352 wt %; 0.00359 wt % to 0.351 wt %; 0.0036 wt % to 0.35 wt %; 0.00361 wt % to 0.349 wt %; 0.00362 wt % to 0.348 wt %; 0.00363 wt % to 0.347 wt %; 0.00364 wt % to 0.346 wt %; 0.00365 wt % to 0.345 wt %; 0.00366 wt % to 0.344 wt %; 0.00367 wt % to 0.343 wt %; 0.00368 wt % to 0.342 wt %; 0.00369 wt % to 0.341 wt %; 0.0037 wt % to 0.34 wt %; 0.00371 wt % to 0.339 wt %; 0.00372 wt % to 0.338 wt %; 0.00373 wt % to 0.337 wt %; 0.00374 wt % to 0.336 wt %; 0.00375 wt % to 0.335 wt %; 0.00376 wt % to 0.334 wt %; 0.00377 wt % to 0.333 wt %; 0.00378 wt % to 0.332 wt %; 0.00379 wt % to 0.331 wt %; 0.0038 wt % to 0.33 wt %; 0.00381 wt % to 0.329 wt %; 0.00382 wt % to 0.328 wt %; 0.00383 wt % to 0.327 wt %; 0.00384 wt % to 0.326 wt %; 0.00385 wt % to 0.325 wt %; 0.00386 wt % to 0.324 wt %; 0.00387 wt % to 0.323 wt %; 0.00388 wt % to 0.322 wt %; 0.00389 wt % to 0.321 wt %; 0.0039 wt % to 0.32 wt %; 0.00391 wt % to 0.319 wt %; 0.00392 wt % to 0.318 wt %; 0.00393 wt % to 0.317 wt %; 0.00394 wt % to 0.316 wt %; 0.00395 wt % to 0.315 wt %; 0.00396 wt % to 0.314 wt %; 0.00397 wt % to 0.313 wt %; 0.00398 wt % to 0.312 wt %; 0.00399 wt % to 0.311 wt %; 0.004 wt % to 0.31 wt %; 0.00401 wt % to 0.309 wt %; 0.00402 wt % to 0.308 wt %; 0.00403 wt % to 0.307 wt %; 0.00404 wt % to 0.306 wt %; 0.00405 wt % to 0.305 wt %; 0.00406 wt % to 0.304 wt %; 0.00407 wt % to 0.303 wt %; 0.00408 wt % to 0.302 wt %; 0.00409 wt % to 0.301 wt %; 0.0041 wt % to 0.3 wt %; 0.00411 wt % to 0.299 wt %; 0.00412 wt % to 0.298 wt %; 0.00413 wt % to 0.297 wt %; 0.00414 wt % to 0.296 wt %; 0.00415 wt % to 0.295 wt %; 0.00416 wt % to 0.294 wt %; 0.00417 wt % to 0.293 wt %; 0.00418 wt % to 0.292 wt %; 0.00419 wt % to 0.291 wt %; 0.0042 wt % to 0.29 wt %; 0.00421 wt % to 0.289 wt %; 0.00422 wt % to 0.288 wt %; 0.00423 wt % to 0.287 wt %; 0.00424 wt % to 0.286 wt %; 0.00425 wt % to 0.285 wt %; 0.00426 wt % to 0.284 wt %; 0.00427 wt % to 0.283 wt %; 0.00428 wt % to 0.282 wt %; 0.00429 wt % to 0.281 wt %; 0.0043 wt % to 0.28 wt %; 0.00431 wt % to 0.279 wt %; 0.00432 wt % to 0.278 wt %; 0.00433 wt % to 0.277 wt %; 0.00434 wt % to 0.276 wt %; 0.00435 wt % to 0.275 wt %; 0.00436 wt % to 0.274 wt %; 0.00437 wt % to 0.273 wt %; 0.00438 wt % to 0.272 wt %; 0.00439 wt % to 0.271 wt %; 0.0044 wt % to 0.27 wt %; 0.00441 wt % to 0.269 wt %; 0.00442 wt % to 0.268 wt %; 0.00443 wt % to 0.267 wt %; 0.00444 wt % to 0.266 wt %; 0.00445 wt % to 0.265 wt %; 0.00446 wt % to 0.264 wt %; 0.00447 wt % to 0.263 wt %; 0.00448 wt % to 0.262 wt %; 0.00449 wt % to 0.261 wt %; 0.0045 wt % to 0.26 wt %; 0.00451 wt % to 0.259 wt %; 0.00452 wt % to 0.258 wt %; 0.00453 wt % to 0.257 wt %; 0.00454 wt % to 0.256 wt %; 0.00455 wt % to 0.255 wt %; 0.00456 wt % to 0.254 wt %; 0.00457 wt % to 0.253 wt %; 0.00458 wt % to 0.252 wt %; 0.00459 wt % to 0.251 wt %; 0.0046 wt % to 0.25 wt %; 0.00461 wt % to 0.249 wt %; 0.00462 wt % to 0.248 wt %; 0.00463 wt % to 0.247 wt %; 0.00464 wt % to 0.246 wt %; 0.00465 wt % to 0.245 wt %; 0.00466 wt % to 0.244 wt %; 0.00467 wt % to 0.243 wt %; 0.00468 wt % to 0.242 wt %; 0.00469 wt % to 0.241 wt %; 0.0047 wt % to 0.24 wt %; 0.00471 wt % to 0.239 wt %; 0.00472 wt % to 0.238 wt %; 0.00473 wt % to 0.237 wt %; 0.00474 wt % to 0.236 wt %; 0.00475 wt % to 0.235 wt %; 0.00476 wt % to 0.234 wt %; 0.00477 wt % to 0.233 wt %; 0.00478 wt % to 0.232 wt %; 0.00479 wt % to 0.231 wt %; 0.0048 wt % to 0.23 wt %; 0.00481 wt % to 0.229 wt %; 0.00482 wt % to 0.228 wt %; 0.00483 wt % to 0.227 wt %; 0.00484 wt % to 0.226 wt %; 0.00485 wt % to 0.225 wt %; 0.00486 wt % to 0.224 wt %; 0.00487 wt % to 0.223 wt %; 0.00488 wt % to 0.222 wt %; 0.00489 wt % to 0.221 wt %; 0.0049 wt % to 0.22 wt %; 0.00491 wt % to 0.219 wt %; 0.00492 wt % to 0.218 wt %; 0.00493 wt % to 0.217 wt %; 0.00494 wt % to 0.216 wt %; 0.00495 wt % to 0.215 wt %; 0.00496 wt % to 0.214 wt %; 0.00497 wt % to 0.213 wt %; 0.00498 wt % to 0.212 wt %; 0.00499 wt % to 0.211 wt %; 0.005 wt % to 0.21 wt %; 0.00501 wt % to 0.209 wt %; 0.00502 wt % to 0.208 wt %; 0.00503 wt % to 0.207 wt %; 0.00504 wt % to 0.206 wt %; 0.00505 wt % to 0.205 wt %; 0.00506 wt % to 0.204 wt %; 0.00507 wt % to 0.203 wt %; 0.00508 wt % to 0.202 wt %; 0.00509 wt % to 0.201 wt %; 0.0051 wt % to 0.2 wt %; 0.00511 wt % to 0.199 wt %; 0.00512 wt % to 0.198 wt %; 0.00513 wt % to 0.197 wt %; 0.00514 wt % to 0.196 wt %; 0.00515 wt % to 0.195 wt %; 0.00516 wt % to 0.194 wt %; 0.00517 wt % to 0.193 wt %; 0.00518 wt % to 0.192 wt %; 0.00519 wt % to 0.191 wt %; 0.0052 wt % to 0.19 wt %; 0.00521 wt % to 0.189 wt %; 0.00522 wt % to 0.188 wt %; 0.00523 wt % to 0.187 wt %; 0.00524 wt % to 0.186 wt %; 0.00525 wt % to 0.185 wt %; 0.00526 wt % to 0.184 wt %; 0.00527 wt % to 0.183 wt %; 0.00528 wt % to 0.182 wt %; 0.00529 wt % to 0.181 wt %; 0.0053 wt % to 0.18 wt %; 0.00531 wt % to 0.179 wt %; 0.00532 wt % to 0.178 wt %; 0.00533 wt % to 0.177 wt %; 0.00534 wt % to 0.176 wt %; 0.00535 wt % to 0.175 wt %; 0.00536 wt % to 0.174 wt %; 0.00537 wt % to 0.173 wt %; 0.00538 wt % to 0.172 wt %; 0.00539 wt % to 0.171 wt %; 0.0054 wt % to 0.17 wt %; 0.00541 wt % to 0.169 wt %; 0.00542 wt % to 0.168 wt %; 0.00543 wt % to 0.167 wt %; 0.00544 wt % to 0.166 wt %; 0.00545 wt % to 0.165 wt %; 0.00546 wt % to 0.164 wt %; 0.00547 wt % to 0.163 wt %; 0.00548 wt % to 0.162 wt %; 0.00549 wt % to 0.161 wt %; 0.0055 wt % to 0.16 wt %; 0.00551 wt % to 0.159 wt %; 0.00552 wt % to 0.158 wt %; 0.00553 wt % to 0.157 wt %; 0.00554 wt % to 0.156 wt %; 0.00555 wt % to 0.155 wt %; 0.00556 wt % to 0.154 wt %; 0.00557 wt % to 0.153 wt %; 0.00558 wt % to 0.152 wt %; 0.00559 wt % to 0.151 wt %; 0.0056 wt % to 0.15 wt %; 0.00561 wt % to 0.149 wt %; 0.00562 wt % to 0.148 wt %; 0.00563 wt % to 0.147 wt %; 0.00564 wt % to 0.146 wt %; 0.00565 wt % to 0.145 wt %; 0.00566 wt % to 0.144 wt %; 0.00567 wt % to 0.143 wt %; 0.00568 wt % to 0.142 wt %; 0.00569 wt % to 0.141 wt %; 0.0057 wt % to 0.14 wt %; 0.00571 wt % to 0.139 wt %; 0.00572 wt % to 0.138 wt %; 0.00573 wt % to 0.137 wt %; 0.00574 wt % to 0.136 wt %; 0.00575 wt % to 0.135 wt %; 0.00576 wt % to 0.134 wt %; 0.00577 wt % to 0.133 wt %; 0.00578 wt % to 0.132 wt %; 0.00579 wt % to 0.131 wt %; 0.0058 wt % to 0.13 wt %; 0.00581 wt % to 0.129 wt %; 0.00582 wt % to 0.128 wt %; 0.00583 wt % to 0.127 wt %; 0.00584 wt % to 0.126 wt %; 0.00585 wt % to 0.125 wt %; 0.00586 wt % to 0.124 wt %; 0.00587 wt % to 0.123 wt %; 0.00588 wt % to 0.122 wt %; 0.00589 wt % to 0.121 wt %; 0.0059 wt % to 0.12 wt %; 0.00591 wt % to 0.119 wt %; 0.00592 wt % to 0.118 wt %; 0.00593 wt % to 0.117 wt %; 0.00594 wt % to 0.116 wt %; 0.00595 wt % to 0.115 wt %; 0.00596 wt % to 0.114 wt %; 0.00597 wt % to 0.113 wt %; 0.00598 wt % to 0.112 wt %; 0.00599 wt % to 0.111 wt %; 0.0056 wt % to 0.11 wt %; 0.00561 wt % to 0.109 wt %; 0.00562 wt % to 0.108 wt %; 0.00563 wt % to 0.107 wt %; 0.00564 wt % to 0.106 wt %; 0.00565 wt % to 0.105 wt %; 0.00566 wt % to 0.104 wt %; 0.00567 wt % to 0.103 wt %; 0.00568 wt % to 0.102 wt %; 0.00569 wt % to 0.101 wt %; 0.0057 wt % to 0.1 wt %; 0.00571 wt % to 0.099 wt %; 0.00572 wt % to 0.098 wt %; 0.00573 wt % to 0.097 wt %; 0.00574 wt % to 0.096 wt %; 0.00575 wt % to 0.095 wt %; 0.00576 wt % to 0.094 wt %; 0.00577 wt % to 0.093 wt %; 0.00578 wt % to 0.092 wt %; 0.00579 wt % to 0.091 wt %; 0.0058 wt % to 0.09 wt %; 0.00581 wt % to 0.089 wt %; 0.00582 wt % to 0.088 wt %; 0.00583 wt % to 0.087 wt %; 0.00584 wt % to 0.086 wt %; 0.00585 wt % to 0.085 wt %; 0.00586 wt % to 0.084 wt %; 0.00587 wt % to 0.083 wt %; 0.00588 wt % to 0.082 wt %; 0.00589 wt % to 0.081 wt %; 0.0059 wt % to 0.08 wt %; 0.00591 wt % to 0.079 wt %; 0.00592 wt % to 0.078 wt %; 0.00593 wt % to 0.077 wt %; 0.00594 wt % to 0.076 wt %; 0.00595 wt % to 0.075 wt %; 0.00596 wt % to 0.074 wt %; 0.00597 wt % to 0.073 wt %; 0.00598 wt % to 0.072 wt %; 0.00599 wt % to 0.071 wt %; 0.006 wt % to 0.07 wt %; 0.00601 wt % to 0.069 wt %; 0.00602 wt % to 0.068 wt %; 0.00603 wt % to 0.067 wt %; 0.00604 wt % to 0.066 wt %; 0.00605 wt % to 0.065 wt %; 0.00606 wt % to 0.064 wt %; 0.00607 wt % to 0.063 wt %; 0.00608 wt % to 0.062 wt %; 0.00609 wt % to 0.061 wt %; 0.0061 wt % to 0.06 wt %; 0.00611 wt % to 0.059 wt %; 0.00612 wt % to 0.058 wt %; 0.00613 wt % to 0.057 wt %; 0.00614 wt % to 0.056 wt %; 0.00615 wt % to 0.055 wt %; 0.00616 wt % to 0.054 wt %; 0.00617 wt % to 0.053 wt %; 0.00618 wt % to 0.052 wt %; 0.00619 wt % to 0.051 wt %; 0.0062 wt % to 0.05 wt %; 0.00621 wt % to 0.049 wt %; 0.00622 wt % to 0.048 wt %; 0.00623 wt % to 0.047 wt %; 0.00624 wt % to 0.046 wt %; 0.00625 wt % to 0.045 wt %; 0.00626 wt % to 0.044 wt %; 0.00627 wt % to 0.043 wt %; 0.00628 wt % to 0.042 wt %; 0.00629 wt % to 0.041 wt %; 0.0063 wt % to 0.04 wt %; 0.00631 wt % to 0.0399 wt %; 0.00632 wt % to 0.0398 wt %; 0.00633 wt % to 0.0397 wt %; 0.00534 wt % to 0.0396 wt %; 0.00535 wt % to 0.0395 wt %; 0.00536 wt % to 0.0394 wt %; 0.00537 wt % to 0.0393 wt %; 0.00538 wt % to 0.0392 wt %; 0.00539 wt % to 0.0391 wt %; 0.0054 wt % to 0.039 wt %; 0.00541 wt % to 0.0389 wt %; 0.00542 wt % to 0.0388 wt %; 0.00543 wt % to 0.0387 wt %; 0.00544 wt % to 0.0386 wt %; 0.00545 wt % to 0.0385 wt %; 0.00546 wt % to 0.0384 wt %; 0.00547 wt % to 0.0383 wt %; 0.00548 wt % to 0.0382 wt %; 0.00549 wt % to 0.0381 wt %; 0.0055 wt % to 0.038 wt %; 0.00551 wt % to 0.0379 wt %; 0.00552 wt % to 0.0378 wt %; 0.00553 wt % to 0.0377 wt %; 0.00554 wt % to 0.0376 wt %; 0.00555 wt % to 0.0375 wt %; 0.00556 wt % to 0.0374 wt %; 0.00557 wt % to 0.0373 wt %; 0.00558 wt % to 0.0372 wt %; 0.00559 wt % to 0.0371 wt %; 0.0056 wt % to 0.037 wt %; 0.00561 wt % to 0.0369 wt %; 0.00562 wt % to 0.0368 wt %; 0.00563 wt % to 0.0367 wt %; 0.00564 wt % to 0.0366 wt %; 0.00565 wt % to 0.0365 wt %; 0.00566 wt % to 0.0364 wt %; 0.00567 wt % to 0.0363 wt %; 0.00568 wt % to 0.0362 wt %; 0.00569 wt % to 0.0361 wt %; 0.0057 wt % to 0.036 wt %; 0.00571 wt % to 0.0359 wt %; 0.00572 wt % to 0.0358 wt %; 0.00573 wt % to 0.0357 wt %; 0.00574 wt % to 0.0356 wt %; 0.00575 wt % to 0.0355 wt %; 0.00576 wt % to 0.0354 wt %; 0.00577 wt % to 0.0353 wt %; 0.00578 wt % to 0.0352 wt %; 0.00579 wt % to 0.0351 wt %; 0.0058 wt % to 0.035 wt %; 0.00581 wt % to 0.0349 wt %; 0.00582 wt % to 0.0348 wt %; 0.00583 wt % to 0.0347 wt %; 0.00584 wt % to 0.0346 wt %; 0.00585 wt % to 0.0345 wt %; 0.00586 wt % to 0.0344 wt %; 0.00587 wt % to 0.0343 wt %; 0.00588 wt % to 0.0342 wt %; 0.00589 wt % to 0.0341 wt %; 0.0059 wt % to 0.034 wt %; 0.00591 wt % to 0.0339 wt %; 0.00592 wt % to 0.0338 wt %; 0.00593 wt % to 0.0337 wt %; 0.00594 wt % to 0.0336 wt %; 0.00595 wt % to 0.0335 wt %; 0.00596 wt % to 0.0334 wt %; 0.00597 wt % to 0.0333 wt %; 0.00598 wt % to 0.0332 wt %; 0.00599 wt % to 0.0331 wt %; 0.0056 wt % to 0.033 wt %; 0.00561 wt % to 0.0329 wt %; 0.00562 wt % to 0.0328 wt %; 0.00563 wt % to 0.0327 wt %; 0.00564 wt % to 0.0326 wt %; 0.00565 wt % to 0.0325 wt %; 0.00566 wt % to 0.0324 wt %; 0.00567 wt % to 0.0323 wt %; 0.00568 wt % to 0.0322 wt %; 0.00569 wt % to 0.0321 wt %; 0.0057 wt % to 0.032 wt %; 0.00571 wt % to 0.0319 wt %; 0.00572 wt % to 0.0318 wt %; 0.00573 wt % to 0.0317 wt %; 0.00574 wt % to 0.0316 wt %; 0.00575 wt % to 0.0315 wt %; 0.00576 wt % to 0.0314 wt %; 0.00577 wt % to 0.0313 wt %; 0.00578 wt % to 0.0312 wt %; 0.00579 wt % to 0.0311 wt %; 0.0058 wt % to 0.031 wt %; 0.00581 wt % to 0.0309 wt %; 0.00582 wt % to 0.0308 wt %; 0.00583 wt % to 0.0307 wt %; 0.00584 wt % to 0.0306 wt %; 0.00585 wt % to 0.0305 wt %; 0.00586 wt % to 0.0304 wt %; 0.00587 wt % to 0.0303 wt %; 0.00588 wt % to 0.0302 wt %; 0.00589 wt % to 0.0301 wt %; 0.0059 wt % to 0.03 wt %; 0.00591 wt % to 0.0299 wt %; 0.00592 wt % to 0.0298 wt %; 0.00593 wt % to 0.0297 wt %; 0.00594 wt % to 0.0296 wt %; 0.00595 wt % to 0.0295 wt %; 0.00596 wt % to 0.0294 wt %; 0.00597 wt % to 0.0293 wt %; 0.00598 wt % to 0.0292 wt %; 0.00599 wt % to 0.0291 wt %; 0.006 wt % to 0.029 wt %; 0.00601 wt % to 0.0289 wt %; 0.00602 wt % to 0.0288 wt %; 0.00603 wt % to 0.0287 wt %; 0.00604 wt % to 0.0286 wt %; 0.00605 wt % to 0.0285 wt %; 0.00606 wt % to 0.0284 wt %; 0.00607 wt % to 0.0283 wt %; 0.00608 wt % to 0.0282 wt %; 0.00609 wt % to 0.0281 wt %; 0.0061 wt % to 0.028 wt %; 0.00611 wt % to 0.0279 wt %; 0.00612 wt % to 0.0278 wt %; 0.00613 wt % to 0.0277 wt %; 0.00614 wt % to 0.0276 wt %; 0.00615 wt % to 0.0275 wt %; 0.00616 wt % to 0.0274 wt %; 0.00617 wt % to 0.0273 wt %; 0.00618 wt % to 0.0272 wt %; 0.00619 wt % to 0.0271 wt %; 0.0062 wt % to 0.027 wt %; 0.00621 wt % to 0.0269 wt %; 0.00622 wt % to 0.0268 wt %; 0.00623 wt % to 0.0267 wt %; 0.00624 wt % to 0.0266 wt %; 0.00625 wt % to 0.0265 wt %; 0.00626 wt % to 0.0264 wt %; 0.00627 wt % to 0.0263 wt %; 0.00628 wt % to 0.0262 wt %; 0.00629 wt % to 0.0261 wt %; 0.0063 wt % to 0.026 wt %; 0.00631 wt % to 0.0259 wt %).
