BIOACTIVE NUT PEPTIDES TO TREAT HAIR LOSS

Abstract

BIOACTIVE WALNUT PEPTIDES FOR TREATING HAIR LOSS Bioactive walnut peptides and their use for topical application to the scalp are described. One or more bioactive walnut peptides may be incorporated into a pharmaceutical or cosmetic composition for application to the scalp. Pharmaceutical or cosmetic compositions containing one or more bioactive walnut peptides are particularly useful for treating or preventing hair loss, and for treating scalp conditions such as dandruff, scalp psoriasis, and acne. Figure for abstract: none

Description

Title of the invention: BIOACTIVE NUT PEPTIDES FOR TREATING HAIR LOSS SCOPE OF DISCLOSURE

[0001] This disclosure relates to bioactive nut peptides and methods for treating and preventing hair loss, and for treating skin conditions of the scalp. CONTEXT

[0002] Hair follicles, particularly in humans, are complex structures with distinct components, each composed of various specialized cells. In addition to the cells responsible for hair production and anchoring, most hair follicles also contain sebaceous glands, which produce sebum. Some follicles have apocrine glands, which are found in specific areas such as the armpit. The structures within a hair follicle include the follicular papilla (FP) and the germinal epithelium (GE), collectively referred to as the bulb.The PF consists of mesenchymal cells and connective tissue, while the epithelial components include at least eight distinct cell lineages: the outer epithelial sheath (ORS), the companion layer (CL), the inner epithelial sheath Henle layer (He), the Huxley layer (Hu), the inner epithelial sheath cuticle (Csth), the capillary stem cuticle (Csft), the capillary stem cortex, and the stem medulla (Med).

[0003] In humans, scalp hair and hair in certain other areas grows in follicular units. A typical follicular unit on the scalp consists of two to four terminal hair follicles, one or rarely two vellus follicles, along with their associated sebaceous glands, a neurovascular plexus, an arrector pili muscle, and a circumferential band of adventitia collagen called the "perifolliculum."

[0004] Hair follicles are thought to produce approximately 20 individual hair shafts throughout their lifetime, going through phases of growth, shedding, involution, and regrowth. Although hair growth and follicular regeneration have been studied in mouse models, there are important differences between the biology of human and mouse hair follicles. In mice, a thick fur coat is vital for thermoregulation and other functions, with the follicles primarily producing terminal hairs (fur). In contrast, human skin has areas covered with hair follicles that produce vellus hair, which is less visible. The hair follicles of Mice have synchronous growth cycles early in life, but these become less synchronized with age, while human hair follicles progress asynchronously through their growth cycles. On an adult human scalp, approximately 80 to 90% of follicles are in the anagen (growth) phase, 10 to 20% in the telogen (resting) phase, and 1 to 2% in the catagen (regression) phase.

[0005] Dermal papilla cells (DPCs) in hair follicles are crucial for hair growth, residing in the hair bulb and regulating the hair follicle growth cycle. Their ability to induce hair follicle formation, known as inductivity, is essential for both preventing hair loss and promoting hair growth. DPCs secrete growth factors and signaling molecules that interact with epithelial cells in the hair follicle, driving cell proliferation and differentiation. This interaction is essential for initiating and sustaining hair growth, as DPCs play a key role in the cyclical phases of hair growth: anagen (growth), catagen (regression), and telogen (resting). DPC activity influences the transitions between these phases, affecting hair length and thickness.

[0006] The transition from telogen to anagen is complex and involves several molecular signals, including Wnt and bone morphogenetic protein (BMP) signaling pathways. During telogen, Wnt signaling is inactive in the hair follicle. The transition to anagen is triggered by signals from CPDs, which activate Wnt signaling in epithelial stem cells located in the follicle bulge region. This activation promotes the proliferation and differentiation of these stem cells, leading to the formation of a new hair shaft and downward growth of the follicle into the dermis. In contrast, BMP signaling generally inhibits hair growth. High BMP signaling during telogen helps maintain the follicle in its resting state.For anagen to begin, PMO signaling must be reduced, often involving inhibitory molecules that block PMO pathways and allow Wnt signaling to prevail, thus initiating hair growth.

