Formulations and methods for skin care

Abstract

Described herein are cosmetic and pharmaceutical preparations that contain specific amino acid blends. Such preparations strengthen and enhance the barrier function of skin epithelial cells, thereby reducing water loss from the skin, restoring the structural integrity of the skin, epithelial differentiation, cell-cell adhesion, and intracellular signaling, and / or promoting the maintenance of these characteristics. Additional preparations described herein contain a therapeutically effective amount of free amino acids and a therapeutically effective amount of a plant extract for repairing the skin barrier. Methods for administering these preparations for cosmetic and / or therapeutic purposes, and their use for the same, for example, in the preparation of medicaments, are also described herein.

Description

[Technical field]

[0001] The present disclosure is directed to formulations and methods of their use in improving skin barrier repair. [Background technology]

[0002] Skin diseases and conditions can cause considerable physical discomfort and mental distress to patients suffering from such diseases and conditions. Common skin diseases and conditions include atopic dermatitis, psoriasis, and ichthyosis. Xeroderma (dry skin) is a very common skin disease that afflicts a significant portion of the population. The skin diseases, conditions, and disorders that afflict patients often vary with age and are influenced by lifestyle and geographical region. Despite the high prevalence of skin diseases, conditions, and disorders in the population and their adverse impact on people's quality of life and associated economic burden, there are few effective treatments available. There is an unmet need for effective formulations to address cosmetic skin care objectives and for treatment against skin diseases, conditions, and disorders. Summary of the Invention

[0003] Cosmetic and pharmaceutical formulations are described herein. Such formulations strengthen and enhance the barrier function of skin epithelial cells, thereby reducing water loss from the skin, restoring the structural integrity, cell-cell adhesion, and biochemistry of the skin, and / or promoting the maintenance of these characteristics. Described herein are formulations that comprise, consist essentially of, or consist of a therapeutically effective amount of the free amino acids alanine, glutamine, glycine, and serine, and optionally a therapeutically effective amount of at least one additional free amino acid valine, isoleucine, arginine, threonine, or tryptophan, or any combination thereof. Also included herein are methods for maintaining and / or improving the barrier integrity of skin cells in a subject in need thereof or for improving the cosmetic skin condition of a human, comprising administering the formulations described herein to the subject. Also included herein are uses of the formulations described herein for maintaining and / or improving the barrier integrity of skin cells in a subject in need thereof or for improving the cosmetic skin condition of a human, comprising administering the formulations described herein to the subject or human. The formulations described herein may be used to prepare a dermatological medicine for treating a cosmetic skin condition in a subject in need thereof. Such methods, uses, and dermatological medicines are administered or used to improve a cosmetic skin condition. In some embodiments, the cosmetic skin condition is associated with skin aging. The cosmetic skin condition may be associated with at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin. Thus, improving at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin reduces the length, depth, and / or other size of lines and / or wrinkles in the skin. Also included herein are methods for treating and / or preventing a skin disease, condition, or disorder in a subject in need thereof, comprising administering the formulations described herein to the subject.Also included is the use of the formulations described herein for the treatment and / or prevention of a skin disease, condition, or disorder in a subject in need thereof, comprising administering to the subject a formulation described herein. The formulations described herein may be used in the preparation of a medicament for the treatment and / or prevention of a skin disease, condition, or disorder in a subject in need thereof.

[0004] Embodiment 1. A formulation for administration to the skin comprising, as free amino acids, a therapeutically effective amount of a blend of alanine, glutamine, glycine, and serine, or salts thereof, and optionally a therapeutically effective amount of at least one additional free amino acid valine, isoleucine, arginine, threonine, or tryptophan, or salts thereof, or any combination thereof, wherein said therapeutically effective amount of the blend of glutamine, glycine, alanine, and serine, or salts thereof, and said therapeutically effective amount of said at least one additional free amino acid or salt thereof improves barrier integrity of skin cells, optionally comprising a dermatologically acceptable carrier, and wherein the barrier integrity of skin cells is improved when tested by a method detecting expression of a barrier marker gene (e.g., CPT2, EGF, FGF2, FLG, MKI67, PDGF, PPARD, TGM4) to determine that said barrier marker gene is modulated with an average fold change of 1.5 to 7 for amino acid treated skin compared to amino acid untreated skin.

[0005] Embodiment 2. The formulation of embodiment 1, wherein the free 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 isoleucine, arginine, threonine, or tryptophan, or salts thereof, or any combination thereof.

[0006] Embodiment 3. The formulation of embodiment 1 or embodiment 2, wherein the free 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 arginine, or threonine, or salts thereof, or any combination thereof.

[0007] Embodiment 4. The formulation of embodiment 1 or embodiment 2, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine, or salts thereof.

[0008] Embodiment 5. The formulation of embodiment 1 or embodiment 2, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine, or salts thereof.

[0009] Embodiment 6. The formulation of any one of embodiments 1-3, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and arginine, or salts thereof.

[0010] Embodiment 7. The formulation of any one of embodiments 1-3, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and threonine, or salts thereof.

[0011] Embodiment 8. The formulation of embodiment 1, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, and serine, or salts thereof, and at least one additional amino acid isoleucine, arginine, threonine, or tryptophan, or salts thereof, or any combination thereof.

[0012] Embodiment 9. The formulation of embodiment 8, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine, or salts thereof.

[0013] Embodiment 10. The formulation of embodiment 8, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine, or salts thereof.

[0014] Embodiment 11. The formulation of embodiment 8, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine, or salts thereof.

[0015] Embodiment 15. The formulation of embodiment 8, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan, or salts thereof.

[0016] Embodiment 16. The formulation of embodiment 8, wherein the free 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 arginine, or threonine, or tryptophan, or salts thereof, or any combination thereof.

[0017] Embodiment 17. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and arginine, or salts thereof.

[0018] Embodiment 18. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and threonine, or salts thereof.

[0019] Embodiment 19. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and tryptophan, or salts thereof.

[0020] Embodiment 20. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine, or salts thereof.

[0021] Embodiment 21. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and tryptophan, or salts thereof.

[0022] Embodiment 22. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, threonine, and tryptophan, or salts thereof.

[0023] Embodiment 23. The formulation of embodiment 16, wherein the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, threonine, and tryptophan, or salts thereof.

[0024] Embodiment 24. The formulation of embodiment 8, wherein the free 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 isoleucine, threonine, or tryptophan, or salts thereof, or any combination thereof.

[0025] Embodiment 25. The formulation of embodiment 8, wherein the free 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 isoleucine, arginine, or tryptophan, or salts thereof, or any combination thereof.

[0026] Embodiment 26. The formulation of embodiment 8, wherein the free 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 isoleucine, arginine, or threonine, or salts thereof, or any combination thereof.

[0027] Embodiment 27. A formulation according to any one of embodiments 1 to 26, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.05 mM to 20 mM.

[0028] Embodiment 28. A formulation according to any one of embodiments 1 to 27, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.05 mM to 10 mM.

[0029] Embodiment 29. A formulation according to any one of embodiments 1 to 28, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.05 mM to 5 mM.

[0030] Embodiment 30. A formulation according to any one of embodiments 1 to 26, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.1 mM to 10 mM.

[0031] Embodiment 31. A formulation according to any one of embodiments 1 to 26, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.1 mM to 5 mM.

[0032] Embodiment 32. A formulation according to any one of embodiments 1 to 11, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.1 mM to 4 mM or less.

[0033] Embodiment 33. A formulation according to any one of embodiments 1 to 26, wherein each of the free amino acids or salts thereof is present at a concentration ranging from 0.5 mM to 4 mM.

[0034] Embodiment 34. A formulation according to any one of embodiments 1 to 33, wherein, if present, the free amino acid cysteine, or a salt thereof, is present at a concentration of 1 mM or less; if present, the free amino acid histidine, or a salt thereof, is present at a concentration of 0.5 mM or less; if present, the free amino acid tyrosine, or a salt thereof, is present at a concentration of 1 mM or less; if present, the free amino acid leucine, or a salt thereof, is present at a concentration of 4 mM or less; or if present, the free amino acid aspartic acid, or a salt thereof, is present at a concentration of 2 mM or less; or any combination thereof.

[0035] Embodiment 35. A formulation according to any one of embodiments 1 to 34, wherein the formulation does not contain the free amino acids cysteine, histidine, tyrosine, leucine, aspartic acid, taurine, taurate, or glutamic acid, or salts thereof, or any combination thereof.

[0036] Embodiment 36. The formulation of any one of embodiments 1 to 35, wherein the formulation is administered to the skin or can be administered transdermally, subcutaneously, or topically.

[0037] Embodiment 37. A method for maintaining and / or improving skin cell barrier integrity in a subject in need thereof, comprising administering to the skin of said subject a formulation according to any one of embodiments 1 to 36.

[0038] Embodiment 38 The method of embodiment 37, wherein the formulation is administered to improve a cosmetic skin condition.

[0039] Embodiment 39. The method of embodiment 38, wherein the cosmetic skin condition is associated with skin aging.

[0040] Embodiment 40. The method of any one of embodiments 37-39, wherein the formulation is administered to improve at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics, of the skin.

[0041] Embodiment 41. The method of embodiment 40, wherein improving at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin reduces the length, depth, and / or other size of lines and / or wrinkles in the skin.

[0042] Embodiment 42. The method of embodiment 37, wherein the subject in need thereof is suffering from a skin condition.

[0043] Embodiment 43 The method of embodiment 42, wherein the skin condition comprises atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, wounds, or burns.

[0044] Embodiment 44. The method of any one of embodiments 37 to 43, wherein the subject is a human.

[0045] Embodiment 45. A method for improving the cosmetic skin condition of a human, comprising administering to the skin of the human a formulation described in any one of embodiments 1 to 36 in an amount effective to improve at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin.

[0046] Embodiment 46 The method of embodiment 45, wherein the cosmetic skin condition is associated with skin aging.

[0047] Embodiment 47. The method of embodiment 45 or embodiment 46, wherein improving at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin reduces the length, depth, and / or other size of lines and / or wrinkles in the skin.

[0048] Embodiment 48. Use of a formulation according to any one of embodiments 1 to 36 for the treatment of a cosmetic skin condition.

[0049] Embodiment 49. The use according to embodiment 48, wherein the cosmetic skin condition is associated with skin aging.

[0050] Embodiment 50. The use of embodiment 48 or embodiment 49, wherein the formulation is administered to improve at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics, of the skin.

[0051] Embodiment 51. The method of embodiment 50, wherein improving at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin reduces the length, depth, and / or other size of lines and / or wrinkles in the skin.

[0052] Embodiment 52. Use of a formulation according to any one of embodiments 1 to 36 for the treatment of a skin condition.

[0053] Embodiment 53. The use of embodiment 52, wherein the skin condition comprises atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, wounds, or burns.

[0054] Embodiment 58. Use of a formulation according to any one of embodiments 1 to 36 in the preparation of a dermatological medicine for treating a cosmetic skin condition in a subject in need thereof.

[0055] Embodiment 59. The dermatological drug product of embodiment 58, wherein the cosmetic skin condition is associated with skin aging.

[0056] Embodiment 60. The dermatological agent of embodiment 58 or embodiment 59, wherein the dermatological agent is capable of being administered to improve at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics, of the skin.

[0057] Embodiment 61. The dermatological pharmaceutical of embodiment 60, wherein improving at least one visual characteristic, or at least one tactile characteristic, or a combination of visual and tactile characteristics of the skin reduces the length, depth, and / or other size of lines and / or wrinkles in the skin.

[0058] Embodiment 62. Use of a formulation according to any one of embodiments 1 to 36 in the preparation of a medicament for treating a skin condition in a subject in need thereof.

[0059] Embodiment 63. The pharmaceutical composition of embodiment 62, wherein the skin condition comprises atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, wounds, or burns.

[0060] In some embodiments, the topical formulation comprises (or consists essentially of, or consists of) a therapeutically effective amount of a formulation comprising (or consisting essentially of, or consisting of) at least one of alanine, glutamine, glycine, and serine as free amino acids or salts thereof; a therapeutically effective amount of a plant extract comprising (or consisting essentially of, or consisting of) a plant extract such as Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense); and, optionally, a dermatologically acceptable carrier and / or additive.

[0061] In an additional embodiment, a topical formulation is provided comprising (or consisting essentially of, or consisting of) a therapeutically effective amount of a formulation comprising (or consisting essentially of, or consisting of) as free amino acids alanine, glutamine, glycine, and serine, or salts thereof; a therapeutically effective amount of a plant extract formulation (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense); texturing agents and / or fillers (e.g., cellulose (microcrystalline)); phospholipids (e.g., lecithin or phosphatidylcholine, ceramides); and anti-caking agents (e.g., silica, absorbs water in hygroscopic applications); and optionally, a dermatologically acceptable carrier and / or additive is provided.

[0062] In further embodiments, the formulation for administration to the skin comprises a therapeutically effective amount of a blend of free amino acids comprising (or consisting essentially of, or consisting of) alanine, glutamine, glycine, and serine, or salts thereof; a therapeutically effective amount of a blend of plant extracts (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); texturing agents and / or fillers (e.g., cellulose (microcrystalline); phospholipids (e.g., lecithin); and anti-caking agents (e.g., silica, absorbs water in hygroscopic applications); and optionally a dermatologically acceptable carrier, comprising (or consisting essentially of, or consisting of) a plant extract formulation, wherein a therapeutically effective amount of the formulation of free amino acids and a therapeutically effective amount of the plant extract formulation demonstrate skin barrier repair (e.g., correcting the skin barrier, strengthening the skin barrier, enhancing ... and (e.g., see Figures 8-14), where such formulations improve skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; epidermal cell, keratinocyte, endothelial cell, and fibroblast cell growth, stimulation, proliferation, differentiation, and migration; dermal regeneration promotion). and improvement in skin barrier repair was 1.5 or higher (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 to 40 (e.g., 1.6 to 39.9, 1.7 to 39.8, 1.8 to 39.7, 1.9 to 39.6, 2 to 39.5, 2.1 to 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).

[0063] In some embodiments, a method of improving skin barrier repair of the skin of a subject is provided, comprising: a topical formulation (as described in any one of the preceding formulations or embodiments 64-66) comprising a therapeutically effective amount of a blend of free amino acids 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 blend (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense), alone or optionally in combination with at least one of a texturing agent and / or filler (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, which adsorbs water in hygroscopic applications), a plant extract blend; and optionally, a dermatologically acceptable carrier and / or additive, whereby skin barrier repair is improved when tested by a method of detecting expression of a barrier marker gene (e.g., EGF) to determine that the barrier marker gene is increased by an average fold change of 1.5 to 40 in amino acid-treated skin compared to amino acid-untreated skin.

[0064] In yet another embodiment of the method or use, the formulation is sterile.

[0065] In certain embodiments, the formulations described herein are in the form of a single unit dose. In one aspect, the formulations described herein have a pH of about 2.0 to about 8.5. According to one embodiment, the formulations described herein are formulated for transdermal, subcutaneous or topical administration.

[0066] In yet another aspect of the method or use, the formulations described in various embodiments herein are administered on a daily dosing schedule.

[0067] Also described herein are kits that include a formulation described herein and instructions for administering the formulation to a subject or contacting the formulation with a biological sample.

[0068] definition All terms used herein are intended to have their ordinary meaning in the art unless otherwise specified. All concentrations are in terms of weight percent of the particular ingredient relative to the total weight of the topical composition unless otherwise defined.

[0069] As used herein, "a" or "an" means one or more. When used herein in conjunction with the word "comprising," the words "a" or "an" mean one or more. As used herein, "another" means at least a second or more instances.

[0070] As used herein, any numerical range includes the endpoints and all possible values ​​disclosed between the disclosed values. All half-integer exact values ​​are also contemplated as specific disclosures and limits for all subsets of the disclosed ranges. For example, the range 0.1%-3% specifically discloses the percentages 0.1%, 1%, 1.5%, 2.0%, 2.5%, and 3%. Furthermore, the range 0.1%-3% includes subsets of the original range, such as 0.5%-2.5%, 1%-3%, 0.1%-2.5%, etc. It is understood that the sum of all weight percentages of the individual components does not exceed 100%.

[0071] The terms "improving a cosmetic skin condition," "improving a skin condition," or "treating a skin condition" include treating a cosmetic skin condition or therapeutically treating a skin condition, which may involve at least one of the following benefits: improved moisture retention, reduced permeability of the skin epithelial cell barrier, thickening of the skin (increasing the depth of the skin epithelial layer), restoring skin elasticity, preventing loss of skin elasticity, reducing skin cell desquamation, improving smoother skin texture, reducing the physical appearance of lines or wrinkles (e.g., as determined by appearance or tactile perception), and any combination thereof.

[0072] The skin comprises three distinct layers: the stratum corneum (the outermost layer), the epidermis, and the dermis.

[0073] The term "skin cell barrier integrity" as used herein refers to the ability of the epidermis, particularly the stratum corneum (SC), to function as a selective permeability barrier that limits transdermal evaporative water loss and allows survival in a dry external environment. The SC is a multi-layered tissue composed of flat, anucleated keratinocytes, surrounded by multiple planar lamellar sheets, rich in ceramides, cholesterol, and free fatty acids (FFAs) that hold the entire matrix together. These components, together with tight junction proteins and antimicrobial peptides, contribute to the skin's ability to act as a permeability barrier. Tight junction proteins contribute to the cell-cell adhesion interface, essentially acting as a seal between cells. Unregulated intercellular leakage from the outermost layer of skin can result in water loss from, for example, the inner layer of the skin. Barrier integrity can be measured using a wide variety of assays, such as transepidermal water loss (TEWL) at the surface of the skin (in situ); transepithelial / transendothelial electrical resistance (TEER), which is also a widely accepted quantitative technique for measuring tight junction dynamic integrity in cell culture models of endothelial and epithelial monolayers; techniques monitoring the diffusion of dyes (X-Gal or Lucifer Yellow, Toluidine Blue) through the upper epidermis, which are also used in skin explants and / or in situ skin; 2D or 3D keratinocyte cell cultures, which are used to evaluate epidermal differentiation processes that are directly related to skin barrier permeability; upregulation of proteins that promote skin barrier function and / or downregulation of genes that reduce skin barrier function; as well as immunohistochemical staining, microscopy and / or other imaging techniques.

[0074] The term "improving" when used in relation to improving "skin cell barrier integrity" refers to an increase in the skin's permeability barrier, for example, by increasing the strength of adhesion between skin cells, as indicated by the ability of the cell-cell adhesion interface to prevent unregulated leakage between cells. Barrier integrity may be detected using assays such as those described for barrier integrity. The term "improving" may be used to describe an increase of 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% or more compared to the starting point measurement before contact with the amino acid formulations described herein.

[0075] The term "maintaining" when used in relation to maintaining "skin cell barrier integrity" refers to preserving the strength of the skin's permeability barrier, for example by preserving the strength of adhesion between skin cells, as indicated by the ability of the cell-cell adhesion interface to prevent unregulated leakage between cells. In relation to maintaining "skin cell barrier integrity", measurements can be taken, for example, on healthy skin to determine the baseline cell-cell adhesion strength, to establish the desired target level of cell-cell adhesion to be maintained. Samples of healthy skin can then be exposed to stressors, such as airborne pollution (e.g., cigarette smoke, harmful smoke, airborne irritants) or physical stressors (e.g., abrasion, harsh cleaning agents, or chemicals), in the presence or absence of the formulations described herein. Amino acid formulations described herein that at least partially prevent the loss of cell-cell adhesion even when stimulated by a stressor are characterized as capable of maintaining a determined degree of barrier integrity. In such context, the term "maintaining" may be used to describe the preservation of baseline intercellular strength at 100% pre-stressor levels, or at 99% or more, 98% or more, 97% or more, 95% or more, 90% or more, 85% or more, 80% or more, 75% or more, 70% or more, 65% or more, 60% or more, 55% or more, 50% or more, 45% or more, 40% or more, 35% or more, 30% or more, 25% or more, 20% or more, 15% or more, 10% or more, 5% or more, 2% or more, or 1% or more pre-stressor levels, due to the presence of an amino acid formulation described herein that attenuates the adverse effects of such stressor.

[0076] The term "topical application," as used herein, means applying or painting the formulations described herein onto the surface of the epidermal tissue.

[0077] The term "dermatologically acceptable," as used herein, means that the formulation or components thereof so described are suitable for use in contact with the epidermal tissue of a mammal without undue toxicity, incompatibility, instability, allergic response, and the like.

