Bioactive nut peptides and compositions for skin treatment

Abstract

Bioactive Walnut Peptides and Compositions for Skin Treatment Bioactive walnut peptides and a use of the bioactive walnut peptides for topical application to the skin are described. The bioactive walnut peptides may be incorporated into a pharmaceutical or cosmetic composition and are useful for improving skin barrier function. Figure for abstract: none

Description

Title of Invention: Bioactive Walnut Peptides and Compositions for Skin Treatment FIELD OF DISCLOSURE

[0001] The present disclosure relates to bioactive walnut peptides and the use of the walnut peptides for treating skin. The walnut peptides and compositions comprising them are particularly useful for improving skin barrier function and treating damaged skin, inflamed skin and skin suffering from damage, disease or aggression. CONTEXT

[0002] The skin is a complex organ composed of several cell types and microstructures that work together to perform critical functions and support the body's homeostasis. It is the outermost cornified layer of our body that is primarily responsible for the permeability barrier, protecting against external aggressions and preventing water loss from within. The understanding of the organization, functionality, and underlying mechanisms of the skin barrier has evolved considerably over the years. The formation of an intact and well-maintained stratum corneum, where the permeability barrier resides, is highly dependent on the differentiation of epidermal keratinocytes and the synthesis, release, localization, and binding of lipids that primarily include ceramides, cholesterol, and free fatty acids.Extensive research on the stratum corneum barrier, its disruption in disease pathogenesis, as well as barrier responses to environmental insults, has enabled the development of modern treatments and topical care routines.

[0003] The epidermis maintains its homeostasis and performs essential functions through a dynamic process of self-renewal in which basal keratinocytes divide and migrate through the stratum spinosum and stratum granulosum while progressively differentiating. When the keratinocytes reach the top of the stratum granulosum, the process of terminal differentiation occurs in which the keratinocytes undergo programmed cell death and flatten to form the stratum corneum. During this process, the lamellar bodies of the stratum granulosum keratinocytes fuse with the plasma membranes and release their primarily lipid contents into the intercellular spaces of the nascent stratum corneum.An interaction of hydrolytic enzymes and their inhibitors, also excreted via the lamellar bodies, participates in the elaboration of the layered intercellular lipid structure and, ultimately, is involved in the . Cellular desquamation on the top of the skin. Simultaneously with the accumulation of extracellular lipids, significant changes occur in keratinocytes during the formation of the stratum corneum. Transglutaminase-1-mediated crosslinking of cytoplasmic proteins at the cell periphery results in the formation of highly insoluble cornified envelopes of the stratum corneum cells, subsequently called corneocytes. This is followed by covalent attachment to these structures of a ceramide monolayer, replacing the phospholipid plasma membranes of living cells. These newly formed cornified lipid envelopes provide the scaffolding for the subsequent stacking and organization of intercellular lipids. The composite structure of the stratum corneum, consisting of corneocytes intercalated by polar lipids, can be compared to a brick-and-mortar wall constituting the permeability barrier of the stratum corneum..

[0004] To fulfill its function as a permeability barrier, the epidermis must remain mechanically strong while being flexible enough to accommodate skin movements and the flow of keratinocytes through successive layers, like a treadmill. Cell-cell and cell-substrate junctions play a central role in maintaining the mechanical properties of the epidermis. Desmosomes, which interconnect individual cell cytoskeletons into a superstructure, evolve throughout the epithelial tissue and modify their location, protein composition, and glycan distribution depending on the stage of cell differentiation and the onset of mechanical stress. In this process, adherens junctions linked to the actin cytoskeleton participate in the dynamics of desmosome and tight junction expression.During stratum corneum formation, these junctions are cross-linked to the comified envelopes and contribute to strengthening the physical strength of the stratum corneum functional barrier. The mechanical properties of the stratum corneum show a significant increase in stiffness between deep and superficial corneocytes. The mechanical integrity of the stratum corneum also depends on the direction of applied shear forces, as lateral, side-to-side adhesion between cells is stronger than that between successive layers of corneocytes.

[0005] The relative impermeability of the stratum corneum and thus its barrier function depend essentially on intercellular lipids, although they represent only 15% of the weight of the stratum corneum. Nearly equimolar proportions of ceramides, cholesterol and free fatty acids appear to be essential for the correct self-assembly of the intercellular lipid multilayers within the stratum corneum. The composition of the stratum corneum lipids is further subdivided into free fatty acids (10%), cholesterol (27%), cholesterol esters (10%), cholesterol sulfate (3%) and ceramides (50%). These lipids, organized in multiple double layers parallel to the corneocyte surfaces, can assemble within the layers into domains with different densities. A dense orthorhombic lateral packing of lipid molecules and a more fluid hexagonal format predominate in normal human skin. Efficient filling of stratum corneum interstices is essential to prevent excessive water loss and the penetration of contaminants / environmental aggressors.

[0006] The formation and restoration of a suppressed stratum corneum barrier is a dynamic, finely regulated process subject to the influences of intrinsic and environmental factors. In addition to diseases and severe environmental exposures to ultraviolet radiation or pollution, events occurring in daily life can also negatively impact the skin barrier. The importance of the stratum corneum in maintaining skin homeostasis, coupled with the prevalence and severity of internal and external factors that can alter its permeability, underscore the need for topical products to support the skin barrier. Understanding the structure, composition, and function of the epidermal permeability barrier provides a solid foundation for the knowledge-based development of topical treatments aimed at maintaining and improving the skin of patients, whether healthy or diseased.The formulation of pharmaceutical and cosmetic compositions to improve barrier integrity has continued as knowledge of the skin barrier continually expands. Efforts are currently being made to address the need to improve barrier function and barrier regeneration. SUMMARY OF DISCLOSURE

[0007] The present disclosure relates to bioactive walnut peptides and the use of the walnut peptides for topical application to the skin. The bioactive walnut peptides, which may be incorporated into a pharmaceutical or cosmetic composition, improve skin barrier function and re-epithelialization of the skin, repair damaged skin, and improve wound healing. For example, the walnut peptides and compositions comprising them are useful for improving the skin barrier function of healthy skin and useful for treating skin that has suffered physical damage, chemical damage, environmental damage, sun damage, and damage caused by disease, including inflammation.

[0008] Walnuts are one of the most widespread and oldest nuts in the world. They have high nutritional value and are rich in oleic acid, linoleic acid, alpha-linolenic acid and other unsaturated fatty acids, vitamins and proteins. Walnuts are commonly used to make walnut oil because they contain a high lipid content. The residue remaining after lipid extraction is considered a by-product, although it contains nut protein and other useful components. Walnut protein is mainly composed of albumin, globulin, gliadin, and glutenin.

[0009] The bioactive nut peptides may be derived from nut proteins or may be synthesized. For example, the nut peptides are obtained by enzymatic hydrolysis, fermentation hydrolysis, or chemical hydrolysis of nut proteins or produced synthetically, for example, by solid-phase synthesis. The bioactive peptides that are particularly useful according to the present disclosure typically have a molecular weight of less than 6,000 Da and often less than 1,000 Da. These nut peptides may have from 2 to 50 amino acid residues, but typically have from 2 or 3 amino acid residues to about 20 amino acid residues.

[0010] Preferably, the bioactive nut peptides have a minimum of 2 or 3 amino acid residues up to about 20 amino acid residues and confer a positive physiological or dermatological effect to skin cells. Bioactive peptides include amino acids joined by covalent bonds, also called amidic or peptide bonds, while proteins are higher molecular weight (MW) polypeptides, i.e., having more than 50 amino acid residues. Bioactive nut peptides usually exhibit hormonal or drug-like activities and are classified according to their mode of action. Many bioactive peptides share certain structural features, for example, an amino acid residue length for 2 to 20 amino acids.

[0011] The bioactive walnut peptides useful according to the present disclosure often include one or more amino acid residues selected from leucine, proline, or combinations thereof. In other embodiments, the walnut peptides include three or more amino acid residues selected from leucine, proline, or combinations thereof. Non-limiting examples of amino acid residues within the walnut peptides that include leucine and proline include Leu-Pro-Leu (LPP), Leu-Leu-Pro, Pro-Pro-Leu (PPL), and Pro-Leu-Pro (PLP). Other non-limiting examples of amino acid sequence that may be included in the bioactive nut peptides include Thr-Trp-Leu-Pro-Leu-Pro-Arg (TWLPLPR), Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys (YVLLPSPK), Lys-Val-Pro-Pro-Leu-Leu-Tyr (KVPPLLY), and combinations thereof.

[0012] In various embodiments, one or more of the following nut peptides are preferred:

[0013] Peptide I Thr-Trp-Leu-Pro-Leu-Pro-Arg (TWLPLPR),

[0014] Peptide II Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys (YVLLPSPK), and / or

[0015] Peptide III Lys-Val-Pro-Pro-Leu-Leu-Tyr (KVPPLLY).

[0016] As already mentioned, bioactive walnut peptides are particularly useful for topical application to the skin. Accordingly, the present disclosure relates to the use of walnut peptides in methods of treating the skin. In various embodiments, the one or more walnut peptides are applied to the skin in a pharmaceutical or cosmetic composition, which typically includes a physiologically acceptable carrier, e.g., water and optionally water-soluble solvents. The pharmaceutical or cosmetic composition includes an amount of the one or more bioactive walnut peptides sufficient to ensure that a therapeutically effective amount of the one or more walnut peptides is delivered to the skin during use.

[0017] Bioactive walnut peptides are useful for treating skin inflammation. For example, bioactive walnut peptides surprisingly and beneficially reduce, treat, or prevent pro-inflammatory cytokines in the skin. In addition, the bioactive walnut peptides are useful in methods of treating skin conditions, diseases or disorders such as psoriasis, dermatitis, atopic dermatitis, allergic dermatitis, eczema, spongiosis, edema, hereditary ichthyosis, senile xerosis, palmar hyperkeratosis, plantar hyperkeratosis, cuts, bruises, pore size, skin cancer, wound healing or re-epithelialization disorders, keloids, hypertrophic scars, cellulite, orange peel, elastosis, actinic elastosis, keratosis, rosacea, telangiectasia, couperose, or combinations thereof.