[0007] Furthermore, hypoxia (low oxygen levels) has been shown to enhance CPD inductivity in 2D models and promote angiogenesis, which is vital for the healthy development of hair follicles and the scalp. A proper balance between these signaling pathways is essential for efficient hair follicle development and cycling. Recent studies have also observed an increase in autophagy at the onset of anagen in the natural hair cycle, and it has been found that activating autophagy in aged mice prevented (or slowed) hair loss. DISCLOSURE SUMMARY

[0008] This disclosure relates to bioactive walnut peptides and the use of walnut peptides for topical application to the scalp for use in the treatment or prevention (or slowing) of hair loss, and the treatment of scalp conditions. The inventors have made a surprising discovery that bioactive walnut peptides positively influence physiological pathways associated with hair growth and hair follicle health, and downregulate physiological pathways detrimental to healthy hair growth and scalp function. For example, bioactive walnut peptides stimulate or activate dermal papilla cells while downregulating genes associated with cell cycle arrest and IL-1 (interleukin-1) signaling, which is associated with inflammation, the immune response, and apoptosis.

[0009] 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 have a high lipid content. The residue remaining after lipid extraction is considered a by-product, although it contains nut proteins and other useful components. Walnut protein is mainly composed of albumin, globulin, gliadin, and glutenin.

[0010] Bioactive nut peptides can be derived from nut proteins or can be synthesized. For example, nut peptides are obtained by enzymatic hydrolysis, fermentation hydrolysis, or chemical hydrolysis of nut proteins or produced synthetically, for example, by solid-phase synthesis. 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.

[0011] Preferably, bioactive walnut 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 on the scalp, hair follicles, or both. Bioactive peptides include amino acids joined by covalent bonds, also called amide or peptide bonds, while proteins are polypeptides of higher molecular weight (MW), i.e., having more than 50 amino acid residues. Bioactive walnut peptides exhibit They typically have hormonal or drug-like activities and are classified according to their mode of action. Many bioactive peptides share certain structural characteristics, for example, an amino acid residue length of 2 to 20 amino acids.

[0012] Useful bioactive nut peptides according to this disclosure often include one or more amino acid residues selected from leucine, proline, or combinations thereof. In preferred embodiments, the one or more bioactive nut peptides include from 2 to about 20 amino acids, in which one or more of the amino acids are leucine, proline, or combinations thereof. Non-limiting examples of bioactive nut peptides comprising from 2 to about 20 amino acid residues, in which one or more amino acid residues are leucine, proline, or combinations thereof, include or consist of the peptides represented by SEQ ID NO: 2-5, 7-9, and 11-46.

[0013] One or more bioactive nut peptides may have at least two amino acid residues (dipeptides) up to approximately 20 amino acid residues. Non-limiting examples of dipeptides include those represented by SEQ ID NOS: 1-12. One or more bioactive nut peptides may have at least three amino acid residues up to approximately 20 amino acid residues. Non-limiting examples of bioactive nut peptides having three amino acid residues include those represented by SEQ ID NOS: 13-23. In addition, one or more bioactive nut peptides may have at least four to approximately 20 amino acid residues. Non-limiting examples of bioactive nut peptides having four amino acids include those represented by SEQ ID NOS: 24-32. In addition, the one or more bioactive nut peptides may include at least five amino acid residues up to about 20 amino acid residues.Non-limiting examples of bioactive nut peptides having five amino acid residues include the peptides represented by SEQ ID NOS: 33-39. In various embodiments, one or more bioactive nut peptides include at least about six amino acid residues. Non-limiting examples of bioactive nut peptides having six amino acid residues include the peptides represented by SEQ ID NOS: 10-44. In still other embodiments, one or more bioactive nut peptides include at least seven amino acid residues up to about 20 amino acid residues. Non-limiting examples of bioactive nut peptides having seven amino acid residues include the peptides represented by SEQ ID NOS: 45 and 46.