[0078] The term "therapeutically effective amount," as used herein, refers to an amount of a compound or formulation sufficient to induce a beneficial benefit, improvement in the appearance and / or texture of the skin, and in some embodiments, the beneficial benefit of amino acid treated skin is an increase of 1% or more, 2% or more, 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, 50% or more, 55% or more, 60% or more, 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, 95% or more, 97% or more, 98% or more, 99% or more, or 100% or more compared to a starting measurement prior to contact with an amino acid formulation described herein; an increase of 1-fold (e.g., 3-fold, 5-fold, 8-fold, 9-fold, 10 ... , 7x, 9x, 11x, 13x, 15x, 17x, 19x, 21x, 23x, 25x, 27x, 29x, 31x, 33x, 35x, 37x, 39x, 41x); 40x or less (e.g., 38x, 36x, 34x, 32x, 30x, 28x, 26x, 24x, 22x, 20x, 18x, 16x, 14x, 12x, 10x, 8x, 6x, 4x, 2x, 0.8x, 0.6 times); or 1 to 40 times (e.g., 2 to 39 times, 3 to 38 times, 4 to 37 times, 5 to 36 times, 6 to 35 times, 7 to 34 times, 8 to 33 times, 9 to 32 times, 10 to 31 times, 11 to 30 times, 12 to 29 times, 13 to 28 times, 14 to 27 times, 15 to 26 times, 16 to 25 times, 17 to 24 times, 18 to 23 times, 19 to 22 times, 20 to 21 times) better. According to the present disclosure, a therapeutically effective amount is an amount of a combination of free amino acids, alone or in combination with other agents, that physically and / or visually regulates and / or improves the skin.

[0079] The term "amelioration" or any grammatical variations thereof (e.g., ameliorate, ameliorating, ameliorating, 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.

[0080] The term "effective amount" or "substantial amount," as used herein, refers to an amount that is capable of treating or ameliorating a disease, condition, or disorder, or that is otherwise capable of producing the intended therapeutic effect.

[0081] The term "healthy functional food" refers to food that has been prepared or processed into tablets, capsules, powders, granules, liquids, pills, or any other form using raw materials or ingredients that have functions that are beneficial to the human body.

[0082] The term "functional" refers to an effect beneficial to human health, for example, regulation of structure or function, such as nutrients, immune system, inflammation, fluid balance, physiological actions, etc.

[0083] The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle used in administration of a compound. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin (23rd edition), which is incorporated herein by reference.

[0084] The term "treatment" or any grammatical variations thereof (e.g., treat, treating, treatment, etc.) as used herein includes, but is not limited to, alleviating the symptoms of a disease, condition, or disorder; and / or reducing, suppressing, inhibiting, reducing, or affecting the progression, severity, and / or extent of a disease, condition, or disorder.

[0085] The term "consisting essentially of," as used herein, limits the scope of ingredients and steps to the particular materials or steps, and those that do not materially affect the basic and novel characteristic(s) of the present invention, namely, the formulations and methods for promoting at least one of the integrity, function, texture, or appearance of skin, or any combination thereof.

[0086] The term "amino acid" includes all known amino acids that contain an amine (-NH2) functional group, a carboxyl (-COOH) functional group, and / or a side chain ("R") group specific to each amino acid. "Amino acid" includes the 21 amino acids encoded by the human genome (i.e., proteinogenic amino acids), amino acids encoded or produced by bacteria or unicellular organisms, and naturally occurring amino acids. For purposes of this disclosure, the conjugate acid form of an amino acid with a basic side chain (arginine, lysine, and histidine) or the conjugate base form of an amino acid with an acidic side chain (aspartic acid and glutamic acid) are essentially the same unless otherwise specified. "Amino acid" also includes derivatives and analogs thereof that retain substantially the same activity in improving the barrier integrity of skin cells, for example, in a TEER assay or a TEWL assay as described above. Derivatives and analogs may be, for example, mirror images, including both D- and L-forms. The derivatives and analogs may be derivatives of "natural" or "unnatural" amino acids (e.g., β-amino acids, homoamino 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), such as selenocysteine, pyrrolysine, iodocysteine, norleucine, or norvaline. The derivatives and analogs may contain protecting groups (α-amino groups, α-carboxylic acid groups, or suitable R groups, where R contains NH2, OH, SH, COOH, or other reactive functionalities). Other amino acid derivatives include, but are not limited to, those synthesized by, for example, acylation, methylation, glycosylation, and / or halogenation of amino acids. These include, for example, β-methyl amino acids, C-methyl amino acids, and N-methyl amino acids. The amino acids described herein may exist as free amino acids. The term "free amino acid" refers to an amino acid that is not part of a peptide or polypeptide (eg, not connected to another amino acid via a peptide bond).A free amino acid is free in solution (as opposed to being linked to at least one other amino acid, e.g., via a dipeptide bond), but may be associated with salts or other components in the solution.

[0087] As used herein, the term "salt" refers to any salt, including pharma- ceutically acceptable salts. For example, embodiments of the present disclosure are directed to amino acids or their salts.

[0088] Exemplary salts for inclusion in the formulations described herein include sodium chloride, potassium chloride, calcium chloride, magnesium chloride, or phosphate buffers of trisodium citrate, sodium bicarbonate, sodium gluconate, using monosodium phosphate, disodium phosphate, or trisodium phosphate, or any combination thereof.

[0089] 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.

[0090] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical formulations described herein include inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents or phospholipids, in some embodiments phospholipids are used as emulsifying agents (e.g., lecithin or phosphatidylcholine), disintegrating agents, binders, preservatives, buffers, lubricants, and / or oils. Excipients such as cocoa butter and suppository waxes, colorants, coating agents, and flavoring agents may also be present in the composition.

[0091] 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.Gene / Product: 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; PPAR-peroxisome proliferator-activated receptors; SMAD-acronym for the fusion of the Caenorhabditis elegans Sma gene and Drosophila Mad, Mothers against decapentaplegic; TGFB1-transforming grown factor beta 1; TGM-transglutaminase, TNFRSF10D-TNF Receptor Superfamily Member 10d (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. [Brief description of the drawings]

[0092] [Figure 1]Figure 1 shows the effect of amino acid formulation treatment on mRNA expression of barrier marker genes in differentiated normal human epidermal keratinocyte (NHEK) cells from five donors. Treatment time -4 hours. 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 were: 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). [Diagram 2] A table listing the amino acid combinations is shown. Combo #1, #2, #4, #6 and #7 amino acids were used at a concentration of 4 mM for each amino acid. Combo #3 and #4 amino acids were used at the lower indicated concentrations (mM) since toxic effects were detected at the 4 mM concentration. [Diagram 3] Figure 2 shows a table showing transcript analysis of skin barrier integrity marker genes in primary keratinocytes. Differentiation genes are designated with * and proliferation genes with +. EGF contributes to both differentiation and proliferation. Scoring system developed: number of points given for relative expression. (mean relative expression >1.5 (50% increase); untreated = 1). [Figure 4] Figure 2 shows a table showing transcript analysis of skin barrier integrity marker genes in primary keratinocytes. Differentiation genes are designated with * and proliferation genes with +. EGF contributes to both differentiation and proliferation. Scoring system developed: number of points given for relative expression. (mean relative expression >1.5 (50% increase); untreated = 1) (n = 5). [Diagram 5]Quantification of Western blot analysis of involucrin (INV) expression at 24 hours 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. [Figure 6] Quantification of Western blot analysis of filaggrin (FLG) expression at 4 and 24 hours 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. [Figure 7] A-D show graphs of EGF expression comparing a four amino acid blend (Ala, Gln, Gly, Ser; 4GAA) and a Boswellia plant extract blend (BSW=BSXL) alone or in combination. A shows EGF expression for donor O cells with 4GAA alone (4 mM); BSW (1%) alone; and a combination of 4GAA (4 mM) and BSW (1%). B shows EGF expression for donor O cells with 4GAA alone (1 mM); BSW (0.25%) alone; and a combination of 4GAA (1 mM) and BSW (0.25%). C shows EGF expression for donor G cells with 4GAA alone (4 mM); BSW (0.5%) alone; and a combination of 4GAA (4 mM) and BSW (0.5%). D shows EGF expression in donor G keratinocyte cells with 4GAA alone (1 mM); BSW (0.25%) alone; and 4GAA (1 mM) and BSW (0.25%) in combination. *p value=0.05; **p value=0.01. "CTR" or "CTR untr" is untreated control. n=3 [Figure 8]A and B show the fold change in EGF mRNA in donor G keratinocyte cells compared with 1 mM or 4 mM of a 4 amino acid blend (Ala, Gln, Gly, Ser; 4GAA) and / or a plant extract blend containing, for example, Boswellia (BSXL), alone (0.1%, 0.25%, 0.5%) or in combination at different concentrations. Data are shown in graphical (A) and tabular (B) formats. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 vs. untreated control (ctr). n=3 [Figure 9] A and B show fold change in EGF mRNA comparing a 4 amino acid blend (Ala, Gln, Gly, Ser; 4GAA) and a Boswellia plant extract blend (BSXL; Bosexil®) alone or in combination at different concentrations using donor O keratinocyte cells. Data are shown in graphical (A) and tabular (B) formats. **p<0.01, ***p≦0.001 vs. untreated CTR control. n=2 or n=3. [Figure 10] A and B show fold change in EGF mRNA comparing a 4 amino acid formulation (Ala, Gln, Gly, Ser; 4GAA; 4 mM), a 3 amino acid formulation (Cys 1 mM, His 0.5 mM, Tyr 1 mM; 3BAA), a 0.5% Boswellia plant extract formulation (BSXL), and piperonylic acid (100 μM; PA) alone or in combination using donor G keratinocyte cells. Data are shown in graphical (A) and tabular (B) formats. *p<0.05, **p≦0.01, ***p≦0.001 vs. untreated CTR control. n=3. [Figure 11]A and B show fold change in EGF mRNA comparing a 4 amino acid formulation (Ala, Gln, Gly, Ser; 4GAA; 4 mM), a 3 amino acid formulation (Cys 1 mM, His 0.5 mM, Tyr 1 mM; 3BAA), a Boswellia plant extract formulation (BSXL), and piperonylic acid (100 μM; PA) alone or in combination using donor T keratinocyte cells. Data are shown in graphical (A) and tabular (B) formats. *p<0.05, **p<0.01, ****p<0.001 vs. untreated CTR control. n=3. [Figure 12] A and B show the mean fold change in EGF mRNA comparing a 4 amino acid blend (Ala, Gln, Gly, Ser; 4GAA; 4mM), a Boswellia plant extract blend (BSXL), and a boswellic acid extract (BA) alone or in combination at various concentrations or weight percent (wt / wt%) using donor O keratinocytes. Data are shown in graphical (A) and tabular (B) formats. *p<0.05, **p≦0.01, ***p≦0.001 vs. untreated CTR control. n=2 or n=3. [Figure 13] A and B show the mean fold change in EGF mRNA comparing a 4 amino acid formulation (Ala, Gln, Gly, Ser; 4GAA; 4 mM), an alternative 4 amino acid formulation (Asp, Ile, Tyr, Val; 4OAA; 4 mM), and a Boswellia plant extract formulation (BSXL; combined at 0.5% or 0.5 w / w%) alone or in combination at various concentrations or weight percent (wt / wt%) using donor A keratinocytes. Data are shown in graphical (A) and tabular (B) formats. ***p≦0.001 vs. untreated CTR control. n=3 [Figure 14A] Figure 1 shows the mean fold change in EGF mRNA comparing a 4 amino acid formulation (Ala, Gln, Gly, Ser; 4GAA; 4 mM) and a Boswellia plant extract formulation (BSXL; 0.5% w / w), alone or in combination, using keratinocytes from donor A, donor G, and donor T. Data is presented in tabular format (Donor A (n=1), Donor G (n=3), and Donor T (n=2)). [Figure 14B] Average fold change in EGF mRNA comparing a 4 amino acid formulation (Ala, Gln, Gly, Ser; 4GAA; 4 mM) and a Boswellia plant extract formulation (BSXL; 0.5% w / w) alone or in combination using keratinocytes from donor A, donor G, and donor T. Data are presented in tabular format (Donor A (n=3), Donor G (n=9), and Donor T (n=6)). [Figure 14C] 14B shows the mean fold change in EGF mRNA comparing a 4 amino acid formulation (Ala, Gln, Gly, Ser; 4GAA; 4 mM) and a Boswellia plant extract formulation (BSXL; 0.5 w / w%) alone or in combination using keratinocytes from donors A, G, and T. Data from FIG. 14B is presented in graphical form. CTR; 4GAA; BSXL; 4GAA+BSXL formulation: Donor G (top three), Donor T (middle), Donor A (bottom three) are shown. *p<0.05 vs. untreated CTR control; #p<0.05 vs. 4GAA and BSXL. Analyzed using repeated measures analysis of variance (RMANOVA) with Tukey post-hoc test. [Figure 15] A and B show the change in transepidermal water loss (TEWL) after 10 days of application of test article (active (4GAA), left bar; placebo, right bar) dissolved in an oil / water (O / W) vehicle in 12 subjects followed by repeated tape stripping (15 tape strips). A shows the change in TEWL from baseline immediately after tape stripping on day 1, and days 2, 3, and 5 (X-axis) after twice-daily treatment with test article. B shows the relative TEWL increase after tape stripping, all values ​​normalized to baseline day 1, which represents TEWL immediately after tape stripping. Active (left bar); placebo (right bar). Mean + / - 95% confidence interval (CI). [Figure 16]Skin barrier improvement as measured by skin hydration (Y-axis) is shown after 10 days of test article application followed by repeated tape stripping (25 times) in 25 subjects. Control represents no treatment (top line on day 3; left bar in inset), placebo represents water / oil / water (W / O / W) vehicle without active drug (middle line on day 3; middle bar in inset), and amino acids represent a 4 amino acid blend (Ala (4%), Gln (4%), Gly (4%), Ser (4%) dissolved in W / O / W vehicle (bottom line on day 3; right bar in inset). *, p-value=0.05. Inset: control (left), placebo (middle), amino acids (right). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0093] Although detailed embodiments of the present disclosure are disclosed herein, it should be understood that the disclosed embodiments are merely exemplary of the present disclosure, which may be embodied in various forms. Moreover, each of the examples provided in connection with the various embodiments of the present disclosure are intended to be illustrative and not limiting.

[0094] Skin functions as a physical barrier that isolates the organism from its surrounding environment, but also serves as an interactive interface with that environment. Skin comprises three distinct layers, namely, the stratum corneum, epidermis, and dermis. The stratum corneum (outermost layer) contains keratinocytes and provides a physical barrier with the external environment. Environmental stressors vary, for example, depending on the climate and / or lifestyle, and can include xenobiotics, UV radiation, pollution (e.g., cigarette smoke or engine exhaust), and pathogens. In addition to acting as a shield to protect the lower layers of the skin from such external stimuli, the stratum corneum also reduces water loss from the skin. Skin keratinocytes are connected through tight junction (TJ) proteins, such as zona occludin (ZO). TJ proteins play a crucial role in barrier function and also contribute to keratinocyte proliferation and differentiation. The integrity of the barrier function prevents excessive water loss from the skin, a key feature of skin, and thereby plays a role in maintaining healthy skin.

[0095] Dysregulation of the skin barrier is involved in a wide variety of common skin disorders such as atopic dermatitis, psoriasis, and ichthyosis. Dysregulation of the skin barrier also contributes to xerosis (dry skin), a common skin disorder that is exacerbated by low humidity, repeated washing, and exposure to harsh chemicals. Such external stressors promote water loss from the skin. Dry skin may result in or be accompanied by itching and / or skin irritation. Dry skin may be considered a symptom of compromised skin barrier function. Dry skin is also a common feature of aging skin.

[0096] Optimal management of many skin diseases, conditions, or disorders includes proper skin care. The incorporation of a properly designed moisturizer / barrier repair formulation may help reduce disease-related signs and symptoms, thereby improving the disease. Described herein is an effective moisturizer / barrier repair formulation for treating dry skin, which enhances skin regeneration and strengthens skin barrier function. The formulation includes a specific amino acid blend that provides cosmetic and / or therapeutic benefits in connection with the treatment of a wide variety of cosmetic skin conditions and skin diseases, conditions, and disorders (e.g., dry skin).

[0097] 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. Described herein, in one aspect, are formulations and methods for improving skin barrier integrity and promoting cell proliferation and / or development. As used herein, reference to "development" may include, for example, skin cell migration, maturation, and / or differentiation. The present disclosure also provides formulations and methods for treating and / or preventing cosmetic skin conditions or skin conditions associated with skin diseases or disorders. Such skin conditions include, but are not limited to, atopic dermatitis, psoriasis, conditions associated with skin aging, pruritus, eczema, and / or cosmetic conditions. The present disclosure also provides formulations and methods for treating wounds or burns.

[0098] For example, atopic dermatitis (AD) is the most common chronic inflammatory skin disease in humans, affecting up to 15 million Americans (17% of children and 6% of adults). Despite its high prevalence, impact on quality of life, and economic burden, there are still no effective treatments for AD, most of which are largely focused on suppressing inflammation. AD is thought to develop in part as a result of acquired or genetic abnormalities in the skin barrier. The epidermis of AD subjects showed alterations in tight junctions (TJs), which were associated with reduced expression of select TJ components (e.g., claudins). TJs seal the intercellular spaces between epithelial cells, and the "tightness" of this structure is dynamically regulated by endogenous or environmental factors. Regulation of TJ sealing is important for a variety of reasons, including proper transport patterns of ions, proteins, and water, as well as immune cell permeability.

[0099] The integrity of tight junction dynamics between endothelial and epithelial cells, which regulates diffusion and maintains homeostasis of organs protected by physiological barriers, can be measured in vitro using transepithelial / transendothelial electrical resistance (TEER). TEER can be used to identify agents that improve physiological barriers and thus have beneficial effects on conditions associated with increased levels of permeability of physiological barriers compared to normal, healthy conditions. Conditions associated with increased levels of permeability of physiological barriers include, for example, atopic dermatitis and dry skin. The results presented herein reveal that certain amino acid formulations improve / enhance skin cell barrier integrity. The improvement of skin cell barrier integrity is believed to be mediated, at least in part, by tight junction dynamics and reflects reduced permeability of tight junctions. See, for example, Figures 1-6.

[0100] The subject may be a patient who needs to improve the barrier integrity of the skin. The patient may have such a need due to, for example, exposure to chemical or physical irritants, age-related predisposition, genetic predisposition, or skin inflammation. In one embodiment, the patient is asymptomatic. The subject may be any animal, including, for example, a human. In addition to humans, the animal may be, for example, a mammal, such as rabbits, cows, horses, sheep, pigs, goats, dogs, and cats.

[0101] As shown above, skin barrier dysfunction is involved in common skin diseases such as atopic dermatitis, psoriasis, and ichthyosis, as well as xerosis (dry skin), a very common skin disease. Dry skin is a condition that most people experience at some point in their lives. Seasonal xerosis is common during cold, dry winter months, and the frequency of xerosis increases with age. Optimal management of many skin diseases involves proper skin care. Incorporating a properly designed moisturizer / barrier repair formulation into a skin care regimen may contribute to the improvement or reduction of symptoms associated with the condition or disease.

[0102] Effective moisturizer / barrier repair formulations for dry, dehydrated skin that enhance the skin's own ability to regenerate, restore and strengthen its own barrier function using amino acid-based technology are desirable. These formulations would provide a new treatment option for dry, scaly skin and would benefit the many people who suffer from dry, scaly skin and are at risk of developing more severe conditions resulting from neglected dry skin.

[0103] Physiologically, amino acids and their derivatives are important elements of the skin, since they are components of the Natural Moisturizing Factor (NMF) of the stratum corneum (SC), the outermost layer of the skin. Free amino acids (FAA) are generated by hydrolysis of keratinocyte proteins filaggrin, corneodesmosomes, and keratin of SC during the normal physiological process of epidermal maturation and terminal differentiation. NMF contributes to maintaining adequate skin moisture. Some amino acids can effectively bind water and therefore may play a role in regulating skin moisture. Therefore, supplementation of skin with certain amino acids is proposed herein to ensure adequate levels of NMF and protein synthesis or production of certain important amino acid metabolites [pyrrolidone carboxylic acid (PCA) and trans-urocanic acid (t-UCA)] that regulate, nourish, and / or replenish proteins in normal and dry skin, as well as in other skin conditions in which the epidermal barrier is compromised.

[0104] Without being bound by theory, by delivering specific amino acid blends to the skin, the skin's natural ability to form an appropriate barrier may be "fertilized." The problem of how to maintain / restore skin barrier function is solved by developing amino acid formulations that target specific intracellular pathways involved in skin barrier function and repair, thereby converting amino acids from mere building blocks into small molecules that can trigger signaling pathways that promote skin barrier function.

[0105] The results presented herein show that supplementation with a specific amino acid blend can regulate the transcription of key regulators of differentiation and proliferation, such as the EGF and PDGF genes, in normal primary human keratinocytes. The specific combinations of amino acids that show this functional ability were determined by analyzing their effects on skin barrier markers. Notably, certain combinations of certain amino acids show beneficial effects on skin barrier function (such as Ser, Ala, Gly and Gln), while other amino acid combinations (such as Cys, His and Tyr) have no effect or even a negative impact on skin barrier function. Additional screening assays to identify the most effective combination(s) of amino acids to improve the barrier integrity of skin / skin barrier function are performed, tested and validated in preclinical studies using different skin models and different administration methods (e.g., systemic and / or topical).