[0018] In various embodiments, the use of two or more bioactive walnut peptides is preferred. The two or more bioactive walnut peptides may have similar activities or may provide different activities beneficial to the skin. In other embodiments, the use of three or more bioactive walnut peptides is preferred. The use of multiple bioactive peptides allows for the modification of more than one physiological mechanism in the treatment of the skin. For example, one or more bioactive walnut peptides may be useful in preventing and / or treating age-related inflammation while another bioactive walnut peptide may be useful in stimulating fibroblasts that produce collagen and elastin to clarify, thicken, and firm the skin.Furthermore, combinations of bioactive nut peptides may interact synergistically and provide benefits that go beyond the sum of the individual contributions of the peptides. For example, the synergistic activity of a combination may be at least 5%, at least 10%, or at least 25% greater than the sum of the individual activities of the corresponding amounts of the bioactive nut peptides.

[0019] The one or more bioactive nut peptides are often incorporated into a pharmaceutical or cosmetic composition for application to the skin. The pharmaceutical and cosmetic composition typically includes one or more nut peptides bioactives and one or more physiologically acceptable carriers, for example, water. Non-limiting examples of physiologically acceptable carriers include water, water-soluble solvents such as alcohols, polyols, and glycols, a fatty compound such as oils, triglycerides, fatty acids, fatty alcohols, and the like. Pharmaceutical and cosmetic compositions include lotions, creams, serums, sprays, emulsions, gels, powders, dispersions, ointments, sticks, pastes, and foams. Brief description of the drawings

[0020] An implementation of the present technology is described, by way of example only, with reference to the appended figures, in which:

[0021] [Fig-1] [Fig.l] is a heat map showing genes with different expression ential in adult human epidermal keratinocytes treated with walnut peptides; and

[0022] [Fig.2] [Fig.2] is a heat map showing differentially expressed genes in reconstructed human epidermis treated with walnut peptides.

[0023] The various aspects of the disclosure are not limited to the results, arrangements and representations shown in the drawings. DETAILED DESCRIPTION OF THE INVENTION

[0024] The present disclosure relates to bioactive walnut peptides and their topical use for treating the skin. The bioactive walnut peptides include two to several dozen amino acids linked together by peptide bonds. Their molecular weight is generally less than 6,000 Da, preferably less than 3,000 Da, and more preferably less than 1,000 Da. The term "peptide" in accordance with the present disclosure is a compound that includes an uninterrupted sequence of at least two amino acids within its structure and has a maximum of about 50 amino acids. The terms "di-peptide" or "dipeptide" as used herein refer to a compound that includes an uninterrupted sequence of two amino acids within its structure. The terms "tri-peptide" or "tripeptide" as used herein refer to a compound that includes an uninterrupted sequence of three amino acids within its structure.As used herein, a "tetrapeptide" or "tetrapeptide" is a compound that includes an uninterrupted sequence of four amino acids within its structure. These amino acids are indicated herein using a traditional one-letter convention from left (N-terminus) to right (C-terminus). In this nomenclature, G is glycine, H is histidine, K is lysine, E is glutamic acid, and the like, according to nomenclature well known and accepted in the art.

[0025] A "bioactive" peptide within the meaning of the present disclosure has a minimum of 2 or 3 amino acid residues up to about 20 amino acid residues in length and has an effect measurable physiological effect on skin cells. Bioactive peptides include amino acids linked by covalent bonds, also called amido or peptide bonds, while proteins are polypeptides with a higher molecular weight (MW) and generally more than 50 amino acid residues. Bioactive peptides usually exhibit hormonal or drug-like activities and are classified according to their mode of action. Many bioactive peptides share certain structural features that include, for example, a peptide residue length of 2 to 20 amino acids.

[0026] The term "amino acid" as used herein includes and encompasses all naturally occurring amino acids, in the D or L configuration if optically active, and known non-native, synthetic and modified amino acids, such as homocysteine, omithine, norleucine and p-valine. A list of non-naturally occurring amino acids can be found in The Peptides, Vol. 5 (1983), Academie Press, Chapter VI, by D.C. Roberts and F. Vellaccio, which is incorporated herein by reference in its entirety. The amino acids in the peptides of the present invention may be present in their naturally occurring L configuration, their non-naturally occurring D configuration or as a racemic mixture.

[0027] As used herein, the term "peptide" also refers to salts, deprotected forms, acylated forms of the peptide, deacylated forms of the peptide, enantiomers, diastereomers, racemates, prodrugs, and hydrates of the aforementioned peptide. Diastereomers of the peptide are obtained when the stereochemical or chiral center of one or more amino acids is changed. The enantiomer exhibits the opposite stereochemistry in all chiral centers. In various embodiments, the C-terminus of a peptide is synthesized as an amide to neutralize a negative charge created by the C-terminal COOH. This modification may be added to help prevent enzymatic degradation.

[0028] The term "prodrug" refers to any precursor compound that can generate or release the above-mentioned peptide under physiological conditions. Such prodrugs are, for example, larger peptides that are selectively cleaved to form the peptide of the invention. Other prodrugs are protected amino acids having protecting groups at the carboxylic acid and / or amino group. Suitable protecting groups for amino groups include, for example, benzyloxycarbonyl, t-butyloxycarbonyl (BOC), formyl, and acetyl or acyl group. Suitable protecting groups for the carboxylic acid group are esters such as benzyl esters or t-butyl esters.

[0029] Epidermal deterioration can result from acute injury or exposure. Virtually all epidermal dysfunctions, whether congenital or acquired, are associated with significant changes in the permeability barrier. This is particularly evident in dermatoses with a significant inflammatory component. In many cases, barrier dysfunction can be the cause of skin disease, as is the case in atopic dermatitis, and contributes to the vicious cycle of a given pathology by inducing an inflammatory response. Deficient expression of an epidermal protein filaggrin, due to loss-of-function genetic mutations, has been shown to be responsible for the onset of atopic dermatitis in up to 50% of Northern European cases.

[0030] Filaggrin is produced in stratum granulosum keratinocytes and its catabolic processing in the stratum corneum leads to the abundance of hydrophilic amino acids constituting the bulk of the so-called natural moisturizing factor (NMF). The absence or marked reduction of the natural moisturizing factor compromises stratum corneum hydration and, consequently, the barrier function. Interestingly, the same filaggrin mutations present on both alleles of the gene result in an ichthyosis vulgaris phenotype, most frequently associated with atopy. In ichthyosis, the epidermis must compensate for the permeable barrier with hyperkeratosis. The accumulation of corneocytes is probably favored by a particularly low degree of stratum corneum hydration, which may prevent the activity of stratum corneum hydrolytic enzymes.This putative mechanism could exacerbate the desquamation-promoting context of serine protease activation due to a more basic (optimal) intracellular pH in the amino acid-deficient tissue. Nanomechanical and ultrastructural studies of the elastic properties of filaggrin-deficient corneocytes demonstrate a significant reduction in cellular stiffness and delayed degradation of corneodesmosomes, both of which are potential indicators of stratum corneum functionality. In addition to impaired filaggrin expression, epidermal atopic dermatitis also exhibits a significant reduction in key TJ proteins and, more importantly, ceramides, including EOS ceramides.

[0031] Regarding ceramide changes, their decreased levels and shortening of their acyl chains have been observed in skin not involved in atopic dermatitis, independently of filaggrin mutations, which may have etiological significance. Altered ceramide expression levels and their lamellar and lateral organization are correlated with disease activity. Even lower levels of free ceramides and sterols have been reported in atopic dermatitis lesions, with a concomitant increase in sphingosine and sphinganine-based ceramides.

[0032] In psoriasis, inflammatory skin lesions induced by interleukin 23-recruited Thl7 lymphocytes are characterized by hyperproliferation of keratinocytes and incomplete terminal differentiation resulting in impaired function. ineffective permeability barrier. Although the immune cell subsets and cytokines involved in atopic dermatitis and psoriasis pathogenesis differ considerably, the deleterious vicious cycle of barrier disruption / inflammation is still present in the latter. Incomplete terminal differentiation of psoriatic lesional keratinocytes is induced by T-cell-mediated cutaneous inflammation, which significantly impacts ceramide expression compared to normal or uninvolved skin. Similar to atopic dermatitis, ceramide species exhibit shorter fatty acid chains in psoriasis lesions.Clinical observations of improvement in psoriasis vulgaris lesions under simple occlusion and in atopic dermatitis lesions with topical emollient therapy alone clearly indicate that restoration or compensation of the stratum corneum barrier helps to interrupt the vicious cycle of pathogenic self-propagation.

[0033] To fulfill its protective functions, the epidermis must continuously adapt to changes in environmental conditions. These include climate / seasonal factors such as relative humidity, ambient temperature and sun exposure, as well as environmental stresses due to the widespread use of chemicals, the presence of air pollutants and changes in the composition and size of the skin surface microbiota, the latter being largely related to the aforementioned factors.

[0034] Humidity influences the renewal of the stratum corneum by changing the desquamation rate of corneocytes. Indeed, it promotes a rapid rise in the pH of the stratum corneum, leading to an increase in the activity of kallikreins, the main serum proteases of the stratum corneum involved in desquamation. In addition, exposure to water facilitates the accessibility of corneodesmosomes to proteolytic enzymes, which otherwise remain encapsulated within the largely hydrophobic extracellular spaces, and thus promotes the release of cells to the skin surface. Conversely, a persistence of corneodesmosomes is observed in the outer stratum corneum of xerotic winter skin compared to normal skin. Furthermore, it is known that cold and dry weather increase the prevalence and risk of new flare-ups in patients with atopic dermatitis.

[0035] Environmental factors leading to a breakdown of skin barrier function include exposure to irritants and allergens. In industrialized societies, the skin barrier is affected by the daily use of detergents and disinfectants, combined with the deleterious action of air pollutants that vary depending on geographic location and source. These pollutants contain airborne solid and liquid particles and various gases such as ozone, nitrogen oxides, volatile organic compounds, and carbon monoxide. Particles vary in number, size, shape, surface area, and chemical composition, while both particles and gases can vary in solubility and toxicity. Occupational factors also play a role, increasing risks in specific subpopulations. In healthcare workers, intensive glove use leads to occlusion, which significantly exacerbates the negative effect of detergents / soaps on skin barrier function. Published data indicate that a dose-response relationship is significant with respect to the duration of occlusion. This is particularly relevant for workplaces where switching between glove use and hand washing is common.