- Embodiment 63. The formulation according to any one of embodiments 57-59, wherein glutamine is present at a weight percent (wt %) to the formulation of:
  - 0.00584 wt % or greater (e.g., 0.0059; 0.006; 0.007; 0.008; 0.009; 0.01; 0.011; 0.012; 0.013; 0.014; 0.015; 0.016; 0.017; 0.018; 0.019; 0.02; 0.021; 0.022; 0.023; 0.024; 0.025; 0.026; 0.027; 0.028; 0.029; 0.03; 0.031; 0.032; 0.033; 0.034; 0.035; 0.036; 0.037; 0.038; 0.039; 0.04; 0.041; 0.042; 0.043; 0.044; 0.045; 0.046; 0.047; 0.048; 0.049; 0.05; 0.051; 0.052; 0.053; 0.054; 0.055; 0.056; 0.057; 0.058; 0.059; 0.06; 0.061; 0.062; 0.063; 0.064; 0.065; 0.066; 0.067; 0.068; 0.069; 0.07; 0.071; 0.072; 0.073; 0.074; 0.075; 0.076; 0.077; 0.078; 0.079; 0.08; 0.081; 0.082; 0.083; 0.084; 0.085; 0.086; 0.087; 0.088; 0.089; 0.09; 0.091; 0.092; 0.093; 0.094; 0.095; 0.096; 0.097; 0.098; 0.099; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.2; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.3; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.4; 0.41; 0.42; 0.43; 0.44; 0.45; 0.46; 0.47; 0.48; 0.49; 0.5; 0.51; 0.52; 0.53; 0.54; 0.55; 0.56; 0.57; 0.58; 0.59; 0.6);
  - 0.584 wt % or less (e.g., 0.583; 0.582; 0.581; 0.58; 0.579; 0.578; 0.577; 0.576; 0.575; 0.574; 0.573; 0.572; 0.571; 0.57; 0.569; 0.568; 0.567; 0.566; 0.565; 0.564; 0.563; 0.562; 0.561; 0.56; 0.559; 0.558; 0.557; 0.556; 0.554; 0.553; 0.552; 0.551; 0.55; 0.549; 0.548; 0.547; 0.546; 0.545; 0.544; 0.543; 0.542; 0.541; 0.54; 0.539; 0.538; 0.537; 0.536; 0.535; 0.534; 0.533; 0.532; 0.531; 0.53; 0.529; 0.528; 0.527; 0.526; 0.525; 0.524; 0.523; 0.522; 0.521; 0.52; 0.519; 0.518; 0.517; 0.516; 0.515; 0.514; 0.513; 0.512; 0.511; 0.51; 0.509; 0.508; 0.507; 0.506; 0.505; 0.504; 0.503; 0.502; 0.501; 0.5; 0.499; 0.498; 0.497; 0.496; 0.495; 0.494; 0.493; 0.492; 0.491; 0.49; 0.489; 0.488; 0.487; 0.486; 0.485; 0.484; 0.483; 0.482; 0.481; 0.48; 0.479; 0.478; 0.477; 0.476; 0.475; 0.474; 0.473; 0.472; 0.471; 0.47; 0.469; 0.468; 0.467; 0.466; 0.465; 0.464; 0.463; 0.462; 0.461; 0.46; 0.459; 0.458; 0.457; 0.456; 0.455; 0.454; 0.453; 0.452; 0.451; 0.45; 0.449; 0.448; 0.447; 0.446; 0.445; 0.444; 0.443; 0.442; 0.441; 0.44; 0.439; 0.438; 0.437; 0.436; 0.435; 0.434; 0.433; 0.432; 0.431; 0.43; 0.429; 0.428; 0.427; 0.426; 0.425; 0.424; 0.423; 0.422; 0.421; 0.42; 0.419; 0.418; 0.417; 0.416; 0.415; 0.414; 0.413; 0.412; 0.411; 0.41; 0.409; 0.408; 0.407; 0.406; 0.405; 0.404; 0.403; 0.402; 0.401; 0.4; 0.3; 0.2; 0.1; 0.09; 0.08; 0.07; 0.06; 0.05; 0.04; 0.03; 0.02; 0.01; 0.009; 0.008; 0.007; 0.006; 0.005; 0.004; 0.003; 0.002; 0.001); or
  - 0.00584 wt % to 0.584 wt % (e.g., 0.00585 wt % to 0.582 wt %; 0.0059 wt % to 0.58 wt %; 0.0065 wt % to 0.578 wt %; 0.007 wt % to 0.576 wt %; 0.0075 wt % to 0.574 wt %; 0.008 wt % to 0.572 wt %; 0.0085 wt % to 0.57 wt %; 0.009 wt % to 0.568 wt %; 0.0095 wt % to 0.566 wt %; 0.01 wt % to 0.564 wt %; 0.015 wt % to 0.562 wt %; 0.02 wt % to 0.56 wt %; 0.025 wt % to 0.558 wt %; 0.03 wt % to 0.556 wt %; 0.035 wt % to 0.554 wt %; 0.04 wt % to 0.552 wt %; 0.045 wt % to 0.55 wt %; 0.05 wt % to 0.548 wt %; 0.055 wt % to 0.546 wt %; 0.06 wt % to 0.544 wt %; 0.065 wt % to 0.542 wt %; 0.07 wt % to 0.54 wt %; 0.075 wt % to 0.538 wt %; 0.08 wt % to 0.536 wt %; 0.085 wt % to 0.534 wt %; 0.09 wt % to 0.532 wt %; 0.095 wt % to 0.53 wt %; 0.1 wt % to 0.528 wt %; 0.15 wt % to 0.526 wt %; 0.2 wt % to 0.524 wt %; 0.25 wt % to 0.522 wt %; 0.3 wt % to 0.52 wt %; 0.35 wt % to 0.518 wt %; 0.4 wt % to 0.516 wt %; 0.45 wt % to 0.514 wt %; 0.5 wt % to 0.512 wt %).
- Embodiment 64. The formulation according to any one of embodiments 57-59, wherein glycine is present at a weight percent (wt %) to the formulation of:
  - 0.003 wt % or greater (e.g., 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.2; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.3; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.4);
  - 0.3 wt % or less (e.g., 0.298; 0.296; 0.294; 0.292; 0.29; 0.288; 0.286; 0.284; 0.282; 0.28; 0.278; 0.276; 0.274; 0.272; 0.27; 0.268; 0.266; 0.264; 0.262; 0.26; 0.258; 0.256; 0.254; 0.252; 0.25; 0.248; 0.246; 0.244; 0.242; 0.24; 0.238; 0.236; 0.234; 0.232; 0.23; 0.228; 0.226; 0.224; 0.222; 0.22; 0.218; 0.216; 0.214; 0.212; 0.21; 0.208; 0.206; 0.204; 0.202; 0.2; 0.198; 0.196; 0.194; 0.192; 0.19; 0.188; 0.186; 0.184; 0.182; 0.18; 0.178; 0.176; 0.174; 0.172; 0.17; 0.168; 0.166; 0.164; 0.162; 0.16; 0.158; 0.156; 0.154; 0.152; 0.15; 0.148; 0.146; 0.144; 0.142; 0.14; 0.138; 0.136; 0.134; 0.132; 0.13; 0.128; 0.126; 0.124; 0.122; 0.12; 0.118; 0.116; 0.114; 0.112; 0.11; 0.108; 0.106; 0.104; 0.102; 0.1; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.008; 0.006; 0.004; 0.002); or
  - 0.003 wt % to 0.3 wt % (e.g., 0.0035 wt % to 0.29 wt %; 0.004 wt % to 0.27 wt %; 0.0045 wt % to 0.25 wt %; 0.005 wt % to 0.23 wt %; 0.0055 wt % to 0.21 wt %; 0.006 wt % to 0.19 wt %; 0.0065 wt % to 0.17 wt %; 0.007 wt % to 0.15 wt %; 0.0075 wt % to 0.13 wt %; 0.008 wt % to 0.11 wt %; 0.0085 wt % to 0.099 wt %; 0.009 wt % to 0.097 wt %; 0.0095 wt % to 0.095 wt %; 0.01 wt % to 0.093 wt %; 0.015 wt % to 0.091 wt %; 0.02 wt % to 0.089 wt %; 0.025 wt % to 0.087 wt %; 0.03 wt % to 0.085 wt %; 0.035 wt % to 0.083 wt %; 0.04 wt % to 0.081 wt %; 0.045 wt % to 0.079 wt %; 0.05 wt % to 0.077 wt %; 0.055 wt % to 0.075 wt %; 0.06 wt % to 0.073 wt %; 0.065 wt % to 0.071 wt %).
- Embodiment 65. The formulation according to any one of embodiments 57-59, wherein serine is present at a weight percent (wt %) to the formulation of:
  - 0.0042 wt % or greater (e.g., 0.0044; 0.0046; 0.0048; 0.005; 0.0052; 0.0054; 0.0056; 0.0058; 0.006; 0.0062; 0.0064; 0.0066; 0.0068; 0.007; 0.0072; 0.0074; 0.0076; 0.0078; 0.008; 0.0082; 0.0084; 0.0086; 0.0088; 0.009; 0.0092; 0.0094; 0.0096; 0.0098; 0.01; 0.0102; 0.0104; 0.0106; 0.0108; 0.011; 0.0112; 0.0114; 0.0116; 0.0118; 0.012; 0.0122; 0.0124; 0.0126; 0.0128; 0.013; 0.0132; 0.0134; 0.0136; 0.0138; 0.014; 0.0142; 0.0144; 0.0146; 0.0148; 0.015; 0.0152; 0.0154; 0.0156; 0.0158; 0.016; 0.0162; 0.0164; 0.0166; 0.0168; 0.017; 0.0172; 0.0174; 0.0176; 0.0178; 0.018; 0.0182; 0.0184; 0.0186; 0.0188; 0.019; 0.0192; 0.0194; 0.0196; 0.0198; 0.02; 0.0202; 0.0204; 0.0206; 0.208; 0.021; 0.0212; 0.0214; 0.0216; 0.0218; 0.022; 0.0222; 0.0224; 0.0226; 0.0228; 0.023; 0.0232; 0.0234; 0.0236; 0.0238; 0.024; 0.0242; 0.0244; 0.0246; 0.0248; 0.025; 0.0252; 0.0254; 0.0256; 0.0258; 0.026; 0.0262; 0.0264; 0.0266; 0.0268; 0.027; 0.0272; 0.0274; 0.0276; 0.0278; 0.028; 0.0282; 0.0284; 0.0286; 0.0288; 0.029; 0.0292; 0.0294; 0.0296; 0.0298; 0.03; 0.0302; 0.0304; 0.0306; 0.0308; 0.031; 0.0312; 0.0314; 0.0316; 0.0318; 0.032; 0.0322; 0.0324; 0.0326; 0.0328; 0.033; 0.0332; 0.0334; 0.0336; 0.0338; 0.034; 0.0342; 0.0344; 0.0346; 0.0348; 0.035; 0.0352; 0.0354; 0.0356; 0.0358; 0.036; 0.0362; 0.0364; 0.0366; 0.0368; 0.037; 0.0372; 0.0374; 0.0376; 0.0378; 0.038; 0.0382; 0.0384; 0.0386; 0.0388; 0.039; 0.0392; 0.0394; 0.0396; 0.0398; 0.04; 0.0402; 0.0404; 0.0406; 0.0408; 0.041; 0.0412; 0.0414; 0.0416; 0.0418; 0.042; 0.0422; 0.0424; 0.0426; 0.0428; 0.043; 0.0432; 0.0434; 0.0436; 0.0438; 0.044; 0.0442; 0.0444; 0.0446; 0.0448; 0.045; 0.0452; 0.0454; 0.0456; 0.0458; 0.046; 0.0462; 0.0464; 0.0466; 0.0468; 0.047; 0.0472; 0.0474; 0.0476; 0.0478; 0.048; 0.0482; 0.0484; 0.0486; 0.0488; 0.049; 0.0492; 0.0494; 0.0496; 0.0498; 0.05; 0.0502; 0.0504; 0.0506; 0.0508; 0.051; 0.0512; 0.0514; 0.0516; 0.0518; 0.052; 0.0522; 0.0524; 0.0526; 0.0528; 0.053; 0.0532; 0.0534; 0.0536; 0.0538; 0.054; 0.0542; 0.0544; 0.0546; 0.0548; 0.055; 0.0552; 0.0554; 0.0556; 0.0558; 0.056; 0.0562; 0.0564; 0.0566; 0.0568; 0.057; 0.0572; 0.0574; 0.0576; 0.0578; 0.058; 0.0582; 0.0584; 0.0586; 0.0588; 0.059; 0.0592; 0.0594; 0.0596; 0.0598; 0.06; 0.0602; 0.0604; 0.0606; 0.0608; 0.061; 0.0612; 0.0614; 0.0616; 0.0618; 0.062; 0.0622; 0.0624; 0.0626; 0.0628; 0.063; 0.0632; 0.0634; 0.0636; 0.0638; 0.064; 0.0642; 0.0644; 0.0646; 0.0648; 0.065; 0.0652; 0.0654; 0.0656; 0.0658; 0.066; 0.0662; 0.0664; 0.0666; 0.0668; 0.067; 0.0672; 0.0674; 0.0676; 0.0678; 0.068; 0.0682; 0.0684; 0.0686; 0.0688; 0.069; 0.0692; 0.0694; 0.0696; 0.0698; 0.07; 0.0702; 0.0704; 0.0706; 0.0708; 0.071; 0.0712; 0.0714; 0.0716; 0.0718; 0.072; 0.0722; 0.0724; 0.0726; 0.0728; 0.073; 0.0732; 0.0734; 0.0736; 0.0738; 0.074; 0.0742; 0.0744; 0.0746; 0.0748; 0.075; 0.0752; 0.0754; 0.0756; 0.0758; 0.076; 0.0762; 0.0764; 0.0766; 0.0768; 0.077; 0.0772; 0.0774; 0.0776; 0.0778; 0.078; 0.0782; 0.0784; 0.0786; 0.0788; 0.079; 0.0792; 0.0794; 0.0796; 0.0798; 0.08; 0.0802; 0.0804; 0.0806; 0.0808; 0.081; 0.0812; 0.0814; 0.0816; 0.0818; 0.082; 0.0822; 0.0824; 0.0826; 0.0828; 0.083; 0.0832; 0.0834; 0.0836; 0.0838; 0.084; 0.0842; 0.0844; 0.0846; 0.0848; 0.085; 0.0852; 0.0854; 0.0856; 0.0858; 0.086; 0.0862; 0.0864; 0.0866; 0.0868; 0.087; 0.0872; 0.0874; 0.0876; 0.0878; 0.088; 0.0882; 0.0884; 0.0886; 0.0888; 0.089; 0.0892; 0.0894; 0.0896; 0.0898; 0.09; 0.0902; 0.0904; 0.0906; 0.0908; 0.091; 0.0912; 0.0914; 0.0916; 0.0918; 0.092; 0.0922; 0.0924; 0.0926; 0.0928; 0.093; 0.0932; 0.0934; 0.0936; 0.0938; 0.094; 0.0942; 0.0944; 0.0946; 0.0948; 0.095; 0.0952; 0.0954; 0.0956; 0.0958; 0.096; 0.0962; 0.0964; 0.0966; 0.0968; 0.097; 0.0972; 0.0974; 0.0976; 0.0978; 0.098; 0.0982; 0.0984; 0.0986; 0.0988; 0.099; 0.0992; 0.0994; 0.0996; 0.0998; 0.1; 0.102; 0.104; 0.106; 0.108; 0.11; 0.112; 0.114; 0.116; 0.118; 0.12; 0.122; 0.124; 0.126; 0.128; 0.13; 0.132; 0.134; 0.136; 0.138; 0.14; 0.142; 0.144; 0.146; 0.148; 0.15; 0.152; 0.154; 0.156; 0.158; 0.16; 0.162; 0.164; 0.166; 0.168; 0.17; 0.172; 0.174; 0.176; 0.178; 0.18; 0.182; 0.184; 0.186; 0.188; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.22; 0.24; 0.26; 0.28; 0.3; 0.32; 0.34; 0.36; 0.38; 0.4; 0.42; 0.44; 0.46; 0.48; 0.5);
  - 0.42 wt % or less (e.g., 0.418; 0.416; 0.414; 0.412; 0.41; 0.408; 0.406; 0.404; 0.402; 0.4; 0.398; 0.396; 0.394; 0.392; 0.39; 0.388; 0.386; 0.384; 0.382; 0.38; 0.378; 0.376; 0.374; 0.372; 0.37; 0.368; 0.366; 0.364; 0.362; 0.36; 0.358; 0.356; 0.354; 0.352; 0.35; 0.348; 0.346; 0.344; 342; 0.34; 0.338; 0.336; 0.334; 0.332; 0.33; 0.328; 0.326; 0.324; 0.322; 0.32; 0.318; 0.316; 0.314; 0.312; 0.31; 0.308; 0.306; 0.304; 0.302; 0.3; 0.298; 0.296; 0.294; 0.292; 0.294; 0.292; 0.29; 0.288; 0.286; 0.284; 0.282; 0.28; 0.278; 0.276; 0.274; 0.272; 0.27; 0.268; 0.266; 0.264; 0.262; 0.26; 0.258; 0.256; 0.254; 0.252; 0.25; 0.248; 0.246; 0.244; 0.242; 0.24; 0.238; 0.236; 0.234; 0.232; 0.23; 0.228; 0.226; 0.224; 0.222; 0.22; 0.218; 0.216; 0.214; 0.212; 0.21; 0.208; 0.206; 0.204; 0.202; 0.2; 0.198; 0.196; 0.194; 0.192; 0.19; 0.188; 0.186; 0.184; 0.182; 0.18; 0.178; 0.176; 0.174; 0.172; 0.17; 0.168; 0.166; 0.164; 0.162; 0.16; 0.158; 0.156; 0.154; 0.152; 0.15; 0.148; 0.146; 0.144; 0.142; 0.14; 0.138; 0.136; 0.134; 0.132; 0.13; 0.128; 0.126; 0.124; 0.122; 0.12; 0.118; 0.116; 0.114; 0.112; 0.11; 0.108; 0.106; 0.104; 0.102; 0.1; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.0088; 0.0086; 0.0084; 0.0082; 0.008; 0.0078; 0.0076; 0.0074; 0.0072; 0.007; 0.0068; 0.0066; 0.0064; 0.0062; 0.006; 0.0058; 0.0056; 0.0054; 0.0052; 0.005; 0.0048; 0.0046; 0.0044; 0.0042; 0.004); or
  - 0.0042 wt % to 0.42 wt % (e.g., 0.0043 wt % to 0.415 wt %; 0.0045 wt % to 0.41 wt %; 0.0047 wt % to 0.405 wt %; 0.0049 wt % to 0.4 wt %; 0.0051 wt % to 0.395 wt %; 0.0053 wt % to 0.39 wt %; 0.0055 wt % to 0.385 wt %; 0.0057 wt % to 0.38 wt %; 0.0059 wt % to 0.375 wt %; 0.0061 wt % to 0.37 wt %; 0.0063 wt % to 0.365 wt %; 0.0065 wt % to 0.36 wt %; 0.0067 wt % to 0.355 wt %; 0.0069 wt % to 0.35 wt %; 0.0061 wt % to 0.345 wt %; 0.0063 wt % to 0.34 wt %; 0.0065 wt % to 0.335 wt %; 0.0067 wt % to 0.33 wt %; 0.0069 wt % to 0.325 wt %; 0.0071 wt % to 0.32 wt %; 0.0073 wt % to 0.315 wt %; 0.0075 wt % to 0.31 wt %; 0.0077 wt % to 0.305 wt %; 0.0079 wt % to 0.3 wt %; 0.0081 wt % to 0.295 wt %; 0.0083 wt % to 0.29 wt %; 0.0085 wt % to 0.285 wt %; 0.0087 wt % to 0.28 wt %; 0.0089 wt % to 0.275 wt %; 0.0091 wt % to 0.27 wt %; 0.0093 wt % to 0.265 wt %; 0.0095 wt % to 0.26 wt %; 0.0097 wt % to 0.255 wt %; 0.0099 wt % to 0.35 wt %; 0.0101 wt % to 0.345 wt %; 0.0103 wt % to 0.34 wt %; 0.0105 wt % to 0.335 wt %; 0.0107 wt % to 0.33 wt %; 0.0109 wt % to 0.325 wt %; 0.0111 wt % to 0.32 wt %; 0.0113 wt % to 0.315 wt %; 0.0115 wt % to 0.31 wt %; 0.0117 wt % to 0.305 wt %; 0.0119 wt % to 0.3 wt %; 0.0121 wt % to 0.295 wt %; 0.0123 wt % to 0.29 wt %; 0.0125 wt % to 0.285 wt %; 0.0127 wt % to 0.28 wt %; 0.0129 wt % to 0.275 wt %; 0.0131 wt % to 0.27 wt %; 0.0133 wt % to 0.265 wt %; 0.0135 wt % to 0.26 wt %; 0.0137 wt % to 0.255 wt %; 0.0139 wt % to 0.25 wt %; 0.0141 wt % to 0.245 wt %; 0.0143 wt % to 0.24 wt %; 0.0145 wt % to 0.235 wt %; 0.0147 wt % to 0.23 wt %; 0.0149 wt % to 0.225 wt %; 0.0151 wt % to 0.22 wt %; 0.0153 wt % to 0.215 wt %; 0.0155 wt % to 0.21 wt %; 0.0157 wt % to 0.205 wt %; 0.0159 wt % to 0.2 wt %; 0.0161 wt % to 0.195 wt %; 0.0163 wt % to 0.19 wt %; 0.0165 wt % to 0.185 wt %; 0.0167 wt % to 0.18 wt %; 0.0169 wt % to 0.175 wt %; 0.0171 wt % to 0.17 wt %; 0.0173 wt % to 0.165 wt %; 0.0175 wt % to 0.16 wt %; 0.0177 wt % to 0.155 wt %; 0.0179 wt % to 0.15 wt %; 0.0181 wt % to 0.145 wt %; 0.0183 wt % to 0.14 wt %; 0.0185 wt % to 0.135 wt %; 0.0187 wt % to 0.13 wt %; 0.0189 wt % to 0.125 wt %; 0.0191 wt % to 0.12 wt %; 0.0193 wt % to 0.115 wt %; 0.0195 wt % to 0.11 wt %; 0.0197 wt % to 0.105 wt %; 0.0199 wt % to 0.1 wt %; 0.0201 wt % to 0.095 wt %; 0.0203 wt % to 0.09 wt %; 0.0205 wt % to 0.085 wt %; 0.0207 wt % to 0.08 wt %; 0.0209 wt % to 0.075 wt %; 0.0211 wt % to 0.07 wt %; 0.0213 wt % to 0.065 wt %; 0.0215 wt % to 0.06 wt %; 0.0217 wt % to 0.055 wt %; 0.0219 wt % to 0.05 wt %; 0.0221 wt % to 0.045 wt %; 0.0223 wt % to 0.04 wt %; 0.0225 wt % to 0.035 wt %; 0.0227 wt % to 0.03 wt %; 0.0229 wt % to 0.025 wt %).