[0106] Specific goals include defining effective amino acid formulations that promote skin barrier function (see Figures 1-6) and further evaluating and developing such amino acid formulations using 2D monolayer cultures (human differentiated keratinocytes) and 3D models (skin equivalents of human epidermis). Determine the effect of adding several amino acid formulations (e.g., the four major amino acid formulations of Ser, Ala, Gly, and Gln described above with and without Ile, Trp, and / or Arg) to the culture medium on the mRNA and protein levels of the skin barrier marker panel identified based on replicate analysis. See, e.g., Example 1.

[0107] In brief, genes and proteins related to epidermal differentiation, proliferation and lipid regulation in skin are repeatedly evaluated and weighted for their structural / functional impact on skin barrier integrity. The determination of the panel of markers used herein that reflects skin barrier integrity is performed by a series of multivariate repeated analyses. The complexity of this task is highlighted by the vast array of potential genes that are suggested to be involved in, for example, barrier function, proliferation, differentiation, and inflammation, each of which biological processes contribute to some degree to skin barrier integrity. In fact, hundreds of genes have been suggested to be involved in these processes. Thus, the starting point for determining a suitable panel of markers that reflects skin barrier integrity is given an unpredictable number of different potential skin barrier marker panels. The selection of a panel from this myriad of options required experiments using multiple sample sources to avoid sample bias, repeated analyses to study different endpoints, weighted importance of different genes in the skin barrier marker panel, and careful analysis of all variables in these analytical methods. The panel of skin barrier markers selected to reflect skin barrier integrity is the result of the analytical approach described above and was therefore determined according to a mathematical and biological approach involving experimental results, their statistical analysis, and algorithmic analysis of all collected inputs.

[0108] After identifying the skin barrier marker panel, a scoring system was developed to evaluate the different amino acid formulations under study. The scoring system was developed to address the many variables that arise from the experimental results. Such variables include, for example, the degree of transcriptional upregulation, the number of different genes that are up-regulated or down-regulated in response to the amino acid formulations investigated, and the variability of the readouts observed between different sample sources. See, for example, Figures 3 and 4.

[0109] Specific goals 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 as well as normal skin tissue cultures (skin explants). To generate an inflammatory skin phenotype, immature epidermal skin equivalents are pretreated with helper T cell (Th-2) derived cytokines in combination with TNFα, and skin explants are degreased by tape stripping. The effect of treatment on barrier function and tight junction (TJ) dynamics integrity is examined by a combination of gene expression, biochemical, morphological, and functional analyses (e.g., transepithelial / transendothelial electrical resistance (TEER)). See, for example, Example 4 and Example 5.

[0110] Therefore, the identification of effective amino acid formulations and their use in barrier repair formulations / moisturizers provides a solution to the problem of how to maintain and restore healthy skin as people age and under duress and / or disease conditions.Therefore, the formulations identified herein fulfill the long-standing need for scientifically verified barrier repair formulations / moisturizers that address the underlying biological causes of skin conditions (e.g., dry skin) rather than simply the symptoms.Proof-of-concept testing is carried out on human subjects using the most promising amino acid-based cosmetic formulations.

[0111] Formulations and methods for treating cosmetic or other skin conditions Dry and sensitive dry skin (also called xerosis, xerosis dermoides, asteatosis) is the most common skin disorder affecting approximately 50% of the world's population. The severity of the disorder increases with age. It affects the quality of life of patients and is a risk factor for the development of atopic or allergic dermatitis and other skin diseases due to impaired skin barrier.

[0112] Dry skin is associated with the development of skin cracks and pressure-related ulcers that can be avoided with proper skin care. Many skin care products used to manage dry skin mimic various components of the skin barrier. They are designed to incorporate lipophilic (lipid replenishing, film forming) and hydrophilic (remoisturizing) ingredients into the skin. Although such products can improve the moisture content of the SC, thereby attenuating inflammation, they do not address the underlying biochemical abnormalities in dry skin. Furthermore, many skin care products available on the market do not provide scientifically proven benefits.

[0113] Therefore, there is a need for topical skin care that is effective against dry skin, which not only mimics skin barrier components, but also enhances the skin's own ability to maintain and / or repair the strength of its barrier.The protective barrier thus formed at least partially restores the structural and functional properties of healthy skin.Thus, a topical skin care product that enhances the skin's ability to maintain and / or repair barrier integrity would address the underlying causes of dry skin, rather than simply addressing the symptoms.A formulation has been developed that aims to address this objective.

[0114] For example, dry aged skin has reduced levels of profilaggrin and filaggrin, which are the main sources of free amino acids (FAA) in SC, and thus the reduction in the levels of these proteins causes the reduction of available FAA pool.The results presented herein show that the specific amino acid blends described herein target-specifically supplement the FAA pool that is reduced by aging.The results presented herein further show that certain combinations of amino acids act synergistically to provide benefits to the structural / functional properties of skin barrier integrity, and surprisingly, certain amino acids can also be detrimental to skin barrier integrity.

[0115] The data presented herein also show that treatment with specific amino acid formulations upregulates a panel of skin barrier marker genes (e.g., EGF, PDGF, CPT2, TGM4) in normal human keratinocytes that play important roles in skin development and homeostasis.

[0116] Described herein are mechanism(s) by which amino acids enhance skin barrier integrity, focusing on their effects on TJs and / or improving skin barrier repair, which, to the extent that they are known, have not previously been scientifically rigorously explored in skin.

[0117] As highlighted above, not all amino acids have the same beneficial effect on skin barrier integrity.In fact, some amino acids, either alone or in combination, are contraindicated with respect to skin barrier integrity.Therefore, there is a need for a targeted approach to identify the most effective amino acids for promoting skin barrier health.This paper proposes a new scientifically proven amino acid-based technology that targets hypofunctional epithelial skin to provide a better solution for people suffering from dry and sensitive skin.

[0118] Thus, in certain embodiments, the present disclosure provides a method for treating and / or preventing a skin condition in a subject in need thereof, comprising administering to the subject a formulation as described herein.In certain aspects, the skin condition is dry skin, atopic dermatitis, dermatitis, psoriasis, skin aging, skin aging-related conditions (e.g., wrinkles, loss of flexibility, increased roughness), pruritus, or eczema.In further aspects, the skin condition is a cosmetic skin condition.

[0119] In certain aspects, the formulation for treating and / or preventing cosmetic skin conditions or for treating and / or preventing skin conditions comprises, as free amino acids, a therapeutically effective amount of a combination of glutamine, glycine, alanine, and serine, and optionally a therapeutically effective amount of at least one additional free amino acid valine, isoleucine, arginine, threonine, or tryptophan, or any combination thereof, and the therapeutically effective amount of the combination of glutamine, glycine, alanine, and serine and the therapeutically effective amount of at least one additional free amino acid improves the barrier integrity of skin cells, and the formulation optionally comprises a dermatologically acceptable carrier and improves the barrier integrity of skin.In certain embodiments, the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine, and optionally a therapeutically effective amount of at least one additional amino acid isoleucine, arginine, threonine, or tryptophan, or any combination thereof. In more particular embodiments, the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine, and at least one additional amino acid arginine or threonine, or any combination thereof. In more particular embodiments, the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine; the free amino acids or peptide combinations of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and arginine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and threonine;The free amino acids or peptide combinations of amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, and serine, and at least one additional amino acid isoleucine, arginine, threonine, or tryptophan, or any combination thereof; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine, and at least one additional amino acid arginine, or threonine, if or tryptophan, or any combination thereof; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and arginine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and threonine; the free amino acids or peptide combinations of amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine; the free amino acids or peptide combinations of amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine; the free amino acids or peptide combinations of amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine; the free amino acids or peptide combinations of amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine;The free amino acids or peptide combinations of amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, threonine, and tryptophan; the free amino acids or peptide combinations of amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, threonine, and tryptophan; the free amino acids or peptide combinations of amino acids include glutamine, glycine, alanine, serine, and arginine, and at least one additional amino acid isoleucine, threonine, or tryptophan, or any combination thereof. or the free amino acids or peptide combination of amino acids comprises, consists essentially of, or consists of glutamine, glycine, alanine, serine, and threonine, and at least one additional amino acid isoleucine, arginine, or tryptophan, or any combination thereof; or the free amino acids or peptide combination of amino acids comprises, consists essentially of, or consists of glutamine, glycine, alanine, serine, and tryptophan, and at least one additional amino acid isoleucine, arginine, or threonine, or any combination thereof;

[0120] In one embodiment, the formulation optionally includes, for example, pharma- ceutically acceptable carriers, adjuvants, other active agents, and additives (eg, sugars, electrolytes, vitamins, minerals, etc.).

[0121] In some embodiments of the formulations described herein, the amino acid cysteine, if present, is present at a concentration of 1 mM or less; the free amino acid histidine, if present, is present at a concentration of 0.5 mM or less; the free amino acid tyrosine, if present, is present at a concentration of 1 mM or less; the free amino acid leucine, if present, is present at a concentration of 4 mM or less; or the free amino acid aspartic acid, if present, is present at a concentration of 2 mM or less; or any combination thereof. In some embodiments of the formulations, the formulation does not include the free amino acids cysteine, histidine, tyrosine, leucine, aspartic acid, taurate, taurine, or glutamic acid, or any combination thereof.

[0122] In some embodiments of the formulations described herein, if present, the free amino acid alanine is 4% or less (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, 0.01 6, 0.014, 0.012); 0.01% or more (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 0.3, 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, the free amino acid glutamine is present at a concentration of 4% or less (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, 0.06, 0.05 ... 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 more (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, the free amino acid glycine is present at a concentration of 4% or less (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, 0.0 18, 0.016, 0.014, 0.012); 0.01% or more (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 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, the free amino acid serine is present at a concentration of 4% or less (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 more (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, 0.1 1, 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.

[0123] Or, in certain embodiments, even if these amino acids are present in the formulation, they are not present in such an amount that they affect the barrier integrity of the skin.By "negligible" it is meant that the particular amino acid present has no effect on the barrier integrity of the skin.By "negligible" it is meant that the particular amino acid present has no effect on the disease or condition related to the barrier integrity of the skin, for example, has no effect on wound healing, the treatment of skin conditions (e.g., atopic dermatitis, psoriasis, or conditions related to skin aging) in the subject who needs it.

[0124] When present in the formulations described herein, the amino acids may be present at a concentration of, for example, 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 embodiments, each of the amino acids in the formulations described herein (4% or 4 mM) is about 100 times higher than the concentration used in tissue culture (TC) experiments.

[0125] In one embodiment, the total osmolality of the formulation is from about 10 mosm to about 280 mosm, from 50 mosm to about 280 mosm, from 100 mosm to about 280 mosm, or from about 150 to about 260 mosm.

[0126] In some embodiments, the formulation has a pH of, for example, about 2.5 to about 8.5. In some embodiments, the formulation has a pH of 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.

[0127] In some embodiments, the formulation is administered systemically or locally. In some embodiments, the formulation is used for treating diseases or conditions related to skin barrier integrity, such as treating dry skin, wound healing, treating skin conditions (e.g., atopic dermatitis, dermatitis, psoriasis, skin aging, conditions related to skin aging, pruritus, or eczema), and / or improving skin barrier integrity. Therapeutic formulations can be administered by enteral route, parenterally or topically, or by inhalation. In certain embodiments, the formulation is therapeutic, cosmetic, or nutritional.

[0128] In some embodiments, the formulation is a solution. The formulation can be administered together with other therapeutic agents.

[0129] Formulations and uses thereof for treating and / or preventing conditions associated with dry skin, atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, wounds, burns, or skin aging In certain embodiments, the present disclosure provides methods for treating and / or preventing conditions associated with dry skin, atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, wounds, burns, or skin aging, comprising administering to a subject a formulation described herein.

[0130] In certain aspects, the formulation for use in the treatment and / or prevention of dry skin, atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, wound, burn, or skin aging-related conditions comprises, as free amino acids, a therapeutically effective amount of a combination of glutamine, glycine, alanine, and serine, and optionally a therapeutically effective amount of at least one additional free amino acid valine, isoleucine, arginine, threonine, or tryptophan, or any combination thereof, and the therapeutically effective amount of the combination of glutamine, glycine, alanine, and serine and the therapeutically effective amount of at least one additional free amino acid improves the barrier integrity of skin cells, and the formulation optionally comprises a dermatologically acceptable carrier and improves the barrier integrity of skin.In certain embodiments, the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine, and optionally a therapeutically effective amount of at least one additional amino acid isoleucine, arginine, threonine, or tryptophan, or any combination thereof. In more particular embodiments, the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine, and at least one additional amino acid arginine or threonine, or any combination thereof. In more particular embodiments, the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, and valine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, and isoleucine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and arginine; the free amino acids comprise, consist essentially of, or consist of glutamine, glycine, alanine, serine, valine, isoleucine, and threonine;The free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, and serine, and at least one additional amino acid isoleucine, arginine, threonine, or tryptophan, or any combination thereof; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and isoleucine, and at least one additional amino acid arginine, or threonine, or tryptophan, or any combination thereof. the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and arginine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and threonine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, and tryptophan. the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and threonine; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, and tryptophan; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, threonine, and tryptophan;The free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, isoleucine, arginine, threonine, and tryptophan; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and arginine, and at least one additional amino acid isoleucine, threonine, or tryptophan, or any combination thereof; the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and threonine, and at least one additional amino acid isoleucine, arginine, or tryptophan, or any combination thereof; or the free amino acids include, consist essentially of, or consist of glutamine, glycine, alanine, serine, and tryptophan, and at least one additional amino acid isoleucine, arginine, or threonine, or any combination thereof;

[0131] In one embodiment, the formulation optionally includes, for example, a pharma- ceutically acceptable carrier, adjuvants, other active agents, and additives (e.g., sugars, electrolytes, vitamins, minerals, etc.) The formulation may further include, for example, a prebiotic or probiotic substance.

[0132] In some embodiments of the formulations described herein, the free amino acid cysteine, if present, is present at a concentration of 1 mM or less; the free amino acid histidine, if present, is present at a concentration of 0.5 mM or less; the free amino acid tyrosine, if present, is present at a concentration of 1 mM or less; the free amino acid leucine, if present, is present at a concentration of 4 mM or less; or the free amino acid aspartic acid, if present, is present at a concentration of 2 mM or less; or any combination thereof. In some embodiments of the formulations, the formulation does not include the free amino acids cysteine, histidine, tyrosine, leucine, aspartic acid, taurate, taurine, or glutamic acid, or any combination thereof.

[0133] Or, in certain embodiments, even if these amino acids are present in the formulation, they are not present in such an amount that they affect the barrier integrity of the skin.By "negligible" it is meant that the particular amino acid present has no effect on the barrier integrity of the skin.By "negligible" it is meant that the particular amino acid present has no effect on the disease or condition related to the barrier integrity of the skin, for example, has no effect on wound healing, the treatment of skin conditions (e.g., atopic dermatitis, psoriasis, or conditions related to skin aging) in the subject who needs it.

[0134] When present in the formulations described herein, the amino acid may be present at a concentration of, for example, 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.

[0135] In one embodiment, the total osmolality of the formulation is from about 10 mosm to about 280 mosm, from 50 mosm to about 280 mosm, from 100 mosm to about 280 mosm, or from about 150 to about 260 mosm.

[0136] In some embodiments, the formulation has a pH of, for example, about 2.5 to about 8.5. In some embodiments, the formulation has a pH of 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.

[0137] In some embodiments, the formulation is administered systemically or locally. In some embodiments, the formulation is used for treating diseases or conditions related to skin barrier integrity, such as treating dry skin, wound healing, treating skin conditions (e.g., atopic dermatitis, dermatitis, psoriasis, skin aging, conditions related to skin aging, pruritus, or eczema), and / or improving skin barrier integrity. Therapeutic formulations can be administered by enteral route, parenterally or topically, or by inhalation, or any combination thereof. In certain embodiments, the formulation is therapeutic, cosmetic, or nutritional.

[0138] In some embodiments, the formulation is a solution. The formulation can be administered together with other therapeutic agents.

[0139] In some embodiments, the formulation is administered systemically or locally. Therapeutic formulations can be administered by enteral route, parenterally or topically, or by inhalation. In certain embodiments, the formulation is therapeutic, cosmetic, or nutritional.

[0140] In certain embodiments, the formulation may include natural amino acids or their derivatives that retain substantially the same or better activity in improving skin barrier integrity. In certain embodiments, the formulation may include natural amino acids or their derivatives that retain substantially the same or better activity in treating dry skin, wound healing, and / or treating skin conditions (e.g., atopic dermatitis, psoriasis, or conditions related to skin aging) in subjects in need of such treatment. Derivatives may be, for example, enantiomers, including both D- and L-forms of amino acids. 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 derivative is at least one of the following derivatives: 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 synthesized by acylation, methylation, and / or halogenation of amino acids.These include, for example, β-methyl amino acids, C-methyl amino acids, and N-methyl amino acids.

[0141] In certain embodiments, the formulation also includes additives (e.g., nutrients, electrolytes, vitamins, minerals, etc.). In certain embodiments, the formulation includes iron or zinc. In certain embodiments, the therapeutic formulation includes, for example, Na + ;K + ;HCO3-;CO3 2- ;Ca 2+ ;Mg 2+ ;Fe2;Cl - ;H2PO4 - , HPO4 2- , and PO4 3-, etc.; zinc; iodine; copper; iron; selenium; chromium; and molybdenum. In an alternative embodiment, the formulation comprises one or more electrolytes selected from HCO3 - CO3 2- In another alternative embodiment, the formulation does not contain HCO3 - and CO3 2- In certain embodiments, the formulation does not contain electrolytes. For example, in certain embodiments, the formulation contains Na + ;K + ;HCO3 - ;CO3 2- ;Ca 2+ ;Mg 2+ ;Fe2;Cl - ;H2PO4 - , HPO4 2- , and PO4 3- and the like; zinc; iodine; copper; iron; selenium; chromium; and molybdenum, or none of them.

[0142] In one embodiment, H2PO4-, HPO4 2- , and PO4 3- Phosphate ions such as phosphate ions are used to buffer the formulations described herein. In one embodiment, the therapeutic formulation contains HCO3 - or CO3 2- In another embodiment, the therapeutic formulation contains HCO3 - or CO3 2- is not used as a buffer.

[0143] In some embodiments, the formulations and methods described herein are useful, for example, in cosmetic applications where rejuvenation of various layers of the skin and / or underlying tissues is desired, which rejuvenation may be facilitated, for example, by improving the barrier integrity of the skin.

[0144] In this embodiment, the disclosed methods generally include the step of topically applying a formulation to the skin (e.g., epidermis) of a patient in need of such treatment, where such formulation is applied in a therapeutically effective amount. In one embodiment, the formulation is applied to the face.

[0145] Advantageously, the present invention provides formulations and methods for combating skin aging, which may include, for example, treating the appearance of wrinkles, fine lines, and other forms of undesirable skin texture. By providing the formulation to the dermis and / or epidermis layer(s) of the skin, the skin's form, strength, and function are enhanced. In certain embodiments, the formulations and methods described herein are useful, for example, in beauty applications where rejuvenation of various layers of the skin and / or underlying tissues is desired.

[0146] In some embodiments, the formulations of the present disclosure include, in addition to amino acids, substances useful for retarding, minimizing, or eliminating skin aging, wrinkling, and / or other histological changes typically associated with intrinsic conditions (e.g., aging, menopause, etc.) and extrinsic conditions (e.g., environmental pollution (e.g., cigarette smoke), wind, heat, sunlight, radiation, low humidity, harsh surfactants, etc.).

[0147] The present invention is useful for therapeutically and / or prophylactically improving the visible and / or tactile characteristics of the skin, for example, in one embodiment, the length, depth, and / or other size of lines and / or wrinkles are reduced.

[0148] In one embodiment, the formulation applied to the skin or other tissues can further include collagen, hyaluronic acid (HA), and / or ceramide. In one embodiment, the HA is cross-linked HA. The formulation can further include ingredients and / or additives such as, but not limited to, dermatologically acceptable carriers, exfoliants, anti-acne agents, anti-wrinkle / anti-atrophy agents, vitamin B3 compounds, retinoids, hydroxyl acids, antioxidants / radical scavengers, chelating agents, flavonoids, anti-inflammatory agents, anti-cellulite agents, local anesthetics, tanning agents, skin lightening agents, skin soothing and healing agents, antibacterial and antifungal agents, sunscreens, conditioning agents, structuring agents, thickening agents (including thickening and gelling agents), formulation preparations, and preservatives. In this regard, International PCT Publication No. WO2008 / 089408 is incorporated herein by reference in its entirety.