[0014] Particularly useful bioactive nut peptides include peptides comprising or consisting of the peptides represented by SEQ ID NOS: 45 and 46, referred to throughout this disclosure as peptide (I) or peptide nut (I) (SEQ ID NO: 45) and the peptide (II) or nut peptide (II) (SEQ ID NO: 46).

[0015] Peptide I Thr-Trp-Leu-Pro-Leu-Pro-Arg (TWLPLPR) (SEQ ID NO: 45), and

[0016] Peptide II Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys (YVLLPSPK) (SEQ ID NO: 46).

[0017] As already mentioned, bioactive walnut peptides are particularly useful for topical application to the scalp in processes for treating or preventing (or slowing down) hair loss, and for treating scalp conditions. Accordingly, this disclosure relates to processes for treating the scalp, hair, or both. For example, bioactive walnut peptides are useful in processes for preventing or slowing down hair loss. With regard to scalp conditions, bioactive walnut peptides are particularly useful in processes for preventing and treating dandruff, scalp psoriasis, acne, or combinations thereof. Typically, one or more walnut peptides are applied to the scalp in a pharmaceutical or cosmetic composition, which includes a physiologically acceptable carrier.Common and useful physically acceptable carriers include water, water-soluble solvents, and mixtures thereof.

[0018] It may be beneficial to use more than one (two or more) bioactive nut peptide in the processes and compositions described throughout this disclosure. Different bioactive nut peptides can act together to provide a variety of beneficial effects on the scalp and hair. For example, one or more bioactive nut peptides may treat or prevent hair loss by stimulating the secretion of growth factors and signaling molecules that interact with epithelial cells in the hair follicle, driving cell proliferation and differentiation. One or more other bioactive nut peptides may combat or prevent chemical changes or natural effects of aging that can shorten the hair growth phase (anagen phase), leading to thinner hair and ultimately hair loss.Due to their different mechanisms of action, two additional bioactive nut peptides can be used together to synergistically prevent hair loss and treat scalp disorders.

[0019] In various embodiments, the use of two or more bioactive nut peptides is preferred. The two or more bioactive nut peptides may have similar activities or may offer different activities that benefit the hair and scalp. In other embodiments, the use of three or more bioactive nut peptides is preferred. The use of multiple bioactive peptides allows for the modification of more than one physiological mechanism in the treatment and prevention of hair loss. Combinations of peptides of Bioactive nuts can interact synergistically and provide benefits that exceed the sum of the individual contributions of the peptides. For example, the synergistic activity of a combination can 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.

[0020] One or more bioactive nut peptides are often incorporated into a pharmaceutical or cosmetic composition for application to the scalp. Pharmaceutical and cosmetic compositions typically include one or more bioactive nut peptides 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, fatty compounds 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.

[0021] The pharmaceutical or cosmetic compositions include a sufficient quantity of one or more bioactive nut peptides to ensure that a therapeutically effective amount of the one or more nut peptides is delivered to the scalp during use. Throughout this disclosure, reference to application or delivery to the scalp includes delivery and application to the hair and hair follicles located within and extending from the scalp. The pharmaceutical and cosmetic compositions typically include one or more bioactive nut peptides and one or more physiologically acceptable carriers, such as water. 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, and the like.Pharmaceutical and cosmetic compositions include lotions, creams, serums, sprays, emulsions, gels, powders, dispersions, ointments, sticks, pastes and mousses.