- Embodiment 66. The formulation according to any one of embodiments 57-59, wherein the therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids comprises (or consists essentially of or consists of):
  - Alanine (Ala),
  - Glutamine (Gln),
  - Glycine (Gly), and
  - Serine (Ser),
  - wherein each amino acid comprises a ratio to each of the other amino acids of:
  - 1:100 [0.1 to 10] or greater (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):1 (Gln):100 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):100 (Gln):1 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):100 (Gln):100 (Gly):1 (Ser) [0.1 to 10]; 1 (Ala):90 (Gln):90 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):1 (Gln):90 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):90 (Gln):1 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):90 (Gln):90 (Gly):1 (Ser) [0.1 to 9]; 1 (Ala):80 (Gln):80 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):1 (Gln):80 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):80 (Gln):1 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):80 (Gln):80 (Gly):1 (Ser) [0.1 to 8]; 1 (Ala):70 (Gln):70 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):1 (Gln):70 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):70 (Gln):1 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):70 (Gln):70 (Gly):1 (Ser) [0.1 to 7]; 1 (Ala):60 (Gln):60 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):1 (Gln):60 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):60 (Gln):1 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):60 (Gln):60 (Gly):1 (Ser) [0.1 to 6]; 1 (Ala):50 (Gln):50 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):1 (Gln):50 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):50 (Gln):1 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):50 (Gln):50 (Gly):1 (Ser) [0.1 to 5]; 1 (Ala):40 (Gln):40 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):1 (Gln):40 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):40 (Gln):1 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):40 (Gln):40 (Gly):1 (Ser) [0.1 to 4]; 1 (Ala):30 (Gln):30 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):1 (Gln):30 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):30 (Gln):1 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):30 (Gln):30 (Gly):1 (Ser) [0.1 to 3]; 1 (Ala):20 (Gln):20 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):1 (Gln):20 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):20 (Gln):1 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):20 (Gln):20 (Gly):1 (Ser) [0.1 to 2]; 1 (Ala):10 (Gln):10 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):1 (Gln):10 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):10 (Gln):1 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):10 (Gln):10 (Gly):1 (Ser) [0.1 to 1]; 1 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):1 (Gln):5 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):5 (Gln):1 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):5 (Gln):5 (Gly):1 (Ser) [0.1 to 0.5]; 2 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):2 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):2 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):5 (Gly):2 (Ser) [0.1 to 0.25]; 1 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.1 to 0.1]; 2 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):2 (Gln):1 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):1 (Gln):2 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):2 (Ser) [0.2 to 0.1]; 3 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):3 (Gln):1 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):1 (Gln):3 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):3 (Ser) [0.3 to 0.1]; 4 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):4 (Gln):1 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):1 (Gln):4 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):4 (Ser) [0.4 to 0.1]; 1 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.5 to 0.1]; 1 (Ala):5 (Gln):1 (Gly):1 (Ser) [0.5 to 0.1]; 1 (Ala):1 (Gln):5 (Gly):1 (Ser) [0.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):5 (Ser) [0.5 to 0.1]; 6 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):6 (Gln):1 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):1 (Gln):6 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):6 (Ser) [0.6 to 0.1]; 7 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):7 (Gln):1 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):1 (Gln):7 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):7 (Ser) [0.7 to 0.1]; 8 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):8 (Gln):1 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):1 (Gln):8 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):8 (Ser) [0.8 to 0.1]; 9 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):9 (Gln):1 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):1 (Gln):9 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):9 (Ser) [0.9 to 0.1]; 10 (Ala):1 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):10 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):10 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):10 (Ser) [1 to 0.1]; 20 (Ala):1 (Gln):1 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):20 (Gln):1 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):1 (Gln):20 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):20 (Ser) [2 to 0.1]; 30 (Ala):1 (Gln):1 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):30 (Gln):1 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):1 (Gln):30 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):30 (Ser) [3 to 0.1]; 40 (Ala):1 (Gln):1 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):40 (Gln):1 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):1 (Gln):40 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):40 (Ser) [4 to 0.1]; 50 (Ala):1 (Gln):1 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):50 (Gln):1 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):1 (Gln):50 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):50 (Ser) [5 to 0.1]; 60 (Ala):1 (Gln):1 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):60 (Gln):1 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):1 (Gln):60 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):60 (Ser) [6 to 0.1]; 70 (Ala):1 (Gln):1 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):70 (Gln):1 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):1 (Gln):70 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):70 (Ser) [7 to 0.1]; 80 (Ala):1 (Gln):1 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):80 (Gln):1 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):1 (Gln):80 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):80 (Ser) [8 to 0.1]; 90 (Ala):1 (Gln):1 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):90 (Gln):1 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):1 (Gln):90 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):90 (Ser) [9 to 0.1]; 100 (Ala):1 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):100 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):1 (Gln):100 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):100 (Ser) [10 to 0.1]);
  - 100:1 [10 to 0.1] or less (e.g., 95 (Ala):1 (Gln):1 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):95 (Gln):1 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):1 (Gln):95 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):95 (Ser) [9.5 to 0.1]; 85 (Ala):1 (Gln):1 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):85 (Gln):1 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):1 (Gln):85 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):85 (Ser) [8.5 to 0.1]; 75 (Ala):1 (Gln):1 (Gly):1 (Ser) [7.5 to 0.1]; 1 (Ala):75 (Gln):1 (Gly):1 (Ser) [7.5 to 0.1]; 1 (Ala):1 (Gln):75 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):55 (Ser) [5.5 to 0.1]; 65 (Ala):1 (Gln):1 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):65 (Gln):1 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):1 (Gln):65 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):65 (Ser) [6.5 to 0.1]; 55 (Ala):1 (Gln):1 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):55 (Gln):1 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):55 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):55 (Ser) [5.5 to 0.1]; 45 (Ala):1 (Gln):1 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):45 (Gln):1 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):1 (Gln):45 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):45 (Ser) [4.5 to 0.1]; 35 (Ala):1 (Gln):1 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):35 (Gln):1 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):1 (Gln):35 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):35 (Ser) [3.5 to 0.1]; 25 (Ala):1 (Gln):1 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):25 (Gln):1 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):1 (Gln):25 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):25 (Ser) [0.1 to 0.5]; 15 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):15 (Gln):1 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):1 (Gln):15 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):15 (Ser) [1.5 to 0.1]; 14 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):14 (Gln):1 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):1 (Gln):14 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):14 (Ser) [1.4 to 0.1]; 13 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):13 (Gln):1 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):1 (Gln):13 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):13 (Ser) [1.3 to 0.1]; 12 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):12 (Gln):1 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):1 (Gln):12 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):12 (Ser) [1.2 to 0.1]; 11 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):11 (Gln):1 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):1 (Gln):11 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):11 (Ser) [1.1 to 0.1]; 10 (Ala):1 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):10 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):10 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):10 (Ser) [1 to 0.1]; 19 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):19 (Gln):2 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):2 (Gln):19 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):19 (Ser) [0.95 to 0.1]; 17 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):17 (Gln):2 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):2 (Gln):17 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):17 (Ser) [0.85 to 0.1]; 15 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):15 (Gln):2 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):2 (Gln):15 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):15 (Ser) [0.75 to 0.1]; 13 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):13 (Gln):2 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):2 (Gln):13 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):13 (Ser) [0.65 to 0.1]; 11 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):11 (Gln):2 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):2 (Gln):11 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):11 (Ser) [0.55 to 0.1]; 9 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):9 (Gln):2 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):2 (Gln):9 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):9 (Ser) [0.45 to 0.1]; 7 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):7 (Gln):2 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):2 (Gln):7 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):7 (Ser) [0.35 to 0.1]; 5 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):5 (Gln):2 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):2 (Gln):5 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):5 (Ser) [0.25 to 0.1]; 3 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):3 (Gln):2 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):2 (Gln):3 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):3 (Ser) [0.15 to 0.1]; 2 (Ala):3 (Gln):3 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):2 (Gln):3 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):3 (Gln):2 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):3 (Gln):3 (Gly):2 (Ser) [0.1 to 0.15]; 2 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):2 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):2 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):5 (Gly):2 (Ser) [0.1 to 0.25]; 2 (Ala):7 (Gln):7 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):2 (Gln):7 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):7 (Gln):2 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):7 (Gln):7 (Gly):2 (Ser) [0.1 to 0.35]; 2 (Ala):9 (Gln):9 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):2 (Gln):9 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):9 (Gln):2 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):9 (Gln):9 (Gly):2 (Ser) [0.1 to 0.45]; 2 (Ala):11 (Gln):11 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):2 (Gln):11 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):11 (Gln):2 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):11 (Gln):11 (Gly):2 (Ser) [0.1 to 0.55]; 2 (Ala):13 (Gln):13 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):2 (Gln):13 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):13 (Gln):2 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):13 (Gln):13 (Gly):2 (Ser) [0.1 to 0.65]; 2 (Ala):15 (Gln):15 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):2 (Gln):15 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):15 (Gln):2 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):15 (Gln):15 (Gly):2 (Ser) [0.1 to 0.75]; 2 (Ala):17 (Gln):17 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):2 (Gln):17 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):17 (Gln):2 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):17 (Gln):17 (Gly):2 (Ser) [0.1 to 0.85]; 2 (Ala):19 (Gln):19 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):2 (Gln):19 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):19 (Gln):2 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):19 (Gln):19 (Gly):2 (Ser) [0.1 to 0.95]); or from 1:100 to 100:1 (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) [0.1 to 10] to 100 (Ala):1 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):1 (Gln):100 (Gly):100 (Ser) [0.1 to 10] to 1 (Ala):100 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):100 (Gln):1 (Gly):100 (Ser) [0.1 to 10] to 1 (Ala):1 (Gln):100 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):100 (Gln):100 (Gly):1 (Ser) [0.1 to 10] to 1 (Ala):1 (Gln):1 (Gly):100 (Ser) [10 to 0.1]).
- Embodiment 67. The formulation according to any one of embodiments 57-59, wherein the therapeutically effective amount of the plant extract or plant extract combination is present at a weight percentage (w/w %) of the formulation of:
  - 0.025 w/w % or greater (e.g., 0.027; 0.029; 0.031; 0.033; 0.035; 0.037; 0.039; 0.041; 0.043; 0.045; 0.047; 0.049; 0.051; 0.053; 0.055; 0.057; 0.059; 0.061; 0.063; 0.065; 0.067; 0.069; 0.071; 0.073; 0.075; 0.077; 0.079; 0.081; 0.083; 0.085; 0.087; 0.089; 0.091; 0.093; 0.095; 0.097; 0.099; 0.1; 0.15; 0.3; 0.35; 0.5; 0.55; 0.7; 0.75; 0.9; 0.95; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3);
  - 5 w/w % or less (e.g., 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.85; 0.8; 0.65; 0.6; 0.45; 0.4; 0.25; 0.2; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018); or
  - 0.025 w/w %-5 w/w % (e.g., 0.026 w/w %-4.9 w/w %; 0.027 w/w %-4.8 w/w %; 0.028 w/w %-4.7 w/w %; 0.029 w/w %-4.6 w/w %; 0.03 w/w %-4.5 w/w %; 0.031 w/w %-4.4 w/w %; 0.032 w/w %-4.3 w/w %; 0.033 w/w %-4.2 w/w %; 0.034 w/w %-4.1 w/w %; 0.035 w/w %-4 w/w %; 0.036 w/w %-3.9 w/w %; 0.037 w/w %-3.8 w/w %; 0.038 w/w %-3.7 w/w %; 0.039 w/w %-3.6 w/w %; 0.04 w/w %-3.5 w/w %; 0.041 w/w %-3.4 w/w %; 0.042 w/w %-3.3 w/w %; 0.043 w/w %-3.2 w/w %; 0.044 w/w %-3.1 w/w %; 0.045 w/w %-3 w/w %; 0.046 w/w %-2.9 w/w %; 0.047 w/w %-2.8 w/w %; 0.048 w/w %-2.7 w/w %; 0.049 w/w %-2.6 w/w %; 0.05 w/w %-2.5 w/w %; 0.051 w/w %-2.4 w/w %; 0.052 w/w %-2.3 w/w %; 0.053 w/w %-2.2 w/w %; 0.054 w/w %-2.1 w/w %; 0.055 w/w %-2 w/w %; 0.056 w/w %-1.9 w/w %; 0.057 w/w %-1.8 w/w %; 0.058 w/w %-1.7 w/w %; 0.059 w/w %-1.6 w/w %; 0.06 w/w %-1.5 w/w %; 0.061 w/w %-1.4 w/w %; 0.062 w/w %-1.3 w/w %; 0.063 w/w %-1.2 w/w %; 0.064 w/w %-1.1 w/w %; 0.065 w/w %-1 w/w %; 0.066 w/w %-0.99 w/w %; 0.067 w/w %-0.98 w/w %; 0.068 w/w %-0.97 w/w %; 0.069 w/w %-0.96 w/w %; 0.07 w/w %-0.95 w/w %; 0.071 w/w %-0.94 w/w %; 0.072 w/w %-0.93 w/w %; 0.073 w/w %-0.92 w/w %; 0.074 w/w %-0.91 w/w %; 0.075 w/w %-0.9 w/w %; 0.076 w/w %-0.89 w/w %; 0.077 w/w %-0.88 w/w %; 0.078 w/w %-0.87 w/w %; 0.079 w/w %-0.86 w/w %; 0.08 w/w %-0.85 w/w %; 0.081 w/w %-0.84 w/w %; 0.082 w/w %-0.83 w/w %; 0.083 w/w %-0.82 w/w %; 0.084 w/w %-0.81 w/w %; 0.085 w/w %-0.8 w/w %; 0.086 w/w %-0.79 w/w %; 0.087 w/w %-0.78 w/w %; 0.088 w/w %-0.77 w/w %; 0.089 w/w %-0.76 w/w %; 0.09 w/w %-0.75 w/w %; 0.091 w/w %-0.74 w/w %; 0.092 w/w %-0.73 w/w %; 0.093 w/w %-0.72 w/w %; 0.094 w/w %-0.71 w/w %; 0.095 w/w %-0.7 w/w %; 0.096 w/w %-0.69 w/w %; 0.097 w/w %-0.68 w/w %; 0.098 w/w %-0.67 w/w %; 0.099 w/w %-0.66 w/w %; 0.1 w/w %-0.65 w/w %; 0.101 w/w %-0.64 w/w %; 0.102 w/w %-0.63 w/w %; 0.103 w/w %-0.62 w/w %; 0.104 w/w %-0.61 w/w %; 0.105 w/w %-0.6 w/w %; 0.106 w/w %-0.59 w/w %; 0.107 w/w %-0.58 w/w %; 0.108 w/w %-0.57 w/w %; 0.109 w/w %-0.56 w/w %; 0.11 w/w %-0.55 w/w %; 0.111 w/w %-0.54 w/w %; 0.112 w/w %-0.53 w/w %; 0.113 w/w %-0.52 w/w %; 0.114 w/w %-0.51 w/w %; 0.115 w/w %-0.5 w/w %; 0.116 w/w %-0.49 w/w %; 0.117 w/w %-0.48 w/w %; 0.118 w/w %-0.47 w/w %; 0.119 w/w %-0.46 w/w %; 0.12 w/w %-0.45 w/w %; 0.121 w/w %-0.44 w/w %; 0.122 w/w %-0.43 w/w %; 0.123 w/w %-0.42 w/w %; 0.124 w/w %-0.41 w/w %; 0.125 w/w %-0.4 w/w %; 0.126 w/w %-0.39 w/w %; 0.127 w/w %-0.38 w/w %; 0.128 w/w %-0.37 w/w %; 0.129 w/w %-0.36 w/w %; 0.13 w/w %-0.35 w/w %; 0.131 w/w %-0.34 w/w %; 0.132 w/w %-0.33 w/w %; 0.133 w/w %-0.32 w/w %; 0.134 w/w %-0.31 w/w %; 0.135 w/w %-0.3 w/w %; 0.136 w/w %-0.29 w/w %; 0.137 w/w %-0.28 w/w %; 0.138 w/w %-0.27 w/w %; 0.139 w/w %-0.26 w/w %; 0.14 w/w %-0.25 w/w %; 0.141 w/w %-0.24 w/w %; 0.142 w/w %-0.23 w/w %; 0.143 w/w %-0.22 w/w %; 0.144 w/w %-0.21 w/w %; 0.145 w/w %-0.2 w/w %; 0.146 w/w %-0.19 w/w %; 0.147 w/w %-0.18 w/w %; 0.148 w/w %-0.17 w/w %; 0.149 w/w %-0.16 w/w %).
- Embodiment 68. The formulation according to any one of embodiments 57-59, wherein the therapeutically effective amount of the combination of amino acids and the therapeutically effective amount of the plant extract or plant extract combination inhibit 5-lipoxygenase (an inflammation-inducing enzyme); decreases mobilization of calcium ions; decreases activation of MAP kinases.
- Embodiment 69. The formulation according to any one of embodiments 57-59, wherein the formulation or the topical formulation comprises, consists essentially of, or consists of: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of) alanine, glutamine, glycine, and serine; a therapeutically effective amount of a plant extract, wherein the plant extract comprises (or consists essentially of or consists of): a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or in combination with at least one of: lecithin, microcrystalline cellulose, and silica; and optionally, a dermatologically acceptable vehicle and/or additive.
- Embodiment 70. The formulation according to any one of embodiments 57-59, wherein the formulation or the topical formulation comprises, consists essentially of, or consists of: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of) an amino acid combination of at least one of alanine, glutamine, glycine, and serine; and a therapeutically effective amount of a plant extract or plant extract combination comprising (or consisting essentially of or consisting of) a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or in combination with at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications), where the amino acid combination and the plant extract combination are in a ratio of: 1:100 or greater (e.g., 2:100; 4:100; 6:100; 8:100; 10:100; 12:100; 14:100; 16:100; 18:100; 20:100; 22:100; 24:100; 26:100; 28:100; 30:100; 32:100; 34:100; 36:100; 38:100; 40:100; 42:100; 44:100; 46:100; 48:100; 50:100; 52:100; 54:100; 56:100; 58:100; 60:100; 62:100; 64:100; 66:100; 68:100; 70:100; 72:100; 74:100; 76:100; 78:100; 80:100; 82:100; 84:100; 86:100; 88:100; 90:100; 92:100; 94:100; 96:100; 98:100; 100:100); 100:1 or less (e.g., 99:100; 97:100; 95:100; 93:100; 91:100; 89:100; 87:100; 85:100; 83:100; 81:100; 79:100; 77:100; 75:100; 73:100; 71:100; 69:100; 67:100; 65:100; 63:100; 61:100; 59:100; 57:100; 55:100; 53:100; 51:100; 49:100; 47:100; 45:100; 43:100; 41:100; 39:100; 37:100; 35:100; 33:100; 31:100; 29:100; 27:100; 25:100; 23:100; 21:100; 19:100; 17:100; 15:100; 13:100; 11:100; 9:100; 7:100; 5:100; 3:100; 1:100); 1:100 to 100:1 (e.g., 1:99 to 99:1; 1:98 to 98:1; 1:97 to 97:1; 1:96 to 96:1; 1:95 to 95:1; 1:94 to 94:1; 1:93 to 93:1; 1:92 to 92:1; 1:91 to 91:1; 1:90 to 90:1; 1:89 to 89:1; 1:88 to 88:1; 1:87 to 87:1; 1:86 to 86:1; 1:85 to 85:1; 1:84 to 84:1; 1:83 to 83:1; 1:82 to 82:1; 1:81 to 81:1; 1:80 to 80:1; 1:79 to 79:1; 1:78 to 78:1; 1:77 to 77:1; 1:76 to 76:1; 1:75 to 75:1; 1:74 to 74:1; 1:73 to 73:1; 1:72 to 72:1; 1:71 to 71:1; 1:70 to 70:1; 1:69 to 69:1; 1:68 to 68:1; 1:67 to 67:1; 1:66 to 66:1; 1:65 to 65:1; 1:64 to 64:1; 1:63 to 63:1; 1:62 to 62:1; 1:61 to 61:1; 1:60 to 60:1; 1:59 to 59:1; 1:58 to 58:1; 1:57 to 57:1; 1:56 to 56:1; 1:55 to 55:1; 1:54 to 54:1; 1:53 to 53:1; 1:52 to 52:1; 1:51 to 51:1; 1:50 to 50:1; 1:49 to 49:1; 1:48 to 48:1; 1:47 to 47:1; 1:46 to 46:1; 1:45 to 45:1; 1:44 to 44:1; 1:43 to 43:1; 1:42 to 42:1; 1:41 to 41:1; 1:40 to 40:1; 1:39 to 39:1; 1:38 to 38:1; 1:37 to 37:1; 1:36 to 36:1; 1:35 to 35:1; 1:34 to 34:1; 1:33 to 33:1; 1:32 to 32:1; 1:31 to 31:1; 1:30 to 30:1; 1:29 to 29:1; 1:28 to 28:1; 1:27 to 27:1; 1:26 to 26:1; 1:25 to 25:1; 1:24 to 24:1; 1:23 to 23:1; 1:22 to 22:1; 1:21 to 21:1; 1:20 to 20:1; 1:19 to 19:1; 1:18 to 18:1; 1:17 to 17:1; 1:16 to 16:1; 1:15 to 15:1; 1:14 to 14:1; 1:13 to 13:1; 1:12 to 12:1; 1:11 to 11:1; 1:10 to 10:1; 1:9 to 9:1; 1:8 to 8:1; 1:7 to 7:1; 1:6 to 6:1; 1:5 to 5:1; 1:4 to 4:1; 1:3 to 3:1; 1:2 to 2:1).
- Embodiment 71. The formulation according to any one of embodiments 57-59, wherein the formulations or the topical formulations described here that optionally comprise, consist essentially of, or consist of a dermatologically acceptable vehicle and/or additive.