[0149] In another aspect, the disclosure provides a method of treating and / or preventing a skin condition (e.g., atopic dermatitis, psoriasis, or a condition associated with skin aging) in a subject in need thereof, comprising administering to the subject a formulation described herein. In certain embodiments, the skin condition is atopic dermatitis, psoriasis, skin aging, or a condition associated with skin aging. In some embodiments, the skin condition is pruritus (itch), psoriasis, eczema, burn, or dermatitis. In certain embodiments, the skin condition is psoriasis. In certain embodiments, the skin condition is pruritus.

[0150] The formulations of the present disclosure can also be administered at the site of surgery, such as the site of minimally invasive surgery, to improve healing and surgical outcomes.

[0151] Formulations for repairing and strengthening the skin barrier and uses thereof In some embodiments, the formulations or topical preparations of the present disclosure comprise (or consist essentially of, or consist of) a therapeutically effective amount of a formulation comprising (or consisting essentially of, or consisting of) at least one of the following free amino acids: alanine, glutamine, glycine, and serine, or salts thereof; a therapeutically effective amount of a plant extract comprising (or consisting essentially of, or consisting of) a plant extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense); and, optionally, a dermatologically acceptable carrier and / or additive. In additional embodiments of the formulation or topical preparation comprising, consisting essentially of, or consisting of a therapeutically effective amount of an amino acid blend, the therapeutically effective amount of the amino acid blend comprises, consists essentially of, or consists of alanine; glutamine; glycine; serine; alanine and glutamine; glycine and serine; alanine and glutamine; alanine and glycine; alanine and serine; glutamine and glycine; glutamine and serine; glycine and serine; alanine, glutamine, and glycine; glutamine, glycine, and serine; alanine, glutamine, and serine; or alanine, glutamine, glycine, and serine, or salts thereof. In further embodiments of the formulations described herein, the mean fold increase in expression of the barrier marker gene EGR is 1.5 or more (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, 3 7.6, 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, 3 2.2, 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 to 40 (e.g., 1.6 to 39.9, 1.7 to 39.8, 1.8 to 39.7, 1.9 to 39.6, 2 to 39.5, 2.1 to 39.4, 2.2 to 39.3, 2.3 to 39.2, 2.4 to 39.1, 2.5 to 39, 2.6 to 38.9, 2.7 to 38.8, 2.8 to 38.7, 2.9 to 38.6, 3 to 38.5, 3.1 to 38.4, 3.2 to 38.3, 3.3 to 38.2, 3.4 to 38.1, 3.5 to 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) (e.g., skin barrier repair, skin barrier strengthening, skin barrier preservation, wound healing; epidermal cell, keratinocyte, endothelial cell, and fibroblast growth, stimulation, proliferation, differentiation, and migration; promotion of dermal regeneration). See, e.g., Figures 8-14.

[0152] In another embodiment, a formulation or topical preparation is provided, comprising as free amino acids a therapeutically effective amount of a formulation 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 formulation (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); texturing agents and / or fillers (e.g., cellulose (microcrystalline)); emulsifiers or phospholipids (in some embodiments, phospholipids are used as emulsifiers) (e.g., lecithin or phosphatidylcholine); and anti-caking agents (e.g., silica, to adsorb water in hygroscopic applications); and optionally comprising (or consisting essentially of, or consisting of) a plant extract or plant extract blend, comprising (or consisting essentially of, or consisting of) a dermatologically acceptable carrier and / or additive.

[0153] In additional embodiments, the formulation or topical preparation for administration to the skin of a subject comprises a therapeutically effective amount of a blend of free amino acids 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 blend (e.g., BOSEXIL™ or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense; texturing agents and / or fillers (e.g., cellulose (microcrystalline); emulsifiers or phospholipids (in some embodiments, phospholipids are used as emulsifiers) (e.g., lecithin or phosphatidylcholine, soy lecithin); and anti-caking agents (e.g., silica, to adsorb water in hygroscopic applications); and optionally, dermatologically acceptable carriers and / or additives, comprising (or consisting essentially of, or consisting of) a plant extract or plant extract blend, wherein a therapeutically effective amount of a blend of free amino acids and a therapeutically effective amount of the plant extract or plant extract blend are The formulation or topical formulation improves skin barrier repair (e.g., skin barrier correction, skin barrier strengthening, skin barrier preservation, wound healing; epidermal cell, keratinocyte, endothelial cell, and fibroblast cell growth, stimulation, proliferation, differentiation, and migration; dermal regeneration promotion) when tested by a method that detects expression of the barrier marker gene, EGF, and determines that the barrier marker gene is increased by an average fold change of 1.5 to 40 for amino acid treated skin compared to amino acid untreated skin (see, e.g., Figures 8-14).

[0154] In further embodiments described herein, there is provided a formulation or topical formulation of the present disclosure, wherein each of the free amino acids (at least one of alanine, glutamine, glycine, and serine, or salts thereof) is at least 0.1 mM (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) concentrations; .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 at concentrations between 0.1 mM and 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, 1.9mM~8.2mM, 2mM~8.1mM, 2.1mM~8mM, 2.2mM~7.9mM, 2.3mM~7.8mM, 2.4mM~ 7.7mM, 2.5mM~7.6mM, 2.6mM~7.5mM, 2.7mM~7.4mM, 2.8mM~7.3mM, 2.9mM~7.2 mM, 3mM~7.1mM, 3.1mM~7mM, 3.2mM~6.9mM, 3.3mM~6.8mM, 3.4mM~6.7mM, 3.5 mM~6.6mM, 3.6mM~6.5mM, 3.7mM~6.4mM, 3.8mM~6.3mM, 3.9mM~6.2mM, 4mM~6.It is present in concentrations ranging from 1mM, 4.1mM-6mM, 4.2mM-5.9mM, 4.3mM-5.8mM, 4.4mM-5.7mM, 4.5mM-5.6mM, 4.6mM-5.5mM, 4.7mM-5.4mM, 4.8mM-5.3mM, 4.9mM-5.2mM, 5mM-5.1mM).

[0155] In some embodiments described herein, a formulation or topical formulation of the present disclosure is provided, wherein each of the free amino acids (at least one of alanine, glutamine, glycine, and serine, or salts thereof) is present in an amount of 0.001% or more by weight of the formulation (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% by weight of the formulation. % 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% to 1% by weight (e.g., 0.0012% to 0.99% by weight, 0.0014% to 0.97% by weight, 0.0016% to 0.95% by weight, 0.0 018wt%~0.93wt%, 0.002wt%~0.91wt%, 0.0022wt%~0.89wt%, 0.0024wt%~0.87wt%, 0.0026wt%~0.85wt%, 0.0028wt%~0.83wt%, 0.003wt%~0.81wt%, 0.0 032wt%~0.79wt%, 0.0034wt%~0.77wt%, 0.0036wt%~0.75wt%, 0.0038wt%~0.73wt%, 0.004wt%~0.71wt%, 0.0042wt%~0.69wt%, 0.0044wt%~0.67wt%, 0. 0046wt%~0.65wt%, 0.0048wt%~0.63wt%, 0.005wt%~0.61wt%, 0.0052wt%~0.59wt%, 0.0054wt%~0.57wt%, 0.0056wt%~0.55wt%, 0.0058wt%~0.53wt%, 0 .006wt%~0.51wt%, 0.0062wt%~0.49wt%, 0.0064wt%~0.47wt%, 0.0066wt%~0.45wt%, 0.0068wt%~0.43wt%, 0.007wt%~0.41wt%, 0.0072wt%~0.39wt%, 0.0.0074 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 0.09 1 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% by weight~0.071% by weight, 0.042% by weight~0.069% by weight, 0.044% by weight~0.067% by weight, 0.046% by weight~0.065% by weight, 0.048% by weight~0 .063% by weight, 0.05% by weight ~ 0.061% by weight, 0.052% by weight ~ 0.059% by weight, 0.054% by weight ~ 0.057% by weight) are not present. .

[0156] In other embodiments, a formulation or topical formulation described herein is provided, wherein alanine is present in an amount of 0.0035% or more by weight of the formulation (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, 0.1 68, 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% by weight or less (e.g., 0.399, 0.397, 0.395, 0.393, 0.391, 0.389, 0.387, 0.3 85, 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 ,0.217,0.215,0.213,0.211,0.209,0.207,0.205,0.203,0.201,0.199,0.197,0.195,0.193,0.191,0.189,0.187,0.185,0.183,0.181,0.179,0.17 7, 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, 0.1 35, 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, 0.01 1, 0.009, 0.007, 0.005, 0.003, 0.002); or 0.0035% by weight to 0.36% by weight (for example, 0.00351% by weight to 0.359% by weight, 0.00352% by weight to 0.358% by weight, 0.00353% by weight to 0.357% by weight, 0.00354% by weight to 0.356% by weight, 0.00355% by weight to 0.355% by weight, 0.00356% by weight to 0.354% by weight, 0.00357% by weight to 0.353% by weight, 0.00358% by weight to 0.352% by weight, 0.00359% by weight to 0.351% by weight, 0.0036% by weight to 0.35wt%, 0.00361wt%~0.349wt%, 0.00362wt%~0.348wt%, 0.00363wt%~0.347wt%, 0.00364wt%~0.346wt%, 0.00365wt%~0.345wt%, 0.00366wt%~0.344 Weight%, 0.00367wt%~0.343wt%, 0.00368wt%~0.342wt%, 0.00369wt%~0.341wt%, 0.0037wt%~0.34wt%, 0.00371wt%~0.339wt%, 0.00372wt%~0.338wt%, 0. 00373wt%~0.337wt%, 0.00374wt%~0.336wt%, 0.00375wt%~0.335wt%, 0.00376wt%~0.334wt%, 0.00377wt%~0.333wt%, 0.00378wt%~0.332wt%, 0.0037 9wt%~0.331wt%, 0.0038wt%~0.33wt%, 0.00381wt%~0.329wt%, 0.00382wt%~0.328wt%, 0.00383wt%~0.327wt%, 0.00384wt%~0.326wt%, 0.00385wt%~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%, 0.00 398 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% ~0.287 wt%, 0.00424 wt%~0.286 wt%, 0.00425 wt%~0.285 wt%, 0.00426 wt%~0.284 wt%, 0.00427 wt%~0.283 wt%, 0.00428 wt%~0.282 wt%, 0.00429 wt%~0.281 wt%, 0.0043 wt%~0.28 wt%, 0.00431 wt%~0.279 wt%, 0.00432 wt%~0.278 wt%, 0.00433 wt%~0.277 wt%, 0.00434 wt%~0.276 wt%, 0.00435 wt%~0.275 wt%, 0.0.00436 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% ~0.23 wt%, 0.00481 wt%~0.229 wt%, 0.00482 wt%~0.228 wt%, 0.00483 wt%~0.227 wt%, 0.00484 wt%~0.226 wt%, 0.00485 wt%~0.225 wt%, 0.00486 wt%~0.224 wt%, 0.00487 wt%~0.223 wt%, 0.00488 wt%~0.222 wt%, 0.00489 wt%~0.221 wt%, 0.0049 wt%~0.22 wt%, 0.00491 wt%~0.219 wt%, 0.00492 wt%~0.218 wt%, 0.0.00493 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 0.2 0.05 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 % 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 0.189 wt%, 0.00532 wt% to 0.188 wt%, 0.00533 wt% to 0.187 wt%, 0.00534 wt% to 0.186 wt%, 0.00535 wt% to 0.186 wt%, 0.00536 wt% to 0.186 wt%, .00531 wt%~0.179 wt%, 0.00532 wt%~0.178 wt%, 0.00533 wt%~0.177 wt%, 0.00534 wt%~0.176 wt%, 0.00535 wt%~0.175 wt%, 0.00536 wt%~0.174 wt%, 0.00537 wt%~0.173 wt%, 0.00538 wt%~0.172 wt%, 0.00539 wt%~0.171 wt%, 0.0054 wt%~0.17 wt%, 0.00541 wt%~0.169 wt%, 0.00542 wt%~0.168 wt%, 0.00543 wt%~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%, 0.00 556 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% ~0.129 wt%, 0.00582 wt%~0.128 wt%, 0.00583 wt%~0.127 wt%, 0.00584 wt%~0.126 wt%, 0.00585 wt%~0.125 wt%, 0.00586 wt%~0.124 wt%, 0.00587 wt%~0.123 wt%, 0.00588 wt%~0.122 wt%, 0.00589 wt%~0.121 wt%, 0.0059 wt%~0.12 wt%, 0.00591 wt%~0.119 wt%, 0.00592 wt%~0.118 wt%, 0.00593 wt%~0.117 wt%, 0.0.00594 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%, 0.0057 9 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 0.0.066 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%, 0.0061 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%, 0.0 0.0617 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. 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 0.03 89 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 0.0.0377 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. 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%, 0.0 0.0572 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%, 0.0 0.0584 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 0.0345 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. 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%, 0.0.00568 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%, 0.0 0.0592 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%, 0.0 0.0604 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%, 0.0061 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%, 0.00616wt%~0.0274wt%, 0.00617wt%~0.0273wt%, 0.00618wt%~0.0272wt%, 0.00619wt%~0.0271wt%, 0.0062 Weight% ~ 0.027% by weight, 0.00621% by weight ~ 0.0269% by weight, 0.00622% by weight ~ 0.0268% by weight, 0.00623% by weight ~ 0.0267% by weight, 0.00624% by weight ~ 0 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%.

[0157] In additional embodiments, the formulations or topical formulations described herein comprise 0.00584% or more by weight of the formulation (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, 0.040, 0.041, 0.042, 0.043, 0.044, 0.045, 0.046, 0.047, 0.048, 0.049, 0.050, 0.051, 0.052, 0.053, 0.054, 0.055, 0.056, 0.057, 0.058, 0.059, 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, .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, 0.0 8, 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, 0.3 7, 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 0.584% by weight or less (For example, 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、0.0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004, 0.003, 0.0 ...01); or 0.00584% by weight to 0.584% by weight (e.g., 0.00585% by weight to 0.582% by weight, 0.0059% by weight to 0.58% by weight, 0.0065% by weight to 0.578% by weight, 0.007% by weight to 0.576% by weight, 0.0075% by weight to 0.574% by weight). Amount%, 0.008% to 0.572% by weight, 0.0085% to 0.57% by weight, 0.009% to 0.568% by weight, 0.0095% to 0.566% by weight, 0.01% to 0.564% by weight, 0.015wt%~0.562wt%, 0.02wt%~0.56wt%, 0.025wt%~0.558wt%, 0.03wt%~0.556wt%, 0.035wt%~0.554wt%, 0.04wt% ~0.552wt%, 0.045wt%~0.55wt%, 0.05wt%~0.548wt%, 0.055wt%~0.546wt%, 0.06wt%~0.544wt%, 0.065wt%~0.542 Weight%, 0.07% to 0.54% by weight, 0.075% to 0.538% by weight, 0.08% to 0.536% by weight, 0.085% to 0.534% by weight, 0.09% to 0.532% by weight, 0.09 and glutamine present in a weight percent (wt%) of 5% to 0.53%, 0.1% to 0.528%, 0.15% to 0.526%, 0.2% to 0.524%, 0.25% to 0.522%, 0.3% to 0.52%, 0.35% to 0.518%, 0.4% to 0.516%, 0.45% to 0.514%, 0.5% to 0.512%.

[0158] In further embodiments described herein, there is provided a formulation or topical formulation as described herein, wherein glycine is present in an amount of 0.003% or more by weight of the formulation (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. 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4); 0.3% by weight 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 ,0.234,0.232,0.23,0.228,0.226,0.224,0.222,0.22,0.218,0.216,0.214,0.212,0.21,0.208,0.206,0.204,0.202,0.2,0.198,0.196,0.194,0.1 92, 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, 0.10 6, 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 weight% to 0.3 weight% (for example, 0.0035 weight% to 0.29 weight%, 0.004 weight% to 0.27 weight%, 0.0045 weight% to 0.25 weight%, 0.005 weight% to 0.23 weight%, 0.0055 weight% to 0.21 weight%, 0.006 weight% to 0.19 weight%, 0.0065 weight% to 0.17 weight%, 0.007 weight% to 0.2 weight%, Amount%~0.15wt%, 0.0075wt%~0.13wt%, 0.008wt%~0.11wt%, 0.0085wt%~0.099wt%, 0.009wt%~0.097wt% , 0.0095wt%~0.095wt%, 0.01wt%~0.093wt%, 0.015wt%~0.091wt%, 0.02wt%~0.089wt%, 0.025wt%~0. 0.087% by weight, 0.03% by weight to 0.085% by weight, 0.035% by weight to 0.083% by weight, 0.04% by weight to 0.081% by weight, 0.045% by weight to 0.079% by weight, 0.05% by weight to 0.077% by weight, 0.055% by weight to 0.075% by weight, 0.06% by weight to 0.073% by weight, 0.065% by weight to 0.071% by weight.

[0159] In some embodiments, the formulations or topical formulations of the present disclosure contain 0.0042% or more by weight of the formulation (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, 0.01 02, 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, 0.028 6, 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, 0.0 948, 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, 0.16 8, 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% by weight 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, 0.3 52, 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、0.0.0054, 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. 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%, 0.0 0.093 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%, 0.0147wt%~0.23wt%, 0.0149wt%~0.225wt%, 0.0151wt%~0.22wt%, 0.0153wt%~0.215wt%, 0.0155wt%~0.21wt%, 0.0157wt% ~0.205wt%, 0.0159wt%~0.2wt%, 0.0161wt%~0.195wt%, 0.0163wt%~0.19wt%, 0.0165wt%~0.185wt%, 0.0167wt%~0.18wt% , 0.0169wt%~0.175wt%, 0.0171wt%~0.17wt%, 0.0173wt%~0.165wt%, 0.0175wt%~0.16wt%, 0.0177wt%~0.155wt%, 0.0179 wt%~0.15wt%, 0.0181wt%~0.145wt%, 0.0183wt%~0.14wt%, 0.0185wt%~0.135wt%, 0.0187wt%~0.13wt%, 0.0189wt%~0.12 5wt%, 0.0191wt%~0.12wt%, 0.0193wt%~0.115wt%, 0.0195wt%~0.11wt%, 0.0197wt%~0.105wt%, 0.0199wt%~0.1wt%, 0.0 201wt%~0.095wt%, 0.0203wt%~0.09wt%, 0.0205wt%~0.085wt%, 0.0207wt%~0.08wt%, 0.0209wt%~0.075wt%, 0.0211wt%~ and serine present in a weight percent (wt%) of 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%, and 0.0229 wt% to 0.025 wt%.

[0160] In additional embodiments described herein, there is provided a formulation or topical formulation as disclosed herein, wherein the amino acid blend comprises, consists essentially of, or consists of 0.0356% alanine (4 mM), 0.0584% glutamine (4 mM), 0.03% glycine (4 mM), and 0.042% serine (4 mM).