[0036] The present disclosure includes methods of improving the barrier function of the skin. Such methods include treating the skin by topically applying a therapeutically effective amount of one or more walnut peptides to the skin. In various embodiments, the one or more bioactive walnut peptides have a molecular weight of less than 10,000 Da. In other embodiments, the one or more bioactive walnut peptides have a molecular weight of less than 8,000 Da, less than 6,000 Da, less than 5,000 Da, less than 4,000 Da, less than 3,000 Da, less than 2,000 Da, less than 1,000 Da, and even less than 500 Da. Preferably, the one or more bioactive nut peptides have a molecular weight of less than 5,000 Da, more preferably less than 2,000 Da, and even more preferably less than 1,000 Da.

[0037] The number of amino acid residues in the one or more bioactive nut peptides will vary and may be limited based on the molecular weights described above. However, in various embodiments, the one or more bioactive nut peptides comprise from 2 to about 50 amino acid residues.In other embodiments, the one or more bioactive peptides have from 2 to about 25 amino acid residues, from about 2 to about 20 amino acid residues, from about 2 to about 18 amino acid residues, from about 2 to about 15 amino acid residues, from about 2 to about 12 amino acid residues, from about 2 to about 10 amino acid residues, from about 3 to about 25 amino acid residues, from about 3 to about 20 amino acid residues, from about 3 to about 18 amino acid residues, from about 3 to about 18 amino acid residues, from about 3 to about 15 amino acid residues, from about 3 to about 12 amino acid residues, or from about 3 to about 10 amino acid residues. of amino acid. Preferably, the one or more bioactive peptides comprise 2 to 20 amino acid residues, more preferably 2 to about 15 amino acid residues, even more preferably 3 to about 12 amino acid residues.

[0038] In various embodiments, the one or more bioactive nut peptides include one or more amino acid residues selected from leucine, proline, and combinations thereof. In another embodiment, the one or more peptides of Bioactive nut peptides include three or more amino acid residues selected from leucine, proline, or a combination thereof. Non-limiting examples of a bioactive peptide of three or more amino acid residues selected from leucine, proline, and combinations thereof include Leu-Pro-Leu, Leu-Leu-Pro, Pro-Pro-Leu, and Pro-Leu-Pro. For example, the one or more bioactive nut peptides may comprise an amino acid sequence selected from Thr-Trp-Leu-Pro-Leu-Pro-Arg, Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys, or Lys-Val-Pro-Pro-Leu-Leu-Tyr. As already mentioned, bioactive nut peptides include salts, deprotected forms, acylated forms, deacylated forms, enantiomers, diastereomers, racemates, prodrugs and hydrates of a particular amino acid sequence.

[0039] In a preferred embodiment, at least one, and preferably all, of the one or more bioactive walnut peptides are selected from peptide I, peptide II and peptide III, including their salts, their deprotected forms, their acylated forms, their deacylated forms, their enantiomers, their diastereomers, their racemates, their prodrugs and their hydrates.

[0040] Peptide I TWLPLPR (Thr-Trp-Leu-Pro-Leu-Pro-Arg),

[0041] Peptide II YVLLPSPK (Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys), and

[0042] Peptide III KVPPLLY (Lys-Val-Pro-Pro-Leu-Leu-Tyr).

[0043] In various embodiments, the use of two bioactive nut peptides or more is desirable. The two or more bioactive walnut peptides may share similar activities or may provide different activities beneficial to the skin. In other embodiments, the use of three or more bioactive walnut peptides is preferred. The use of multiple bioactive peptides allows for more than one physiological mechanism to be utilized for skin treatment. For example, one or more bioactive walnut peptides may be useful for preventing and / or treating inflammation while another bioactive walnut peptide may be useful for stimulating fibroblasts that produce collagen and elastin to clarify, thicken, and firm the skin. Furthermore, as shown below, combinations of bioactive walnut peptides interact synergistically to provide benefits that go beyond the sum of the individual contributions of the peptides.For example, the synergistic activity of a combination of bioactive nut peptides may be at least 5%, preferably at least 10%, more preferably at least 25% greater than the sum of the activity of the corresponding individual amounts of the bioactive nut peptides.

[0044] Combinations of peptide I, peptide II, and peptide III are particularly useful. For example, a combination of peptide I and peptide II, a combination of peptide I and peptide III, a combination of peptide II and peptide III, or a combination of peptide I and peptide III. combinations of peptide I, peptide II, and peptide III may be used. The peptides in the combinations may be included in various weight ratios to each other as described below.

[0045] Peptide I and peptide II may be used together in a weight ratio of about 1:10 to about 10:1. In other embodiments, peptide I and peptide II may be used together in a weight ratio of about 8:1 to about 1:8, about 5:1 to about 1:5, about 4:1 to about 1:4, about 3:1 to about 1:3, about 2:1 to about 1:2, or about 1:1.

[0046] Peptide I and peptide III may be used together in a weight ratio of about 1:10 to about 10:1. In other embodiments, peptide I and peptide III may be used together in a weight ratio of about 8:1 to about 1:8, about 5:1 to about 1:5, about 4:1 to about 1:4, about 3:1 to about 1:3, about 2:1 to about 1:2, or about 1:1.

[0047] Peptide II and peptide III may be used together in a weight ratio of about 1:10 to about 10:1. In other embodiments, peptide II and peptide III may be used together in a weight ratio of about 8:1 to about 1:8, about 5:1 to about 1:5, about 4:1 to about 1:4, about 3:1 to about 1:3, about 2:1 to about 1:2, or about 1:1.

[0048] In various embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for improving skin barrier function or re-epithelialization of skin. Accordingly, the disclosure encompasses methods of improving skin barrier function or re-epithelialization of skin comprising applying a therapeutically effective amount of one or more walnut peptides or a composition comprising them to skin, including damaged skin and undamaged skin.

[0049] In various embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for treating or repairing damaged skin. Accordingly, the disclosure encompasses methods of treating or repairing damaged skin, comprising applying a therapeutically effective amount of one or more walnut peptides or a composition comprising them to skin in need thereof. In other embodiments, the damaged skin has been physically damaged, chemically damaged, environmentally damaged, sun damaged, or damaged due to disease.

[0050] In still other embodiments, the disclosure relates to the use of one or more nut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for wound healing. For example, the disclosure relates to methods of enhancing wound healing and methods of treating or improving the appearance of skin due to the wound healing properties imparted by the walnut peptides and compositions comprising them. Such methods include: (i) damaging skin tissue using a chemical, laser, or physical force; and (ii) applying a therapeutically effective amount of one or more bioactive walnut peptides to the damaged skin.Non-limiting examples of cosmetic procedures that damage the skin include an ablative laser procedure, a non-ablative laser procedure, a microneedling procedure, a cryotherapy procedure, a radiofrequency microneedling procedure, dermabrasion, a chemical peel, an exfoliant or a mechanical or energy-based device, or any chemical procedure that damages the skin (e.g., chemical peels, etc.).

[0051] In various embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for improving skin barrier function. Accordingly, the disclosure encompasses methods of improving skin barrier function, comprising applying a therapeutically effective amount of one or more walnut peptides or a composition comprising them to skin in need thereof. In various embodiments, the skin barrier function of healthy, undamaged skin is improved. In other embodiments, the skin barrier function of damaged skin is improved.

[0052] In various embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for treating dry or itchy skin. Accordingly, the disclosure encompasses methods of treating dry or itchy skin, comprising applying a therapeutically effective amount of one or more walnut peptides or a composition comprising them to skin in need thereof.

[0053] In various embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for treating a skin condition, disease or disorder selected from psoriasis, dermatitis, atopic dermatitis, allergic dermatitis, eczema, spongiosis, edema, hereditary ichthyosis, senile xerosis, palmar hyperkeratosis, plantar hyperkeratosis, cuts, bruises, pore size, skin cancer, wound healing or re-epithelialization disorders, keloids, hypertrophic scars, cellulite, orange peel skin, elastosis, actinic elastosis, keratosis, rosacea, telangiectasia, couperose, or combinations thereof. Accordingly, the disclosure encompasses methods of treating a skin condition, disease, or disorder selected from psoriasis, dermatitis, atopic dermatitis, allergic dermatitis, eczema, spongiosis, edema, hereditary ichthyosis, senile xerosis, palmar hyperkeratosis, plantar hyperkeratosis, cuts, bruises, pore size, skin cancer, wound healing or re-epithelialization disorders, keloids, hypertrophic scars, cellulite, orange peel, elastosis, actinic elastosis, keratosis, rosacea, telangiectasia, couperose, or combinations thereof, comprising applying a therapeutically effective amount of one or more walnut peptides or a composition comprising them to skin in need thereof.

[0054] In other embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for treating skin suffering from inflammation or an inflammatory disorder and methods of treating skin suffering from inflammation or an inflammatory disorder. Non-limiting examples of inflammatory disorders include atopic dermatitis, psoriasis, or a combination thereof.

[0055] In various embodiments, the present disclosure relates to the use of one or more walnut peptides or a pharmaceutical or cosmetic composition comprising the one or more walnut peptides for potentiating terminal differentiation and keratinization of skin cells. Similarly, the disclosure relates to methods of potentiating terminal differentiation and keratinization of skin cells comprising applying a therapeutically effective amount of one or more walnut peptides to skin in need thereof.

[0056] The one or more walnut peptides are often combined with one or more additional skin active agents. Non-limiting examples of skin active agents include anti-atrophy agents, antioxidants, depigmenting agents, or combinations thereof. More specific but non-limiting examples include ceramides, alpha-hydroxy acids, beta-hydroxy acids, vitamin A, vitamin C, vitamin D, vitamin E, niacinamide, caffeine, ferulic acid, salicylic acid, madecassoside, retinoic acid, benzoyl peroxide, or a combination thereof.In a preferred embodiment, the one or more nut peptides are combined with one or more ceramides, for example, one or more ceramides selected from free ceramides, for example, Ceramide EOP, Ceramide AS, Ceramide AP, Ceramide NS, Ceramide NP, Ceramide NH, Ceramide AH, Ceramide EOH, Ceramide EOS, Ceramide AdS, Ceramide NdS and Ceramide EOdS, protein-bound ceramides, phytosphingosine, sphingosine, ceramide precursors, fatty acids, fatty alcohols, cho- . esterol, cholesterol sulfate, or combinations thereof.

[0057] The one or more nut peptides may be combined with cholesterol.

[0058] The one or more nut peptides may be combined with one or more Sphingolipids. Sphingolipids are a class of lipids containing a backbone of sphingoid bases, which are a set of aliphatic amino alcohols that include sphingosine. One or more nut peptides can be combined with sphingosine-1-phosphate.