- Embodiment 72. The formulation according to any one of embodiments 57-59, wherein the dermatologically acceptable vehicle and/or additive can be selected from the group consisting of: water, sodium phytate, xanthan gum, kaolin, glycerin, polyglycerides (e.g., oleic polyglycerides, linoleic polyglycerides, linolenic polyglycerides), triglycerides (e.g., caprylic triglycerides, capric triglycerides), sucrose (e.g., sucrose stearate, sucrose distearate), alcohol (e.g., behenyl alcohol, benzoic alcohol, benzyl alcohol, cetearyl alcohol, oleyl alcohol), cetearyl glucoside, sodium stearoyl lactylate, Cera alba (beeswax), Butyrospermum parkii (shea butter), Prunus amigdalus sulcis (sweet almond oil), Simmondsia chinenesis (jojoba oil) phenoxyethanol, lecithin, tocopherol, tocopherol acetate, ascorbyl palmitate, citric acid, potassium sorbate, lactic acid, UV protectant, hydrogenated olive oil decyl esters, decyl cocaote, astrocaryum, murumuru seed butter, Zanthoxylum bungeanum fruit extract, squalene, pentylene glycol, dehydroacetic acid, tamarinds indica seed polysaccharide, sodium benzoate, curcumin, Zanthoxylum bungeanume fruit extract, Passiflora edulis seed oil, cetearyl ethylhexanoate, dimethicone, isopropyl myristate, acrylates/C10-C30 alkyl acrylate crosspolymer, allantoin, sodium phytate, PEG-90 stearate/glyceryl stearate; menthol, sodium hydroxide, sodium benzoate, dehydroacetic acid, benzoic alcohol, hexyldecanol, cetaryl ethylhexanoate, dimethicone, cetearyl ethylhexanoate, PEG-90 stearate/glyceryl stearate, ascorbyl palmitate, citric acid, Zanthoxylum bungeanum fruit extract, allantoin, disodium EDTA, phenoxyethanol, dehydroacetic acid, allantoin, XILOGEL®, Vaccinium myrtillus fruit extract, Oryza sativa (e.g. rice bran oil), isoamyl laurate, polymethyl methacrylate, fragrance, nutrients, antioxidants, water, or combinations thereof.
- Embodiment 73. The formulation according to any one of embodiments 57-59, wherein the formulation or the topical formulation is prepared by:
- mixing and/or stirring (concurrently, simultaneously, sequentially):
- (a) a therapeutically effective amount of a combination of amino acids comprising, consisting essentially of at least one of alanine, glutamine, glycine, and serine;
- (b) a therapeutically effective amount of a plant extract or a plant extract combination comprising, consisting essentially of, or consisting of: a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); and
- optionally, (c) a dermatologically acceptable vehicle and/or additive;
- heating (a), (b), and optionally (c) (either together or separately) to a temperature sufficient to completely mix, homogenize, or emulsify (a), (b), and optionally (c),
- wherein the temperature is 20° C. or greater (e.g., 22; 24; 26; 28; 30; 32; 34; 36; 38; 40; 42; 44; 46; 48; 50; 52; 54; 56; 58; 60; 62; 64; 66; 68; 70; 72; 74; 76; 78; 80; 82; 84; 86; 88; 90; 92; 94; 96; 98; 100; 102; 104; 106; 108; 110); 100° C. or less (e.g., 99; 97; 95; 93; 91; 89; 87; 85; 83; 81; 79; 77; 75; 73; 71; 69; 67; 65; 63; 61; 59; 57; 55; 53; 51; 49; 47; 45; 43; 41; 39; 37; 35; 33; 31; 29; 27; 25; 23; 21; 19; 17); or 20° C.-100° C. (e.g., 21° C.-99° C.; 22° C.-98° C.; 23° C.-97° C.; 24° C.-96° C.; 25° C.-95° C.; 26° C.-94° C.; 27° C.-93° C.; 28° C.-92° C.; 29° C.-91° C.; 30° C.-90° C.; 31° C.-89° C.; 32° C.-88° C.; 33° C.-87° C.; 34° C.-86° C.; 35° C.-85° C.; 36° C.-84° C.; 37° C.-83° C.; 38° C.-82° C.; 39° C.-81° C.; 40° C.-80° C.; 41° C.-79° C.; 42° C.-78° C.; 43° C.-77° C.; 44° C.-76° C.; 45° C.-75° C.; 46° C.-74° C.; 47° C.-73° C.; 48° C.-72° C.; 49° C.-71° C.; 50° C.-70° C.; 51° C.-69° C.; 52° C.-68° C.; 53° C.-67° C.; 54° C.-66° C.; 55° C.-65° C.; 56° C.-64° C.; 57° C.-63° C.; 58° C.-62° C.; 59° C.-61° C., for a time sufficient to completely mix, homogenize, or emulsify (e.g., seconds to minutes).
- Embodiment 74. The formulation according to any one of embodiments 57-59, wherein the plant extract or the plant extract combination of the topical formulation, further comprises (or consists essentially of or consists of) at least one of: a texturing and/or a bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications).
- Embodiment 75. A topical formulation, comprising (or consisting essentially of or consisting of):
  - as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of, or consisting of):
    - at least one of: alanine, glutamine, glycine, and serine, or salts thereof;
  - a therapeutically effective amount of a retinoid,
    - wherein the retinoid comprises (or consists essentially of or consists of): a retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof; and
  - optionally, a dermatologically acceptable vehicle and/or additive.
- Embodiment 76. A formulation for administration to skin comprising (or consisting essentially of or consisting of):
  - a therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids comprising (or consisting essentially of or consisting of): alanine, glutamine, glycine, and serine, or salts thereof;
  - a therapeutically effective amount of a retinoid,
    - wherein the retinoid is retinol;
    - and
  - optionally, a dermatologically acceptable vehicle and/or additive, wherein the therapeutically effective amount of the combination of free amino acids or the peptide combination of amino acids and the therapeutically effective amount of the retinol improves skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))) when tested by a method detecting expression of a skin moisture marker gene or an HA marker gene (e.g., HAS1, HAS2, HAS3, or EGF) and determining that the skin moisture marker gene or the HA marker gene is increased by a 1.4-450 fold increase of amino acid treated skin cells as compared to amino acid untreated skin cells,
- wherein the formulation improves skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))),
- wherein an improvement in skin moisture is indicated by an average fold increase or fold increase of a skin moisture marker gene or an HA marker gene (e.g., HAS1, HAS2, HAS3, or EGF) expression of: 1.4 or greater (e.g., 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8.1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5; 10.7; 10.9; 11.1; 11.3; 11.5; 11.7; 11.9; 12.1; 12.3; 12.5; 12.7; 12.9; 13.1; 13.3; 13.5; 13.7; 13.9; 14.1; 14.3; 14.5; 14.7; 14.9; 15.1; 15.3; 15.5; 15.7; 15.9; 16.1; 16.3; 16.5; 16.7; 16.9; 17.1; 17.3; 17.5; 17.7; 17.9; 18.1; 18.3; 18.5; 18.7; 18.9; 19.1; 19.3; 19.5; 19.7; 19.9; 20.1; 20.3; 20.5; 20.7; 20.9; 21.1; 21.3; 21.5; 21.7; 21.9; 22.1; 22.3; 22.5; 22.7; 22.9; 23.1; 23.3; 23.5; 23.7; 23.9; 23.1; 23.3; 23.5; 23.7; 23.9; 24.1; 24.3; 24.5; 24.7; 24.9; 25.1; 25.3; 25.5; 25.7; 25.9; 26.1; 26.3; 26.5; 26.7; 26.9; 27.1; 27.3; 27.5; 27.7; 27.9; 28.1; 28.3; 28.5; 28.7; 28.9; 29.1; 29.3; 29.5; 29.7; 29.9; 30.1; 30.3; 30.5; 30.7; 30.9; 31.1; 31.3; 31.5; 31.7; 31.9; 32.1; 32.3; 32.5; 32.7; 32.9; 33.1; 33.3; 33.5; 33.7; 33.9; 34.1; 34.3; 34.5; 34.7; 34.9; 35.1; 35.3; 35.5; 35.7; 35.9; 36.1; 36.3; 36.5; 36.7; 36.9; 37.1; 37.3; 37.5; 37.7; 37.9; 38.1; 38.3; 38.5; 38.7; 38.9; 39.1; 39.3; 39.5; 39.7; 39.9; 40.1; 40.3; 40.5; 40.7; 40.9; 41.1; 41.3; 41.5; 55; 70; 90; 105; 120; 170; 195; 205; 255; 270; 295; 305; 320; 355; 370; 405; 420; 455); 450 or less (e.g., 425; 405; 400; 375; 350; 325; 300; 290; 275; 250; 225; 200; 190; 175; 150; 125; 100; 95; 75; 50; 39.8; 39.6; 39.4; 39.2; 39; 38.8; 38.6; 38.4; 38.2; 38; 37.8; 37.6; 37.4; 37.2; 37; 36.8; 36.6; 36.4; 36.2; 36; 35.8; 35.6; 35.4; 35.2; 35; 34.8; 34.6; 34.4; 34.2; 34; 33.8; 33.6; 33.4; 33.2; 33; 32.8; 32.6; 32.4; 32.2; 32; 31.8; 31.6; 31.4; 31.2; 31; 30.8; 30.6; 30.4; 30.2; 30; 29.8; 29.6; 29.4; 29.2; 29; 28.8; 28.6; 28.4; 28.2; 28; 27.8; 27.6; 27.4; 27.2; 27; 26.8; 26.6; 26.4; 26.2; 26; 25.8; 25.6; 25.4; 25.2; 25; 24.8; 24.6; 24.4; 24.2; 24; 23.8; 23.6; 23.4; 23.2; 23; 22.8; 22.6; 22.4; 22.2; 22; 21.8; 21.6; 21.4; 21.2; 21; 20.8; 20.6; 20.4; 20.2; 20; 19.8; 19.6; 19.4; 19.2; 19; 18.8; 18.6; 18.4; 18.2; 18; 17.8; 17.6; 17.4; 17.2; 17.2; 17; 16.8; 16.6; 16.4; 16.2; 16; 15.8; 15.6; 15.4; 15.2; 15; 14.8; 14.6; 14.4; 14.2; 14; 13.8; 13.6; 13.4; 13.2; 13; 12.8; 12.6; 12.4; 12.2; 12; 11.8; 11.6; 11.4; 11.2; 11; 10.8; 10.6; 10.4; 10.2; 10; 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0.6); or 1.4-450 (e.g., 1.5-440; 1.6-445; 1.7-430; 1.8-420; 1.9-410; 2-400; 2.1-390; 2.2-380; 2.3-370; 2.4 360; 2.5-350; 2.6-340; 2.7-330; 2.8-320; 2.9-310; 3-300; 3.1-290; 3.2-280; 3.3-270; 3.4-260; 3.5-250; 3.6-240; 3.7-230; 3.8-220; 3.9-210; 4-200; 4.1-190; 4.2-180; 4.3-170; 4.4-160; 4.5-150; 4.6-140; 4.7-130; 4.8-120; 4.9-110; 5-100; 5.1-90; 5.2-80; 5.3-70; 5.4-60; 5.5-50; 5.6-40; 5.7-35.8; 5.8-35.7; 5.9-35.6; 6-35.5; 6.1-35.4; 6.2-35.3; 6.3-35.2; 6.4-35.1; 6.5-35; 6.6-34.9; 6.7-34.8; 6.8-34.7; 6.9-34.6; 7-34.5; 7.1-34.4; 7.2-34.3; 7.3-34.2; 7.4-34.1; 7.5-34; 7.6-33.9; 7.7-33.8; 7.8-33.7; 7.9-33.6; 8-33.5; 8.1-33.4; 8.2-33.3; 8.3-33.2; 8.4-33.1; 8.5-33; 8.6-32.9; 8.7-32.8; 8.8-32.7; 8.9-32.6; 9-32.5; 9.1-32.4; 9.2-32.3; 9.3-32.2; 9.4-32.1; 9.5-32; 9.6-31.9; 9.7-31.8; 9.8-31.7; 9.9-31.6; 10-31.5; 10.1 31.4; 10.2-31.3; 10.3-31.2; 10.4-31.1; 10.5-31; 10.6-30.9; 10.7-30.8; 10.8-30.7; 10.9-30.6; 11-30.5; 11.1-30.4; 11.2-30.3; 11.3-30.2; 11.4-30.1; 11.5-30; 11.6-29.9; 11.7-29.8; 11.8-29.7; 11.9-29.6; 12-29.5; 12.1-29.4; 12.2-29.3; 12.3-29.2; 12.4-29.1; 12.5-29; 12.6-28.9; 12.7-28.8; 12.8-28.7; 12.9-28.6; 13-28.5; 13.1-28.4; 13.2-28.3; 13.3-28.2; 13.4-28.1; 13.5-28; 13.6-27.9; 13.7-27.8; 13.8-27.7; 13.9-27.6; 14-27.5; 14.1-27.4; 14.2-27.3; 14.3-27.2; 14.4-27.1; 14.5-27; 14.6-26.9; 14.7-26.8; 14.8-26.7; 14.9-26.6; 15-26.5; 15.1-26.4; 15.2-26.3; 15.3-26.2; 15.4-26.1; 15.5-26; 15.6-25.9; 15.7-25.8; 15.8-25.7; 15.9-25.6; 16-25.5; 16.1-25.4; 16.2-25.3; 16.3-25.2; 16.4-25.1; 16.5-25; 16.6-24.9; 16.7-24.8; 16.8-24.7; 16.9-24.6; 17-24.5; 17.1-24.4; 17.2-24.3; 17.3-24.2; 17.4-24.1; 17.5-24; 17.6-23.9; 17.7-23.8; 17.8-23.7; 17.9-23.6; 18-23.5; 18.1-23.4; 18.2-23.3; 18.3-23.2; 18.4-23.1; 18.5-23; 18.6-22.9; 18.7-22.8; 18.8-22.7; 18.9-22.6; 19-22.5; 19.1-22.4; 19.2-22.3; 19.3-22.2; 19.4 22.1; 19.5-22; 19.6-21.9; 19.7-21.8; 19.8-21.7; 19.9-21.6; 20-21.5; 20.1-21.4; 20.2-21.3; 20.3-21.2; 20.4-21.1; 20.5-21; 20.6-20.9; 20.7-20.8).
- Embodiment 77. The formulation according to any one of embodiments 75-76, wherein each of the amino acids is present at a concentration:
  - of 0.1 mM or greater (e.g., 0.3, 0.5, 0.7; 0.9; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3.1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3; 5.5; 5.7; 5.9; 6.1; 6.3; 6.5; 6.7; 6.9; 7.1; 7.3; 7.5; 7.7; 7.9; 8, 1; 8.3; 8.5; 8.7; 8.9; 9.1; 9.3; 9.5; 9.7; 9.9; 10.1; 10.3; 10.5);
  - of 10 mM or less (e.g., 9.8; 9.6; 9.4; 9.2; 9; 8.8; 8.6; 8.4; 8.2; 8; 7.8; 7.6; 7.4; 7.2; 7; 6.8; 6.6; 6.4; 6.2; 6; 5.8; 5.6; 5.4; 5.2; 5; 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.8; 0, 6; 0.4; 0.2; 0.08; 0.06; 0.04; 0.02); or
  - ranging from 0.1 mM to 10 mM (e.g., 0.2 mM to 9.9 mM; 0.3 mM to 9.8 mM; 0.4 mM to 9.7 mM; 0.5 mM to 9.6 mM; 0.6 mM to 9.5 mM; 0.7 mM to 9.4 mM; 0.8 mM to 9.3 mM; 0.9 mM to 9.2 mM; 1 mM to 9.1 mM; 1.1 mM to 9 mM; 1.2 mM to 8.9 mM; 1.3 mM to 8.8 mM; 1.4 mM to 8.7 mM; 1.5 mM to 8.6 mM; 1.6 mM to 8.5 mM; 1.7 mM to 8.4 mM; 1.8 mM to 8.3 mM; 1.9 mM to 8.2 mM; 2 mM to 8.1 mM; 2.1 mM to 8 mM; 2.2 mM to 7.9 mM; 2.3 mM to 7.8 mM; 2.4 mM to 7.7 mM; 2.5 mM to 7.6 mM; 2.6 mM to 7.5 mM; 2.7 mM to 7.4 mM; 2.8 mM to 7.3 mM; 2.9 mM to 7.2 mM; 3 mM to 7.1 mM; 3.1 mM to 7 mM; 3.2 mM to 6.9 mM; 3.3 mM to 6.8 mM; 3.4 mM to 6.7 mM; 3.5 mM to 6.6 mM; 3.6 mM to 6.5 mM; 3.7 mM to 6.4 mM; 3.8 mM to 6.3 mM; 3.9 mM to 6.2 mM; 4 mM to 6.1 mM; 4.1 mM to 6 mM; 4.2 mM to 5.9 mM; 4.3 mM to 5.8 mM; 4.4 mM to 5.7 mM; 4.5 mM to 5.6 mM; 4.6 mM to 5.5 mM; 4.7 mM to 5.4 mM; 4.8 mM to 5.3 mM; 4.9 mM to 5.2 mM; 5 mM to 5.1 mM),
- Embodiment 78. The formulation according to one of embodiments 75-76, wherein each of the amino acids is present at a weight percent to the formulation (wt/wt %) of:
  - 0.001 wt % or greater (e.g., 0.002; 0.003; 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.2; 0.3; 0.4; 0.5; 0.6; 0.7; 0.8; 0.9; 1);
  - 1 wt % or less (e.g., 0.95; 0.85; 0.75; 0.65; 0.55; 0.45; 0.35; 0.25; 0.15; 0.095; 0.085; 0.075; 0.065; 0.055; 0.045; 0.035; 0.025; 0.015; 0.0095; 0.0085; 0.0075; 0.0065; 0.0055; 0.0045; 0.0035; 0.0025; 0.0015); or
  - 0.001 wt % to 1 wt % (e.g., 0.0012 wt % to 0.99 wt %; 0.0014 wt % to 0.97 wt %; 0.0016 wt % to 0.95 wt %; 0.0018 wt % to 0.93 wt %; 0.002 wt % to 0.91 wt %; 0.0022 wt % to 0.89 wt %; 0.0024 wt % to 0.87 wt %; 0.0026 wt % to 0.85 wt %; 0.0028 wt % to 0.83 wt %; 0.003 wt % to 0.81 wt %; 0.0032 wt % to 0.79 wt %; 0.0034 wt % to 0.77 wt %; 0.0036 wt % to 0.75 wt %; 0.0038 wt % to 0.73 wt %; 0.004 wt % to 0.71 wt %; 0.0042 wt % to 0.69 wt %; 0.0044 wt % to 0.67 wt %; 0.0046 wt % to 0.65 wt %; 0.0048 wt % to 0.63 wt %; 0.005 wt % to 0.61 wt %; 0.0052 wt % to 0.59 wt %; 0.0054 wt % to 0.57 wt %; 0.0056 wt % to 0.55 wt %; 0.0058 wt % to 0.53 wt %; 0.006 wt % to 0.51 wt %; 0.0062 wt % to 0.49 wt %; 0.0064 wt % to 0.47 wt %; 0.0066 wt % to 0.45 wt %; 0.0068 wt % to 0.43 wt %; 0.007 wt % to 0.41 wt %; 0.0072 wt % to 0.39 wt %; 0.0074 wt % to 0.37 wt %; 0.0076 wt % to 0.35 wt %; 0.0078 wt % to 0.33 wt %; 0.008 wt % to 0.31 wt %; 0.0082 wt % to 0.29 wt %; 0.0084 wt % to 0.27 wt %; 0.0086 wt % to 0.25 wt %; 0.0088 wt % to 0.23 wt %; 0.009 wt % to 0.21 wt %; 0.0092 wt % to 0.19 wt %; 0.0094 wt % to 0.17 wt %; 0.0096 wt % to 0.15 wt %; 0.0098 wt % to 0.13 wt %; 0.01 wt % to 0.11 wt %; 0.012 wt % to 0.099 wt %; 0.014 wt % to 0.097 wt %; 0.016 wt % to 0.095 wt %; 0.018 wt % to 0.093 wt %; 0.02 wt % to 0.091 wt %; 0.022 wt % to 0.089 wt %; 0.024 wt % to 0.087 wt %; 0.026 wt % to 0.085 wt %; 0.028 wt % to 0.083 wt %; 0.03 wt % to 0.081 wt %; 0.032 wt % to 0.079 wt %; 0.034 wt % to 0.077 wt %; 0.036 wt % to 0.075 wt %; 0.038 wt % to 0.073 wt %; 0.04 wt % to 0.071 wt %; 0.042 wt % to 0.069 wt %; 0.044 wt % to 0.067 wt %; 0.046 wt % to 0.065 wt %; 0.048 wt % to 0.063 wt %; 0.05 wt % to 0.061 wt %; 0.052 wt % to 0.059 wt %; 0.054 wt % to 0.057 wt %).