[0161] In other embodiments described herein, there is provided a formulation or topical formulation as disclosed herein, wherein the therapeutically effective amount of the blend of free amino acids comprises (or consists essentially of, or consists of) alanine (Ala), glutamine (Gln), glycine (Gly), and serine (Ser), each free amino acid being arranged in the following ratio to each of the other free amino acids: 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):10 0(Gly):100(Ser)[0.1 vs. 10], 100(Ala):100(Gln):1(Gly):100(Ser)[0.1 vs. 10], 100(Ala):100(Gln):100(Gly):1(Ser)[0.1 vs. 10], 1(Ala):90(Gln):90(Gly):90(Ser)[0.1 vs. 9], 90(Ala):1(Gln):90(Gly):90(Ser)[0.1 vs. 9], 90(Ala):90(Gln):1(Gly):90(Ser)[0.1 vs. 9], 90(Ala):90(Gln):90(Gly):1(Se r) [0.1 vs. 9], 1(Ala):80(Gln):80(Gly):80(Ser) [0.1 vs. 8], 80(Ala):1(Gln):80(Gly):80(Ser) [0.1 vs. 8], 80(Ala):80(Gln):1(Gly):80(Ser) [0.1 vs. 8], 80(Ala):80(Gln):80(Gly):1(Ser) [0.1 vs. 8], 1(Ala):70(Gln):70(Gly):70(Ser) [0.1 vs. 7], 70(Ala):1(Gln):70(Gly):70(Ser) [0.1 vs. 7], 70(Ala):70( Gln):1(Gly):70(Ser)[0.1 vs. 7], 70(Ala):70(Gln):70(Gly):1(Ser)[0.1 vs. 7], 1(Ala):60(Gln):60(Gly):60(Ser)[0.1 vs. 6], 60(Ala):1(Gln):60(Gly):60(Ser)[0.1 vs. 6], 60(Ala):60(Gln):1(Gly):60(Ser)[0.1 vs. 6], 60(Ala):60(Gln):1(Gly):60(Ser)[0.1 vs. 6], 60(Ala):60(Gln):60(Gly):1(Ser)[0.1 vs. 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):1(Gly):40(Ser)[0.1 to 4], 40(Ala):40(Gln): n):40(Gly):1(Ser)[0.1 vs. 4], 1(Ala):30(Gln):30(Gly):30(Ser)[0.1 vs. 3], 30(Ala):1(Gln):30(Gly):30(Ser)[0.1 vs. 3], 30(Ala):30(Gln):1(Gly):30(Ser)[0.1 vs. 3], 30(Ala):30(Gln):1(Gly):30(Ser)[0.1 vs. 3], 30(Ala):30(Gln):30(Gly):1(Ser)[0.1 vs. 3], 1(Ala):20(Gln):20(Gly):20(Ser)[0.1 vs. 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):1(Gly):10(Ser)[0.1 to 1], 10(Ala):10(Gln):10(Gly):1(Ser)[0.1 to 1], 1(Ala):5(Gl n):5(Gly):5(Ser)[0.1 vs. 0.5], 5(Ala):1(Gln):5(Gly):5(Ser)[0.1 vs. 0.5], 5(Ala):5(Gln):1(Gly):5(Ser)[0.1 vs. 0.5], 5(Ala):5(Gln):5(Gly):1(Ser)[0.1 vs. 0.5], 2(Ala):5(Gln):5(Gly):5(Ser)[0.1 vs. 0.25], 5(Ala):2(Gln):5(Gly):5(Ser)[0.1 vs. 0.25], 5(Ala):5(Gln):2(Gly):5(Ser)[0.1 vs. 0.25], 5(Ala):5(Gln):5(Gly):2(Ser) [0.1 vs. 0.25], 1(Ala):1(Gln):1(Gly):1(Ser) [0.1 vs. 0.1], 2(Ala):1(Gln):1(Gly):1(Ser) [0.2 vs. 0.1], 1(Ala):2(Gln):1(Gly):1(Ser) [0.2 vs. 0.1], 1(Ala):1(Gln):2(Gly):1(Ser) [0.2 vs. 0.1], 1(Ala):1(Gln):2(Gly):1(Ser) [0.2 vs. 0.1], 1(Ala):1(Gln):1(Gly):2(Ser) [0.2 vs. 0.1], 3(Ala):1(Gln):1(Gly):1( Ser) [0.3 vs. 0.1], 1(Ala):3(Gln):1(Gly):1(Ser) [0.3 vs. 0.1], 1(Ala):1(Gln):3(Gly):1(Ser) [0.3 vs. 0.1], 1(Ala):1(Gln):1(Gly):3(Ser) [0.3 vs. 0.1], 4(Ala):1(Gln):1(Gly):1(Ser) [0.4 vs. 0.1], 1(Ala):4(Gln):1(Gly):1(Ser) [0.4 vs. 0.1], 1(Ala):1(Gln):4(Gly):1(Ser) [0.4 vs. 0.1], 1(Ala):1(Gln): 1(Gly):4(Ser)[0.4 vs. 0.1], 1(Ala):5(Gln):5(Gly):5(Ser)[0.5 vs. 0.1], 1(Ala):5(Gln):1(Gly):1(Ser)[0.5 vs. 0.1], 1(Ala):1(Gln):5(Gly):1(Ser)[0.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):5(Ser)[0.5 vs. 0.1], 6(Ala):1(Gln):1(Gly):1(Ser)[0.6 vs. 0.1], 1(Ala):6(Gln):1(Gly):1(Ser)[0.6 vs. 0.1], 1(Ala ):1(Gln):6(Gly):1(Ser)[0.6 vs. 0.1], 1(Ala):1(Gln):1(Gly):6(Ser)[0.6 vs. 0.1], 7(Ala):1(Gln):1(Gly):1(Ser)[0.7 vs. 0.1], 1(Ala):7(Gln):1(Gly):1(Ser)[0.7 vs. 0.1], 1(Ala):1(Gln):7(Gly):1(Ser)[0.7 vs. 0.1], 1(Ala):1(Gln):7(Gly):1(Ser)[0.7 vs. 0.1], 1(Ala):1(Gln):1(Gly):7(Ser)[0.7 vs. 0.1], 8(Ala):1(Gln):1(Gly):1(Ser)[0.8 vs. 0.1], 1(Ala):8(Gln):1(Gly):1(Ser) [0.8 vs. 0.1], 1(Ala):1(Gln):8(Gly):1(Ser) [0.8 vs. 0.1], 1(Ala):1(Gln):1(Gly):8(Ser) [0.8 vs. 0.1], 9(Ala):1(Gln):1(Gly):1(Ser) [0.9 vs. 0.1], 1(Ala):9(Gln):1(Gly):1(Ser) [0.9 vs. 0.1], 1(Ala):1(Gln):9(Gly):1(Ser) [0.9 vs. 0.1], 1(Ala):1(Gln):1(Gly ):9(Ser)[0.9 vs. 0.1], 10(Ala):1(Gln):1(Gly):1(Ser)[1 vs. 0.1], 1(Ala):10(Gln):1(Gly):1(Ser)[1 vs. 0.1], 1(Ala):1(Gln):10(Gly):1(Ser)[1 vs. 0.1], 1(Ala):1(Gln):1(Gly):10(Ser)[1 vs. 0.1], 20(Ala):1(Gln):1(Gly):1(Ser)[2 vs. 0.1], 1(Ala):20(Gln):1(Gly):1(Ser)[2 vs. 0.1], 1(Ala):1(Gln) n):20(Gly):1(Ser)[2 vs. 0.1], 1(Ala):1(Gln):1(Gly):20(Ser)[2 vs. 0.1], 30(Ala):1(Gln):1(Gly):1(Ser)[3 vs. 0.1], 1(Ala):30(Gln):1(Gly):1(Ser)[3 vs. 0.1], 1(Ala):1(Gln):30(Gly):1(Ser)[3 vs. 0.1], 1(Ala):1(Gln):1(Gly):30(Ser)[3 vs. 0.1], 40(Ala):1(Gln):1(Gly):1(Ser)[4 vs. 0.1], 1(Al a):40(Gln):1(Gly):1(Ser)[4 vs. 0.1], 1(Ala):1(Gln):40(Gly):1(Ser)[4 vs. 0.1], 1(Ala):1(Gln):1(Gly):40(Ser)[4 vs. 0.1], 50(Ala):1(Gln):1(Gly):1(Ser)[5 vs. 0.1], 1(Ala):50(Gln):1(Gly):1(Ser)[5 vs. 0.1], 1(Ala):1(Gln):50(Gly):1(Ser)[5 vs. 0.1], 1(Ala):1(Gln):1(Gly):50(Ser)[5 vs. 0.1], 60(Ala):1(Gln):1(Gly):1(Ser)[6 vs. 0.1], 1(Ala):60(Gln):1(Gly):1(Ser)[6 vs. 0.1], 1(Ala):1(Gln):60(Gly):1(Ser)[6 vs. 0.1], 1(Ala):1(Gln):1(Gly):60(Ser)[6 vs. 0.1], 1(Ala):1(Gln):1(Gly):60(Ser)[6 vs. 0.1], 70(Ala):1(Gln):1(Gly):1(Ser)[7 vs. 0.1], 1(Ala):70(Gln):1(Gly):1(Ser)[7 vs. 0.1], 1(Ala):1(Gln):70(Gly):1(S er) [7 vs. 0.1], 1(Ala):1(Gln):1(Gly):70(Ser) [7 vs. 0.1], 80(Ala):1(Gln):1(Gly):1(Ser) [8 vs. 0.1], 1(Ala):80(Gln):1(Gly):1(Ser) [8 vs. 0.1], 1(Ala):1(Gln):80(Gly):1(Ser) [8 vs. 0.1], 1(Ala):1(Gln):1(Gly):80(Ser) [8 vs. 0.1], 1(Ala):1(Gln):1(Gly):80(Ser) [8 vs. 0.1], 90(Ala):1(Gln):1(Gly):1(Ser) [9 vs. 0.1], 1(Ala):90(Gln):1( Gly):1(Ser)[9 vs. 0.1], 1(Ala):1(Gln):90(Gly):1(Ser)[9 vs. 0.1], 1(Ala):1(Gln):1(Gly):90(Ser)[9 vs. 0.1], 100(Ala):1(Gln):1(Gly):1(Ser)[10 vs. 0.1], 1(Ala):100(Gln):1(Gly):1(Ser)[10 vs. 0.1], 1(Ala):1(Gln):100(Gly):1(Ser)[10 vs. 0.1], 1(Ala):1(Gln):100(Gly):1(Ser)[10 vs. 0.1], 1(Ala):1(Gln):1(Gly):100(Ser)[10 vs. 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], 1(Ala):1(Gln):85(Gly):1(Ser) [8.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):85(Ser) [8.5 vs. 0.1], 75(Ala):1(Gln):1(Gly):1(Ser) [7.5 vs. 0.1], 1(Ala):75(Gln):1(Gly):1(Ser) [7.5 vs. 0.1], 1(Ala):1(Gln):75 ... er) [5.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):55(Ser) [5.5 vs. 0.1], 65(Ala):1(Gln):1(Gly):1(Ser) [6.5 vs. 0.1], 1(Ala):65(Gln):1(Gly):1(Ser) [6.5 vs. 0.1], 1(Ala):1(Gln):65(Gly):1(Ser) [6.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):65(Ser) [6.5 vs. 0.1], 55(Ala):1(Gln):1(Gly):1(Ser) [5.5 vs. 0.1], 1(Ala):5 5(Gln):1(Gly):1(Ser)[5.5 vs. 0.1], 1(Ala):1(Gln):55(Gly):1(Ser)[5.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):55(Ser)[5.5 vs. 0.1], 45(Ala):1(Gln):1(Gly):1(Ser)[4.5 vs. 0.1], 1(Ala):45(Gln):1(Gly):1(Ser)[4.5 vs. 0.1], 1(Ala):1(Gln):45(Gly):1(Ser)[4.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):45(Ser)[4 .5 vs. 0.1], 35(Ala):1(Gln):1(Gly):1(Ser)[3.5 vs. 0.1], 1(Ala):35(Gln):1(Gly):1(Ser)[3.5 vs. 0.1], 1(Ala):1(Gln):35(Gly):1(Ser)[3.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):35(Ser)[3.5 vs. 0.1], 25(Ala):1(Gln):1(Gly):1(Ser)[2.5 vs. 0.1], 1(Ala):25(Gln):1(Gly):1(Ser)[2.5 vs. 0.1], 1(Ala):1(Gln :25(Gly):1(Ser)[2.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):25(Ser)[0.1 vs. 0.5], 15(Ala):1(Gln):1(Gly):1(Ser)[1.5 vs. 0.1], 1(Ala):15(Gln):1(Gly):1(Ser)[1.5 vs. 0.1], 1(Ala):1(Gln):15(Gly):1(Ser)[1.5 vs. 0.1], 1(Ala):1(Gln):15(Gly):1(Ser)[1.5 vs. 0.1], 1(Ala):1(Gln):1(Gly):15(Ser)[1.5 vs. 0.1], 14(Ala):1(Gln):1(Gly):1(Ser)[1.4 vs. 0.1], 1(Ala):14(Gln):1(Gly):1(Ser) [1.4 vs. 0.1], 1(Ala):1(Gln):14(Gly):1(Ser) [1.4 vs. 0.1], 1(Ala):1(Gln):1(Gly):14(Ser) [1.4 vs. 0.1], 13(Ala):1(Gln):1(Gly):1(Ser) [1.3 vs. 0.1], 1(Ala):13(Gln):1(Gly):1(Ser) [1.3 vs. 0.1], 1(Ala):1(Gln):13(Gly):1(Ser) [1.3 vs. 0.1], 1(Ala):1(Gln):1 (Gly):13(Ser)[1.3 vs. 0.1], 12(Ala):1(Gln):1(Gly):1(Ser)[1.2 vs. 0.1], 1(Ala):12(Gln):1(Gly):1(Ser)[1.2 vs. 0.1], 1(Ala):1(Gln):12(Gly):1(Ser)[1.2 vs. 0.1], 1(Ala):1(Gln):12(Gly):1(Ser)[1.2 vs. 0.1], 1(Ala):1(Gln):1(Gly):12(Ser)[1.2 vs. 0.1], 11(Ala):1(Gln):1(Gly):1(Ser)[1.1 vs. 0.1], 1(Ala):11(Gln):1(Gly):1(Ser)[1.1 vs. 0.1] .1], 1(Ala):1(Gln):11(Gly):1(Ser)[1.1 vs. 0.1], 1(Ala):1(Gln):1(Gly):11(Ser)[1.1 vs. 0.1], 10(Ala):1(Gln):1(Gly):1(Ser)[1 vs. 0.1], 1(Ala):10(Gln):1(Gly):1(Ser)[1 vs. 0.1], 1(Ala):1(Gln):10(Gly):1(Ser)[1 vs. 0.1], 1(Ala):1(Gln):10(Gly):1(Ser)[1 vs. 0.1], 19(Ala):2(Gln):2(Gly):2 (Ser) [0.95 vs. 0.1], 2(Ala):19(Gln):2(Gly):2(Ser) [0.95 vs. 0.1], 2(Ala):2(Gln):19(Gly):2(Ser) [0.95 vs. 0.1], 2(Ala):2(Gln):2(Gly):19(Ser) [0.95 vs. 0.1], 17(Ala):2(Gln):2(Gly):2(Ser) [0.85 vs. 0.1], 2(Ala):17(Gln):2(Gly):2(Ser) [0.85 vs. 0.1], 2(Ala):2(Gln):17(Gly):2(Ser) [0.85 vs. 0.1], 2(Ala):2(Gln):17(Gly):2(Ser) [0.85 vs. 0.1]1], 2(Ala):2(Gln):2(Gly):17(Ser) [0.85 vs. 0.1], 15(Ala):2(Gln):2(Gly):2(Ser) [0.75 vs. 0.1], 2(Ala):15(Gln):2(Gly):2(Ser) [0.75 vs. 0.1], 2(Ala):2(Gln):15(Gly):2(Ser) [0.75 vs. 0.1], 2(Ala):2(Gln):2(Gly):15(Ser) [0.75 vs. 0.1], 13(Ala):2(Gln):2(Gly):2(Ser) [0.65 vs. 0.1], 2(Ala):13(Gln ):2(Gly):2(Ser)[0.65 vs. 0.1], 2(Ala):2(Gln):13(Gly):2(Ser)[0.65 vs. 0.1], 2(Ala):2(Gln):2(Gly):13(Ser)[0.65 vs. 0.1], 11(Ala):2(Gln):2(Gly):2(Ser)[0.55 vs. 0.1], 2(Ala):11(Gln):2(Gly):2(Ser)[0.55 vs. 0.1], 2(Ala):2(Gln):11(Gly):2(Ser)[0.55 vs. 0.1], 2(Ala):2(Gln):2(Gly):11(Ser)[ 0.55 vs. 0.1], 9(Ala):2(Gln):2(Gly):2(Ser) [0.45 vs. 0.1], 2(Ala):9(Gln):2(Gly):2(Ser) [0.45 vs. 0.1], 2(Ala):2(Gln):9(Gly):2(Ser) [0.45 vs. 0.1], 2(Ala):2(Gln):9(Gly):2(Ser) [0.45 vs. 0.1], 2(Ala):2(Gln):2(Gly):9(Ser) [0.45 vs. 0.1], 7(Ala):2(Gln):2(Gly):2(Ser) [0.35 vs. 0.1], 2(Ala):7(Gln):2(Gly):2(Ser) [0.35 vs. 0.1], 2(Ala):2(Gln ):7(Gly):2(Ser)[0.35 vs. 0.1], 2(Ala):2(Gln):2(Gly):7(Ser)[0.35 vs. 0.1], 5(Ala):2(Gln):2(Gly):2(Ser)[0.25 vs. 0.1], 2(Ala):5(Gln):2(Gly):2(Ser)[0.25 vs. 0.1], 2(Ala):2(Gln):5(Gly):2(Ser)[0.25 vs. 0.1], 2(Ala):2(Gln):2(Gly):5(Ser)[0.25 vs. 0.1], 3(Ala):2(Gln):2(Gly):2(Ser)[0.15 vs. 0.1], 2(Ala):3(Gln):2(Gly):2(Ser) [0.15 vs. 0.1], 2(Ala):2(Gln):3(Gly):2(Ser) [0.15 vs. 0.1], 2(Ala):2(Gln):2(Gly):3(Ser) [0.15 vs. 0.1], 2(Ala):2(Gln):2(Gly):3(Ser) [0.15 vs. 0.1], 2(Ala):3(Gln):3(Gly):3(Ser) [0.1 vs. 0.15], 3(Ala):2(Gln):3(Gly):3(Ser) [0.1 vs. 0.15], 3(Ala):3(G ln):3(Gly):2(Ser) [0.1 vs. 0.15], 2(Ala):5(Gln):5(Gly):5(Ser) [0.1 vs. 0.25], 5(Ala):2(Gln):5(Gly):5(Ser) [0.1 vs. 0.25], 5(Ala):5(Gln):2(Gly):5(Ser) [0.1 vs. 0.25], 5(Ala):5(Gln):2(Gly):5(Ser) [0.1 vs. 0.25], 5(Ala):5(Gln):5(Gly):2(Ser) [0.1 vs. 0.25], 2(Ala):7(Gln):7(Gly):7(Ser) [0.1 vs. 0.35], 7(Ala):2(Gln):7(Gly):7(Ser) er) [0.1 vs. 0.35], 7(Ala):7(Gln):2(Gly):7(Ser) [0.1 vs. 0.35], 7(Ala):7(Gln):7(Gly):2(Ser) [0.1 vs. 0.35], 2(Ala):9(Gln):9(Gly):9(Ser) [0.1 vs. 0.45], 9(Ala):2(Gln):9(Gly):9(Ser) [0.1 vs. 0.45], 9(Ala):9(Gln):2(Gly):9(Ser) [0.1 vs. 0.45], 9(Ala):9(Gln):9(Gly):2(Ser) [0.1 vs. 0.45], 2(Ala):11(Gln):11(Gly):11(Ser) [0.1 vs. 0.55], 11(Ala):2(Gln):11(Gly):11(Ser) [0.1 vs. 0.55], 11(Ala):11(Gln):2(Gly):11(Ser) [0.1 vs. 0.55], 11(Ala):11(Gln):11(Gly):2(Ser) [0.1 vs. 0.55], 2(Ala):13(Gln):13(Gly):13(Ser) [0.1 vs. 0.65], 13(Ala):2(Gln):13(Gly):13(Ser) [0.1 vs. 0.65], 13(Ala):13(Gln):2(Gly):13(Ser) [0.1 vs. 0.65], 13(Ala):13(Gln):13(Gly):2(Ser) [0.1 vs. 0.65], 2(Ala):15(Gln):15(Gly):15(Ser) [0.1 vs. 0.75], 15(Ala):2(Gln):15(Gly):15(Ser) [0.1 vs. 0.75], 15(Ala):15(Gln):2(Gly):15(Ser) [0.1 vs. 0.75], 15(Ala):15(Gln):15(Gly) ly):2(Ser)[0.1 vs. 0.75], 2(Ala):17(Gln):17(Gly):17(Ser)[0.1 vs. 0.85], 17(Ala):2(Gln):17(Gly):17(Ser)[0.1 vs. 0.85], 17(Ala):17(Gln):2(Gly):17(Ser)[0.1 vs. 0.85], 17(Ala):17(Gln):2(Gly):17(Ser)[0.1 vs. 0.85], 17(Ala):17(Gln):17(Gly):2(Ser)[0.1 vs. 0.85], 2(Ala):19(Gln):19(Gly):19(Ser)[0.1 vs. 0.95], 19(Ala):17(Gln):17(Gly):17(Ser)[0.1 vs. 0.95] 19(Ala):19(Gln):2(Gly):19(Ser)[0.1 vs. 0.95], 19(Ala):19(Gln):2(Gly):19(Ser)[0.1 vs. 0.95], 19(Ala):19(Gln):19(Gly):2(Ser)[0.1 vs. 0.95]; or 1:100 to 100:1 (e.g., 1(Ala):100(Gln):100(Gly):100(Ser)[0.1 vs. 10] to 100(Ala):1(Gln):1(Gly):1(Ser)[10 vs. 0.1], 100(Ala):1( Gln):100(Gly):100(Ser)[0.1 vs. 10]~1(Ala):100(Gln):1(Gly):1(Ser)[10 vs. 0.1], 100(Ala):100(Gln):1(Gly):100(Ser)[0.1 vs. 10]~1(Ala):1(Gln):100(Gly):1(Ser)[10 vs. 0.1], 100(Ala):100(Gln):100(Gly):1(Ser)[0.1 vs. 10]~1(Ala):1(Gln):1(Gly):100(Ser)[10 vs. 0.1]).