[0059] The one or more bioactive nut peptides are often incorporated into a pharmaceutical or cosmetic composition for application to the skin. Pharmaceutical and cosmetic compositions typically include one or more bioactive nut peptides and a physiologically acceptable carrier. A "physiological carrier" as used herein is a carrier suitable and safe for application to the skin of a human. A particularly common physiologically acceptable carrier is water. However, physiologically acceptable carriers may be oil, fats, organic solvents, and the like, provided they are suitable and safe for application to the skin.Non-limiting examples of physiologically acceptable carriers include water, water-soluble solvents such as alcohols, polyols and glycols, fatty compounds such as oils, triglycerides, fatty acids, fatty alcohols, petrolatum, and the like. The pharmaceutical and cosmetic compositions of the present disclosure may be lotions, creams, serums, sprays, emulsions, gels, powders, dispersions, ointments, sticks, pastes or mousses.

[0060] Bioactive nut peptides may be derived from the hydrolysis of nut proteins into small molecular peptides with molecular weights between the molecular weights of individual amino acids and the molecular weights of proteins. This may be achieved using biological or chemical processes. For example, bioactive peptides may be prepared by enzymatic procedures, fermentation, and chemical processes.

[0061] Bioactive walnut peptides can be synthesized by coupling the carboxyl group or C-terminus of one amino acid to the amino group or N-terminus of another. Due to the possibility of unforeseen reactions, protecting groups are sometimes required. The chemical synthesis of peptides begins at the C-terminus of the peptide and ends at the N-terminus. Peptides can be synthesized either by solid-phase peptide synthesis, liquid-phase peptide synthesis, or fragment condensation. In principle, the seemingly simple formation of a peptide bond can be accomplished using all procedures available in organic chemistry for the synthesis of carboxylic acid amides.

[0062] The general process of solid-phase peptide synthesis (e.g., of a resin) begins with the attachment of the first amino acid, the C-terminal residue, to the resin. To prevent polymerization of the amino acid, the alpha-amino group and reactive side chains are protected by a temporary protecting group. Once the amino acid is attached to the resin, the resin is filtered and washed to remove by-products and excess reagents. Next, the N-alpha protecting group is removed in a deprotection process, and the resin is washed again to remove by-products and excess reagents. Next, the next amino acid is coupled to the attached amino acid. This is followed by another washing procedure, which leaves the resin-peptide assembly ready for the next coupling cycle. The cycle is repeated until the peptide sequence is completed.Then, typically, all protecting groups are removed, the peptide resin is washed, and the peptide is cleaved from the resin.

[0063] Enzymatic hydrolysis involves the use of commercial enzymes to obtain bioactive peptides. Enzymes are responsible for cleaving the peptide bonds established in the protein, thereby releasing the encrypted peptide. For the enzyme to carry out its activities, it is important that the enzyme binds to the substrate and continues the enzymatic catalysis. The enzyme has specific active sites containing residues that form temporary bonds with the substrate and residues that catalyze the reaction with the substrate. In this way, binding sites and catalytic sites are formed respectively. The bonds forming the enzyme-substrate complex are usually hydrogen bonds, hydrophobic bonds or Van der Waals interactions.Enzymatic hydrolysis can generally be performed in three ways: (i) under traditional batch conditions; (ii) using immobilized enzymes; or (iii) using ultrafiltration membranes. Many proteolytic enzymes are known and include those described, for example, in Cruz-Casas et al., Enzymatic Hydrolysis and Microbial Fermentation: The Most Favorable Biotechnological Methods for the Release of Bioactive Peptides (Food Chem (Oxf). 3:100047, Dec. 30, 2021), which is incorporated herein by reference in its entirety.

[0064] Microbial fermentation is a biotechnological process that allows the production of bioactive peptides. This process involves the use of microorganisms capable of producing proteolytic enzymes with the objective that these enzymes hydrolyze proteins into shorter peptides. The microorganisms generally used are bacteria, fungi or yeasts, which can be present in the substrate indigenously or added as a starter culture. The microbial fermentation process can be divided into several systems. However, immersion fermentation and solid-state fermentation are the most widely used.

[0065] Immersion fermentation uses a culture of microorganisms in a liquid medium, which contains nutrients. This system is suitable for microorganisms with high water level activities, such as bacteria, and offers the advantage that the generated bioactive peptides are easy to purify. Solid-state fermentation uses microbial growth on nutrient-rich solid substrates. It has the advantage of releasing nutrients in a controlled manner and is suitable for fungi and microorganisms with lower moisture requirements.

[0066] The bioactive walnut peptides may be produced using any method known to those skilled in the art such as those disclosed in Merrifield, R.B., Solid Phase Peptide Synthesis I., J. AM. CHEM. SOC. 85:2149-2154 (1963); Carpino, L.A. et al., [(9-Fluorenylmethyl)Oxy] Carbonyl (Fmoc) Amino Acid Chlorides: Synthesis, Characterization, And Application To The Rapid Synthesis Of Short Peptides, J. ORG. CHEM. 37:51:3732-3734; Merrifield, R.B. et al., Instrument For Automated Synthesis Of Peptides, ANAL. CHEM. 38:1905-1914 (1966); or Kent, SBH et al., High Yield Chemical Synthesis Of Biologically Active Peptides On An Automated Peptide Syn-thesizer Of Novel Design, IN: PEPTIDES 1984 (Ragnarsson U., ed.) Almqvist and Wiksell Int., Stockholm (Sweden), pp. 185-188, all of which are incorporated herein by reference in their entirety.

[0067] The therapeutically effective amount of the one or more bioactive walnut peptides will vary depending on the bioactive walnut peptide and the combination of bioactive walnut peptides. However, in various embodiments, the therapeutically effective amount may be from about 1 μg to about 50 mg (50,000 μg) per cm2 of skin. In other embodiments, the therapeutically effective amount of the one or more walnut peptides is from about 1 pg to about 40 mg, from about 1 pg to about 30 mg, from about 1 to about 20 mg, from about 1 pg to about 10 mg, from about 1 pg to about 8,000 pg, from about 1 pg to about 5,000 pg, from about 1 pg to about 2,000 pg, from about 1 pg to about 1,000 pg, from about 10 pg to about 40 mg, from about 10 pg to about 30 mg, from about 10 to about 20 mg, from about 10 pg to about 10 mg, from about 10 pg to about 8,000 pg, from about 10 pg to about 5,000 pg, from about 10 pg to about 2,000 pg, from about 10 pg to about 1,000 pg,from about 100 pg to about 50 mg, from about 100 pg to about 40 mg, from about 100 pg to about 30 mg, from about 100 pg to about 20 mg, from about 100 pg to about 10 mg, from about 100 pg to about 8,000 pg, from about 100 pg to about 5,000 pg, from about 100 pg to about 2,000 pg, from about 100 pg to about 1,000 pg, from about 500 pg to about 50 mg, from about 500 pg to about 40 mg, from about 500 pg to about 30 mg, from about 500 pg to about 30 mg, from about 500 pg to about 20 mg, from about 500 pg to about 10 mg, from about 500 to about 8,000 pg, from about 500 pg to about 5,000 pg, from about 500 pg to about 2,000 pg or from about 500 pg to about 1,000 pg, per cm2 of skin.

[0068] The bioactive walnut peptides are typically formulated with a physiologically acceptable carrier and applied to the skin as a pharmaceutical or cosmetic composition. The amount of the one or more bioactive walnut peptides will vary depending on their activity, use, and interaction with additional bioactive walnut peptides that may optionally be included in the pharmaceutical or cosmetic composition. However, in various embodiments, the pharmaceutical or cosmetic composition includes from about 0.01 to about 10% by weight of the one or more bioactive walnut peptides, based on the total weight of the pharmaceutical or cosmetic composition.

[0069] In other embodiments, the pharmaceutical or cosmetic composition includes from about 0.01 to about 8% by weight, from about 0.01 to about 5% by weight, from about 0.01 to about 3% by weight, from about 0.01 to about 1% by weight, from about 0.01 to about 0.5% by weight, from about 0.1 to about 10% by weight, from about 0.1 to about 8% by weight, from about 0.1 to about 5% by weight, from about 0.1 to about 3% by weight, from about 0.1 to about 1% by weight, from about 0.5 to about 10% by weight, from about 0.5 to about 8% by weight, from about 0.5 to about 5% by weight, from about 0.5 to about 3% by weight, from about 0.5 to about 2% by weight, about 1 to about 10% by weight, about 1 to about 8% by weight, about 1 to about 5% by weight, about 1 to about 3% by weight, or about 0.1 to about 2% by weight of the one or more bioactive nut peptides, based on the total weight of the pharmaceutical or cosmetic composition.

[0070] The pharmaceutical and cosmetic compositions including the one or more bioactive nut peptides may be formulated in various forms, for example, lotions, creams, serums, sprays, emulsions, gels, powders, dispersions, ointments, sticks, pastes and mousses. In addition to a physiologically acceptable carrier, the pharmaceutical and cosmetic composition may optionally include one or more of the following: • Active desquamating agents, keratolytic agents and exfoliating agents, • Suspending agents, • Emulsifying agents, • Thickeners, and / or • Antihistamines. Skin active agents

[0071] Non-limiting examples of skin active agents include rosacea inhibitory agents (e.g., metronidazole, sulfacetamide, sodium sulfacetamide, sulfur, dapson, doxycycline, minocycline, clindamycin, clindamycin phosphate, erythromycin, tetracyclines, azelaic acid, calcium dobesilate, maleic acid and any compatible combination thereof); a-adrenergic receptor agonists (e.g., clonidine, amphetamine, doxtroamphetamine, apra-clonidine, dipivefrine, a-methyldopa, oxymetazoline, oxymetazoline hydrochloride, methoxamine, metaraminol, medetomidine, dexmedetomidine, ethylnorepinephrine, guanfacine, guanabenz, phenylephrine, phenylephrine hydrochloride, ephedrine, epinine, epinephrine, ethylnorepinephrine, levarterenol, lofexidine, norepinephrine, norphenylephrine, norephredine, phenylpropanolamine, pemoline, propylhexadrine, pseudoephedrine, methamphetamine, a-methylnorepinephrine, methylphenidate, me-phentermine, midodrine, mivazerol, moxonidine, desglymidodrine, tetrahydrozoline, tetrahydrozoline hydrochloride, cirazoline, amidephrine, brimonidine, bri-monidine tartrate, naphazoline, isoproterenol, xylazine, xylometazoline and / or tizanidine);chemicals and botanical extracts with vasoconstrictor properties, including, but not limited to, corticosteroids, ephedrine, pseudoephedrine, caffeine and / or escin; extracts of ephedra, phedra sinica, hamamelis virginiana, hydrastis ca-nadensis, lycopus virginicus, aspidosperma quebracho, cytisus scoparius, raphanus sativus linn (radish leaf extracts), horse chestnut, etc.;

[0072] In a preferred embodiment, the skin active agents are selected from ceramides, ceramide precursors, free fatty acids, linoleic / oleic acids, cholesterol, cholesterol sulfate, b-glucan, carob seed extract, eperua falcata extract, amino acids, niacinamide and its derivatives, hyaluronic acid and its derivatives, glycerin, allantoin, squalane, omega fatty acids, or combinations thereof.