- Embodiment 79. The formulation according to any one of embodiments 75-76, wherein alanine is present at a weight percent to the formulation (wt/wt %) of:
  - 0.0035 wt % or greater (e.g., 0.00352; 0.00354; 0.00356; 0.00358; 0.0036; 0.00362; 0.00364; 0.00366; 0.00368; 0.0037; 0.00372; 0.00374; 0.00376; 0.00378; 0.0038; 0.00382; 0.00384; 0.00386; 0.00388; 0.004; 0.0042; 0.0044; 0.0046; 0.0048; 0.005; 0.0052; 0.0054; 0.0056; 0.0058; 0.006; 0.0062; 0.0064; 0.0066; 0.0068; 0.007; 0.0072; 0.0074; 0.0076; 0.0078; 0.008; 0.0082; 0.0084; 0.0086; 0.0088; 0.009; 0.0092; 0.0094; 0.0096; 0.0098; 0.01; 0.012; 0.014; 0.016; 0.018; 0.02; 0.022; 0.024; 0.026; 0.028; 0.03; 0.032; 0.034; 0.036; 0.038; 0.04; 0.042; 0.044; 0.046; 0.048; 0.05; 0.052; 0.054; 0.056; 0.058; 0.06; 0.062; 0.064; 0.066; 0.068; 0.07; 0.072; 0.074; 0.076; 0.078; 0.08; 0.082; 0.084; 0.086; 0.088; 0.09; 0.092; 0.094; 0.096; 0.098; 0.1; 0.102; 0.104; 0.106; 0.108; 0.11; 0.112; 0.114; 0.116; 0.118; 0.12; 0.122; 0.124; 0.126; 0.128; 0.13; 0.132; 0.134; 0.136; 0.138; 0.14; 0.142; 0.144; 0.146; 0.148; 0.15; 0.152; 0.154; 0.156; 0.158; 0.16; 0.162; 0.164; 0.166; 0.168; 0.17; 0.172; 0.174; 0.176; 0.178; 0.18; 0.182; 0.184; 0.186; 0.188; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.202; 0.204; 0.206; 0.208; 0.21; 0.212; 0.214; 0.216; 0.218; 0.22; 0.222; 0.224; 0.226; 0.228; 0.23; 0.232; 0.234; 0.236; 0.238; 0.24; 0.242; 0.244; 0.246; 0.248; 0.25; 0.252; 0.254; 0.256; 0.258; 0.26; 0.262; 0.264; 0.266; 0.268; 0.27; 0.272; 0.274; 0.276; 0.278; 0.28; 0.282; 0.284; 0.286; 0.288; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.202; 0.204; 0.206; 0.208; 0.21; 0.212; 0.214; 0.216; 0.218; 0.22; 0.222; 0.224; 0.226; 0.228; 0.23; 0.232; 0.234; 0.236; 0.238; 0.24; 0.242; 0.244; 0.246; 0.248; 0.25; 0.252; 0.254; 0.256; 0.258; 0.26; 0.262; 0.264; 0.266; 0.268; 0.27; 0.272; 0.274; 0.276; 0.278; 0.28; 0.282; 0.284; 0.286; 0.288; 0.3; 0.302; 0.304; 0.306; 0.308; 0.31; 0.312; 0.314; 0.316; 0.318; 0.32; 0.322; 0.324; 0.326; 0.328; 0.33; 0.332; 0.334; 0.336; 0.338; 0.34; 0.342; 0.344; 0.346; 0.348; 0.35; 0.352; 0.354; 0.356; 0.358; 0.36; 0.362; 0.364; 0.366; 0.368; 0.37; 0.372; 0.374; 0.376; 0.378; 0.38; 0.382; 0.384; 0.386; 0.388; 0.39; 0.392; 0.394; 0.396; 0.398; 0.4; 0.402; 0.404; 0.406; 0.408; 0.41; 0.412; 0.414; 0.416; 0.418; 0.42; 0.422; 0.424; 0.426; 0.428; 0.43; 0.432; 0.434; 0.436; 0.438; 0.44; 0.442; 0.444; 0.446; 0.448; 0.45; 0.452; 0.454; 0.456; 0.458; 0.46; 0.462; 0.464; 0.466; 0.468; 0.47; 0.472; 0.474; 0.476; 0.478; 0.48; 0.482; 0.484; 0.486; 0.488; 0.5);
  - 0.4 wt % or less (e.g., 0.399; 0.397; 0.395; 0.393; 0.391; 0.389; 0.387; 0.385; 0.383; 0.381; 0.379; 0.377; 0.375; 0.373; 0.371; 0.369; 0.367; 0.365; 0.363; 0.361; 0.359; 0.357; 0.355; 0.353; 0.351; 0.349; 0.347; 0.345; 0.343; 0.341; 0.339; 0.337; 0.335; 0.333; 0.331; 0.329; 0.327; 0.325; 0.323; 0.321; 0.319; 0.317; 0.315; 0.313; 0.311; 0.309; 0.307; 0.305; 0.303; 0.301; 0.299; 0.297; 0.295; 0.293; 0.291; 0.289; 0.287; 0.285; 0.283; 0.281; 0.279; 0.277; 0.275; 0.273; 0.271; 0.269; 0.267; 0.265; 0.263; 0.261; 0.259; 0.257; 0.255; 0.253; 0.251; 0.249; 0.247; 0.245; 0.243; 0.241; 0.239; 0.237; 0.235; 0.233; 0.231; 0.229; 0.227; 0.225; 0.223; 0.221; 0.219; 0.217; 0.215; 0.213; 0.211; 0.209; 0.207; 0.205; 0.203; 0.201; 0.199; 0.197; 0.195; 0.193; 0.191; 0.189; 0.187; 0.185; 0.183; 0.181; 0.179; 0.177; 0.175; 0.173; 0.171; 0.169; 0.167; 0.165; 0.163; 0.161; 0.159; 0.157; 0.155; 0.153; 0.151; 0.149; 0.147; 0.145; 0.143; 0.141; 0.139; 0.137; 0.135; 0.133; 0.131; 0.129; 0.127; 0.125; 0.123; 0.121; 0.119; 0.117; 0.115; 0.113; 0.111; 0.109; 0.107; 0.105; 0.103; 0.101; 0.099; 0.097; 0.095; 0.093; 0.091; 0.089; 0.087; 0.085; 0.083; 0.081; 0.079; 0.077; 0.075; 0.073; 0.071; 0.069; 0.067; 0.065; 0.063; 0.061; 0.059; 0.057; 0.055; 0.053; 0.051; 0.049; 0.047; 0.045; 0.043; 0.041; 0.039; 0.037; 0.035; 0.033; 0.031; 0.029; 0.027; 0.025; 0.023; 0.021; 0.019; 0.017; 0.015; 0.013; 0.011; 0.009; 0.007; 0.005; 0.003; 0.002); or
  - 0.0035 wt % to 0.36 wt % (e.g., 0.00351 wt % to 0.359 wt %; 0.00352 wt % to 0.358 wt %; 0.00353 wt % to 0.357 wt %; 0.00354 wt % to 0.356 wt %; 0.00355 wt % to 0.355 wt %; 0.00356 wt % to 0.354 wt %; 0.00357 wt % to 0.353 wt %; 0.00358 wt % to 0.352 wt %; 0.00359 wt % to 0.351 wt %; 0.0036 wt % to 0.35 wt %; 0.00361 wt % to 0.349 wt %; 0.00362 wt % to 0.348 wt %; 0.00363 wt % to 0.347 wt %; 0.00364 wt % to 0.346 wt %; 0.00365 wt % to 0.345 wt %; 0.00366 wt % to 0.344 wt %; 0.00367 wt % to 0.343 wt %; 0.00368 wt % to 0.342 wt %; 0.00369 wt % to 0.341 wt %; 0.0037 wt % to 0.34 wt %; 0.00371 wt % to 0.339 wt %; 0.00372 wt % to 0.338 wt %; 0.00373 wt % to 0.337 wt %; 0.00374 wt % to 0.336 wt %; 0.00375 wt % to 0.335 wt %; 0.00376 wt % to 0.334 wt %; 0.00377 wt % to 0.333 wt %; 0.00378 wt % to 0.332 wt %; 0.00379 wt % to 0.331 wt %; 0.0038 wt % to 0.33 wt %; 0.00381 wt % to 0.329 wt %; 0.00382 wt % to 0.328 wt %; 0.00383 wt % to 0.327 wt %; 0.00384 wt % to 0.326 wt %; 0.00385 wt % to 0.325 wt %; 0.00386 wt % to 0.324 wt %; 0.00387 wt % to 0.323 wt %; 0.00388 wt % to 0.322 wt %; 0.00389 wt % to 0.321 wt %; 0.0039 wt % to 0.32 wt %; 0.00391 wt % to 0.319 wt %; 0.00392 wt % to 0.318 wt %; 0.00393 wt % to 0.317 wt %; 0.00394 wt % to 0.316 wt %; 0.00395 wt % to 0.315 wt %; 0.00396 wt % to 0.314 wt %; 0.00397 wt % to 0.313 wt %; 0.00398 wt % to 0.312 wt %; 0.00399 wt % to 0.311 wt %; 0.004 wt % to 0.31 wt %; 0.00401 wt % to 0.309 wt %; 0.00402 wt % to 0.308 wt %; 0.00403 wt % to 0.307 wt %; 0.00404 wt % to 0.306 wt %; 0.00405 wt % to 0.305 wt %; 0.00406 wt % to 0.304 wt %; 0.00407 wt % to 0.303 wt %; 0.00408 wt % to 0.302 wt %; 0.00409 wt % to 0.301 wt %; 0.0041 wt % to 0.3 wt %; 0.00411 wt % to 0.299 wt %; 0.00412 wt % to 0.298 wt %; 0.00413 wt % to 0.297 wt %; 0.00414 wt % to 0.296 wt %; 0.00415 wt % to 0.295 wt %; 0.00416 wt % to 0.294 wt %; 0.00417 wt % to 0.293 wt %; 0.00418 wt % to 0.292 wt %; 0.00419 wt % to 0.291 wt %; 0.0042 wt % to 0.29 wt %; 0.00421 wt % to 0.289 wt %; 0.00422 wt % to 0.288 wt %; 0.00423 wt % to 0.287 wt %; 0.00424 wt % to 0.286 wt %; 0.00425 wt % to 0.285 wt %; 0.00426 wt % to 0.284 wt %; 0.00427 wt % to 0.283 wt %; 0.00428 wt % to 0.282 wt %; 0.00429 wt % to 0.281 wt %; 0.0043 wt % to 0.28 wt %; 0.00431 wt % to 0.279 wt %; 0.00432 wt % to 0.278 wt %; 0.00433 wt % to 0.277 wt %; 0.00434 wt % to 0.276 wt %; 0.00435 wt % to 0.275 wt %; 0.00436 wt % to 0.274 wt %; 0.00437 wt % to 0.273 wt %; 0.00438 wt % to 0.272 wt %; 0.00439 wt % to 0.271 wt %; 0.0044 wt % to 0.27 wt %; 0.00441 wt % to 0.269 wt %; 0.00442 wt % to 0.268 wt %; 0.00443 wt % to 0.267 wt %; 0.00444 wt % to 0.266 wt %; 0.00445 wt % to 0.265 wt %; 0.00446 wt % to 0.264 wt %; 0.00447 wt % to 0.263 wt %; 0.00448 wt % to 0.262 wt %; 0.00449 wt % to 0.261 wt %; 0.0045 wt % to 0.26 wt %; 0.00451 wt % to 0.259 wt %; 0.00452 wt % to 0.258 wt %; 0.00453 wt % to 0.257 wt %; 0.00454 wt % to 0.256 wt %; 0.00455 wt % to 0.255 wt %; 0.00456 wt % to 0.254 wt %; 0.00457 wt % to 0.253 wt %; 0.00458 wt % to 0.252 wt %; 0.00459 wt % to 0.251 wt %; 0.0046 wt % to 0.25 wt %; 0.00461 wt % to 0.249 wt %; 0.00462 wt % to 0.248 wt %; 0.00463 wt % to 0.247 wt %; 0.00464 wt % to 0.246 wt %; 0.00465 wt % to 0.245 wt %; 0.00466 wt % to 0.244 wt %; 0.00467 wt % to 0.243 wt %; 0.00468 wt % to 0.242 wt %; 0.00469 wt % to 0.241 wt %; 0.0047 wt % to 0.24 wt %; 0.00471 wt % to 0.239 wt %; 0.00472 wt % to 0.238 wt %; 0.00473 wt % to 0.237 wt %; 0.00474 wt % to 0.236 wt %; 0.00475 wt % to 0.235 wt %; 0.00476 wt % to 0.234 wt %; 0.00477 wt % to 0.233 wt %; 0.00478 wt % to 0.232 wt %; 0.00479 wt % to 0.231 wt %; 0.0048 wt % to 0.23 wt %; 0.00481 wt % to 0.229 wt %; 0.00482 wt % to 0.228 wt %; 0.00483 wt % to 0.227 wt %; 0.00484 wt % to 0.226 wt %; 0.00485 wt % to 0.225 wt %; 0.00486 wt % to 0.224 wt %; 0.00487 wt % to 0.223 wt %; 0.00488 wt % to 0.222 wt %; 0.00489 wt % to 0.221 wt %; 0.0049 wt % to 0.22 wt %; 0.00491 wt % to 0.219 wt %; 0.00492 wt % to 0.218 wt %; 0.00493 wt % to 0.217 wt %; 0.00494 wt % to 0.216 wt %; 0.00495 wt % to 0.215 wt %; 0.00496 wt % to 0.214 wt %; 0.00497 wt % to 0.213 wt %; 0.00498 wt % to 0.212 wt %; 0.00499 wt % to 0.211 wt %; 0.005 wt % to 0.21 wt %; 0.00501 wt % to 0.209 wt %; 0.00502 wt % to 0.208 wt %; 0.00503 wt % to 0.207 wt %; 0.00504 wt % to 0.206 wt %; 0.00505 wt % to 0.205 wt %; 0.00506 wt % to 0.204 wt %; 0.00507 wt % to 0.203 wt %; 0.00508 wt % to 0.202 wt %; 0.00509 wt % to 0.201 wt %; 0.0051 wt % to 0.2 wt %; 0.00511 wt % to 0.199 wt %; 0.00512 wt % to 0.198 wt %; 0.00513 wt % to 0.197 wt %; 0.00514 wt % to 0.196 wt %; 0.00515 wt % to 0.195 wt %; 0.00516 wt % to 0.194 wt %; 0.00517 wt % to 0.193 wt %; 0.00518 wt % to 0.192 wt %; 0.00519 wt % to 0.191 wt %; 0.0052 wt % to 0.19 wt %; 0.00521 wt % to 0.189 wt %; 0.00522 wt % to 0.188 wt %; 0.00523 wt % to 0.187 wt %; 0.00524 wt % to 0.186 wt %; 0.00525 wt % to 0.185 wt %; 0.00526 wt % to 0.184 wt %; 0.00527 wt % to 0.183 wt %; 0.00528 wt % to 0.182 wt %; 0.00529 wt % to 0.181 wt %; 0.0053 wt % to 0.18 wt %; 0.00531 wt % to 0.179 wt %; 0.00532 wt % to 0.178 wt %; 0.00533 wt % to 0.177 wt %; 0.00534 wt % to 0.176 wt %; 0.00535 wt % to 0.175 wt %; 0.00536 wt % to 0.174 wt %; 0.00537 wt % to 0.173 wt %; 0.00538 wt % to 0.172 wt %; 0.00539 wt % to 0.171 wt %; 0.0054 wt % to 0.17 wt %; 0.00541 wt % to 0.169 wt %; 0.00542 wt % to 0.168 wt %; 0.00543 wt % to 0.167 wt %; 0.00544 wt % to 0.166 wt %; 0.00545 wt % to 0.165 wt %; 0.00546 wt % to 0.164 wt %; 0.00547 wt % to 0.163 wt %; 0.00548 wt % to 0.162 wt %; 0.00549 wt % to 0.161 wt %; 0.0055 wt % to 0.16 wt %; 0.00551 wt % to 0.159 wt %; 0.00552 wt % to 0.158 wt %; 0.00553 wt % to 0.157 wt %; 0.00554 wt % to 0.156 wt %; 0.00555 wt % to 0.155 wt %; 0.00556 wt % to 0.154 wt %; 0.00557 wt % to 0.153 wt %; 0.00558 wt % to 0.152 wt %; 0.00559 wt % to 0.151 wt %; 0.0056 wt % to 0.15 wt %; 0.00561 wt % to 0.149 wt %; 0.00562 wt % to 0.148 wt %; 0.00563 wt % to 0.147 wt %; 0.00564 wt % to 0.146 wt %; 0.00565 wt % to 0.145 wt %; 0.00566 wt % to 0.144 wt %; 0.00567 wt % to 0.143 wt %; 0.00568 wt % to 0.142 wt %; 0.00569 wt % to 0.141 wt %; 0.0057 wt % to 0.14 wt %; 0.00571 wt % to 0.139 wt %; 0.00572 wt % to 0.138 wt %; 0.00573 wt % to 0.137 wt %; 0.00574 wt % to 0.136 wt %; 0.00575 wt % to 0.135 wt %; 0.00576 wt % to 0.134 wt %; 0.00577 wt % to 0.133 wt %; 0.00578 wt % to 0.132 wt %; 0.00579 wt % to 0.131 wt %; 0.0058 wt % to 0.13 wt %; 0.00581 wt % to 0.129 wt %; 0.00582 wt % to 0.128 wt %; 0.00583 wt % to 0.127 wt %; 0.00584 wt % to 0.126 wt %; 0.00585 wt % to 0.125 wt %; 0.00586 wt % to 0.124 wt %; 0.00587 wt % to 0.123 wt %; 0.00588 wt % to 0.122 wt %; 0.00589 wt % to 0.121 wt %; 0.0059 wt % to 0.12 wt %; 0.00591 wt % to 0.119 wt %; 0.00592 wt % to 0.118 wt %; 0.00593 wt % to 0.117 wt %; 0.00594 wt % to 0.116 wt %; 0.00595 wt % to 0.115 wt %; 0.00596 wt % to 0.114 wt %; 0.00597 wt % to 0.113 wt %; 0.00598 wt % to 0.112 wt %; 0.00599 wt % to 0.111 wt %; 0.0056 wt % to 0.11 wt %; 0.00561 wt % to 0.109 wt %; 0.00562 wt % to 0.108 wt %; 0.00563 wt % to 0.107 wt %; 0.00564 wt % to 0.106 wt %; 0.00565 wt % to 0.105 wt %; 0.00566 wt % to 0.104 wt %; 0.00567 wt % to 0.103 wt %; 0.00568 wt % to 0.102 wt %; 0.00569 wt % to 0.101 wt %; 0.0057 wt % to 0.1 wt %; 0.00571 wt % to 0.099 wt %; 0.00572 wt % to 0.098 wt %; 0.00573 wt % to 0.097 wt %; 0.00574 wt % to 0.096 wt %; 0.00575 wt % to 0.095 wt %; 0.00576 wt % to 0.094 wt %; 0.00577 wt % to 0.093 wt %; 0.00578 wt % to 0.092 wt %; 0.00579 wt % to 0.091 wt %; 0.0058 wt % to 0.09 wt %; 0.00581 wt % to 0.089 wt %; 0.00582 wt % to 0.088 wt %; 0.00583 wt % to 0.087 wt %; 0.00584 wt % to 0.086 wt %; 0.00585 wt % to 0.085 wt %; 0.00586 wt % to 0.084 wt %; 0.00587 wt % to 0.083 wt %; 0.00588 wt % to 0.082 wt %; 0.00589 wt % to 0.081 wt %; 0.0059 wt % to 0.08 wt %; 0.00591 wt % to 0.079 wt %; 0.00592 wt % to 0.078 wt %; 0.00593 wt % to 0.077 wt %; 0.00594 wt % to 0.076 wt %; 0.00595 wt % to 0.075 wt %; 0.00596 wt % to 0.074 wt %; 0.00597 wt % to 0.073 wt %; 0.00598 wt % to 0.072 wt %; 0.00599 wt % to 0.071 wt %; 0.006 wt % to 0.07 wt %; 0.00601 wt % to 0.069 wt %; 0.00602 wt % to 0.068 wt %; 0.00603 wt % to 0.067 wt %; 0.00604 wt % to 0.066 wt %; 0.00605 wt % to 0.065 wt %; 0.00606 wt % to 0.064 wt %; 0.00607 wt % to 0.063 wt %; 0.00608 wt % to 0.062 wt %; 0.00609 wt % to 0.061 wt %; 0.0061 wt % to 0.06 wt %; 0.00611 wt % to 0.059 wt %; 0.00612 wt % to 0.058 wt %; 0.00613 wt % to 0.057 wt %; 0.00614 wt % to 0.056 wt %; 0.00615 wt % to 0.055 wt %; 0.00616 wt % to 0.054 wt %; 0.00617 wt % to 0.053 wt %; 0.00618 wt % to 0.052 wt %; 0.00619 wt % to 0.051 wt %; 0.0062 wt % to 0.05 wt %; 0.00621 wt % to 0.049 wt %; 0.00622 wt % to 0.048 wt %; 0.00623 wt % to 0.047 wt %; 0.00624 wt % to 0.046 wt %; 0.00625 wt % to 0.045 wt %; 0.00626 wt % to 0.044 wt %; 0.00627 wt % to 0.043 wt %; 0.00628 wt % to 0.042 wt %; 0.00629 wt % to 0.041 wt %; 0.0063 wt % to 0.04 wt %; 0.00631 wt % to 0.0399 wt %; 0.00632 wt % to 0.0398 wt %; 0.00633 wt % to 0.0397 wt %; 0.00534 wt % to 0.0396 wt %; 0.00535 wt % to 0.0395 wt %; 0.00536 wt % to 0.0394 wt %; 0.00537 wt % to 0.0393 wt %; 0.00538 wt % to 0.0392 wt %; 0.00539 wt % to 0.0391 wt %; 0.0054 wt % to 0.039 wt %; 0.00541 wt % to 0.0389 wt %; 0.00542 wt % to 0.0388 wt %; 0.00543 wt % to 0.0387 wt %; 0.00544 wt % to 0.0386 wt %; 0.00545 wt % to 0.0385 wt %; 0.00546 wt % to 0.0384 wt %; 0.00547 wt % to 0.0383 wt %; 0.00548 wt % to 0.0382 wt %; 0.00549 wt % to 0.0381 wt %; 0.0055 wt % to 0.038 wt %; 0.00551 wt % to 0.0379 wt %; 0.00552 wt % to 0.0378 wt %; 0.00553 wt % to 0.0377 wt %; 0.00554 wt % to 0.0376 wt %; 0.00555 wt % to 0.0375 wt %; 0.00556 wt % to 0.0374 wt %; 0.00557 wt % to 0.0373 wt %; 0.00558 wt % to 0.0372 wt %; 0.00559 wt % to 0.0371 wt %; 0.0056 wt % to 0.037 wt %; 0.00561 wt % to 0.0369 wt %; 0.00562 wt % to 0.0368 wt %; 0.00563 wt % to 0.0367 wt %; 0.00564 wt % to 0.0366 wt %; 0.00565 wt % to 0.0365 wt %; 0.00566 wt % to 0.0364 wt %; 0.00567 wt % to 0.0363 wt %; 0.00568 wt % to 0.0362 wt %; 0.00569 wt % to 0.0361 wt %; 0.0057 wt % to 0.036 wt %; 0.00571 wt % to 0.0359 wt %; 0.00572 wt % to 0.0358 wt %; 0.00573 wt % to 0.0357 wt %; 0.00574 wt % to 0.0356 wt %; 0.00575 wt % to 0.0355 wt %; 0.00576 wt % to 0.0354 wt %; 0.00577 wt % to 0.0353 wt %; 0.00578 wt % to 0.0352 wt %; 0.00579 wt % to 0.0351 wt %; 0.0058 wt % to 0.035 wt %; 0.00581 wt % to 0.0349 wt %; 0.00582 wt % to 0.0348 wt %; 0.00583 wt % to 0.0347 wt %; 0.00584 wt % to 0.0346 wt %; 0.00585 wt % to 0.0345 wt %; 0.00586 wt % to 0.0344 wt %; 0.00587 wt % to 0.0343 wt %; 0.00588 wt % to 0.0342 wt %; 0.00589 wt % to 0.0341 wt %; 0.0059 wt % to 0.034 wt %; 0.00591 wt % to 0.0339 wt %; 0.00592 wt % to 0.0338 wt %; 0.00593 wt % to 0.0337 wt %; 0.00594 wt % to 0.0336 wt %; 0.00595 wt % to 0.0335 wt %; 0.00596 wt % to 0.0334 wt %; 0.00597 wt % to 0.0333 wt %; 0.00598 wt % to 0.0332 wt %; 0.00599 wt % to 0.0331 wt %; 0.0056 wt % to 0.033 wt %; 0.00561 wt % to 0.0329 wt %; 0.00562 wt % to 0.0328 wt %; 0.00563 wt % to 0.0327 wt %; 0.00564 wt % to 0.0326 wt %; 0.00565 wt % to 0.0325 wt %; 0.00566 wt % to 0.0324 wt %; 0.00567 wt % to 0.0323 wt %; 0.00568 wt % to 0.0322 wt %; 0.00569 wt % to 0.0321 wt %; 0.0057 wt % to 0.032 wt %; 0.00571 wt % to 0.0319 wt %; 0.00572 wt % to 0.0318 wt %; 0.00573 wt % to 0.0317 wt %; 0.00574 wt % to 0.0316 wt %; 0.00575 wt % to 0.0315 wt %; 0.00576 wt % to 0.0314 wt %; 0.00577 wt % to 0.0313 wt %; 0.00578 wt % to 0.0312 wt %; 0.00579 wt % to 0.0311 wt %; 0.0058 wt % to 0.031 wt %; 0.00581 wt % to 0.0309 wt %; 0.00582 wt % to 0.0308 wt %; 0.00583 wt % to 0.0307 wt %; 0.00584 wt % to 0.0306 wt %; 0.00585 wt % to 0.0305 wt %; 0.00586 wt % to 0.0304 wt %; 0.00587 wt % to 0.0303 wt %; 0.00588 wt % to 0.0302 wt %; 0.00589 wt % to 0.0301 wt %; 0.0059 wt % to 0.03 wt %; 0.00591 wt % to 0.0299 wt %; 0.00592 wt % to 0.0298 wt %; 0.00593 wt % to 0.0297 wt %; 0.00594 wt % to 0.0296 wt %; 0.00595 wt % to 0.0295 wt %; 0.00596 wt % to 0.0294 wt %; 0.00597 wt % to 0.0293 wt %; 0.00598 wt % to 0.0292 wt %; 0.00599 wt % to 0.0291 wt %; 0.006 wt % to 0.029 wt %; 0.00601 wt % to 0.0289 wt %; 0.00602 wt % to 0.0288 wt %; 0.00603 wt % to 0.0287 wt %; 0.00604 wt % to 0.0286 wt %; 0.00605 wt % to 0.0285 wt %; 0.00606 wt % to 0.0284 wt %; 0.00607 wt % to 0.0283 wt %; 0.00608 wt % to 0.0282 wt %; 0.00609 wt % to 0.0281 wt %; 0.0061 wt % to 0.028 wt %; 0.00611 wt % to 0.0279 wt %; 0.00612 wt % to 0.0278 wt %; 0.00613 wt % to 0.0277 wt %; 0.00614 wt % to 0.0276 wt %; 0.00615 wt % to 0.0275 wt %; 0.00616 wt % to 0.0274 wt %; 0.00617 wt % to 0.0273 wt %; 0.00618 wt % to 0.0272 wt %; 0.00619 wt % to 0.0271 wt %; 0.0062 wt % to 0.027 wt %; 0.00621 wt % to 0.0269 wt %; 0.00622 wt % to 0.0268 wt %; 0.00623 wt % to 0.0267 wt %; 0.00624 wt % to 0.0266 wt %; 0.00625 wt % to 0.0265 wt %; 0.00626 wt % to 0.0264 wt %; 0.00627 wt % to 0.0263 wt %; 0.00628 wt % to 0.0262 wt %; 0.00629 wt % to 0.0261 wt %; 0.0063 wt % to 0.026 wt %; 0.00631 wt % to 0.0259 wt %).