[0162] In additional embodiments, the formulations or topical preparations described herein comprise a therapeutically effective amount of a plant extract (e.g., a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense) or a plant extract blend (e.g., BOSEXIL® (INDENA SpA) or Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense; texturing agents and / or fillers (e.g., cellulose (microcrystalline)); emulsifiers or phospholipids (in some embodiments, phospholipids are used as emulsifiers) (e.g., lecithin or phosphatidylcholine); and anti-caking agents (e.g., silica, to adsorb water in hygroscopic applications); and optionally, a plant extract or plant extract blend comprising (or consisting essentially of, or consisting of) a dermatologically acceptable carrier and / or additive, wherein the plant extract or plant extract blend comprises 0.025% w / w or more (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, 0.0 53, 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); 5w / 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.<h2 style=";text-align:left;direction:ltr">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 ); .028w / w%~4.7w / w%、0.029w / w%~4.6w / w%、0.03w / w%~4.5w / w%、0.031w / w%~ 4.4w / w%, 0.032w / w%~4.3w / w%, 0.033w / w%~4.2w / w%, 0.034w / w%~4.1w / w%, 0.035w / w%~4w / w%, 0.036w / w%~3.9w / w%, 0.037w / w%~3.8w / w%, 0.038w / w%~ 3.7w / w%, 0.039w / w%~3.6w / w%, 0.04w / w%~3.5w / w%, 0.041w / w%~3.4w / w%, 0.042w / w%~3.3w / w%, 0.043w / w%~3.2w / w%, 0.044w / w%~3.1w / w%, 0.045w / w% ~3w / w%、0.046w / w%~2.9w / w%、0.047w / w%~2.8w / w%、0.048w / w%~2.7w / w%、0 .049w / w%~2.6w / w%、0.05w / w%~2.5w / w%、0.051w / w%~2.4w / w%、0.052w / w%~ 2.3w / w%, 0.053w / w%~2.2w / w%, 0.054w / w%~2.1w / w%, 0.055w / w%~2w / w%, 0.056w / w%~1.9w / w%, 0.057w / w%~1.8w / w%, 0.058w / w%~1.7w / w%, 0.059w / w%~ 1.6w / w%、0.06w / w%~1.5w / w%、0.061w / w%~1.4w / w%、0.062w / w%~1.3w / w%、0 0.063w / w%~1.2w / w%、0.064w / w%~1.1w / w%、0.065w / w%~1w / w%、0.066w / w%~0.99w / w%、0.067w / w%~0.98w / w%、0.068w / w%~0.97w / w%、0.069w / w%~0.96w / w%、0.07w / w%~0.95w / w%、0.071w / w%~0.94w / w%、0.072w / w%~0.93w / w%、0.07 3w / w%~0.92w / w%、0.074w / w%~0.91w / w%、0.075w / w%~0.9w / w%、0.076w / w%~0.89w / w%、0.077w / w%~0.88w / w%、0.078w / w%~0.87w / w%、0.079w / w%~0.86w / w%、0.08w / w%~0.85w / w%、0.081w / w%~0.84w / w%、0.082w / w%~0.83w / w%、0.083w / w%~0.82w / w%、0.084w / w%~0.81w / w%、0.085w / w%~0.8w / w%、0.086w / w% ~0.79w / w%、0.087w / w%~0.78w / w%、0.088w / w%~0.77w / w%、0.089w / w%~0.76w / w%、0.09w / w%~0.75w / w%、0.091w / w%~0.74w / w%、0.092w / w%~0.73w / w%、0. 093w / w%~0.72w / w%、0.094w / w%~0.71w / w%、0.095w / w%~0.7w / w%、0.096w / w%~0.69w / w%、0.097w / w%~0.68w / w%、0.098w / w%~0.67w / w%、0.099w / w%~0.6 6w / w%、0.1w / w%~0.65w / w%、0.101w / w%~0.64w / w%、0.102w / w%~0.63w / w%、0.103w / w%~0.62w / w%、0.104w / w%~0.61w / w%、0.105w / w%~0.6w / w%、0.106w / w %~0.59w / w%、0.107w / w%~0.58w / w%、0.108w / w%~0.57w / w%、0.109w / w%~0.56w / w%、0.11w / w%~0.55w / w%、0.111w / w%~0.54w / w%、0.112w / w%~0.53w / w%、 0.113w / w%~0.52w / w%、0.114w / w%~0.51w / w%、0.115w / w%~0.5w / w%、0.116w / w%~0.49w / w%、0.117w / w%~0.48w / w%、0.118w / w%~0.47w / w%、0.119w / w%~0.<h2 style=";text-align:left;direction:ltr">46w / w%、0.12w / w%~0.45w / w%、0.121w / w%~0.44w / w%、0.122w / w%~0.43w / w%、0.123w / w%~0.42w / w%、0.124w / w%~0.41w / w%、0.125w / w%~0.4w / w%、0.126w / w%~0.39w / w%、0.1 27w / w%~0.38w / w%、0.128w / w%~0.37w / w%、0.129w / w%~0.36w / w%、0.13w / w%~0.35w / w %、0.131w / w%~0.34w / w%、0.132w / w%~0.33w / w%、0.133w / w%~0.32w / w%、0.134w / w%~0. 31w / w%、0.135w / w%~0.3w / w%、0.136w / w%~0.29w / w%、0.137w / w%~0.28w / w%、0.138w / w%~0.27w / w%、0.139w / w%~0.26w / w%、0.14w / w%~0.25w / w%、0.141w / w%~0.24w / w%、0.1 42w / w%~0.23w / w%、0.143w / w%~0.22w / w%、0.144w / w%~0.21w / w%、0.145w / w%~0.2w / w %、0.146w / w%~0.19w / w%、0.147w / w%~0.18w / w%、0.148w / w%~0.17w / w%、0.149w / w%~0.16 w / w%). In some embodiments, the plant extract comprises, consists essentially of, or consists of Boswellia extract, including Boswellia serrata, Burseraceae family, olibanum, or frankincense. Other embodiments are directed to plant extract formulations comprising, consisting essentially of, or consisting of Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense); texturing agents and / or bulking agents (e.g., cellulose (microcrystalline); phospholipids (e.g., lecithin or phosphatidylcholine); and anti-caking agents (e.g., silica, absorbs water in hygroscopic applications). In additional embodiments, the plant extract formulation comprises, consists essentially of, or consists of Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, and silica.

[0163] In further embodiments, there is provided a formulation or topical formulation as described herein, wherein the therapeutically effective amount of the combination of free amino acids and the therapeutically effective amount of the plant extract or plant extract combination inhibits 5-lipoxygenase (an inflammation-inducing enzyme); reduces calcium ion mobilization; and reduces MAP kinase activation. Without wishing to be bound by theory, the Boswellia plant extract or plant extract combination, including Boswellia extract, contains anti-inflammatory properties, which, for example, reduce inflammation induced by 5-lipoxygenase. See, for example, D. Poeckel and O. Werz (see, for example, Curr Med Chem. 13(28):3359-3369, 2006; Singh et al. Phytomedicine. 15(6-7):400-407, 2008).

[0164] In some of the embodiments, the final formulation or topical formulation comprises a therapeutically effective amount of a formulation comprising (or consisting essentially of, or consisting of) alanine, glutamine, glycine, and serine as free amino acids; a therapeutically effective amount of a plant extract comprising (or consisting essentially of, or consisting of) Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense); and optionally, a dermatologically acceptable carrier and / or additive, such as, but not limited to, lecithin, microcrystalline cellulose, and silica.

[0165] In other embodiments, a topical formulation or topical preparation is provided that comprises a therapeutically effective amount of a combination comprising (or consisting essentially of, or consisting of) a combination of at least one of the following amino acids as free amino acids: alanine, glutamine, glycine, and serine; and a therapeutically effective amount of a plant extract or combination of plant extracts, comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense), alone or in combination with at least one of a texturing agent and / or filler (e.g., cellulose (microcrystalline); phospholipids (e.g., lecithin); and an anti-caking agent (e.g., silica, which adsorbs water in hygroscopic applications), a plant extract or a plant extract blend, wherein the amino acid blend and the plant extract blend are in a ratio of 1:100 or more (e.g., 2:100, 4:100, 6:100, 8:100, 10:100, 12:100, 14:1 00, 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);1:100-100:1 (e.g. 1:99-99:1, 1:98-98:1, 1:97-97:1, 1:96-96:1, 1:95-95:1, 1:94-94:1, 1:93-93:1, 1:92-92:1, 1:91-91:1, 1:90-90:1, 1:89-89:1, 1:88-88:1, 1:87-87:1, 1:86-86:1, 1:85-85:1, 1:84-84:1, 1:83-83:1, 1:82-82:1, 1:81-81:1, 1:80-80:1, 1:79-79:1, 1:78-78:1, 1:77-77:1, 1:76~76:1, 1:75~75:1, 1:74~74:1, 1:73~73:1, 1:72~72:1, 1:71~71:1, 1:70~70:1, 1:69~69:1, 1:68~68:1, 1:67~67:1, 1:66~66:1, 1:65~65:1, 1:6 4~64:1, 1:63~63:1, 1:62~62:1, 1:61~61:1, 1:60~60:1, 1:59~59:1, 1:58~58:1, 1:57~57:1, 1:56~56:1, 1:55~55:1, 1:54~54:1, 1:53~53:1, 1:52~52 :1, 1:51~51:1, 1:50~50:1, 1:49~49:1, 1:48~48:1, 1:47~47:1, 1:46~46:1, 1:45~45:1, 1:44~44:1, 1:43~43:1, 1:42~42:1, 1:41~41:1, 1:40~40:1, 1:39~39:1, 1:38~38:1, 1:37~37:1, 1:36~36:1, 1:35~35:1, 1:34~34:1, 1:33~33:1, 1:32~32:1, 1:31~31:1, 1:30~30:1, 1:29~29:1, 1:28~28:1, 1:2 7~27:1, 1:26~26:1, 1:25~25:1, 1:24~24:1, 1:23~23:1, 1:22~22:1, 1:21~21:1, 1:20~20:1, 1:19~19:1, 1:18~18:1, 1:17~17:1, 1:16~16:1, 1:15~1 5:1, 1:14-14:1, 1:13-13:1, 1:12-12:1, 1:11-11:1, 1:10-10:1, 1:9-9:1, 1:8-8:1, 1:7-7:1, 1:6-6:1, 1:5-5:1, 1:4-4:1, 1:3-3:1, 1:2-2:1);

[0166] In other embodiments, any of the formulations or topical formulations described herein are provided that optionally comprise, consist essentially of, or consist of dermatologically acceptable carriers and / or excipients. Non-limiting examples of carriers and additives useful in any of the formulations or topical formulations described herein may be selected from the group consisting of water, sodium phytate, xanthan gum, kaolin, glycerin, polyglycerides (e.g., oleic acid polyglyceride, linoleic acid polyglyceride, linoleic acid polyglyceride), triglycerides (e.g., caprylic acid triglyceride, capric acid triglyceride), sucrose (e.g., sucrose stearate, sucrose distearate), alcohols (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 chinensis (Sweet almond oil), glyceryl stearate, ... chinenesis (jojoba oil), phenoxyethanol, lecithin, tocopherol, tocopheryl acetate, ascorbyl palmitate, citric acid, potassium sorbate, lactic acid, UV protection agent, hydrogenated olive oil decyl esters, decyl coconut, astrocaryum, murumuru seed oil, 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, polymethylmethacrylate, fragrance, nutrients, antioxidants, water, or combinations thereof. In some embodiments, a formulation or topical formulation of the present disclosure comprises (or consists essentially of, or consists of) a therapeutically effective amount of a formulation comprising (or consisting essentially of, or consisting of) alanine, glutamine, glycine, and serine as free amino acids; a therapeutically effective amount of a plant extract or plant extract formulation; a texturing agent and / or bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (in some embodiments, a phospholipid is used as an emulsifier) ​​(e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, which adsorbs water in hygroscopic applications); and, optionally, a dermatologically acceptable carrier and / or additive. Non-limiting examples of at least one phospholipid useful in the formulations of the present disclosure include any phospholipid containing two fatty acids per phosphate backbone, including, but not limited to, phosphatidic acid (phosphatidate) (PA); phosphatidylethanolamine (cephalin) (PE); phosphatidylcholine (lecithin) (PC); phosphatidylserine (PS); phosphatidylinositol (PI); phosphatidylinositol phosphate (PIP); phosphatidylinositol diphosphate (PIP2); and phosphatidylinositol triphosphate (PIP3); ceramides containing a phosphate group; ceramide phosphorylcholine (sphingomyelin) (SPH);Ceramide phosphorylethanolamine (Sphingomelin) (Cer-PE); phospholipids, including 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); acid); vaccenic acid (trans-11-octadecenoic acid); elaidic acid (trans-9-octadecenoic acid (trans-oleic acid); linoleic acid; linolenic acid; paulic 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.

[0167] In additional embodiments, the formulations or topical formulations described herein may be prepared by mixing and / or stirring (concurrently, simultaneously, sequentially) (a) a therapeutically effective amount of a blend of free amino acids, including, consisting essentially of, at least one of alanine, glutamine, glycine, and serine; (b) a therapeutically effective amount of a plant extract or plant extract blend, including, consisting essentially of, or consisting of Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense); and, optionally, (c) a dermatologically acceptable carrier and / or additive.The mixture of (a), (b), and optionally (c) is heated to a temperature sufficient to thoroughly mix, homogenize, or emulsify (a), (b), and optionally (c), and is preferably at least 20° C. (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); or at most 100° C. (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℃~100℃ (e.g., 21℃~99℃, 22℃~98℃, 23℃~97℃, 24℃~96℃, 25℃~95℃, 26℃~94℃, 27℃~93℃, 28℃~92℃, 29℃~91℃, 30℃~90℃, 31℃~89℃, 32℃~88℃, 33℃ 3. Thoroughly mix, homogenize or milk until the temperature is between 87°C, 34°C to 86°C, 35°C to 85°C, 36°C to 84°C, 37°C to 83°C, 38°C to 82°C, 39°C to 81°C, 40°C to 80°C, 41°C to 79°C, 42°C to 78°C, 43°C to 77°C, 44°C to 76°C, 45°C to 75°C, 46°C to 74°C, 47°C to 73°C, 48°C to 72°C, 49°C to 71°C, 50°C to 70°C, 51°C to 69°C, 52°C to 68°C, 53°C to 67°C, 54°C to 66°C, 55°C to 65°C, 56°C to 64°C, 57°C to 63°C, 58°C to 62°C, 59°C to 61°C. The plant extract blends of the formulations or topical formulations described herein can be heated (together or separately) for a time sufficient to melt the emulsion (e.g., seconds to minutes). In some embodiments, the plant extract blends of the formulations or topical formulations described herein further comprise (or consist essentially of, or consist of) texturing agents and / or fillers (e.g., cellulose (microcrystalline)); phospholipids (e.g., lecithin or phosphatidylcholine); and anti-caking agents (e.g., silica, which adsorbs water in hygroscopic applications). See, e.g., Example 7.

[0168] Use of formulations for repairing and strengthening the skin barrier In some embodiments of the disclosure, a formulation or topical formulation as described herein is provided, comprising a therapeutically effective amount of a blend of free amino acids, 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 blend (e.g., Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense; texturing agents and / or fillers (e.g., cellulose (microcrystalline); phospholipids (e.g., lecithin or phosphatidylcholine); and anti-caking agents (e.g., silica, absorbs water in hygroscopic applications). In additional aspects, a therapeutically effective amount of the combination of free amino acids and a therapeutically effective amount of a plant extract or combination of plant extracts improves skin barrier repair. Improvements in skin barrier repair include skin barrier correction, skin barrier enhancement, skin barrier preservation, skin barrier integrity, wound healing; epidermal cells The effects of the present invention include, but are not limited to, the growth, stimulation, proliferation, differentiation, and migration of keratinocytes, endothelial cells, and fibroblasts; promotion of dermal regeneration, and combinations thereof. Further aspects are directed to the modulation of epidermal growth factor (EGF) using a therapeutically effective amount of an amino acid blend or formulation comprising (or consisting essentially of, or consisting of) an amino acid blend described herein, such amino acid blend comprising, consisting essentially of, or consisting of at least one of alanine, glutamine, glycine, and serine, or salts thereof, for example in a 1:1:1:1 ratio.

[0169] In an additional embodiment, a method of improving skin barrier repair of the skin of a subject includes a topical formulation comprising a therapeutically effective amount of a blend of free amino acids 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 blend (e.g., Boswellia serrata resin extract, cellulose (microcrystalline), lecithin, silica), comprising a Boswellia extract (e.g., Boswellia serrata; Burseraceae family; olibanum; frankincense), alone or optionally in combination with at least one of a texturing agent and / or filler (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, which adsorbs water in hygroscopic applications); and optionally a dermatologically acceptable carrier and / or additive; and determining that the barrier marker gene is increased by an average fold change of 1.5 to 40 for amino acid treated skin compared to amino acid untreated skin when tested by a method that detects expression of a barrier marker gene (such as, but not limited to, EGF) (see, e.g., Figures 8-14). Improved skin barrier repair is measured or indicated by an average fold increase in skin barrier marker gene expression, such as EGF, of 1.5 to 40 when comparing skin treated with any of the formulations described herein to untreated skin, when barrier marker gene expression is tested by expression detection methods. Methods of improving skin barrier repair include, but are not limited to, correcting the skin barrier, strengthening the skin barrier, preserving the skin barrier, improving skin barrier integrity, wound healing; growth, stimulation, proliferation, differentiation, and migration of epidermal cells, keratinocytes, endothelial cells, and fibroblasts; promoting dermal regeneration, and any combination thereof.

[0170] Use of formulations for improving skin cell barrier integrity Formulations and kits The present disclosure provides a therapeutic or pharmaceutical formulation comprising a therapeutically effective amount of the formulation and, optionally, one or more pharma- ceutically acceptable carriers. The present disclosure provides a therapeutic, pharmaceutical, cosmetic, or nutritional formulation comprising a therapeutically effective amount of the formulation and, optionally, one or more pharma- ceutically acceptable carriers. Such pharmaceutical carriers may be liquids, such as water. The therapeutic formulation may also include excipients, adjuvants, flavorings, etc., that facilitate processing of the active compound into a medicament that can be used pharma- ceutically. Suitable formulations vary depending on the route of administration selected. In one embodiment, the therapeutic formulation and all components contained therein are sterile. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin. Such formulations contain a therapeutically effective amount of the therapeutic formulation together with a suitable amount of carriers to provide a form for proper administration to the patient. The formulation should be suitable for enteral administration methods.

[0171] In one embodiment, administration of the formulation may be systemic. Oral, intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, intracerebroventricular, intranasal, transmucosal, subcutaneous, topical, rectal, and other modes of administration are all contemplated, as are any combinations thereof. In certain embodiments, the formulations described herein are administered to the skin (e.g., administered topically, transdermally, and / or subcutaneously, or any combination thereof). The formulations described herein may be incorporated into gauzes, pads (adhesive or non-adhesive), bandages, and / or dressings for chronic application to the skin. In certain embodiments, the formulations described herein are administered to the skin in combination with oral administration of the formulations described herein. In certain such embodiments, oral administration of the formulations described herein is performed before, during, or after administration of the formulation to the skin. As described herein, administration to the skin may be achieved by topical, transdermal, and / or subcutaneous administration.

[0172] In one embodiment, for injection, the active ingredient can be formulated in an aqueous solution, in one example, a physiologically compatible buffer solution. For transmucosal administration, a penetrant suitable for the barrier to be permeated is used in the formulation. For oral administration, the active ingredient can be combined with a suitable carrier for inclusion in tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. The formulation 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 from a pressurized pack or nebulizer using a suitable propellant. The formulation can also be administered as a powder by inhalation or other routes.

[0173] The therapeutically effective dose of the formulations described herein can be determined by those skilled in the art to achieve the desired strength of cell-cell adhesion that reflects healthy barrier integrity of the skin. Skin cell proliferation, skin cell differentiation, and / or increased expression of marker genes indicative of cell-cell adhesion can be evaluated using the assays described herein. Immunohistochemical staining, behavioral evaluation, and / or electrophysiological techniques can also be used to evaluate skin barrier integrity.

[0174] In some embodiments, the method according to the present disclosure includes administration of the therapeutic formulation by a sustained release system. Examples of suitable sustained release systems include suitable polymeric materials (semipermeable polymer matrices in the form of shaped articles, such as films, or microcapsules), suitable hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, and sparingly soluble derivatives (e.g., as a sparingly soluble salt, etc.). The sustained release formulations can be administered orally, parenterally, intracisternally, intraperitoneally, topically (as a powder, ointment, gel, drop, or transdermal patch), or as a buccal or nasal spray. Sustained release matrices include polylactic acid, copolymers of L-glutamic acid and gamma-ethyl-L-glutamate, poly(2-hydroxyethyl methacrylate), ethylene vinyl acetate, or poly-D-(-)-3-hydroxybutyric acid.