[0073] Additional non-limiting examples include chemicals or botanical extracts with anti-inflammatory properties (e.g., corticosteroids (for short-term use)), non-steroidal anti-inflammatory drugs, linoleic acid, linolenic acid, bisabolol, glycyrrhetinic acid, glycerin, plant extracts with anti-inflammatory properties (e.g., tea extracts, chamomile extracts), anti-inflammatory interleukins (e.g., II-Ira); isoprenylcysteine ​​analogs (i.e., N-acetyl-S-famesyl-L-cysteine), aromatic aldehydes with anti-inflammatory properties (e.g., 4-ethoxybenzaldehyde), etc., as well as any compatible combination thereof); chemicals or botanical extracts with antimicrobial properties (e.g., antibiotics including, but not limited to, gentamicin, penicillins, cephalosporins, quinolones, cipro-floxacin and / or novobiocin); chemicals or botanical extracts with antifungal properties (e.g., ketoconazole, naphtifin hydrochloride, oxyconazole nitrate, sulconazole nitrate, urea, ter-binafine hydrochloride, selenium sulfide, etc.); chemicals or botanical extracts with . antidandruff properties; chemicals or botanical extracts with antiseborrheic properties; keratolytic agents or botanical extracts with keratolytic properties (i.e., alpha-hydroxy acids; beta-hydroxy acids, polyhydroxy acids, urea, salicylic acid, etc.); chemicals with astringent properties; serine protease inhibitors; saturated dicarboxylic acids; alpha-hydroxy acids (e.g., glycolic acids, lactic acid, malic acid, citric acid, tartaric acid, etc.); beta-hydroxy acids (e.g., carnitine, 3-hydroxybutyric acid, 3-hydroxypropionic acid, [3-hydroxy-[3-methylbutyric acid, salicylic acid, etc.).

[0074] Additional non-limiting examples of skin active agents include retinoic acid, tretinoin, isotretinoin, adapalene, retinol and / or their derivatives; benzoyl peroxide; dapsone; kinetin (N6-furfuryladenine) and their derivatives (e.g., furfurylaminotetrahydropyranyladenine); niacinamide (nicotinamide), and combinations thereof.

[0075] Additional non-limiting examples of skin active agents include metronidazole, sulfacetamide, sodium sulfacetamide, sulfur, tetracyclines, doxycycline, clindamycin, clindamycin phosphate, erythromycin and / or minocycline, azelaic acid, calcium dobesylate, caffeine, theobromine, theophylline and / or a derivative thereof (i.e., xanthines), vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D, vitamin E and vitamin K, creatine, carnitine, and essential fatty acids such as linoleic acid and / or linolenic acid, zinc salts such as, for example, zinc sulfate, zinc chloride, zinc glycinate, zinc gluconate, zinc histidine, zinc L-2-pyrrolidone-5-carboxylate (zinc PCA), zinc salt of linoleic acid,a zinc salt of linolenic acid, a zinc salt of azelaic acid, zinc peptides, zinc oxide, copper salts including, but not limited to, copper sulfate, copper chloride, copper glycinate, copper gluconate, copper histidine, copper L-2-pyrrolidone-5-carboxylate (copper PCA), a copper salt of linoleic acid, a copper salt of linolenic acid, a copper salt of azelaic acid, copper peptides, ,

[0076] Non-limiting examples of skin active agents include madecassoside, retinoic acid, benzoyl peroxide, sulfur, vitamin B6 (pyridoxine or chloride), selenium, samphire, cinnamon extract mixtures, zinc gluconate, zinc pyrrolidonecarboxylate (or zinc pidolate), zinc lactate, zinc aspartate, zinc carboxylate, zinc salicylate, zinc cysteate, copper and copper pidolate as Cuivridone Solabia, plant extracts of Arnica montana, Cinchona succirubra, Eugenia caryophyllata, Humulus lupulus, of Hypericum perforatum, Mentha piperita, Rosmarinus officinalis, Salvia officinalis and Thymus vulgaris, extracts of meadowsweet (Spiraea ulmaria), and mixtures thereof.

[0077] In various embodiments, one or more skin active agents may be selected from adenosine, 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES), hyaluronic acid, lanolin, citric acid, malic acid, lactic acid, tartaric acid, salicylic acid, vitamin C, a vitamin, a retinoid, a retinal, a retinoic acid, a carotenoid, an amino acid, a protein, an enzyme, and a coenzyme. In some cases, the active ingredient is adenosine.

[0078] In one embodiment, the composition includes one or more skin active agents selected from humectants and moisturizing ingredients, a depigmenting agent, or an agent that treats oily skin.

[0079] The humectants and moisturizing ingredients may be in particular glycerol and its derivatives, urea and its derivatives, in particular Hydrovance marketed by National Starch, lactic acid, hyaluronic acid, AHA, BHA, sodium pidolate, xylitol, serine, sodium lactate, ectoin and its derivatives, chitosan and its derivatives, collagen, plankton, an extract of Imperata cylindra sold under the name Moist 24 by Sederma, homopolymers of acrylic acid such as Lipidure-HM from NOF Corporation, beta-glucan and in particular sodium carboxymethyl beta-glucan from Mibelle-AG-Biochemistry, a mixture of passion flower, apricot, corn and rice bran oils sold by Nestlé under the name NutraLipids, a C-glycoside derivative,in particular C-13-D-xylopyranoside-2-hydroxypropane in the form of a 30% by weight solution of active ingredient in a water / propylene glycol mixture (60 / 40% by weight) such as the product manufactured by the company Chimex under the trade name “Mexoryl SBB”, a rosehip oil marketed by Nestle, an extract of Porphyridium cruentum microalgae enriched with zinc, marketed under the name Vincience Algualane Zinc, the spheres of collagen and chondroitin sulfate of marine origin (Atélocollagen) sold by the company Engelhard Lyon under the name Marine Filling Spheres, the spheres of hyaluronic acid such as those marketed by Engelhard Lyon, and arginine.

[0080] Depigmenting agents include vitamin C and its derivatives and in particular vitamin CG, CP and 3-O ethyl vitamin C, alpha and beta-arbutin, ferulic acid, lucinol and its derivatives, kojic acid, resorcinol and its derivatives, tranexamic acid and its derivatives, gentisic acid, homogentisic acid, methyl gentisate or homo-gentisate, dioic acid, D-panthetin calcium sulfonate, lipoic acid, ellagic acid, vitamin B3, linoleic acid and its derivatives, ceramides and their homologues, derived from plants such as chamomile, bearberry, the aloe family (vera, ferox, bardensis), blackberry, skullcap, a kiwi fruit (Actinidia chinensis) marketed by Gattefosse, a root extract of Paeonia suffruticosa, such as that sold by Ichimaru Pharcos under the name Liquid Botanpi Be, a brown sugar extract (Saccharum officinarum) such as the molasses extract marketed by Taiyo Kagaku under the name Liquid Molasses, but this list is not exhaustive. Particular depigmenting agents include vitamin C and its derivatives, including vitamin CG, CP and 3-O ethyl vitamin C, alpha and beta-arbutin, ferulic acid, kojic acid, resorcinol and its derivatives, panthetin D calcium sulfonate, lipoic acid, ellagic acid, vitamin B3, a kiwi fruit (Actinidiadia chinensis) marketed by Gattefosse, a Paeonia suffruticosa root extract, such as that sold by the company Ichimaru Pharcos under the name Botanpi Liquid B.

[0081] Useful skin active agents include adenosine and its derivatives and retinol and its derivatives, such as retinyl palmitate, ascorbic acid and its derivatives such as magnesium ascorbyl phosphate and ascorbyl glucoside; tocopherol and its derivatives such as tocopheryl acetate, nicotinic acid and its precursors such as nicotinamide; ubiquinone; glutathione and its precursors such as L-2-oxothiazolidine-4-carboxylic acid, C-glycoside compounds and their derivatives as described in particular in EP-1345919, in particular C-beta-D-xylopyranoside-2-hydroxy-propane as described in particular in EP-1345919, plant extracts including sea fennel and olive leaf extracts, as well as their plants and hydrolysates such as rice protein hydrolysates or soy proteins;algae extracts and in particular laminaria, bacterial extracts, sapogenins such as diogenin and Dioscorea plant extracts, in particular wild yam, including: α-hydroxy acids, β-hydroxy acids, such as salicylic acid and n-octanoyl-5-salicylic oligopeptides and pseudodipeptides and their acyl derivatives, in particular acid; {2-[acetyl-(3-trifluoromethyl-phenyl)-amino]-3-methyl-]acetic acid and lipopeptides marketed by the company under the trade names SEDERMA Matrixyl 500 and Matrixyl 3000; lycopene, manganese salts and magnesium salts, including gluconates, and mixtures thereof.

[0082] Adenosine derivatives include in particular non-phosphate derivatives of adenosine, such as in particular, 2'-deoxyadenosine, 2',3'-adenosine isopro-pylidene; toyocamycin, 1-methyladenosine, N-6-methyladenosine; adenosine N-oxide, 6-methylmercaptopurine riboside and 6-chloropurine riboside.

[0083] Other derivatives include adenosine receptor agonists such as phenylisopropyl adenosine ("PIA"), 1-methylisoguanosine, N6-cyclohexyladenosine (CHA), N6-cyclopentyladenosine (CPA), 2-chloro-N6-cyclopentyladenosine, 2-chloroadenosine, N6-phenyladenosine, 2-phenylaminoadenosine, MECA, N6-phenethyladenosine, 2-p-(2-carboxy-ethyl) phenethyl-amino-5'- -N-ethylcarboxamido adenosine (CGS-21680), N-ethylcarboxamido-adenosine (NECA), 5'(N-cyclopropyl)-carboxamidoadenosine, DPMA (PD 129.944) and metrifudil.