- Embodiment 80. The formulation according to any one of embodiments 75-76, wherein glutamine is present at a weight percent (wt %) to the formulation of:
  - 0.00584 wt % or greater (e.g., 0.0059; 0.006; 0.007; 0.008; 0.009; 0.01; 0.011; 0.012; 0.013; 0.014; 0.015; 0.016; 0.017; 0.018; 0.019; 0.02; 0.021; 0.022; 0.023; 0.024; 0.025; 0.026; 0.027; 0.028; 0.029; 0.03; 0.031; 0.032; 0.033; 0.034; 0.035; 0.036; 0.037; 0.038; 0.039; 0.04; 0.041; 0.042; 0.043; 0.044; 0.045; 0.046; 0.047; 0.048; 0.049; 0.05; 0.051; 0.052; 0.053; 0.054; 0.055; 0.056; 0.057; 0.058; 0.059; 0.06; 0.061; 0.062; 0.063; 0.064; 0.065; 0.066; 0.067; 0.068; 0.069; 0.07; 0.071; 0.072; 0.073; 0.074; 0.075; 0.076; 0.077; 0.078; 0.079; 0.08; 0.081; 0.082; 0.083; 0.084; 0.085; 0.086; 0.087; 0.088; 0.089; 0.09; 0.091; 0.092; 0.093; 0.094; 0.095; 0.096; 0.097; 0.098; 0.099; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.2; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.3; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.4; 0.41; 0.42; 0.43; 0.44; 0.45; 0.46; 0.47; 0.48; 0.49; 0.5; 0.51; 0.52; 0.53; 0.54; 0.55; 0.56; 0.57; 0.58; 0.59; 0.6);
  - 0.584 wt % or less (e.g., 0.583; 0.582; 0.581; 0.58; 0.579; 0.578; 0.577; 0.576; 0.575; 0.574; 0.573; 0.572; 0.571; 0.57; 0.569; 0.568; 0.567; 0.566; 0.565; 0.564; 0.563; 0.562; 0.561; 0.56; 0.559; 0.558; 0.557; 0.556; 0.554; 0.553; 0.552; 0.551; 0.55; 0.549; 0.548; 0.547; 0.546; 0.545; 0.544; 0.543; 0.542; 0.541; 0.54; 0.539; 0.538; 0.537; 0.536; 0.535; 0.534; 0.533; 0.532; 0.531; 0.53; 0.529; 0.528; 0.527; 0.526; 0.525; 0.524; 0.523; 0.522; 0.521; 0.52; 0.519; 0.518; 0.517; 0.516; 0.515; 0.514; 0.513; 0.512; 0.511; 0.51; 0.509; 0.508; 0.507; 0.506; 0.505; 0.504; 0.503; 0.502; 0.501; 0.5; 0.499; 0.498; 0.497; 0.496; 0.495; 0.494; 0.493; 0.492; 0.491; 0.49; 0.489; 0.488; 0.487; 0.486; 0.485; 0.484; 0.483; 0.482; 0.481; 0.48; 0.479; 0.478; 0.477; 0.476; 0.475; 0.474; 0.473; 0.472; 0.471; 0.47; 0.469; 0.468; 0.467; 0.466; 0.465; 0.464; 0.463; 0.462; 0.461; 0.46; 0.459; 0.458; 0.457; 0.456; 0.455; 0.454; 0.453; 0.452; 0.451; 0.45; 0.449; 0.448; 0.447; 0.446; 0.445; 0.444; 0.443; 0.442; 0.441; 0.44; 0.439; 0.438; 0.437; 0.436; 0.435; 0.434; 0.433; 0.432; 0.431; 0.43; 0.429; 0.428; 0.427; 0.426; 0.425; 0.424; 0.423; 0.422; 0.421; 0.42; 0.419; 0.418; 0.417; 0.416; 0.415; 0.414; 0.413; 0.412; 0.411; 0.41; 0.409; 0.408; 0.407; 0.406; 0.405; 0.404; 0.403; 0.402; 0.401; 0.4; 0.3; 0.2; 0.1; 0.09; 0.08; 0.07; 0.06; 0.05; 0.04; 0.03; 0.02; 0.01; 0.009; 0.008; 0.007; 0.006; 0.005; 0.004; 0.003; 0.002; 0.001); or
  - 0.00584 wt % to 0.584 wt % (e.g., 0.00585 wt % to 0.582 wt %; 0.0059 wt % to 0.58 wt %; 0.0065 wt % to 0.578 wt %; 0.007 wt % to 0.576 wt %; 0.0075 wt % to 0.574 wt %; 0.008 wt % to 0.572 wt %; 0.0085 wt % to 0.57 wt %; 0.009 wt % to 0.568 wt %; 0.0095 wt % to 0.566 wt %; 0.01 wt % to 0.564 wt %; 0.015 wt % to 0.562 wt %; 0.02 wt % to 0.56 wt %; 0.025 wt % to 0.558 wt %; 0.03 wt % to 0.556 wt %; 0.035 wt % to 0.554 wt %; 0.04 wt % to 0.552 wt %; 0.045 wt % to 0.55 wt %; 0.05 wt % to 0.548 wt %; 0.055 wt % to 0.546 wt %; 0.06 wt % to 0.544 wt %; 0.065 wt % to 0.542 wt %; 0.07 wt % to 0.54 wt %; 0.075 wt % to 0.538 wt %; 0.08 wt % to 0.536 wt %; 0.085 wt % to 0.534 wt %; 0.09 wt % to 0.532 wt %; 0.095 wt % to 0.53 wt %; 0.1 wt % to 0.528 wt %; 0.15 wt % to 0.526 wt %; 0.2 wt % to 0.524 wt %; 0.25 wt % to 0.522 wt %; 0.3 wt % to 0.52 wt %; 0.35 wt % to 0.518 wt %; 0.4 wt % to 0.516 wt %; 0.45 wt % to 0.514 wt %; 0.5 wt % to 0.512 wt %).
- Embodiment 81. The formulation according to any one of embodiments 75-76, wherein glycine is present at a weight percent (wt %) to the formulation of:
  - 0.003 wt % or greater (e.g., 0.004; 0.005; 0.006; 0.007; 0.008; 0.009; 0.01; 0.02; 0.03; 0.04; 0.05; 0.06; 0.07; 0.08; 0.09; 0.1; 0.11; 0.12; 0.13; 0.14; 0.15; 0.16; 0.17; 0.18; 0.19; 0.2; 0.21; 0.22; 0.23; 0.24; 0.25; 0.26; 0.27; 0.28; 0.29; 0.3; 0.31; 0.32; 0.33; 0.34; 0.35; 0.36; 0.37; 0.38; 0.39; 0.4);
  - 0.3 wt % or less (e.g., 0.298; 0.296; 0.294; 0.292; 0.29; 0.288; 0.286; 0.284; 0.282; 0.28; 0.278; 0.276; 0.274; 0.272; 0.27; 0.268; 0.266; 0.264; 0.262; 0.26; 0.258; 0.256; 0.254; 0.252; 0.25; 0.248; 0.246; 0.244; 0.242; 0.24; 0.238; 0.236; 0.234; 0.232; 0.23; 0.228; 0.226; 0.224; 0.222; 0.22; 0.218; 0.216; 0.214; 0.212; 0.21; 0.208; 0.206; 0.204; 0.202; 0.2; 0.198; 0.196; 0.194; 0.192; 0.19; 0.188; 0.186; 0.184; 0.182; 0.18; 0.178; 0.176; 0.174; 0.172; 0.17; 0.168; 0.166; 0.164; 0.162; 0.16; 0.158; 0.156; 0.154; 0.152; 0.15; 0.148; 0.146; 0.144; 0.142; 0.14; 0.138; 0.136; 0.134; 0.132; 0.13; 0.128; 0.126; 0.124; 0.122; 0.12; 0.118; 0.116; 0.114; 0.112; 0.11; 0.108; 0.106; 0.104; 0.102; 0.1; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.008; 0.006; 0.004; 0.002); or
  - 0.003 wt % to 0.3 wt % (e.g., 0.0035 wt % to 0.29 wt %; 0.004 wt % to 0.27 wt %; 0.0045 wt % to 0.25 wt %; 0.005 wt % to 0.23 wt %; 0.0055 wt % to 0.21 wt %; 0.006 wt % to 0.19 wt %; 0.0065 wt % to 0.17 wt %; 0.007 wt % to 0.15 wt %; 0.0075 wt % to 0.13 wt %; 0.008 wt % to 0.11 wt %; 0.0085 wt % to 0.099 wt %; 0.009 wt % to 0.097 wt %; 0.0095 wt % to 0.095 wt %; 0.01 wt % to 0.093 wt %; 0.015 wt % to 0.091 wt %; 0.02 wt % to 0.089 wt %; 0.025 wt % to 0.087 wt %; 0.03 wt % to 0.085 wt %; 0.035 wt % to 0.083 wt %; 0.04 wt % to 0.081 wt %; 0.045 wt % to 0.079 wt %; 0.05 wt % to 0.077 wt %; 0.055 wt % to 0.075 wt %; 0.06 wt % to 0.073 wt %; 0.065 wt % to 0.071 wt %).
- Embodiment 82. The formulation according to any one of embodiments 75-76, wherein serine is present at a weight percent (wt %) to the formulation of:
  - 0.0042 wt % or greater (e.g., 0.0044; 0.0046; 0.0048; 0.005; 0.0052; 0.0054; 0.0056; 0.0058; 0.006; 0.0062; 0.0064; 0.0066; 0.0068; 0.007; 0.0072; 0.0074; 0.0076; 0.0078; 0.008; 0.0082; 0.0084; 0.0086; 0.0088; 0.009; 0.0092; 0.0094; 0.0096; 0.0098; 0.01; 0.0102; 0.0104; 0.0106; 0.0108; 0.011; 0.0112; 0.0114; 0.0116; 0.0118; 0.012; 0.0122; 0.0124; 0.0126; 0.0128; 0.013; 0.0132; 0.0134; 0.0136; 0.0138; 0.014; 0.0142; 0.0144; 0.0146; 0.0148; 0.015; 0.0152; 0.0154; 0.0156; 0.0158; 0.016; 0.0162; 0.0164; 0.0166; 0.0168; 0.017; 0.0172; 0.0174; 0.0176; 0.0178; 0.018; 0.0182; 0.0184; 0.0186; 0.0188; 0.019; 0.0192; 0.0194; 0.0196; 0.0198; 0.02; 0.0202; 0.0204; 0.0206; 0.208; 0.021; 0.0212; 0.0214; 0.0216; 0.0218; 0.022; 0.0222; 0.0224; 0.0226; 0.0228; 0.023; 0.0232; 0.0234; 0.0236; 0.0238; 0.024; 0.0242; 0.0244; 0.0246; 0.0248; 0.025; 0.0252; 0.0254; 0.0256; 0.0258; 0.026; 0.0262; 0.0264; 0.0266; 0.0268; 0.027; 0.0272; 0.0274; 0.0276; 0.0278; 0.028; 0.0282; 0.0284; 0.0286; 0.0288; 0.029; 0.0292; 0.0294; 0.0296; 0.0298; 0.03; 0.0302; 0.0304; 0.0306; 0.0308; 0.031; 0.0312; 0.0314; 0.0316; 0.0318; 0.032; 0.0322; 0.0324; 0.0326; 0.0328; 0.033; 0.0332; 0.0334; 0.0336; 0.0338; 0.034; 0.0342; 0.0344; 0.0346; 0.0348; 0.035; 0.0352; 0.0354; 0.0356; 0.0358; 0.036; 0.0362; 0.0364; 0.0366; 0.0368; 0.037; 0.0372; 0.0374; 0.0376; 0.0378; 0.038; 0.0382; 0.0384; 0.0386; 0.0388; 0.039; 0.0392; 0.0394; 0.0396; 0.0398; 0.04; 0.0402; 0.0404; 0.0406; 0.0408; 0.041; 0.0412; 0.0414; 0.0416; 0.0418; 0.042; 0.0422; 0.0424; 0.0426; 0.0428; 0.043; 0.0432; 0.0434; 0.0436; 0.0438; 0.044; 0.0442; 0.0444; 0.0446; 0.0448; 0.045; 0.0452; 0.0454; 0.0456; 0.0458; 0.046; 0.0462; 0.0464; 0.0466; 0.0468; 0.047; 0.0472; 0.0474; 0.0476; 0.0478; 0.048; 0.0482; 0.0484; 0.0486; 0.0488; 0.049; 0.0492; 0.0494; 0.0496; 0.0498; 0.05; 0.0502; 0.0504; 0.0506; 0.0508; 0.051; 0.0512; 0.0514; 0.0516; 0.0518; 0.052; 0.0522; 0.0524; 0.0526; 0.0528; 0.053; 0.0532; 0.0534; 0.0536; 0.0538; 0.054; 0.0542; 0.0544; 0.0546; 0.0548; 0.055; 0.0552; 0.0554; 0.0556; 0.0558; 0.056; 0.0562; 0.0564; 0.0566; 0.0568; 0.057; 0.0572; 0.0574; 0.0576; 0.0578; 0.058; 0.0582; 0.0584; 0.0586; 0.0588; 0.059; 0.0592; 0.0594; 0.0596; 0.0598; 0.06; 0.0602; 0.0604; 0.0606; 0.0608; 0.061; 0.0612; 0.0614; 0.0616; 0.0618; 0.062; 0.0622; 0.0624; 0.0626; 0.0628; 0.063; 0.0632; 0.0634; 0.0636; 0.0638; 0.064; 0.0642; 0.0644; 0.0646; 0.0648; 0.065; 0.0652; 0.0654; 0.0656; 0.0658; 0.066; 0.0662; 0.0664; 0.0666; 0.0668; 0.067; 0.0672; 0.0674; 0.0676; 0.0678; 0.068; 0.0682; 0.0684; 0.0686; 0.0688; 0.069; 0.0692; 0.0694; 0.0696; 0.0698; 0.07; 0.0702; 0.0704; 0.0706; 0.0708; 0.071; 0.0712; 0.0714; 0.0716; 0.0718; 0.072; 0.0722; 0.0724; 0.0726; 0.0728; 0.073; 0.0732; 0.0734; 0.0736; 0.0738; 0.074; 0.0742; 0.0744; 0.0746; 0.0748; 0.075; 0.0752; 0.0754; 0.0756; 0.0758; 0.076; 0.0762; 0.0764; 0.0766; 0.0768; 0.077; 0.0772; 0.0774; 0.0776; 0.0778; 0.078; 0.0782; 0.0784; 0.0786; 0.0788; 0.079; 0.0792; 0.0794; 0.0796; 0.0798; 0.08; 0.0802; 0.0804; 0.0806; 0.0808; 0.081; 0.0812; 0.0814; 0.0816; 0.0818; 0.082; 0.0822; 0.0824; 0.0826; 0.0828; 0.083; 0.0832; 0.0834; 0.0836; 0.0838; 0.084; 0.0842; 0.0844; 0.0846; 0.0848; 0.085; 0.0852; 0.0854; 0.0856; 0.0858; 0.086; 0.0862; 0.0864; 0.0866; 0.0868; 0.087; 0.0872; 0.0874; 0.0876; 0.0878; 0.088; 0.0882; 0.0884; 0.0886; 0.0888; 0.089; 0.0892; 0.0894; 0.0896; 0.0898; 0.09; 0.0902; 0.0904; 0.0906; 0.0908; 0.091; 0.0912; 0.0914; 0.0916; 0.0918; 0.092; 0.0922; 0.0924; 0.0926; 0.0928; 0.093; 0.0932; 0.0934; 0.0936; 0.0938; 0.094; 0.0942; 0.0944; 0.0946; 0.0948; 0.095; 0.0952; 0.0954; 0.0956; 0.0958; 0.096; 0.0962; 0.0964; 0.0966; 0.0968; 0.097; 0.0972; 0.0974; 0.0976; 0.0978; 0.098; 0.0982; 0.0984; 0.0986; 0.0988; 0.099; 0.0992; 0.0994; 0.0996; 0.0998; 0.1; 0.102; 0.104; 0.106; 0.108; 0.11; 0.112; 0.114; 0.116; 0.118; 0.12; 0.122; 0.124; 0.126; 0.128; 0.13; 0.132; 0.134; 0.136; 0.138; 0.14; 0.142; 0.144; 0.146; 0.148; 0.15; 0.152; 0.154; 0.156; 0.158; 0.16; 0.162; 0.164; 0.166; 0.168; 0.17; 0.172; 0.174; 0.176; 0.178; 0.18; 0.182; 0.184; 0.186; 0.188; 0.19; 0.192; 0.194; 0.196; 0.198; 0.2; 0.22; 0.24; 0.26; 0.28; 0.3; 0.32; 0.34; 0.36; 0.38; 0.4; 0.42; 0.44; 0.46; 0.48; 0.5);
  - 0.42 wt % or less (e.g., 0.418; 0.416; 0.414; 0.412; 0.41; 0.408; 0.406; 0.404; 0.402; 0.4; 0.398; 0.396; 0.394; 0.392; 0.39; 0.388; 0.386; 0.384; 0.382; 0.38; 0.378; 0.376; 0.374; 0.372; 0.37; 0.368; 0.366; 0.364; 0.362; 0.36; 0.358; 0.356; 0.354; 0.352; 0.35; 0.348; 0.346; 0.344; 342; 0.34; 0.338; 0.336; 0.334; 0.332; 0.33; 0.328; 0.326; 0.324; 0.322; 0.32; 0.318; 0.316; 0.314; 0.312; 0.31; 0.308; 0.306; 0.304; 0.302; 0.3; 0.298; 0.296; 0.294; 0.292; 0.294; 0.292; 0.29; 0.288; 0.286; 0.284; 0.282; 0.28; 0.278; 0.276; 0.274; 0.272; 0.27; 0.268; 0.266; 0.264; 0.262; 0.26; 0.258; 0.256; 0.254; 0.252; 0.25; 0.248; 0.246; 0.244; 0.242; 0.24; 0.238; 0.236; 0.234; 0.232; 0.23; 0.228; 0.226; 0.224; 0.222; 0.22; 0.218; 0.216; 0.214; 0.212; 0.21; 0.208; 0.206; 0.204; 0.202; 0.2; 0.198; 0.196; 0.194; 0.192; 0.19; 0.188; 0.186; 0.184; 0.182; 0.18; 0.178; 0.176; 0.174; 0.172; 0.17; 0.168; 0.166; 0.164; 0.162; 0.16; 0.158; 0.156; 0.154; 0.152; 0.15; 0.148; 0.146; 0.144; 0.142; 0.14; 0.138; 0.136; 0.134; 0.132; 0.13; 0.128; 0.126; 0.124; 0.122; 0.12; 0.118; 0.116; 0.114; 0.112; 0.11; 0.108; 0.106; 0.104; 0.102; 0.1; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.0088; 0.0086; 0.0084; 0.0082; 0.008; 0.0078; 0.0076; 0.0074; 0.0072; 0.007; 0.0068; 0.0066; 0.0064; 0.0062; 0.006; 0.0058; 0.0056; 0.0054; 0.0052; 0.005; 0.0048; 0.0046; 0.0044; 0.0042; 0.004); or
  - 0.0042 wt % to 0.42 wt % (e.g., 0.0043 wt % to 0.415 wt %; 0.0045 wt % to 0.41 wt %; 0.0047 wt % to 0.405 wt %; 0.0049 wt % to 0.4 wt %; 0.0051 wt % to 0.395 wt %; 0.0053 wt % to 0.39 wt %; 0.0055 wt % to 0.385 wt %; 0.0057 wt % to 0.38 wt %; 0.0059 wt % to 0.375 wt %; 0.0061 wt % to 0.37 wt %; 0.0063 wt % to 0.365 wt %; 0.0065 wt % to 0.36 wt %; 0.0067 wt % to 0.355 wt %; 0.0069 wt % to 0.35 wt %; 0.0061 wt % to 0.345 wt %; 0.0063 wt % to 0.34 wt %; 0.0065 wt % to 0.335 wt %; 0.0067 wt % to 0.33 wt %; 0.0069 wt % to 0.325 wt %; 0.0071 wt % to 0.32 wt %; 0.0073 wt % to 0.315 wt %; 0.0075 wt % to 0.31 wt %; 0.0077 wt % to 0.305 wt %; 0.0079 wt % to 0.3 wt %; 0.0081 wt % to 0.295 wt %; 0.0083 wt % to 0.29 wt %; 0.0085 wt % to 0.285 wt %; 0.0087 wt % to 0.28 wt %; 0.0089 wt % to 0.275 wt %; 0.0091 wt % to 0.27 wt %; 0.0093 wt % to 0.265 wt %; 0.0095 wt % to 0.26 wt %; 0.0097 wt % to 0.255 wt %; 0.0099 wt % to 0.35 wt %; 0.0101 wt % to 0.345 wt %; 0.0103 wt % to 0.34 wt %; 0.0105 wt % to 0.335 wt %; 0.0107 wt % to 0.33 wt %; 0.0109 wt % to 0.325 wt %; 0.0111 wt % to 0.32 wt %; 0.0113 wt % to 0.315 wt %; 0.0115 wt % to 0.31 wt %; 0.0117 wt % to 0.305 wt %; 0.0119 wt % to 0.3 wt %; 0.0121 wt % to 0.295 wt %; 0.0123 wt % to 0.29 wt %; 0.0125 wt % to 0.285 wt %; 0.0127 wt % to 0.28 wt %; 0.0129 wt % to 0.275 wt %; 0.0131 wt % to 0.27 wt %; 0.0133 wt % to 0.265 wt %; 0.0135 wt % to 0.26 wt %; 0.0137 wt % to 0.255 wt %; 0.0139 wt % to 0.25 wt %; 0.0141 wt % to 0.245 wt %; 0.0143 wt % to 0.24 wt %; 0.0145 wt % to 0.235 wt %; 0.0147 wt % to 0.23 wt %; 0.0149 wt % to 0.225 wt %; 0.0151 wt % to 0.22 wt %; 0.0153 wt % to 0.215 wt %; 0.0155 wt % to 0.21 wt %; 0.0157 wt % to 0.205 wt %; 0.0159 wt % to 0.2 wt %; 0.0161 wt % to 0.195 wt %; 0.0163 wt % to 0.19 wt %; 0.0165 wt % to 0.185 wt %; 0.0167 wt % to 0.18 wt %; 0.0169 wt % to 0.175 wt %; 0.0171 wt % to 0.17 wt %; 0.0173 wt % to 0.165 wt %; 0.0175 wt % to 0.16 wt %; 0.0177 wt % to 0.155 wt %; 0.0179 wt % to 0.15 wt %; 0.0181 wt % to 0.145 wt %; 0.0183 wt % to 0.14 wt %; 0.0185 wt % to 0.135 wt %; 0.0187 wt % to 0.13 wt %; 0.0189 wt % to 0.125 wt %; 0.0191 wt % to 0.12 wt %; 0.0193 wt % to 0.115 wt %; 0.0195 wt % to 0.11 wt %; 0.0197 wt % to 0.105 wt %; 0.0199 wt % to 0.1 wt %; 0.0201 wt % to 0.095 wt %; 0.0203 wt % to 0.09 wt %; 0.0205 wt % to 0.085 wt %; 0.0207 wt % to 0.08 wt %; 0.0209 wt % to 0.075 wt %; 0.0211 wt % to 0.07 wt %; 0.0213 wt % to 0.065 wt %; 0.0215 wt % to 0.06 wt %; 0.0217 wt % to 0.055 wt %; 0.0219 wt % to 0.05 wt %; 0.0221 wt % to 0.045 wt %; 0.0223 wt % to 0.04 wt %; 0.0225 wt % to 0.035 wt %; 0.0227 wt % to 0.03 wt %; 0.0229 wt % to 0.025 wt %).