[0175] In one embodiment, implantable drug infusion devices may be used to provide a patient with a constant and long-term dosage or infusion of a therapeutic formulation. Such devices may be classified as either active or passive.

[0176] In one embodiment, polymers can be used for ion-controlled release. A variety of degradable and non-degradable polymer matrices can be used for controlled drug delivery, such as block copolymers, polaxamer 407, hydroxyapatite, and liposomes.

[0177] The pharmaceutical formulation of the present invention may be used alone or in combination with one or more drugs to be effective in treating disease.The formulation may also be formulated in combination with at least one other agent, such as a stabilizing compound or a buffering compound, and may be dissolved and administered in any sterile biocompatible pharmaceutical carrier, including but not limited to saline, buffered saline, dextrose, and water.In addition to the key components, cells, or influencing factors of the formulation discussed herein, the formulation may contain suitable pharmaceutically acceptable carriers, including excipients and auxiliaries, which facilitate the processing of the active compound into a preparation 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 multi-dose container.

[0178] In one embodiment, the formulation may further contain other proliferation and / or differentiation inducers, examples include fibroblast growth factor (FGF), epidermal growth factor (EGF), and retinoic acid.

[0179] The preparation may further contain other commonly used additives such as antioxidants, buffers, bacteriostatic agents, etc., and may be formulated into injection preparations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, tablets, etc., by further adding diluents, dispersants, surfactants, binders, lubricants, etc.

[0180] The food preparation of the present invention can be contained in a health functional food. The health functional food can be prepared according to the method commonly used in the art, and commonly used raw materials and ingredients can be added when preparing the health functional food. When the preparation described herein is included in the health functional food, the preparation can be added alone or together with another health functional food or other food ingredient(s) according to the method commonly used. The amount of active ingredient can be appropriately determined according to the purpose of use (e.g., prevention, health improvement, or therapeutic intervention). The food preparation can further include, for example, a prebiotic or probiotic substance.

[0181] The type of food is not limited. Examples of foods to which the formulation can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products such as ice cream, soup, beverages, tea, drinks, alcoholic beverages, multivitamins, etc.

[0182] Kits (e.g., pharmaceutical, therapeutic, cosmetic, or nutritional packs) are also encompassed by the present disclosure. The kits provided may include a pharmaceutical formulation or compound described herein and a container (e.g., a vial, an ampoule, a bottle, a syringe, and / or a dispenser package, or other suitable container). In some embodiments, the kits provided optionally further include a second container that includes a pharmaceutical excipient for diluting or suspending the pharmaceutical formulation or compound described herein. In some embodiments, the pharmaceutical formulation or compound provided in the first container and the second container combined to form a unit dosage form.

[0183] Thus, in one aspect, a kit is provided, comprising a first container comprising the formulation described herein.In certain embodiments, the kit is useful for treating disorders (e.g., skin disorders) in a subject in need of the treatment.In certain embodiments, the kit is useful for preventing disorders (e.g., skin disorders) in a subject in need of the treatment.

[0184] In certain embodiments, the kits described herein further include instructions for using the formulations included in the kit. The kits described herein may also include information required by regulatory agencies such as the US Food and Drug Administration (FDA). In certain embodiments, the information included in the kit is prescription information. In certain embodiments, the kit and instructions provide for the treatment and / or prevention of disorders (e.g., skin disorders) in subjects in need thereof. The kits described herein may include one or more additional pharmaceutical or other agents as described herein in separate formulations.

[0185] Method of administration The formulations can be administered by any of a number of routes, including, but not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracerebroventricular, transdermal, subcutaneous, intraperitoneal, intranasal, parenteral, topical, sublingual, or rectal means.

[0186] For example, by using "consisting essentially of," the therapeutic formulation does not contain any unspecified ingredients, including, but not limited to, free amino acids, dipeptides, oligopeptides, or polypeptides or proteins; and monosaccharides, disaccharides, oligosaccharides, polysaccharides, and carbohydrates, that have a direct beneficial or detrimental therapeutic effect on improving skin cell barrier integrity. Also, by using the term "consisting essentially of," the formulation may include substances that do not have a therapeutic effect on improving skin cell barrier integrity, such as carriers, excipients, adjuvants, flavorings, and the like, that do not affect skin cell barrier integrity.

[0187] In some embodiments, the formulations (e.g., topical formulations) described herein can be applied topically to the skin of a subject in need of skin barrier repair, such formulations comprising a therapeutically effective amount of an amino acid blend comprising, 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 (e.g., a plant extract blend) comprising, consisting essentially of, or consisting of Boswellia extract (e.g., Boswellia serrata; Burseraceae family; Olibanum; Frankincense), alone or in combination with at least one of a texturing agent and / or bulking agent (e.g., cellulose (microcrystalline)); a phospholipid (e.g., lecithin or phosphatidylcholine); and an anti-caking agent (e.g., silica, which adsorbs water in hygroscopic applications). and optionally a dermatologically acceptable carrier and / or additive, wherein the concentration of the therapeutically effective amount of amino acid is 0.1 mM or more (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 0.8, 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 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 9.9 mM, 0.8 ...4mM, 0.8mM~9.3mM, 0.9mM~9.2mM, 1mM~9.1mM), 1.1mM~9mM, 1.2mM~8.9mM, 1 .3mM~8.8mM, 1.4mM~8.7mM, 1.5mM~8.6mM, 1.6mM~8.5mM, 1.7mM~8.4mM, 1.8 mM~8.3mM, 1.9mM~8.2mM, 2mM~8.1mM, 2.1mM~8mM, 2.2mM~7.9mM, 2.3mM~7.8 mM, 2.4mM~7.7mM, 2.5mM~7.6mM, 2.6mM~7.5mM, 2.7mM~7.4mM, 2.8mM~7.3mM, 2.9mM~7.2mM, 3mM~7.1mM, 3.1mM~7mM, 3.2mM~6.9mM, 3.3mM~6.8mM, 3.4mM~ 6.7mM, 3.5mM~6.6mM, 3.6mM~6.5mM, 3.7mM~6.4mM, 3.8mM~6.3mM, 3.9mM~6. 2mM, 4mM~6.1mM, 4.1mM~6mM, 4.2mM~5.9mM, 4.3mM~5.8mM, 4.4mM~5.7mM, 4. 5mM~5.6mM, 4.6mM~5.5mM, 4.7mM~5.4mM, 4.8mM~5.3mM, 4.9mM~5.2mM, 5mM~5 0.1 mM), and a therapeutically effective amount of the plant extract (or plant extract concentration) is 0.01% by weight or more (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, 0.70, 0.75, 0.76, 0.77, 0.78, 0.79, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.99, 0.99, 10 ... 1, 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% to 10% (e.g., 0.02% to 9.9%, 0.03% to 9.8%, 0.04% to 9.7%, 0.05% to 9.6%, 0.06% to 9.5%, 0.07% to 9.4%, 0.08% to 9.3%, 0.09% to 9.2%, 0.1% to 9.1%, 0.2% to 9%, 0.3% to 8.9%, 0.4% to 8.8%, 0.5% to 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% to 6.4%, 2.9% to 6.3%, 3% to 6.2%, 3.1% to 6.1%, 3.2% to 6%, 3.3% to 5.9%, 3.4% to 5.8%, 3.5% to 5.7%, 3.6% to 5.6%, 3.7% to 5.5%, 3.8% to 5.4%, 3.9% to 5.3%, 4% to 5.2%, 4.1% to 5.1%, 4.2% to 5%, 4.3% to 4.9%, 4.4% to 4.8%, 4.5% to 4.7%). EXAMPLES

[0188] The following examples are illustrative of certain aspects of the present description. The examples and embodiments described herein are for illustrative purposes only, and various modifications or changes in light thereof will be suggested to those skilled in the art and are within the spirit and scope of the present application. Furthermore, any element or limitation of any of the embodiments disclosed herein can be combined (individually or in any combination) with any and / or all other elements or limitations disclosed herein or with any other embodiments thereof, and all such combinations are contemplated with the scope of the embodiments described herein without being limited thereto. The examples should not be construed as limiting, as they merely provide a specific understanding and implementation of the embodiments and their various aspects.

[0189] Example 1: Amino acid testing To test the effect of amino acid treatment on the epithelial skin barrier, a cell model of differentiated normal human epidermal keratinocytes (NHEK) was used. Commercially available NHEKs (ATCC) derived from three different healthy donors were used. Based on multivariate analysis, three amino acids were selected for treatment: Ser, Gln, and Gly.

[0190] Using microarray gene expression and RNA sequencing (RNA-seq) analysis, the transcriptome of a panel of differentiated NHEKs was evaluated, focusing on a specific set of marker genes whose upregulation or downregulation (depending on the marker gene) reflects a tendency toward improved skin barrier function. For example, upregulation of marker genes that promote keratinocyte differentiation and / or proliferation, and downregulation of marker genes that promote, for example, apoptosis / cell death and / or inflammation, reflects a tendency toward improved skin barrier function. 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, reflects a tendency toward impaired skin barrier function.

[0191] Using gene arrays, a comparative analysis of the two data sets was performed in three different donors, focusing on genes that were consistently differentially expressed [filter criteria: fold change 1.5 (positive or negative) and p-value <0.05)] after treatment with various amino acids and their combinations. qPCR was used to evaluate the mRNA expression results. The qPCR results confirmed the array results associated with differential expression, thereby validating the panel of genes.

[0192] Marker genes: EGF, FGF, CPT2, TGFB1, SMAD1, MKI67, PPARGC1B, CASP8, and TNFRSF10D. PPARA and PPARD were also included in the marker gene panel investigated, in part because the transcription factor PPARGC1B encodes the coactivators PPARα, PPARδ, and PPARγ. The significant upregulation of CPT2, which is a major target of PPARα, solidified the inclusion of PPARA and PPARD in the panel.

[0193] Each of the amino acids was also adjusted to arrive at a suitable concentration for use in the amino acid blend. Thus, the suitable (e.g., non-toxic, non-transforming, therapeutically effective) concentration for each single amino acid was determined in the course of developing a suitable amino acid blend.

[0194] Comparative analysis of qPCR data from two donors, including the scoring system (as described above), consistently identified two sets of most (Gly, Gln, Ser, Ala) and least effective (Asp, Thr, Cys, Pro, Tyr) amino acid combinations. See, e.g., Figure 2.

[0195] The two most effective amino acid combinations were tested in five different donors as an initial set of ingredients for skin formulations [Combo#1 (Gln, Gly, Ala, Ser) and Combo#2 (Gln, Gly, Ala, Ser, Ile, Val)] and their effects on ten genes, namely EGF, FGF2, PDGF, CPT2, TGM4, PPARD, TGFB1, FLG, MKI67, and TNFRSF10D, were evaluated. For comparison, the least effective set of amino acids (Combo#3) was used in qPCR analysis. See, for example, FIG. 1.

[0196] The conclusion from the experiment was that the most effective amino acid combinations (Combo#1 and Combo#2) significantly increased gene expression of 6 out of 10 tested genes (Figure 1), whereas Combo#3 had no effect on the transcripts of the majority of genes or even downregulated them, as in the case of EGF. In contrast, the mRNA expression of TNFRSF10D, which encodes a member of the TNF receptor superfamily, was significantly downregulated by Combo#1, suggesting the anti-inflammatory / anti-apoptotic functionality of Combo#1. The most prominent effect of Combo#1 and #2 was detected on the upregulation of the transcription of EGF. Taken together, these results indicate that specific combinations of specific amino acid blends, but not all or random amino acid blends, upregulate keratinocyte genes important for normal barrier function.

[0197] These results further indicate that careful testing of amino acid formulations, taking into account multivariate interdependent data points, is necessary to obtain the best beneficial effects. It is noteworthy that in the context of healthy skin, as provided by keratinocytes from three different healthy donors, each of Combo#1 and Combo#2 conferred a statistically significant increase in the expression of many of the identified marker genes compared to the control. Results determined in the context of damaged / compromised skin may even provide more meaningful results that demonstrate the efficacy of Combo#1 and Combo#2 as well as other amino acid formulations described herein. The results presented herein clearly demonstrate the efficacy of Combo#1 and Combo#2 and are reasonably predictive of their potential for skin barrier enhancement, especially in the context of damaged / compromised skin.

[0198] The results presented herein also show that proteins are upregulated to provide beneficial contributions to skin barrier integrity.For example, involucrin is upregulated to a statistically significant extent 24 hours after treatment with either Combo #1 or Combo #2 compared to control.See, for example, Figure 5. Filaggrin is also upregulated to a statistically significant extent 24 hours after treatment with either Combo #1 or Combo #2 compared to control.See, for example, Figure 6.

[0199] Specific combinations of amino acids in suitable ratios have synergistic effects and the ability to modulate to improve and / or restore skin barrier integrity. Specific amino acid blends trigger several intracellular pathways that collectively provide the desired synergistic effect.

[0200] Example 2: Experimental Method Amino acids are mixed in water at previously established concentrations and added to the amino acid deficient medium.

[0201] Gene Expression: mRNA is extracted, converted to cDNA, and analyzed by qPCR using a custom TaqMan array 384-well plate (Thermo Fisher Scientific) containing 34 of the above genes. Four control housekeeping genes are included to normalize mRNA expression: GAPDH, POLR2A, YWHAZ, and PGK1. The array plate contains a total of 38 genes, allowing the analysis of 10 samples in one qPCR run.

[0202] Protein Analysis: Western Blot. Keratinocytes or epidermal skin equivalents are homogenized in lysis buffer, proteins are extracted and Western Blots are performed according to standard protocols using the following commercial antibodies: filaggrin (#sc-66192, Santa Cruz Bio), loricrin (#ab183646), involucrin (#ab68), TGM2 (#ab2386) (all from Abcam), β-actin (#SAB3500350, Sigma).

[0203] Western blot analysis is labor intensive and not a high throughput assay, therefore ELISA is used instead for some proteins, using previously published and validated assays for protein ELISA. Immunoassays for ANGPTL4 and Involucrin are performed on the Luminex xMAP platform using Milliplex MapHuman Liver Protein Magnetic Bead Panel hANGPTL4-MAG and Milliplex MapHuman Skin Magnetic Bead INVOL-MAG (EMD Millipore, Billerica, MA).

[0204] statistics: Differences between control and treatment groups are assessed by one-way ANOVA. Statistically significant variation is defined as 20% or greater variation from control for protein analysis and a positive or negative fold change threshold of 1.5 for gene expression with a p-value of p<0.05.

[0205] Example 3: Experimental design and methods to define additional effective combinations of amino acids that promote skin barrier function In addition to the amino acid combinations described above, several other combinations, such as including Trp and / or Arg, will be tested to determine whether the inclusion of these amino acids confers additional dermatological benefits. These two amino acids have shown positive effects in preliminary screening, making them attractive candidates for inclusion in the amino acid formulations described herein. Initially, the following amino acid combinations will be 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.

[0206] Experiments are performed using differentiated normal primary keratinocytes from the five different donors mentioned above, as well as a commercially available epidermal skin equivalent (3D model) (EPI-200, MatTech, Ashland, MA). Such 3D cultures are close to the human epidermis and are commonly used in skin research. To ensure proper experimental conditions, the experiments are performed in amino acid-deficient medium during the time of treatment with the specific amino acid formulation. Since amino acids constitute an important component of the formulations described herein, the presence of amino acids normally present in the medium may obscure the results, thus leading to the choice of amino acid-deficient medium as a control. The cells or equivalents are treated with the amino acid combinations for 4 or 24 hours. The differentiation of the keratinocytes is induced by culturing them in the presence of 1.2 mM CaCl2 for 48 hours before treatment in FBS-free medium (amino acid-deficient medium).

[0207] First, a series of classical skin barrier-related genes is investigated, including the previously tested panel. The following genes encoding key components of the barrier or barrier formation process are included in the array: growth factors EGF, TGFB1, FGF2, PDGF, terminal differentiation markers such as Filaggrin (FLG), Loricrin (LOR), TGM4, CORIN, KRT6, KRT16, SFN, SPRR2H and late cornified envelope genes (LCE1, LCE3, S100A8 / A9); lipid metabolism regulators such as PPARA, PPARD, CPT2, PNPLA1, FASN, UGCG, HMGCR, PLA2G5, ELOVL; adhesion 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 percentage of co-expressed genes after treatment with different amino acid formulations is determined. Expression levels of cross-linked proteins such as involucrin, loricrin, TGM2 and ANGPTL are assessed by Western blot analysis or ELISA using standard protocols.

[0208] The method is carried out as described in Example 2.

[0209] Example 4: Additional model systems for evaluating and identifying effective amino acid formulations To evaluate the effect of the amino acid formulations described herein, alternative models of keratinocytes with immature or affected barriers are tested. Such model systems include, for example, undifferentiated keratinocytes (cultured at low calcium concentration (0.03 mM)). These cells have immature epithelial barriers, and differentiated keratinocytes are switched to low Ca2+-containing medium (0.03 mM) for 24 hours before treatment. It has been shown that epithelial cells are deprived of Ca2+, opening TJs and thus disrupting the permeability barrier. This process is restored by Ca2+ supplementation, and the expression of filaggrin is downregulated in differentiated keratinocytes treated with Th2 cytokines, namely IL-4 (50 ng / mL) and IL-13 (50 ng / mL) for 24 hours.

[0210] Example 5: Experimental design and methods to evaluate the efficacy of topical administration of amino acid formulations on skin barrier repair in in vitro and ex vivo models of normal and compromised skin The next step in evaluating the efficacy of the amino acid formulations described herein is tested by topical administration, which is achieved by applying directly to the stratum corneum surface. Normal and immature epidermal skin equivalents (EPI-200 and EPI-20, MatTek Corporation, Ashland, MA) as well as normal and delipidated (tape-stripped) skin explants are used as normal and impaired skin models. Full-thickness human skin obtained from normal human adults undergoing abdominoplasty is commercially available for research and is obtained from Zen Bio (Durham, NC). In addition to evaluating the expression of key barrier markers, for example, as described in Example 1, TJ proteins are also investigated. TJ proteins are involved in the normal barrier function of preventing transepidermal water loss (TEWL) in healthy skin. Therefore, the expression of TJ proteins is evaluated by Western blot analysis and immunohistochemical staining. Transepithelial electrical resistance (TEER) measurements are also included as an indicator of barrier integrity and TJ dynamics. Expression of filaggrin, involucrin, and loricrin is also performed as in Example 1.

[0211] method: Ex vivo model: Ex vivo studies are performed using human skin explants. Subcutaneous fat is removed from abdominal skin (Zen Bio) and 0.93 cm 2 Skin biopsies are prepared under sterile conditions and placed in keratinocyte growth medium in a 5% CO2 humidified atmosphere. To artificially induce scaly delipidated skin, abdominal skin is first tape-stripped according to standard protocols and then biopsies are prepared. After 24 hours of acclimation, amino acid formulations are applied topically every 24 hours for a total of 7 days. Skin explants are analyzed for gene and protein expression levels at 24 hours, 48 ​​hours, 72 hours, and at the end of the experiment.

[0212] Measurement of transepithelial electrical resistance (TEER) in a three-dimensional model. For TEER, immature epidermal skin equivalents (EPI-20, MatTek Corporation, Ashland, MA) are transferred to medium containing 100ng / ml IL-4, 100ng / ml IL-13, 50ng / ml IL-31, and 30ng / ml TNFα (R&D Systems, Minneapolis, MN) as described above. Amino acid formulations are applied topically every 24 hours, 4 hours after cytokine pretreatment. TEER is measured at 0, 24, and 48 hours using a 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: (TEER 24 hours / TEER 0 hours) x 100. At the end of the experiment, the skin equivalents are used for mRNA and protein expression analysis.

[0213] qPCR analysis. Expression of the 38 gene array is performed as described in Examples 1 and 2 using custom TaqMan array plates.

[0214] Protein Analysis: Western Blot: Expression of filaggrin, involucrin, loricrin, ANGPTL4 and TGM-2 is performed as described in Examples 1 and 2.

[0215] Immunohistochemical staining. Skin explants treated topically with amino acid formulations are incubated for 7 days, re-treated every 24 hours, and harvested for staining at the end of the experiment. Tissues are fixed in 10% buffered neutral formalin solution. Fixed tissues are embedded and 5 μm thick sections are prepared. Immunohistochemical analysis is performed using standard protocols. Sections are deparaffinized in xylene, hydrated in ethanol, and washed in water. Non-specific binding is blocked with 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 controls as indicated. Tissue sections are then incubated with the appropriate biotinylated secondary antibodies, and antibody binding is visualized using the DAB Peroxidase Substrate Kit (Vector Laboratories). Staining is assessed by microscopy.

[0216] Example 6: Materials and Methods Cell culture and therapeutic agents To prepare experimental samples for RNA analysis, human primary keratinocytes were treated with a panel of amino acid(s), either single amino acids and / or amino acid combinations.