[0084] In one embodiment, the composition comprises an active ingredient that treats oily skin. These active ingredients may be sebum-regulating or antiseborrheic agents capable of regulating the activity of the sebaceous glands. These include: retinoic acid, benzoyl peroxide, sulfur, vitamin B6 (pyridoxine chloride or), selenium, samphire—mixtures of cinnamon, tea and octanoylglycine extract such as— Seppic's Se-picontrol A5 TEA—the mixture of cinnamon, sarcosine and octanoylglycine marketed in particular by Seppic under the trade name Sepicontrol A5—zinc salts such as zinc gluconate, zinc pyrrolidonecarboxylate (or zinc pidolate), zinc lactate, zinc aspartate, zinc carboxylate, zinc salicylate, zinc cysteate derivatives in particular copper and copper pidolate such as Cuivridone Solabia—plant extracts of Arnica montana, Cinchona succirubra, Eugenia caryophyllata, Humulus lupulus, Hypericum perforatum,of Mentha piperita, Rosmarinus officinalis, Salvia officinalis and Thymus vulgaris, all marketed, for example, by Maruzen—extracts of meadowsweet (Spiraea ulmaria), such as those sold under the name Sebonormine by Silab—extracts of the seaweed Laminaria saccharina, such as those sold under the name Phlorogine by Biotechmarine—extracts of the roots of mixtures of burnet (Sanguisorba officinalis / Poterium officinale), ginger rhizomes (Zingiber officinalis) and cinnamon bark (Cinnamomum cassia), such as those sold under the name Sebustop by Solabia—extracts of flax seeds such as those sold under the name Linumine by Lucas Meyer—extracts of phelodendron such as those sold under the name Phellodendron extract BG by Maruzen or Oubaku liquid B by Ichimaru Pharcos—the extract of argan oil blends,Serenoa serrulata (Saw palmetto) and sesame seed extracts such as those sold under the name Regu SEB by Pentapharm—blends of extracts of willowherb, Terminalia chebula, nasturtium and bioavailable zinc (microalgae), such as that sold under the name Seborilys Green Tech; —Pygeum afrianum extracts such as that sold under the name Pygeum afrianum sterolic lipid extract by Euromed—Serenoa serrulata extracts such as those sold under the name Viapure Sabal by Actives International, and those sold by the company Euromed—blended extracts of plantain, Berberis aquifolium and sodium salicylate such as that sold under the name Seboclear Rahn—clove extract such as that sold under the name Clove extract powder by Maruzen—argan oil such as , that sold under the name Lipofructyl from Laboratoires Sérobiologiques lactic protein filtrates, such as that sold under the name Normaseb by Sederma—laminaria algae extracts, such as that sold under the name Laminarghane by Bio-techmarine—oligosaccharides from the algae Laminaria digitata, such as that sold under the name Phycosaccharide AC by the company Codif—sugar cane extracts such as that sold under the name Policosanol by the company Sabinsa, sulfonated shale oil, such as that sold under the name Ichtyol Pale by Ichthyol—meadowsweet extracts (Spiraea ulmaria) such as that sold under the name Cytobiol Ulmaire by the company Libiol—sebacic acid, notably sold in the form of sodium polyacrylate gel under the name Sebosoft by Sederma—glucommanans extracted from the konjac tuber and modified by alkylsulfonate chains such as that sold under the name Biopol Beta by Arch Chemical—extracts of Sophora angustifolia,such as that sold under the name Sophora powder or Sophora extract by Bioland—cinchona succirubra bark extracts such as that sold under the name Red Bark HS by Alban Muller—Quillaja saponaria extracts such as that sold under the name Panama wood HS by Alban Muller—glycine grafted onto an undecylenic chain, such as that sold under the name Lipacide UG OR by SEPPIC—the mixture of oleanolic acid and nordihydroguaiaretic acid, such as that sold in gel form under the name AC. Net by Sederma; —phthalimidoperoxyhexanoic acid—tri(C12-C13) citrate sold under the name COSMACOL ECI by Sasol; trialkyl citrate (C14-C15) sold as COSMACOL ECL by Sasol--hydroxydecanoic acid, including mixtures of hydroxydecanoic acid, sebacic acid and 1,10-decanediol such as that sold as Acnacidol BG by Vincience, and mixtures thereof.

[0085] Other skin active agents useful herein include those selected from N-acetyl D-glucosamine, panthenol (e.g., DL panthenol available from Alps Pharmaceutical Inc.), tocopheryl nicotinate, benzoyl peroxide, 3-hydroxybenzoic acid, flavonoids (e.g., flavanone, chalcone), farnesol, phytantriol, glycolic acid, lactic acid, 4-hydroxybenzoic acid, acetylsalicylic acid, 2-hydroxybutanoic acid, 2-hydroxypentanoic acid, 2-hydroxyhexanoic acid, cis-retinoic acid, trans-retinoic acid, retinol, retinyl esters (e.g., retinyl propionate), phytic acid, N-acetyl-L-cysteine, lipoic acid, tocopherol and its esters (e.g., tocopheryl acetate: DL-a-tocopheryl acetate available from Eisai), azelaic acid, arachidonic acid, tetracycline, ibuprofen, naproxen, ketoprofen, hydrocortisone, acetaminophen, resorcinol,phenoxyethanol, phenoxypropanol, phenoxyisopropanol, 2,4,4'-trichloro-2'-hydroxydiphenyl ether, 3,4,4'-trichlorocarbanilide, octopirox, lidocaine hydrochloride, clotrimazole, miconazole, ketoconazole, neomycin sulfate, theophylline, and mixtures thereof.

[0086] In various embodiments, the total amount of skin active agents in the pharmaceutical or cosmetic composition of the disclosure, other than the one or more bioactive nut peptides may be included in an amount of greater than zero to about 9% by weight, greater than zero to about 8% by weight, greater than zero to about 7% by weight, greater than zero to about 6% by weight, greater than zero to about 5% by weight, greater than zero to about 4% by weight, greater than zero to about 3% by weight, greater than zero to about 2% by weight;from about 10 ppm to about 10 wt% (100,000 ppm), from about 10 ppm to about 5 wt% (50,000 ppm), from about 10 ppm to about 2.5 wt% (25,000 ppm), from about 10 ppm to about 1 wt% (10,000 ppm), from about 10 ppm to about 0.5 wt% (5,000 ppm), from about 10 ppm to about 0.3 wt% (3,000 ppm), from about 10 ppm to about 0.2 wt% (2,000 ppm), from about 10 ppm to about 0.1 wt% (1,000 ppm), from about 10 ppm to 500 ppm; from about 0.1 to about 10% by weight, from about 0.1 to about 5% by weight, from about 0.1 to about 2.5% by weight, from about 0.1 to about 1% by weight, from about 0.1 to about 0.5% by weight; from about 1 to about 10% by weight, from about 1 to about 8% by weight, from about 1 to about 6% by weight, from about 1 to about 5% by weight, from about 1 to about 4% by weight, from about 1 to about 3% by weight;from about 2 to about 10% by weight, from about 2 to about 8% by weight, from about 2 to about 6% by weight, from about 2 to about 5% by weight, from about 2 to about 4% by weight; from about 3 to about 10% by weight, from about 3 to about 8% by weight, from about 3 to about 6% by weight, from about 3 to about 5% by weight; from about 4 to about 10% by weight, from about 4 to about 8% by weight or from about 4 to about 6% by weight, based on the total weight of the pharmaceutical or cosmetic composition.; Emollients

[0087] One or more emollients may also optionally be included in the pharmaceutical or cosmetic composition described herein. An emollient generally refers to an ingredient that can help maintain a soft, smooth, and supple appearance of the skin. Emollients generally remain on the surface of the skin or in the stratum corneum and act as a moisturizer or lubricant and reduce delamination. Non-limiting examples of emollients include acetylarginine, acetylated lanolin, algae extract, polyethylene glycol-6 esters from apricot kernel oil, polyethylene glycol-11 esters from avocado oil, bis-polyethylene glycol-4 dimethicone, butoxyethyl stearate, glycol esters, alkyl glycol esters, cetyl laurate, polyethylene glycol-10 esters from coco, alkyl tartrates, diethyl sebacate, dihydrocholesteryl butyrate, dimethiconol, Dimyristyl tartrate, lauroyl glutamate distearate-5, etilavokadate, ethylhexyl myristate, glyceryl isostearate, glyceryl oleate, geksildetsilstearate, geksilizostearate, hydrogenated palm glycerides, hydrogenated soy glycerides, hydrogenated fat glycerides isostearilneopentanoate, isostearyl palmitate, isostearylsilizononanoate, laureth-2 acetate, lauryl polyglyceryl-6 cetearyl glycol ether, methylglyutset-20 benzoate, mineral oil, palm oil, coconut oil, miret-3 palmitate, octyldecanol, octyldodecanol, Odontella aurita oil, 2-oleamido-l,3 octadecanediol, pal commercial glycerides, avocado polyethylene glycol glycerides, castor oil polyethylene glycol, polyethylene glycol-2 / dodecyl glycol copolymer, shea butter polyethylene glycol glycerides, phytol, raffinose, stearyl citrate,sunflower seed oil glycerides, non-ointment, small tocopherol glucoside. , Suspension agents

[0088] The pharmaceutical or cosmetic composition of the present invention may optionally include one or more suspending agents, preferably in a concentration effective to suspend the water-insoluble material in a dispersed form in the compositions or to modify the viscosity of the composition. These concentrations will vary. However, in certain embodiments, the pharmaceutical and cosmetic composition includes from about 0.1% to about 10%, more preferably from about 0.25% to about 5.0% of the one or more suspending agents, based on the total weight of the composition. Non-limiting examples include vinyl polymers, such as crosslinked acrylic acid polymers, referred to as carbomer, cellulose derivatives and modified cellulose polymers such as methylcellulose, ethylcellulose, nitrocellulose, carboxymethylcellulose, crystalline cellulose, cellulose powder,polyvinylpyrrolidone, polyvinyl alcohol, guar gum, hydroxypropyl guar gum, gum arabic, galactan, locust bean gum, pectin, agar, starch (rice, corn, potato, wheat), seaweed colloids (seaweed extract), microbiological polymers such as dextran, succinoglycan, pullulan, starch-based polymers such as carboxymethyl starch, methyl starch, alginic acid polymers such as sodium alginate, propylene glycol esters of alginic acid, acrylate polymers such as sodium polyacrylate, polyacrylate, polyacrylamide, polyethyleneimine and water-soluble inorganic materials such as bentonite, magnesium aluminum silicate, laponite, hectorite and acid anhydrous silicic acid. ,