- Embodiment 83. The formulation according to any one of embodiments 75-76, wherein the therapeutically effective amount of a combination of free amino acids or a peptide combination of amino acids comprises (or consists essentially of or consists of):
  - Alanine (Ala),
  - Glutamine (Gln),
  - Glycine (Gly), and
  - Serine (Ser),
  - wherein each amino acid comprises a ratio to each of the other amino acids of:
  - 1:100 [0.1 to 10] or greater (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):1 (Gln):100 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):100 (Gln):1 (Gly):100 (Ser) [0.1 to 10]; 100 (Ala):100 (Gln):100 (Gly):1 (Ser) [0.1 to 10]; 1 (Ala):90 (Gln):90 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):1 (Gln):90 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):90 (Gln):1 (Gly):90 (Ser) [0.1 to 9]; 90 (Ala):90 (Gln):90 (Gly):1 (Ser) [0.1 to 9]; 1 (Ala):80 (Gln):80 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):1 (Gln):80 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):80 (Gln):1 (Gly):80 (Ser) [0.1 to 8]; 80 (Ala):80 (Gln):80 (Gly):1 (Ser) [0.1 to 8]; 1 (Ala):70 (Gln):70 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):1 (Gln):70 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):70 (Gln):1 (Gly):70 (Ser) [0.1 to 7]; 70 (Ala):70 (Gln):70 (Gly):1 (Ser) [0.1 to 7]; 1 (Ala):60 (Gln):60 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):1 (Gln):60 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):60 (Gln):1 (Gly):60 (Ser) [0.1 to 6]; 60 (Ala):60 (Gln):60 (Gly):1 (Ser) [0.1 to 6]; 1 (Ala):50 (Gln):50 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):1 (Gln):50 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):50 (Gln):1 (Gly):50 (Ser) [0.1 to 5]; 50 (Ala):50 (Gln):50 (Gly):1 (Ser) [0.1 to 5]; 1 (Ala):40 (Gln):40 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):1 (Gln):40 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):40 (Gln):1 (Gly):40 (Ser) [0.1 to 4]; 40 (Ala):40 (Gln):40 (Gly):1 (Ser) [0.1 to 4]; 1 (Ala):30 (Gln):30 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):1 (Gln):30 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):30 (Gln):1 (Gly):30 (Ser) [0.1 to 3]; 30 (Ala):30 (Gln):30 (Gly):1 (Ser) [0.1 to 3]; 1 (Ala):20 (Gln):20 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):1 (Gln):20 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):20 (Gln):1 (Gly):20 (Ser) [0.1 to 2]; 20 (Ala):20 (Gln):20 (Gly):1 (Ser) [0.1 to 2]; 1 (Ala):10 (Gln):10 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):1 (Gln):10 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):10 (Gln):1 (Gly):10 (Ser) [0.1 to 1]; 10 (Ala):10 (Gln):10 (Gly):1 (Ser) [0.1 to 1]; 1 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):1 (Gln):5 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):5 (Gln):1 (Gly):5 (Ser) [0.1 to 0.5]; 5 (Ala):5 (Gln):5 (Gly):1 (Ser) [0.1 to 0.5]; 2 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):2 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):2 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):5 (Gly):2 (Ser) [0.1 to 0.25]; 1 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.1 to 0.1]; 2 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):2 (Gln):1 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):1 (Gln):2 (Gly):1 (Ser) [0.2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):2 (Ser) [0.2 to 0.1]; 3 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):3 (Gln):1 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):1 (Gln):3 (Gly):1 (Ser) [0.3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):3 (Ser) [0.3 to 0.1]; 4 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):4 (Gln):1 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):1 (Gln):4 (Gly):1 (Ser) [0.4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):4 (Ser) [0.4 to 0.1]; 1 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.5 to 0.1]; 1 (Ala):5 (Gln):1 (Gly):1 (Ser) [0.5 to 0.1]; 1 (Ala):1 (Gln):5 (Gly):1 (Ser) [0.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):5 (Ser) [0.5 to 0.1]; 6 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):6 (Gln):1 (Gly):1 (Ser) [0.6 to 0.1];
- 1 (Ala):1 (Gln):6 (Gly):1 (Ser) [0.6 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):6 (Ser) [0.6 to 0.1]; 7 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):7 (Gln):1 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):1 (Gln):7 (Gly):1 (Ser) [0.7 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):7 (Ser) [0.7 to 0.1]; 8 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):8 (Gln):1 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):1 (Gln):8 (Gly):1 (Ser) [0.8 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):8 (Ser) [0.8 to 0.1]; 9 (Ala):1 (Gln):1 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):9 (Gln):1 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):1 (Gln):9 (Gly):1 (Ser) [0.9 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):9 (Ser) [0.9 to 0.1]; 10 (Ala):1 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):10 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):10 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):10 (Ser) [1 to 0.1]; 20 (Ala):1 (Gln):1 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):20 (Gln):1 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):1 (Gln):20 (Gly):1 (Ser) [2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):20 (Ser) [2 to 0.1]; 30 (Ala):1 (Gln):1 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):30 (Gln):1 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):1 (Gln):30 (Gly):1 (Ser) [3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):30 (Ser) [3 to 0.1]; 40 (Ala):1 (Gln):1 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):40 (Gln):1 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):1 (Gln):40 (Gly):1 (Ser) [4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):40 (Ser) [4 to 0.1]; 50 (Ala):1 (Gln):1 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):50 (Gln):1 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):1 (Gln):50 (Gly):1 (Ser) [5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):50 (Ser) [5 to 0.1]; 60 (Ala):1 (Gln):1 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):60 (Gln):1 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):1 (Gln):60 (Gly):1 (Ser) [6 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):60 (Ser) [6 to 0.1]; 70 (Ala):1 (Gln):1 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):70 (Gln):1 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):1 (Gln):70 (Gly):1 (Ser) [7 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):70 (Ser) [7 to 0.1]; 80 (Ala):1 (Gln):1 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):80 (Gln):1 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):1 (Gln):80 (Gly):1 (Ser) [8 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):80 (Ser) [8 to 0.1]; 90 (Ala):1 (Gln):1 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):90 (Gln):1 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):1 (Gln):90 (Gly):1 (Ser) [9 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):90 (Ser) [9 to 0.1]; 100 (Ala):1 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):100 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):1 (Gln):100 (Gly):1 (Ser) [10 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):100 (Ser) [10 to 0.1]);
  - 100:1 [10 to 0.1] or less (e.g., 95 (Ala):1 (Gln):1 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):95 (Gln):1 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):1 (Gln):95 (Gly):1 (Ser) [9.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):95 (Ser) [9.5 to 0.1]; 85 (Ala):1 (Gln):1 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):85 (Gln):1 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):1 (Gln):85 (Gly):1 (Ser) [8.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):85 (Ser) [8.5 to 0.1]; 75 (Ala):1 (Gln):1 (Gly):1 (Ser) [7.5 to 0.1]; 1 (Ala):75 (Gln):1 (Gly):1 (Ser) [7.5 to 0.1]; 1 (Ala):1 (Gln):75 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):55 (Ser) [5.5 to 0.1]; 65 (Ala):1 (Gln):1 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):65 (Gln):1 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):1 (Gln):65 (Gly):1 (Ser) [6.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):65 (Ser) [6.5 to 0.1]; 55 (Ala):1 (Gln):1 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):55 (Gln):1 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):55 (Gly):1 (Ser) [5.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):55 (Ser) [5.5 to 0.1]; 45 (Ala):1 (Gln):1 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):45 (Gln):1 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):1 (Gln):45 (Gly):1 (Ser) [4.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):45 (Ser) [4.5 to 0.1]; 35 (Ala):1 (Gln):1 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):35 (Gln):1 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):1 (Gln):35 (Gly):1 (Ser) [3.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):35 (Ser) [3.5 to 0.1]; 25 (Ala):1 (Gln):1 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):25 (Gln):1 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):1 (Gln):25 (Gly):1 (Ser) [2.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):25 (Ser) [0.1 to 0.5]; 15 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):15 (Gln):1 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):1 (Gln):15 (Gly):1 (Ser) [1.5 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):15 (Ser) [1.5 to 0.1]; 14 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):14 (Gln):1 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):1 (Gln):14 (Gly):1 (Ser) [1.4 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):14 (Ser) [1.4 to 0.1]; 13 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):13 (Gln):1 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):1 (Gln):13 (Gly):1 (Ser) [1.3 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):13 (Ser) [1.3 to 0.1]; 12 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):12 (Gln):1 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):1 (Gln):12 (Gly):1 (Ser) [1.2 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):12 (Ser) [1.2 to 0.1]; 11 (Ala):1 (Gln):1 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):11 (Gln):1 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):1 (Gln):11 (Gly):1 (Ser) [1.1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):11 (Ser) [1.1 to 0.1]; 10 (Ala):1 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):10 (Gln):1 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):10 (Gly):1 (Ser) [1 to 0.1]; 1 (Ala):1 (Gln):1 (Gly):10 (Ser) [1 to 0.1]; 19 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):19 (Gln):2 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):2 (Gln):19 (Gly):2 (Ser) [0.95 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):19 (Ser) [0.95 to 0.1]; 17 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):17 (Gln):2 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):2 (Gln):17 (Gly):2 (Ser) [0.85 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):17 (Ser) [0.85 to 0.1]; 15 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):15 (Gln):2 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):2 (Gln):15 (Gly):2 (Ser) [0.75 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):15 (Ser) [0.75 to 0.1]; 13 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):13 (Gln):2 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):2 (Gln):13 (Gly):2 (Ser) [0.65 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):13 (Ser) [0.65 to 0.1]; 11 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):11 (Gln):2 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):2 (Gln):11 (Gly):2 (Ser) [0.55 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):11 (Ser) [0.55 to 0.1]; 9 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):9 (Gln):2 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):2 (Gln):9 (Gly):2 (Ser) [0.45 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):9 (Ser) [0.45 to 0.1]; 7 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):7 (Gln):2 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):2 (Gln):7 (Gly):2 (Ser) [0.35 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):7 (Ser) [0.35 to 0.1]; 5 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):5 (Gln):2 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):2 (Gln):5 (Gly):2 (Ser) [0.25 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):5 (Ser) [0.25 to 0.1]; 3 (Ala):2 (Gln):2 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):3 (Gln):2 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):2 (Gln):3 (Gly):2 (Ser) [0.15 to 0.1]; 2 (Ala):2 (Gln):2 (Gly):3 (Ser) [0.15 to 0.1]; 2 (Ala):3 (Gln):3 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):2 (Gln):3 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):3 (Gln):2 (Gly):3 (Ser) [0.1 to 0.15]; 3 (Ala):3 (Gln):3 (Gly):2 (Ser) [0.1 to 0.15]; 2 (Ala):5 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):2 (Gln):5 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):2 (Gly):5 (Ser) [0.1 to 0.25]; 5 (Ala):5 (Gln):5 (Gly):2 (Ser) [0.1 to 0.25]; 2 (Ala):7 (Gln):7 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):2 (Gln):7 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):7 (Gln):2 (Gly):7 (Ser) [0.1 to 0.35]; 7 (Ala):7 (Gln):7 (Gly):2 (Ser) [0.1 to 0.35]; 2 (Ala):9 (Gln):9 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):2 (Gln):9 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):9 (Gln):2 (Gly):9 (Ser) [0.1 to 0.45]; 9 (Ala):9 (Gln):9 (Gly):2 (Ser) [0.1 to 0.45]; 2 (Ala):11 (Gln):11 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):2 (Gln):11 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):11 (Gln):2 (Gly):11 (Ser) [0.1 to 0.55]; 11 (Ala):11 (Gln):11 (Gly):2 (Ser) [0.1 to 0.55]; 2 (Ala):13 (Gln):13 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):2 (Gln):13 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):13 (Gln):2 (Gly):13 (Ser) [0.1 to 0.65]; 13 (Ala):13 (Gln):13 (Gly):2 (Ser) [0.1 to 0.65]; 2 (Ala):15 (Gln):15 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):2 (Gln):15 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):15 (Gln):2 (Gly):15 (Ser) [0.1 to 0.75]; 15 (Ala):15 (Gln):15 (Gly):2 (Ser) [0.1 to 0.75]; 2 (Ala):17 (Gln):17 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):2 (Gln):17 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):17 (Gln):2 (Gly):17 (Ser) [0.1 to 0.85]; 17 (Ala):17 (Gln):17 (Gly):2 (Ser) [0.1 to 0.85]; 2 (Ala):19 (Gln):19 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):2 (Gln):19 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):19 (Gln):2 (Gly):19 (Ser) [0.1 to 0.95]; 19 (Ala):19 (Gln):19 (Gly):2 (Ser) [0.1 to 0.95]); or from 1:100 to 100:1 (e.g., 1 (Ala):100 (Gln):100 (Gly):100 (Ser) [0.1 to 10] to 100 (Ala):1 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):1 (Gln):100 (Gly):100 (Ser) [0.1 to 10] to 1 (Ala):100 (Gln):1 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):100 (Gln):1 (Gly):100 (Ser) [0.1 to 10] to 1 (Ala):1 (Gln):100 (Gly):1 (Ser) [10 to 0.1]; 100 (Ala):100 (Gln):100 (Gly):1 (Ser) [0.1 to 10] to 1 (Ala):1 (Gln):1 (Gly):100 (Ser) [10 to 0.1]).