[0217] Primary normal human adult keratinocytes (ATCC, Catalog No. PCS-200-011) were cultured in Dermal Cell Basal Medium (ATCC, Catalog No. PCS-200-030) containing growth supplements (Keratinocyte Growth Kit, ATCC Catalog No. PCS-200-040). The medium was changed three times a week for cell growth. For the experiment, cells were seeded at a density of 20,000 cells / well in 6-well plates. When cells reached 100% confluence, CaCl2 was added to a final concentration of 1.8 mM for 24 hours to induce differentiation. All treatments were performed for 4 h in amino acid deficient medium (choline chloride 0.0285 mM, calcium D-pantothenate 0.00838 mM, folic acid 0.009 mM, nicotinamide 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, monosodium phosphate 0.9 mM, D-glucose 25 mM, sodium pyruvate 1 mM) in the presence of 1.8 mM CaCl2. The 4GAA formulation contained glycine, glutamine, serine, and alanine and was used at a total concentration of 4 mM (1:1:1:1). The 3BAA formulation contained cysteine ​​(1 mM), histidine (0.5 mM), and tyrosine (1 mM). A plant extract formulation containing Boswellia serrata resin extract, lecithin, microcrystalline cellulose, and silica was dissolved in amino acid deficient medium, filtered, and used at concentrations of 0.1% w / w to 1% w / w of the formulation.

[0218] Real-time RT-PCR analysis: Cellular RNA was isolated using the NucleoSpin RNA Plus kit (Macherey Nagel). cDNA synthesis was performed using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific) with 1 μg of total RNA. RT products were amplified using TaqMan Gene Expression Assays for EGF, MYC, GAPDH, and YWHAZ (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.

[0219] Equipment, materials, reagents, and consumables Confluent differentiated primary human keratinocytes were used in 6-well plates. The following were used in the experiments described herein:

[0220] Sterile RNase-free microcentrifuge tubes (1.7 mL) and conical (50 mL and 15 mL) tubes.

[0221] Pipettes, multichannel pipettes, pipette controllers (Aid), serological pipettes and pipette tips, aspirating pipettes.

[0222] Sterile isopropyl alcohol (SIPA, 70%).

[0223] Dulbecco's medium.

[0224] Amino acids: serine, glycine, glutamine, histidine, valine, tyrosine, threonine, aspartic acid, or other amino acids based on experimental needs.

[0225] Vacuum lines / vacuum pump and pump waste collection container.

[0226] Syringe (30 mL), syringe filter (25 mm PES; 0.22 micron), syringe needle (18 G).

[0227] Weighing consumables: paper, spatula, etc.

[0228] Experimental equipment: Biosafety cabinet (BSC), water bath (37 °C), 5% CO2 incubator (37 °C) refrigerator (4 °C), analytical balance, heating / stirring plate. [Table 1]

[0229] Preparation of Amino Acid Therapeutics Treatments were prepared within one week of the start of the experiment. A water bath was pre-heated to 37°C. DMEM was removed and aseptically transferred to the BSC. The required volume of DMEM (2 ml / well) was pipetted into a conical tube (50 mL) which was pre-warmed in a heated water bath for 30 minutes. Other required materials were aseptically transferred to the BSC: conical tubes (50 mL), syringes (30 mL), needles (18G), and micron syringe filters (25 mm; 0.22 microns). Conical tubes (50 mL) were labeled as either single amino acids (4 mM, 20 mM), 4 amino acid blends (4 mM, 20 mM), or ENTERADE™, and amino acids required for treatment were prepared (see Table 2).

[0230] The appropriate amount of each amino acid (see Table 2) was weighed out and then transferred to their respective conical tubes. The amino acids for the 4AA formulation and "ENTERADE®" should be weighed out individually and then transferred into designated tubes 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. A serological pipette was used to transfer the DMEM into the tubes according to Table 2.

[0231] All tubes were vortexed until the mixture was debris-free and homogenous, after which the homogenous mixture was aseptically returned to the BSC.

[0232] If the mixture does not come into solution completely by vortexing alone, place a small stir bar into a conical tube (50 mL), then place the conical tube into a beaker half-filled with water and use a heating / stirring plate (50 °C / 800 rpm) for approximately 10 minutes until the solution is homogenous.

[0233] In a separate rack, a new set of conical tubes (50 mL) were labeled with the AA name, concentration, and date for each amino acid or amino acid blend.

[0234] A needle (18G) was attached to a syringe (10 mL) and the entire contents of one of the AA "media" tubes were aspirated. The needle was removed and a syringe filter (13 mm, 0.22 micron) was attached to the syringe (10 mL). A syringe filter (25 mm, 0.22 micron) was placed over the tube labeled with the corresponding amino acid (AA) "treatment" and the mixture was pushed through the syringe and into the tube, which was immediately capped. This was repeated as necessary, changing the filter after every 100 mL filtration or between samples of each amino acid.

[0235] The syringe filtration step was repeated for each AA or AA formulation, and syringes, needles, and filters were changed between each AA or AA formulation to avoid cross-contamination. [Table 2]

[0236] Amino Acid Therapy PBS and AA treatments were pre-warmed in a water bath (37°C) for 30 minutes. 6-well plates containing "differentiated keratinocytes" were aseptically transferred from a 5% CO2 incubator (37°C) to the BSC. The 6-well plate lids were labeled with the date, initials, passage number, cell family, and targeted treatment. The well prints on the plate lids were labeled with T# (treated wells) or C# (PBS only wells).

[0237] The medium from each well was aspirated and washed with 2.0 mL of PBS.

[0238] The PBS from each well was aspirated and 2.0 mL of AA treatment was added to the wells, making sure that the treatment added corresponded to the label on the plate (Serine treatment was only added to wells of the plate labeled Serine).

[0239] The remaining control wells were filled with 2 mL of DMEM in each plate and the 6-well plate in the secondary container was transferred to a 5% CO2 incubator (37°C) and incubated as per the experiment.

[0240] Once the incubation period was completed, the "treated" cells were removed and transferred to a BSC for RNA / protein extraction.

[0241] Example 7: Preparation of formulations An exemplary method for preparing the formulations or topical formulations disclosed herein included the following steps:

[0242] Xanthan gum was dispersed in glycerin.

[0243] Mix until uniform and then add water.

[0244] Mix until uniform.

[0245] The ingredients of Step A in Table 3 were heated to a temperature of 80°C.

[0246] The ingredients of Step B in Table 3 were heated separately to a temperature of 80°C.

[0247] With both stages at 80° C., add Stage B to Stage A with vigorous mixing, then remove from heat to form a Stage A and Stage B batch.

[0248] Once the batch temperature was below 40° C., the ingredients from Table 3, Step C, were added to the batch.

[0249] The batch is then brought to 100% with deionized water. The various amino acids tested here are combined with the formulations prepared here, including alanine, glutamine, glycine, and serine. [Table 3]

[0250] Example 8: Comparison of EGF expression of amino acid and plant extract formulations, alone or together Keratinocytes from different donors were cultured for 4 hours in amino acid-free medium after full differentiation and treated in parallel with 1 mM or 4 mM amino acid blend (4GAA; alanine, glutamine, glycine, serine) and / or Boswellia plant extract blend (BSW or BSXL) at concentrations of 0.25%, 0.5% or 1% (w / w%) (the combinations are then 4GAA 4mM / 1% BSW (Figure 7A); 4GAA 1mM / 0.25% BSW (Figure 7B); 4GAA 4mM / 0.5% BSW (Figure 7C); 4GAA 1mM / 0.5% BSW (Figure 7D)).

[0251] Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed using GAPDH as a housekeeping gene, see Figures 7A-7D. * p<0.05, ** p<0.01 versus untreated controls (CTR or CTR untr).

[0252] Example 9: Comparison of EGF mRNA fold change of amino acid and plant extract formulations, alone or together, from Donor G Keratinocytes from donor G were cultured in amino acid-free medium for 4 hours after full differentiation and treated in parallel with an amino acid blend (4GAA; alanine, glutamine, glycine, serine) at 1 mM or 4 mM and / or a plant extract blend (BSXL) at concentrations of 0.1%, 0.25% or 0.5% (w / w%) (the combinations in this case are 4GAA 1 mM / 0.25% BSXL and 4GAA 4 mM / 0.5% BSXL).

[0253] Cells were harvested, RNA (EGF mRNA) was extracted and qPCR analysis was performed using GAPDH as a housekeeping gene, see Figures 8A and 8B. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus untreated control (ctr).

[0254] Example 10: Comparison of EGF mRNA fold change of donor O with amino acid and plant extract formulations, alone or together Keratinocytes from donor O were cultured in amino acid-free medium for 4 hours after full differentiation and treated in parallel with an amino acid blend (4GAA; alanine, glutamine, glycine, serine) at 1 mM or 4 mM and / or a plant extract blend (BSXL) at a concentration of 0.25% or 1% (w / w%) (the combinations in that case are BSXL 0.25% / 1 mM 4GAA and BSXL 1% / 4 mM 4GAA).

[0255] Cells were harvested, RNA (EGF mRNA) was extracted and qPCR analysis was performed using GAPDH as a housekeeping gene, see Figures 9A and 9B. ** p<0.01, *** p≦0.001 versus untreated control (CTR untr).

[0256] Example 11: Comparison of EGF mRNA fold change of amino acid and plant extract formulations, alone or together Keratinocytes from donor G were cultured for 4 hours in amino acid-free medium after full differentiation and treated in parallel with amino formulations (4mM 4GAA: alanine, glutamine, glycine, serine; 3BAA: cysteine ​​(Cys), histidine (His), tyrosine (Tyr): Cys 1mM, His 0.5mM, Tyr 1mM) and / or 0.5% plant extract formulation (BSXL) and / or piperonylic acid (100μM; PA) (in that case the combinations are 4GAA 4mM / 0.5% BSXL; 3BAA 2.5mM / 0.5% BSXL; 4GAA 4mM / 100μM PA; 3BAA 2.5mM / 100μM PA).

[0257] The same experiment was performed with similar cells and different amino acid combinations as well as a compound (PA) that upregulates EGFR (EGF receptor).

[0258] Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed using GAPDH as a housekeeping gene, see Figures 10A and 10B. * p<0.05, ** p ≤ 0.01, *** p≦0.001 versus untreated control (CTR).

[0259] Example 12: Comparison of EGF mRNA fold change of amino acid and plant extract formulations, alone or together, from Donor G Keratinocytes from donor G were cultured for 4 hours in amino acid-free medium after full differentiation and treated in parallel with amino formulations (4mM 4GAA: alanine, glutamine, glycine, serine; 3BAA: cysteine ​​(Cys), histidine (His), tyrosine (Tyr): Cys 1mM, His 0.5mM, Tyr 1mM) and / or 0.5% plant extract formulation (BSXL) and / or piperonylic acid (200μM; PA) (in that case the combinations are 4GAA 4mM / 0.5% BSXL; 3BAA 2.5mM / 0.5% BSXL; 4GAA 4mM / 200μM PA; 3BAA 2.5mM / 200μM PA).

[0260] The same experiment was performed with similar cells and different amino acid combinations as well as a compound (PA) that upregulates EGFR (EGF receptor).

[0261] Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed using GAPDH as a housekeeping gene. See Figures 11A and 11B. * p<0.05, ** p ≤ 0.01, *** p≦0.001 versus untreated control (CTR).

[0262] Example 13: Comparison of EGF mRNA fold change of donor O's amino acid blends, plant extracts, and plant extract blends, alone or together Keratinocytes from donor O were cultured for 4 hours in amino acid-free medium after full differentiation and treated in parallel with 1mM or 4mM amino preparation (4GAA; alanine, glutamine, glycine, serine) and / or 0.25% plant extract preparation (BSXL) and / or boswellic acid (BA) at concentrations of 2μM, 10μM and 50μM (in which the combinations are 4GAA 1mM / 0.25% BSXL; 4GAA 4mM / 0.25% BSXL; 4GAA 1mM / 10μM BA; 4GAA 1mM / 2μM BA; 4GAA 1mM / 50μM BA).

[0263] Cells were harvested, RNA (EGF mRNA) was extracted and qPCR analysis was performed using GAPDH as a housekeeping gene, see Figures 12A and 12B. * p<0.05, ** p ≤ 0.01, *** p≦0.001 versus untreated control (CTR).

[0264] Example 14: Comparison of EGF mRNA fold change of amino acid and plant extract formulations, alone or together, from Donor A Keratinocytes from donor A were cultured for 4 hours in amino acid-free medium after full differentiation and treated in parallel with 4 mM amino blend (4GAA: alanine, glutamine, glycine, serine; 4OAA: aspartic acid, isoleucine, tyrosine, valine) and / or 0.5% plant extract blend (BSXL) (in that case the combinations are 4GAA 4mM / 0.5% BSXL; 4OAA 4mM / 0.5% BSXL).

[0265] Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed using GAPDH as a housekeeping gene. See Figures 13A and 13B. *** p≦0.001 versus untreated control (CTR).

[0266] Example 15: Comparison of EGF mRNA fold change of amino acid and plant extract formulations, alone or together Keratinocytes from donor A (n=1), donor G (n=3), and donor T (n=2) were cultured in amino acid-free medium for 4 hours after full differentiation and treated in parallel with 4 mM amino blend (4GAA: alanine, glutamine, glycine, serine) and / or 0.5% plant extract blend (BSXL) (in that case the combination is 4GAA 4mM / 0.5% BSXL).

[0267] Cells were harvested, RNA (EGF mRNA) was extracted, and qPCR analysis was performed using GAPDH as a housekeeping gene. See averages (Figures 14A-14C).* p ≤ 0.05 versus untreated control (CTR).

[0268] Example 16: Testing changes in transepidermal water loss (TEWL) To determine changes in transepidermal water loss (TEWL), test articles (active (4GAA), left bar; placebo, right bar) were applied to the skin of 12 subjects for 5 days, after which subjects underwent 15 repeated tape strips. See Figures 15A and 15B. The active 4GAA amino acid formulation contained alanine (4%), glutamine (4%), glycine (4%), serine (4%) in an oil / water (O / W) vehicle at a total concentration of 12%, and placebo. Baseline, day 1, was immediately after tape stripping. Days 2, 3, and 5 were after twice-daily treatment with the test article, respectively.

[0269] Example 17: Measurement of skin moisture content To determine skin barrier improvement, test articles (untreated control; placebo; active amino acid (4GAA)) were applied to the skin of 25 subjects for 5 days, after which subjects received 15 replicate tape strips. See FIG. 16. Control represents no treatment (top line on day 3; left bar in inset), placebo represents water / oil / water (W / O / W) vehicle without active drug (middle line on day 3; middle bar in inset), and amino acid represents a 4 amino acid blend (Ala (4%), Gln (4%), Gly (4%), Ser (4%) dissolved in W / O / W vehicle (bottom line on day 3; right bar inset). * , p-value ≦ 0.05. Inset: control (left), placebo (middle), amino acid (right).

[0270] Corneometry was used to assess skin moisture content in the surface layer of the subjects' skin (stratum corneum).

[0271] Since various modifications may be made to the above subject matter without departing from the scope and spirit of the disclosure, it is intended that all subject matter contained in the above description or defined in the appended claims be construed as illustrating and illustrating the disclosure. Many modifications and variations of the disclosure are possible in light of the above teachings. Accordingly, the description is intended to embrace all such alternatives, modifications and variations that fall within the scope of the appended claims.

[0272] All documents cited or referenced herein, and all documents cited or referenced in the documents cited herein, 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 used in the practice of this disclosure.

Claims

1. A topical formulation comprising: (a) a formulation consisting essentially of, as free amino acids, a therapeutically effective amount of glutamine, glycine, alanine, and serine, or salts thereof, wherein each free amino acid has a ratio of 1:100 or greater to each of the other free amino acids; (b) a therapeutically effective amount of a plant extract comprising a Boswellia extract; the therapeutically effective amount of the combination and the therapeutically effective amount of the plant extract improve skin cell barrier integrity when tested by a method that detects EGF expression and determines that the EGF is upregulated with an average fold change of 1.5 to 40 for treated formulations compared to untreated skin; The topical formulation, wherein the therapeutically effective amount of the combination and the therapeutically effective amount of the plant extract improves the skin barrier integrity and / or skin barrier repair.

2. 10. The topical formulation of claim 1, wherein each of the free amino acids is present at a concentration of 0.1 mM or more, 10 mM or less, or between 0.1 mM and 10 mM.

3. The topical formulation of claim 1, wherein each of the free amino acids is present in a weight percent (wt / wt%) of the formulation of 0.001% or more, 1% or less, or 0.001% to 1% by weight.

4. The topical formulation of claim 1, wherein the therapeutically effective amount of the plant extract is present in a weight percent (w / w%) of the formulation of 0.025% or more, 5% or less, or 0.025% to 5%.

5. The topical formulation of claim 1, wherein the amino acid blend and the plant extract are in a ratio of 1:100 or more, 100:1 or less, or 1:100 to 100:

1.

6. The topical formulation of claim 1, wherein the plant extract is selected from the group consisting of Boswellia serrata resin extract, Boswellia serrata extract, Burseraceae extract, olibanum, frankincense, and any combination thereof.

7. The topical formulation of claim 1, wherein the therapeutically effective amount of the combination of free amino acids and the therapeutically effective amount of the plant extract inhibits 5-lipoxygenase, reduces calcium ion mobilization, decreases activation of MAP kinase, or any combination thereof.

8. A texturing agent and / or filler, phospholipids, anti-caking agents, a dermatologically acceptable carrier; additives, or 10. The topical formulation of claim 1, further comprising any combination thereof.

9. The topical formulation of claim 8, wherein the filler comprises cellulose, microcrystalline cellulose, or a combination thereof.

10. The topical formulation of claim 8, wherein the phospholipid comprises lecithin, phosphatidylcholine, or a combination thereof.

11. The topical formulation of claim 8, wherein the anti-caking agent comprises silica.

12. The topical formulation of claim 8, wherein the additive is selected from the group consisting of vitamins, minerals, preservatives, exfoliants, anti-acne agents, anti-wrinkle / anti-atrophy agents, hydroxyl acids, antioxidants / radical scavengers, chelating agents, flavonoids, anti-inflammatory agents, anti-cellulite agents, local anesthetics, tanning agents, skin lightening agents, skin soothing and healing agents, antibacterial and antifungal agents, sunscreens, conditioning agents, structuring agents, thickening agents, preservatives, and any combination thereof.

13. The topical formulation of claim 1, wherein the at least one additional free amino acid is selected from the group consisting of valine, isoleucine, arginine, threonine, tryptophan, or salts thereof, or any combination thereof.

14. The topical formulation of claim 1, wherein the free amino acid is in the L-form.

15. The topical formulation of claim 1, wherein the formulation does not contain any of the free amino acids cysteine, histidine, tyrosine, leucine, aspartic acid, taurine, taurate, glutamic acid, or any combination thereof.

16. A topical formulation comprising: A combination of therapeutically effective amounts of glutamine, glycine, alanine, and serine, or salts thereof, as free amino acids; and optionally, at least one additional free amino acid selected from the group consisting of valine, isoleucine, arginine, threonine, or tryptophan, or a salt thereof, or any combination thereof; each free amino acid having a ratio of 1:100 or greater to each of the other free amino acids; the therapeutically effective amount improves skin cell barrier integrity as tested by a method of detecting EGF skin barrier marker gene expression and determining that the EGF skin barrier marker gene is upregulated by an average fold change of 1.5 to 40 for the treated formulation compared to untreated skin; The topical formulation, wherein the therapeutically effective amount improves the skin's barrier integrity and / or skin barrier repair.

17. At least one additional free amino acid of valine, isoleucine, arginine, threonine, or tryptophan, or a salt thereof, or any combination thereof; a therapeutically effective amount of a plant extract comprising a Boswellia extract; texturing agents and / or fillers, phospholipids, anti-caking agents, a dermatologically acceptable carrier, or 17. The topical formulation of claim 16, further comprising any combination thereof.

18. If present, the free amino acid cysteine ​​is present at a concentration of 1 mM or less; If present, the free amino acid histidine is present at a concentration of 0.5 mM or less; If present, the free amino acid tyrosine is present at a concentration of 1 mM or less; If present, the free amino acid leucine is present at a concentration of 4 mM or less; If present, the free amino acid aspartic acid is present at a concentration of 2 mM or less; or The topical formulation of claim 1 , any combination thereof.

19. A method comprising administering the topical formulation of claim 1 to the skin of a subject suffering from a skin condition, said method comprising: Maintaining skin cell barrier integrity, Improves skin cell barrier integrity, Improves skin condition, Treating a skin condition in a subject in need thereof; or The method is a combination thereof.

20. administering the topical formulation improves at least one visual property, or at least one tactile property, or a combination of visual and tactile properties of the skin; 20. The method of claim 19, wherein the skin condition comprises a cosmetic skin condition, atopic dermatitis, dermatitis, psoriasis, pruritus, eczema, a wound, or a burn.

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