[0089] Other optional suspending agents include crystalline suspending agents which can be resolved into acyl derivatives, long chain amine oxides, acyl derivatives long chain, and mixtures thereof. Said preferred suspending agents include fatty acid ethylene glycol esters, fatty acid alkanolamides, long chain fatty acid esters (e.g., stearyl stearate, cetyl palmitate, etc.); long chain esters of long chain alkanolamides (e.g., stearamide distearate diethanolamide, stearamide stearate monoethanol amide); and glyceryl esters (e.g., glyceryl distearate, trihydroxystearin, tribhengene). Other suitable suspending agents include primary amines containing a fatty alkyl moiety containing at least about 16 carbon atoms, examples of which include palmitamine or stearamine, and secondary amines containing two fatty alkyl moieties, each of which contains at least about 12 carbon atoms, examples of which include dipalmitoylamine or di(hydrogenated fat) amine.Other suitable suspending agents include phthalic acid diamide (hydrogenated fat) and a crosslinked copolymer of maleic anhydride / methyl vinyl ether. Emulsifying agents

[0090] Non-limiting examples of emulsifying agents include condensation products of alkylene oxides with fatty acids (i.e., fatty acid alkylene oxide esters), condensation products of alkylene oxides with 2 moles of fatty acids (i.e., fatty acid alkylene oxide diesters), condensation products of alkylene oxides with fatty alcohols (i.e., fatty alcohol alkylene oxide esters), condensation products of alkylene oxides with both fatty acids and fatty alcohols [i.e., where a portion of the polyalkylene oxide is esterified at one end with a fatty acid and esterified (i.e., via an ether linkage) at the other end with a fatty alcohol].Non-limiting examples of non-ionic surfactants derived from said alkylene oxide include ceteth-6, ceteth-10, ceteth-12, ceteareth-6, ceteareth-10, ceteareth-12, steareth-6, steareth-10, steareth-12, steareth-21, PEG-6 stearate, PEG-10 stearate, PEG-100 stearate, PEG-12 stearate, PEG-20 glyceryl stearate, PEG-80 glyceryl tallowate, PEG-10 glyceryl stearate, PEG-30 glyceryl cocoate, PEG-80 glyceryl cocoate, PEG-200 glyceryl tallowate, PEG-8 dilaurate, PEG-10 distearate, and mixtures thereof. Other applicable nonionic surfactants include polyhydroxyamide fatty acid surfactants. A particularly preferred surfactant corresponding to the above structure is coco N-methylglucoside alkylamide.Preferred surfactants among the nonionic surfactants are surfactants selected from the group consisting of steareth-21, ceteareth-20, ceteareth-12, sucrose cocoate, steareth-100, PEG-100 stearate, and mixtures thereof. Other nonionic surfactants suitable for use in this application include esters and . sugar polyesters, alkoxylated sugar esters and polyesters, C1-C30 fatty acid esters, C1-C30 fatty alcohols, alkoxylated C1-C30 ester derivatives, C1-C30 fatty alcohol fatty acids, alkoxylated C1-C30 fatty alcohol esters, polyglyceryl C1-C30 fatty acid esters, C1-C30 polyol esters, C1-C30 polyol esters, alkyl phosphates, polyoxyalkylene fatty ether phosphates, fatty acid amides, acyl lactylates, and mixtures thereof.Non-limiting examples of such emulsifiers include: polyethylene glycol 20 sorbitan monolaurate (polysorbate 20), polyethylene glycol 5 soy sterol, steareth-20, ceteareth-20, PPG-2 methyl glucose ether distearate, ceteth-10, polysorbate 80, cetyl phosphate, cetyl phosphate, cetyl phosphate, cetyl phosphate, cetyl phosphate, polysorbate 60, glyceryl stearate, polyoxyethylene 20 sorbitan triolcat (polysorbate 85), sorbitan monolaurate, polyoxyethylene 4 lauryl ether sodium stearate, polyglyceryl-4 isostearate, hexyl laurate, PPG-2 methyl glucose ether distearate, PEG-100 and their PEG-100. Another group of nonionic surfactants useful herein is a mixture of fatty acid esters based on a mixture of sorbitan or a sorbitol fatty acid ester and a sucrose fatty acid ester, wherein the fatty acid in each example is preferably C8-C24, more preferably C10-C20. Thickeners

[0091] Thickeners suitable for inclusion in the pharmaceutical or cosmetic composition are described herein.Non-limiting examples include acrylamide copolymer, agarose, amylopectin, bentonite, calcium alginate, calcium carboxymethylcellulose, carbomer, carboxymethylchitin, cellulose gum, dextrin, gelatin, hydroxymethyl hydroxycellulose hydroxypropyl, hydroxyethyl hydroxypropyl hydroxypropyl, hydroxyethyl hydroxypropyl, hydroxyethyl hydroxypropyl, hydroxyethyl hydroxypropyl, hydroxyethyl hydroxypropyl, hydroxyethyl hydroxypropyl, hydroxypropyl; magnesium alginate, methylcellulose, microcrystalline cellulose, pectin, various polyethylene glycols, polyacrylic acid, polymethacrylic acid, polyvinyl alcohol, various isopropylene glycols, sodium acrylate copolymers, sodium carrageenan, xanthan gum and / or yeast beta-glucan, or mixtures thereof.

[0092] More generally, carboxylic acid polymers are useful thickeners. Carboxylic acid polymers are crosslinked compounds containing one or more monomers derived from acrylic acid, substituted acrylic acids, and salts and esters of said acrylic acids and substituted acrylic acids, wherein the crosslinking agent contains two or more carbon-carbon double bonds and is derived from a polyhydric alcohol. Examples of carboxylic acid polymers Commercially available compounds useful here include carbomers, which are homopolymers of acrylic acid crosslinked with allyl ethers of sucrose or pentaerythritol. Carbomers are available in BF's Carbopol® 900 range Goodrich (e.g., Carbopol® 954). In addition, other suitable carboxylic acid-based polymeric agents include copolymers of C10-30 alkyl acrylates with one or more monomers of acrylic acid, methacrylic acid, or esters of one of its short chains (i.e., a C1-4 alcohol), the crosslinking agent being an allyl ether of sucrose or pentaerythritol. These copolymers are known as acrylate / C10-C30 alkyl acrylate crosslinked polymers and are commercially available as Carbopol® 1342, Carbopol® 1382, Pemulen TR-1, and Pemulen TR-2. Goodrich. Examples of preferred carboxylic acid polymer-based thickeners useful in this application include thickeners selected from carbomers, acrylate / C10-30 alkyl acrylate crosslinked polymers, and mixtures thereof.

[0093] Further, in some embodiments, the thickeners are selected from polysaccharides. Non-limiting examples of polysaccharide thickeners include cellulose, carboxymethyl hydroxyethyl cellulose, cellulose acetate propionate carboxylate, hydroxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium methyl hydroxyethyl cellulose, and hydroxyethyl methyl hydroxyethyl hydroxyethyl cellulose, hydroxyethyl methyl hydroxyethyl hydroxyethyl cellulose, hydroxyethyl hydroxyethyl cellulose hydroxyethyl methyl hydroxyethyl hydroxyethyl. Alkyl-substituted celluloses are also useful. In these polymers, the hydroxy groups of the cellulose polymer are hydroxylated (preferably hydroxyethylated or hydroxypropylated) to form hydroxylated cellulose, which is then further modified with a straight-chain or branched C10-30 alkyl group via an ether linkage.Typically, these polymers are straight-chain or branched-chain esters of C10-30 alcohols and hydroxy alkyl celluloses. Examples of alkyl groups applicable in this application include groups selected from stearyl, isostearyl, lauryl, myristyl, cetyl, isocetyl, cocoyl (e.g., an alkyl group derived from coconut oil alcohols), palmityl, oleyl, linoleyl, linoleyl, retinoleyl, behenyl, and mixtures thereof. A preferred alkyl hydroxyalkyl cellulose ester is a material called cetyl hydroxyethyl cellulose, which is an ester of cetyl alcohol and hydroxyethyl cellulose, according to the Perfume and Cosmetics and Perfume Association (CTFA). The material indicated is sold under the trade name Natrosol® CS Plus by Aqualon Corporation (Wilmington, Delaware).Other examples can be found in the International Cosmetic Ingredient Dictionary and Handbook, the Cosmetic Bench Reference - Directory of Cosmetic Ingredients, offered . by the United States Pharmacopeia (USP) and the National Formulary (NF), and other references to cosmetic and pharmaceutical ingredients known in the art. Other useful polysaccharides include scleroglucans, which are straight-chain (1-3) linked (1-6) glucose units, where every third glucose unit is linked, a commercially available example of which is Clearogel™ CS11 from Michel Mercier Products Inc. (Mountainside, New Jersey).

[0094] Other useful thickeners include those derived from natural sources. Non-limiting examples include gum arabic, agar-agar, algin, alginic acid, ammonium alginate, amylopectin, calcium alginate, calcium carrageenan, carnitine, carrageenan, dextrin, gelatin, gellan gum, guar gum hydrochloride, gidro hydrochloride, gidro hydrochloride, gidro hydrochloride, gidro hydrochloride, gidro hydrochloride, hyaluronic acid, hydrated silicon dioxide, hydroxypropyl chitosan, hydroxypropyl gum, karaya gum, kelp, fruit tree resin, natto gum, potassium alginate, potassium carrageenan, propylene glycol alginate, sclerotium gum, carboxymethyl sodium dextran, dextran, sodium carrageenan, tragacanth gum, xanthan gum and / or mixtures thereof.