- Embodiment 84. The formulation according to any one of embodiments 75-76, wherein the therapeutically effective amount of the retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof) is present at a weight percentage (w/w %) of the formulation of:
  - 0.01 w/w % or greater (e.g., 0.011; 0.013; 0.015; 0.017; 0.019; 0.021; 0.023; 0.025; 0.027; 0.029; 0.031; 0.033; 0.035; 0.037; 0.039; 0.041; 0.043; 0.045; 0.047; 0.049; 0.051; 0.053; 0.055; 0.057; 0.059; 0.061; 0.063; 0.065; 0.067; 0.069; 0.071; 0.073; 0.075; 0.077; 0.079; 0.081; 0.083; 0.085; 0.087; 0.089; 0.091; 0.093; 0.095; 0.097; 0.099; 0.1; 0.15; 0.3; 0.35; 0.5; 0.55; 0.7; 0.75; 0.9; 0.95; 1.1; 1.3; 1.5; 1.7; 1.9; 2.1; 2.3; 2.5; 2.7; 2.9; 3, 1; 3.3; 3.5; 3.7; 3.9; 4.1; 4.3; 4.5; 4.7; 4.9; 5.1; 5.3);
  - 5 w/w % or less (e.g., 4.8; 4.6; 4.4; 4.2; 4; 3.8; 3.6; 3.4; 3.2; 3; 2.8; 2.6; 2.4; 2.2; 2; 1.8; 1.6; 1.4; 1.2; 1; 0.85; 0.8; 0.65; 0.6; 0.45; 0.4; 0.25; 0.2; 0.098; 0.096; 0.094; 0.092; 0.09; 0.088; 0.086; 0.084; 0.082; 0.08; 0.078; 0.076; 0.074; 0.072; 0.07; 0.068; 0.066; 0.064; 0.062; 0.06; 0.058; 0.056; 0.054; 0.052; 0.05; 0.048; 0.046; 0.044; 0.042; 0.04; 0.038; 0.036; 0.034; 0.032; 0.03; 0.028; 0.026; 0.024; 0.022; 0.02; 0.018; 0.016; 0.014; 0.012; 0.01; 0.008; 0.006; 0.004); or
  - 0.01 w/w %-5 w/w % (e.g., 0.011 w/w %-4.9 w/w %; 0.012 w/w %-4.8 w/w %; 0.013 w/w %-4.7 w/w %; 0.014 w/w %-4.6 w/w %; 0.015 w/w %-4.5 w/w %; 0.016 w/w %-4.4 w/w %; 0.017 w/w %-4.3 w/w %; 0.018 w/w %-4.2 w/w %; 0.019 w/w %-4.1 w/w %; 0.02 w/w %-4 w/w %; 0.021 w/w %-3.9 w/w %; 0.022 w/w %-3.8 w/w %; 0.023 w/w %-3.7 w/w %; 0.024 w/w %-3.6 w/w %; 0.025 w/w %-3.5 w/w %; 0.026 w/w %-3.4 w/w %; 0.027 w/w %-3.3 w/w %; 0.028 w/w %-3.2 w/w %; 0.029 w/w %-3.1 w/w %; 0.03 w/w %-3 w/w %; 0.031 w/w %-2.9 w/w %; 0.032 w/w %-2.8 w/w %; 0.033 w/w %-2.7 w/w %; 0.034 w/w %-2.6 w/w %; 0.035 w/w %-2.5 w/w %; 0.036 w/w %-2.4 w/w %; 0.037 w/w %-2.3 w/w %; 0.038 w/w %-2.2 w/w %; 0.039 w/w %-2.1 w/w %; 0.04 w/w %-2 w/w %; 0.041 w/w %-1.9 w/w %; 0.042 w/w %-1.8 w/w %; 0.043 w/w %-1.7 w/w %; 0.044 w/w %-1.6 w/w %; 0.045 w/w %-1.5 w/w %; 0.046 w/w %-1.4 w/w %; 0.047 w/w %-1.3 w/w %; 0.048 w/w %-1.2 w/w %; 0.049 w/w %-1.1 w/w %; 0.05 w/w %-1 w/w %; 0.051 w/w %-0.99 w/w %; 0.052 w/w %-0.98 w/w %; 0.053 w/w %-0.97 w/w %; 0.054 w/w %-0.96 w/w %; 0.055 w/w %-0.95 w/w %; 0.056 w/w %-0.94 w/w %; 0.057 w/w %-0.93 w/w %; 0.058 w/w %-0.92 w/w %; 0.059 w/w %-0.91 w/w %; 0.06 w/w %-0.9 w/w %; 0.061 w/w %-0.89 w/w %; 0.062 w/w %-0.88 w/w %; 0.063 w/w %-0.87 w/w %; 0.064 w/w %-0.86 w/w %; 0.065 w/w %-0.85 w/w %; 0.066 w/w %-0.84 w/w %; 0.067 w/w %-0.83 w/w %; 0.068 w/w %-0.82 w/w %; 0.069 w/w %-0.81 w/w %; 0.07 w/w %-0.8 w/w %; 0.071 w/w %-0.79 w/w %; 0.072 w/w %-0.78 w/w %; 0.073 w/w %-0.77 w/w %; 0.074 w/w %-0.76 w/w %; 0.075 w/w %-0.75 w/w %; 0.076 w/w %-0.74 w/w %; 0.077 w/w %-0.73 w/w %; 0.078 w/w %-0.72 w/w %; 0.079 w/w %-0.71 w/w %; 0.08 w/w %-0.7 w/w %; 0.081 w/w %-0.69 w/w %; 0.082 w/w %-0.68 w/w %; 0.083 w/w %-0.67 w/w %; 0.084 w/w %-0.66 w/w %; 0.085 w/w %-0.65 w/w %; 0.086 w/w %-0.64 w/w %; 0.087 w/w %-0.63 w/w %; 0.088 w/w %-0.62 w/w %; 0.089 w/w %-0.61 w/w %; 0.09 w/w %-0.6 w/w %; 0.091 w/w %-0.59 w/w %; 0.092 w/w %-0.58 w/w %; 0.093 w/w %-0.57 w/w %; 0.094 w/w %-0.56 w/w %; 0.095 w/w %-0.55 w/w %; 0.096 w/w %-0.54 w/w %; 0.097 w/w %-0.53 w/w %; 0.098 w/w %-0.52 w/w %; 0.099 w/w %-0.51 w/w %; 0.1 w/w %-0.5 w/w %; 0.101 w/w %-0.49 w/w %; 0.102 w/w %-0.48 w/w %; 0.103 w/w %-0.47 w/w %; 0.104 w/w %-0.46 w/w %; 0.105 w/w %-0.45 w/w %; 0.106 w/w %-0.44 w/w %; 0.107 w/w %-0.43 w/w %; 0.108 w/w %-0.42 w/w %; 0.109 w/w %-0.41 w/w %; 0.110 w/w %-0.4 w/w %; 0.111 w/w %-0.39 w/w %; 0.112 w/w %-0.38 w/w %; 0.113 w/w %-0.37 w/w %; 0.114 w/w %-0.36 w/w %; 0.115 w/w %-0.35 w/w %; 0.116 w/w %-0.34 w/w %; 0.117 w/w %-0.33 w/w %; 0.118 w/w %-0.32 w/w %; 0.119 w/w %-0.31 w/w %; 0.120 w/w %-0.3 w/w %; 0.121 w/w %-0.29 w/w %; 0.122 w/w %-0.28 w/w %; 0.123 w/w %-0.27 w/w %; 0.124 w/w %-0.26 w/w %; 0.125 w/w %-0.25 w/w %; 0.126 w/w %-0.24 w/w %; 0.127 w/w %-0.23 w/w %; 0.128 w/w %-0.22 w/w %; 0.129 w/w %-0.21 w/w %; 0.130 w/w %-0.2 w/w %; 0.0131 w/w %-0.19 w/w %; 0.132 w/w %-0.18 w/w %; 0.133 w/w %-0.17 w/w %; 0.134 w/w %-0.16 w/w %; 0.135 w/w %-0.15 w/w %; 0.136 w/w %-0.14 w/w %; 0.137 w/w %-0.139 w/w %).
- Embodiment 85. The formulation according to any one of embodiments 75-76, wherein the formulation or the topical formulation comprises, consists essentially of, or consists of: as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of) an amino acid combination of at least one of alanine, glutamine, glycine, and serine; and a therapeutically effective amount of a retinoid, wherein the amino acid combination and the retinoid are in a ratio of: 1:100 or greater (e.g., 2:100; 4:100; 6:100; 8:100; 10:100; 12:100; 14:100; 16:100; 18:100; 20:100; 22:100; 24:100; 26:100; 28:100; 30:100; 32:100; 34:100; 36:100; 38:100; 40:100; 42:100; 44:100; 46:100; 48:100; 50:100; 52:100; 54:100; 56:100; 58:100; 60:100; 62:100; 64:100; 66:100; 68:100; 70:100; 72:100; 74:100; 76:100; 78:100; 80:100; 82:100; 84:100; 86:100; 88:100; 90:100; 92:100; 94:100; 96:100; 98:100; 100:100); 100:1 or less (e.g., 99:100; 97:100; 95:100; 93:100; 91:100; 89:100; 87:100; 85:100; 83:100; 81:100; 79:100; 77:100; 75:100; 73:100; 71:100; 69:100; 67:100; 65:100; 63:100; 61:100; 59:100; 57:100; 55:100; 53:100; 51:100; 49:100; 47:100; 45:100; 43:100; 41:100; 39:100; 37:100; 35:100; 33:100; 31:100; 29:100; 27:100; 25:100; 23:100; 21:100; 19:100; 17:100; 15:100; 13:100; 11:100; 9:100; 7:100; 5:100; 3:100; 1:100); 1:100 to 100:1 (e.g., 1:99 to 99:1; 1:98 to 98:1; 1:97 to 97:1; 1:96 to 96:1; 1:95 to 95:1; 1:94 to 94:1; 1:93 to 93:1; 1:92 to 92:1; 1:91 to 91:1; 1:90 to 90:1; 1:89 to 89:1; 1:88 to 88:1; 1:87 to 87:1; 1:86 to 86:1; 1:85 to 85:1; 1:84 to 84:1; 1:83 to 83:1; 1:82 to 82:1; 1:81 to 81:1; 1:80 to 80:1; 1:79 to 79:1; 1:78 to 78:1; 1:77 to 77:1; 1:76 to 76:1; 1:75 to 75:1; 1:74 to 74:1; 1:73 to 73:1; 1:72 to 72:1; 1:71 to 71:1; 1:70 to 70:1; 1:69 to 69:1; 1:68 to 68:1; 1:67 to 67:1; 1:66 to 66:1; 1:65 to 65:1; 1:64 to 64:1; 1:63 to 63:1; 1:62 to 62:1; 1:61 to 61:1; 1:60 to 60:1; 1:59 to 59:1; 1:58 to 58:1; 1:57 to 57:1; 1:56 to 56:1; 1:55 to 55:1; 1:54 to 54:1; 1:53 to 53:1; 1:52 to 52:1; 1:51 to 51:1; 1:50 to 50:1; 1:49 to 49:1; 1:48 to 48:1; 1:47 to 47:1; 1:46 to 46:1; 1:45 to 45:1; 1:44 to 44:1; 1:43 to 43:1; 1:42 to 42:1; 1:41 to 41:1; 1:40 to 40:1; 1:39 to 39:1; 1:38 to 38:1; 1:37 to 37:1; 1:36 to 36:1; 1:35 to 35:1; 1:34 to 34:1; 1:33 to 33:1; 1:32 to 32:1; 1:31 to 31:1; 1:30 to 30:1; 1:29 to 29:1; 1:28 to 28:1; 1:27 to 27:1; 1:26 to 26:1; 1:25 to 25:1; 1:24 to 24:1; 1:23 to 23:1; 1:22 to 22:1; 1:21 to 21:1; 1:20 to 20:1; 1:19 to 19:1; 1:18 to 18:1; 1:17 to 17:1; 1:16 to 16:1; 1:15 to 15:1; 1:14 to 14:1; 1:13 to 13:1; 1:12 to 12:1; 1:11 to 11:1; 1:10 to 10:1; 1:9 to 9:1; 1:8 to 8:1; 1:7 to 7:1; 1:6 to 6:1; 1:5 to 5:1; 1:4 to 4:1; 1:3 to 3:1; 1:2 to 2:1).
- Embodiment 86. The formulation according to any one of embodiments 75-76, wherein the formulation or the topical formulation described here further comprises, consists essentially of, or consists of a dermatologically acceptable vehicle and/or additive.
- Embodiment 87. The formulation according to any one of embodiments 75-76, wherein the dermatologically acceptable vehicle and/or additive can be selected from the group consisting of: water, sodium phytate, xanthan gum, kaolin, glycerin, polyglycerides (e.g., oleic polyglycerides, linoleic polyglycerides, linolenic polyglycerides), triglycerides (e.g., caprylic triglycerides, capric triglycerides), sucrose (e.g., sucrose stearate, sucrose distearate), alcohol (e.g., behenyl alcohol, benzoic alcohol, benzyl alcohol, cetearyl alcohol, oleyl alcohol), cetearyl glucoside, sodium stearoyl lactylate, Cera alba (beeswax), Butyrospermum parkii (shea butter), Prunus amigdalus sulcis (sweet almond oil), Simmondsia chinenesis (jojoba oil) phenoxyethanol, lecithin, tocopherol, tocopherol acetate, ascorbyl palmitate, citric acid, potassium sorbate, lactic acid, UV protectant, hydrogenated olive oil decyl esters, decyl cocaote, astrocaryum, murumuru seed butter, Zanthoxylum bungeanum fruit extract, squalene, pentylene glycol, dehydroacetic acid, tamarinds indica seed polysaccharide, sodium benzoate, curcumin. Zanthoxylum bungeanume fruit extract, Passiflora edulis seed oil, cetearyl ethylhexanoate, dimethicone, isopropyl myristate, acrylates/C10-C30 alkyl acrylate crosspolymer, allantoin, sodium phytate, PEG-90 stearate/glyceryl stearate; menthol, sodium hydroxide, sodium benzoate, dehydroacetic acid, benzoic alcohol, hexyldecanol, cetaryl ethylhexanoate, dimethicone, cetearyl ethylhexanoate, PEG-90 stearate/glyceryl stearate, ascorbyl palmitate, citric acid, Zanthoxylum bungeanum fruit extract, allantoin, disodium EDTA, phenoxyethanol, dehydroacetic acid, allantoin, XILOGEL®, Vaccinium myrtillus fruit extract, Oryza sativa (e.g. rice bran oil), isoamyl laurate, polymethyl methacrylate, fragrance, nutrients, antioxidants, water, or combinations thereof,
- Embodiment 88. The formulation according to any one of embodiments 75-76, wherein the formulation or the topical formulation is prepared by:
  - mixing and/or stirring (concurrently, simultaneously, sequentially):
    - (a) a therapeutically effective amount of a combination of amino acids comprising, consisting essentially of at least one of alanine, glutamine, glycine, and serine;
    - (b) a therapeutically effective amount of a retinoid; and
    - optionally, (c) a dermatologically acceptable vehicle and/or additive;
  - heating (a), (b), and optionally (c) (either together or separately) to a temperature sufficient to completely mix, homogenize, or emulsify (a), (b), and optionally (c),
    - wherein the temperature is 20° C. or greater (e.g., 22; 24; 26; 28; 30; 32; 34; 36; 38; 40; 42; 44; 46; 48; 50; 52; 54; 56; 58; 60; 62; 64; 66; 68; 70; 72; 74; 76; 78; 80; 82; 84; 86; 88; 90; 92; 94; 96; 98; 100; 102; 104; 106; 108; 110); 100° C. or less (e.g., 99; 97; 95; 93; 91; 89; 87; 85; 83; 81; 79; 77; 75; 73; 71; 69; 67; 65; 63; 61; 59; 57; 55; 53; 51; 49; 47; 45; 43; 41; 39; 37; 35; 33; 31; 29; 27; 25; 23; 21; 19; 17); or 20° C.-100° C. (e.g., 21° C.-99° C.; 22° C.-98° C.; 23° C.-97° C.; 24° C.-96° C.; 25° C.-95° C.; 26° C.-94° C.; 27° C.-93° C.; 28° C.-92° C.; 29° C.-91° C.; 30° C.-90° C.; 31° C.-89° C.; 32° C.-88° C.; 33° C.-87° C.; 34° C.-86° C.; 35° C.-85° C.; 36° C.-84° C.; 37° C.-83° C.; 38° C.-82° C.; 39° C.-81° C.; 40° C.-80° C.; 41° C.-79° C.; 42° C.-78° C.; 43° C.-77° C.; 44° C.-76° C.; 45° C.-75° C.; 46° C.-74° C.; 47° C.-73° C.; 48° C.-72° C.; 49° C.-71° C.; 50° C.-70° C.; 51° C.-69° C.; 52° C.-68° C.; 53° C.-67° C.; 54° C.-66° C.; 55° C.-65° C.; 56° C.-64° C.; 57° C.-63° C.; 58° C.-62° C.; 59° C.-61° C., for a time sufficient to completely mix, homogenize, or emulsify (e.g., seconds to minutes).
- Embodiment 89. A formulation, comprising (or consisting essentially of or consisting of): as free amino acids or a peptide combination of amino acids, a therapeutically effective amount of:
  - (a) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine; or salts thereof, or any combination thereof); or
  - (b) at least one of: alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and a therapeutically effective amount of a retinoid (e.g., a retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof); and
  - optionally, a pharmaceutically acceptable vehicle and/or additive,
  - wherein the formulation is an injectable formulation.
- Embodiment 90. The formulation of embodiment 89, wherein the therapeutically effective amount of free amino acids or peptide combinations of amino acids is 0.05 mM or greater; 20 mM or less; or 0.05 mM to 20 mM.
- Embodiment 91. The formulation of embodiment 89, wherein the therapeutically effective amount of the retinoid comprises 0.01% or greater by weight of the formulation; 10% or less by weight of the formulation; or 0.01%-10% by weight of the formulation.
- Embodiment 92. A method of improving skin regeneration, including skin barrier repair of skin of a subject, comprises (or consists essentially of or consists of):
  - administering a therapeutically effective amount of a topical formulation (according to any one of the preceding formulations or embodiments 57-59), comprising (or consisting essentially of or consisting of):
    - a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising (or consisting essentially of or consisting of) at least one of alanine, glutamine, glycine, and serine, or salts thereof; and
    - a therapeutically effective amount of a plant extract combination comprising (consisting essentially of or consisting of) (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica): Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense) alone or optionally in combination with at least one of:
      - a texturing and/or a bulking agent (e.g., cellulose (microcrystalline));
      - a phospholipid (e.g., lecithin or phosphatidylcholine); and
      - an anti-caking agent (e.g., silica, adsorbs water in hygroscopic applications);
    - and optionally, a dermatologically acceptable vehicle and/or additive,
- wherein, when tested by a method detecting expression of a skin regeneration, including skin barrier, marker gene (e.g., EGF) is increased by a 1.5-40 average fold change in formulation treated skin as compared to untreated skin, thereby improving skin regeneration, including skin barrier repair.
- Embodiment 93. A method of improving skin moisture of skin of a subject, comprises (or consists essentially of or consists of):
  - administering a therapeutically effective amount of a topical formulation (according to any one of the preceding formulations or embodiments 75-76), comprising (or consisting essentially of or consisting of):
    - a therapeutically effective amount of a combination of free amino acids or a peptide combination thereof comprising (or consisting essentially of or consisting of) at least one of alanine, glutamine, glycine, serine, or salts thereof, or any combination thereof; and
    - a therapeutically effective amount of a retinoid (e.g., retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof); and optionally, a dermatologically acceptable vehicle and/or additive,
  - wherein, when tested by a method detecting expression of a skin moisture marker gene or an HA marker gene, e.g., HAS1, HAS2, HAS3, or EGF, expression is increased by a 1.4-450 fold change in formulation treated skin as compared to untreated skin, thereby improving skin moisture (e.g., increasing skin moisture, decreasing loss of moisture, increasing hyaluronic acid (HA), increasing hyaluronic acid synthase (HAS; e.g., HAS1 (in fibroblasts), HAS2 and HAS3 (in keratinocytes))).
- Embodiment 94. A method of treating a subject suffering from osteoarthritis, wherein the method comprises (or consists essentially of or consists of):
  - administering at a joint of a subject suffering from osteoarthritis (e.g., joints of the hands, hips, or knees) by injecting in the subject a therapeutically effective amount of a formulation comprising (or consisting essentially of or consisting of):
    - as free amino acids or a peptide combination, a therapeutically effective amount of a combination comprising (or consisting essentially of or consisting of):
      - (a) alanine, glutamine, glycine, and serine, or salts thereof; and optionally, a therapeutically effective amount of at least one of: arginine, isoleucine, threonine, tryptophan, or valine (e.g., alanine, glutamine, glycine, and serine and; arginine or isoleucine or threonine or tryptophan or valine; arginine and isoleucine; arginine and threonine; arginine and tryptophan; arginine and valine; isoleucine and threonine; isoleucine and tryptophan; isoleucine and valine; threonine and tryptophan; threonine and valine; tryptophan and valine; arginine, isoleucine, and threonine; arginine, isoleucine, and tryptophan; arginine, isoleucine, and valine; arginine, threonine, and tryptophan; arginine, threonine, and valine; arginine, tryptophan, and valine; isoleucine, threonine, and tryptophan; isoleucine, threonine, and valine; isoleucine, tryptophan, and valine; threonine, tryptophan, and valine; arginine, isoleucine, threonine, and tryptophan; arginine, isoleucine, threonine, and valine; arginine, threonine, tryptophan, and valine; isoleucine, threonine, tryptophan, and valine; arginine, isoleucine, tryptophan, and valine; arginine, isoleucine, threonine, tryptophan, and valine), or any combination thereof; or
      - (b) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; or
      - (c) (i) at least one of alanine, glutamine, glycine, and serine (e.g., alanine; glutamine; glycine; serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine and glutamine and glycine; glutamine and glycine and serine; alanine and glycine and serine; alanine and glutamine and serine; and alanine and glutamine and glycine and serine), or salts thereof; and
      - (ii) a therapeutically effective amount of a retinoid comprising (or consisting essentially of, or consisting of): at least one of retinyl ester (e.g., retinyl palmitate, retinyl oleate, retinyl stearate, retinyl linoleate, retinyl palmitoleate); retinol or vitamin A; retinaldehyde or retinal; retinoic acid; or combinations thereof; and
    - optionally, a pharmaceutically acceptable vehicle and/or additive,
  - thereby treating the subject suffering from osteoarthritis.
- Embodiment 95. A formulation comprising:
  - as free amino acids, a combination of free amino acids, or a peptide combination of amino acids, a therapeutically effective amount comprising, consisting essentially of, or consisting of one or more of: alanine, arginine, glutamine, glycine, isoleucine, serine, threonine, tryptophan, and valine, or derivatives thereof, or salts thereof;
    - wherein the therapeutically effective amount improves skin barrier integrity, improves skin moisture, treats or ameliorates osteoarthritis or its symptoms thereof, or any combination thereof, when tested by a method detecting expression of a barrier marker gene (e.g., EGF, FGF2, PDGF, PPARD, FLG, MKI67, CPT2, TGM4) and determining that the barrier marker gene is modulated by a 1-7 average fold change of formulation-treated cells as compared to untreated cells; and
  - optionally, a dermatologically acceptable or pharmaceutically acceptable carrier;
- wherein the formulation improves barrier integrity of the skin, improves skin moisture, or treats or ameliorates musculoskeletal diseases or disorders (e.g., osteoarthritis), or its symptoms thereof.

As various changes can be made in the above-described subject matter without departing from the scope and spirit of the present disclosure, it is intended that all subject matter contained in the above description, or defined in the appended embodiments, be interpreted as descriptive and illustrative of the present disclosure. Many modifications and variations of the present disclosure are possible in light of the above teachings. Accordingly, the present description is intended to embrace all such alternatives, modifications and variances which fall within the scope of the appended embodiments.

All documents cited or referenced herein and all documents cited or referenced in the herein cited documents, together with any manufacturer's instructions, descriptions, product specifications, and product sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated by reference, and may be employed in the practice of the disclosure.

FORMULATIONS AND METHODS FOR SKIN REGENERATION

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