[0095] Additionally, the compositions may optionally contain polyacrylamide polymers, particularly nonionic polyacrylamide polymers, including substituted branched or unbranched polymers. Other polyacrylamide polymers useful herein include multiblock copolymers of acrylamides and acrylamides substituted with acrylic acids and substituted acrylic acids. Antihistamines

[0096] Antihistamines, also called histamine antagonists, are substances that inhibit the action of histamine by blocking its binding to histamine receptors or by inhibiting the enzymatic activity of histidine decarboxylase, which catalyzes the conversion of histidine to histamine and the like.Non-limiting examples of antihistamines are acrivastine, azelastine, brompheniramine, buclizine, bromodiphenhydramine, carbinoxamine, cetirizine, chlorpromazine, cyclizine, chlorpheniramine, chlordiphenhydramine, cimetidine, clemastine, cyproheptadine, desloratine, dimend mirtazapine, nizatidine, olopadadine, orphenadrine, phenindamine, pheniramine, phenyltoloxamine, . promethazine, pyrilamine, quetiapine, ranitidine, roxatidine, rupatadine, tripelennamine and triprolidine. EXAMPLES

[0097] Various changes may be made to the compositions and methods described above without departing from the scope of the invention. Accordingly, it is intended that any disclosure contained in the above description and in the examples given below, should be interpreted in an illustrative and not limiting sense. Example 1 Differential gene expression

[0098] Peptides I, II and III were studied for their ability to influence gene expression at a concentration of 100 pM to 500 pM for cells and 250 pM for reconstructed human epidermis (RHE).

[0099] Normal adult human epidermal keratinocytes

[0100] Normal adult human epidermal keratinocytes (lot 80819221, passage 3, ATCC) were expanded and seeded in a 6-well plate at a density of 300,000 cells / well. The cells were cultured in dermal cell basal medium (ATCC PCS-200-030) supplemented with a keratinocyte growth kit (ATCC PCS-200-040). The cells were fixed overnight, then the culture medium was aspirated and replaced with medium containing either 100 pM or 500 pM of peptide I, peptide II, or peptide III. The cells were cultured for 4 days with treatment and renewal of the treatment medium every other day except for weekends. After the 7-day incubation period, the cells were trypsinized and the cell pellet was snap-frozen using liquid nitrogen. The cells then underwent RNA sequencing.The log2 relative change for each treatment group compared to untreated control samples for differentially expressed genes (DEGs) related to the skin barrier was analyzed. The relative change was plotted via a heatmap to show the up- or down-regulation of these genes. Microsoft Excel and GraphPad Prism were used for further analysis and data presentation. The results are shown in the heatmap in [Fig.l]. The data show that peptide II and peptide III provide potent benefit to the skin barrier at the cellular level at both concentrations tested, while peptide I showed influence at the higher concentration. Reconstructed human epidermis

[0101] Reconstructed human epidermis (RHE) (EPISKIN / S / 13, Episkin) was received and samples were prepared for use. Upon arrival, the inserts containing the RHE were removed from the multiplate and placed into a 12-well plate containing the preservation medium. Tissues serving as untreated control were placed in routine preservation medium; treatment groups were placed in preservation medium containing 250 pM of peptide I, II, or III in combination. The medium was aspirated and replaced every other day except for weekends. RHE was cultured at the air-liquid interface at 37 °C, 5% CO2, and saturated humidity for 7 days. Following culture, samples were snap-frozen in liquid nitrogen and sent for RNA sequencing. The log2 relative change for each treatment group compared to untreated control samples for differentially expressed genes (DEGs) related to the skin barrier was analyzed. The relative change was plotted via a heat map to show the up- or down-regulation of these genes.Microsoft Excel and GraphPad Prism were used for further analysis and data presentation, shown in the heat map in [Fig.2]. The data show that a combination of the three peptides provides improved skin barrier function for EHR. Example 2 Facial treatment

[0102] An exemplary facial treatment containing about 0.01 to about 1% by weight of peptide I, peptide II, peptide III, or combinations thereof is shown below.

[0103] [Tableauxl] Table 6 INGREDIENTS % by weight PEPTIDE I, II, III OR THEIR COMBINATIONS 0.01 - 10 METHYLPARABEN 0.3 2-PHENOXYETHANOL 0.5 SODIUM POLYACRYLATE 0.8 XANTHAN GUM 0.2 WATER / AQUA 98.2

[0104] The foregoing description illustrates and describes the disclosure. In addition, the disclosure shows and describes only the preferred embodiments. However, as mentioned above, it is to be understood that it is capable of use in many other combinations, modifications, and environments and is capable of undergoing changes or modifications within the scope of the inventive concepts as expressed herein, consistent with the above teachings and / or the skill or knowledge of the relevant art. The embodiments described herein are further intended to explain the best modes known to the applicant and to enable others skilled in the art to use the disclosure in such or other embodiments, and with the various modifications required by the particular applications or uses thereof. Accordingly, the description is not intended to limit the invention to the form disclosed herein. Likewise, the appended claims are intended to be construed as including alternative embodiments.

[0105] As used herein, the terms "comprising," "having," and "including" are used in their broad and non-limiting sense.

[0106] The terms "a," "an," "the," and "the" are understood to encompass both the plural and the singular. Thus, the expression "a mixture thereof" also relates to "mixtures thereof." Throughout the disclosure, the expression "a mixture thereof" is used, following a list of elements as shown in the following example where the letters A through F represent the elements: "one or more elements selected from the group consisting of A, B, C, D, E, F, and a mixture thereof." The expression "a mixture thereof" does not require that the mixture include all of A, B, C, D, E, and F (although all of A, B, C, D, E, and F may be included). Rather, it indicates that a mixture of two or more of A, B, C, D, E, and F may be included. In other words, it is equivalent to the formulation "one or more elements selected from the group consisting of A, B, C, D, E, F, and a mixture of two or more of A, B, C, D, E and F".

[0107] Similarly, the expression "a salt thereof" also relates to "salts thereof". Thus, when the disclosure refers to "an element selected from the group consisting of A, B, C, D, E, F, a salt thereof, and a mixture thereof", it indicates that one or more of A, B, C, D and F may be included, one or more of a salt of A, a salt of B, a salt of C, a salt of D, a salt of E and a salt of F may be included, or a mixture of any two of A, B, C, D, E, F, a salt of A, a salt of B, a salt of C, a salt of D, a salt of E and a salt of F may be included.

[0108] Salts referred to throughout the disclosure may include salts having a counterion such as an alkali metal, alkaline earth metal, or ammonium counterion. This list of counterions, however, is not limiting. Suitable counterions for the components described herein are known in the art.

[0109] The expression “one or more” means “at least one” and therefore includes individual components as well as mixtures / combinations.

[0110] The term “plurality” means “more than one” or “two or more”.

[0111] Except in operational examples, or unless otherwise indicated, all numbers expressing quantities of ingredients and / or reaction conditions may be modified in all cases by the term “approximately”, meaning to within + / - 5% of the number noted.

[0112] All percentages, parts and ratios herein are based on the total weight of the compositions of the present invention, unless otherwise indicated.

[0113] Some of the various categories of components identified may overlap. In such cases where an overlap may exist and the composition includes both components (or the composition includes more than two overlapping components), an overlapping compound does not represent more than one component. For example, some compounds may be considered both an emollient and a nonionic surfactant. If a particular composition includes both an emollient and a nonionic surfactant, a single compound will serve solely as an emollient or solely as a nonionic surfactant (the single compound does not simultaneously serve as both an emollient and a nonionic surfactant).

[0114] As used herein, all ranges provided are intended to include each specific range within the given ranges, as well as each combination of intermediate subranges. Thus, a range of 1 to 5 specifically includes 1, 2, 3, 4, and 5, as well as subranges such as 2 to 5, 3 to 5, 2 to 3, 2 to 4, 1 to 4, etc. All ranges and values ​​disclosed herein are inclusive and combinable. For example, any value or point described herein that falls within a range described herein may serve as a minimum or maximum value to infer a subrange, etc.

[0115] The term "substantially free" or "essentially free" as used herein means that less than about 2% by weight of a specific material is added to a composition, based on the total weight of the compositions. However, the compositions may include less than about 1% by weight, less than about 0.5% by weight, less than about 0.1% by weight, or none at all of the specified material.

[0116] Any components positively presented in the present disclosure may be negatively excluded from the claims, for example, a claimed composition may be "free", "essentially free" (or "substantially free") of one or more components that are positively presented in the present disclosure.

[0117] All publications and patent applications cited in this patent specification are incorporated herein by reference, and for all purposes, as if each individual publication or patent application were specifically and individually indicated as being incorporated by reference. In the event of any inconsistency between this disclosure and any publication or patent application incorporated herein by reference, this disclosure shall prevail.

Claims

Claims

1. A method of improving skin barrier function or improving re-epithelialization or improving wound healing of the skin, comprising topically applying a therapeutically effective amount of one or more bioactive walnut peptides or a composition comprising the one or more bioactive walnut peptides to the skin.

2. The method of claim 1, wherein said wound results from damage to skin tissue using a chemical, laser, or physical force.

3. The method of claim 2, wherein the skin tissue is damaged using an ablative laser procedure, a non-ablative laser procedure, a microneedle procedure, a cryotherapy procedure, a radiofrequency microneedle procedure, dermabrasion, a chemical peel, an exfoliant, or a mechanical device.

4. A method according to any preceding claim, wherein the skin is inflamed or suffering from an inflammatory condition.

5. The method of claim 4, wherein the inflammatory condition is selected from atopic dermatitis, psoriasis or a combination thereof.

6. The method of claim 1, wherein the method treats a skin condition, disease, or disorder selected from psoriasis, dermatitis, atopic dermatitis, allergic dermatitis, eczema, spongiosis, edema, hereditary ichthyosis, senile xerosis, palmar hyperkeratosis, plantar hyperkeratosis, cuts, bruises, pore size, skin cancer, wound healing or re-epithelialization disorders, keloids, hypertrophic scars, cellulite, orange peel skin, elastosis, actinic elastosis, keratosis, rosacea, telangiectasia, couperose, or combinations thereof.

7. A method according to any preceding claim, wherein the one or more nut peptides are combined with one or more additional skin active agents.

8. The method of claim 7, wherein the one or more additional skin active agents are selected from anti-atrophy agents, antioxidants, depigmentation agents, or combinations thereof.

9. A method according to any preceding claim, wherein the one or more bioactive nut peptides have a molecular weight of less than 5,000 Da, preferably less than 2,000 Da, more preferably less than 1,000 Da.

10. A method according to any preceding claim, wherein the one or more bioactive nut peptides have 2 to 20 amino acid residues, preferably 3 to 15 amino acid residues, more preferably 4 to about 10 amino acid residues.

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