[0022] It may be useful to combine one or more bioactive nut peptides with one or more additional active agents, for example, active agents to treat or prevent hair loss, and to treat scalp conditions. Accordingly, in various embodiments, one or more bioactive nut peptides are combined and / or administered in conjunction with one or more hair growth-promoting agents or one or more active agents to treat scalp conditions. Non-limiting examples of hair growth-promoting agents include androgen receptor inhibitors, androgen antagonists and antiandrogens. More specific but not limiting examples of hair growth-promoting agents include episteride, finasteride, cyproterone acetate, alpharadiol, minoxidil, bimatoprost, bicalcutamide, spironolactone, flutamide, lantanoprost, dutasteride, ketoconazole, tofacitinib, ruxolitinib, tacrolimus, bimatoprost, latanoprost, spironolactone, aldactone, kenalog-10, kenalog-40, triamcinolone, azofidine, sulfasalazine, and sulfazine.

[0023] Non-limiting examples of active agents for the treatment of scalp conditions include anti-dandruff agents, agents for the treatment of psoriasis and / or scalp inflammation, anti-acne agents, and combinations thereof. Non-limiting examples of anti-dandruff agents include zinc pyrithione, ketoxonazole, selenium sulfide, coal tar, and salicylic acid. Non-limiting examples of agents for the treatment of psoriasis and / or inflammation include tacrolimus, calcipotriene, corticosteroids, salicyclic acid, and coal tar. Non-limiting examples of anti-acne agents include salicylic acid, benzoyl peroxide, alpha-hydroxy acids (AHAs), tea tree oil, retinoids, and ceramides. Brief description of the drawings

[0024] An implementation of this technology is described, by way of example only, with reference to the accompanying figures, in which:

[0025] [Fig-1] [Fig.1] is a pie chart showing relative gene scores of three bioactive walnut peptides (walnut peptides (I), (II) and (III)) tested at two different concentrations (100 pM and 250 pM); and

[0026] [Fig.2] [Fig.2] is an illustration showing a hair follicle passing from the telogen phase to anagen phase highlighting where different bioactive nut peptides influence various physiological pathways of the anagen and telogen phases.

[0027] The various aspects of disclosure are not limited to the results, arrangements and representations shown in the drawings. DETAILED DESCRIPTION OF THE DISCLOSURE

[0028] This disclosure relates to bioactive walnut peptides and their topical use for treating and preventing hair loss, and for treating scalp disorders. The bioactive walnut peptides comprise 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 this disclosure is a compound that includes an unbroken sequence of at least two amino acids within its structure and has a maximum of about 50 amino acids. amino acids. The terms "di-peptide" or "dipeptide" as used here refer to a compound that includes an unbroken sequence of two amino acids within its structure. The terms "tri-peptide" or "tripeptide" as used here refer to a compound that includes an unbroken sequence of three amino acids within its structure. As used here, a "tetrapeptide" or "tetrapeptide" is a compound that includes an unbroken sequence of four amino acids within its structure. These amino acids are indicated here using a traditional one-letter convention from left (N-terminal end) to right (C-terminal end). In this nomenclature, G is glycine, H is histidine, K is lysine, E is glutamic acid, and so on, according to a well-known and accepted nomenclature in art.

[0029] A “bioactive” peptide, as defined in this disclosure, is a minimum of 2 or 3 amino acid residues up to approximately 20 amino acid residues long and has a measurable physiological effect on the scalp, hair follicles, and / or hair growth. Bioactive peptides include amino acids linked by covalent bonds, also known as amide or peptide bonds, whereas proteins are polypeptides with a higher molecular weight (MW) and generally more than 50 amino acid residues. Bioactive peptides typically exhibit hormonal or drug-like activities and are classified according to their mechanism of action. Many bioactive peptides share certain structural features, which include, for example, a peptide residue length of 2 to 20 amino acids.

[0030] The term "amino acid" as used herein includes and encompasses all natural 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-natural amino acids can be found in *The Peptides*, Vol. 5 (1983), Academy Press, Chapter VI, by D.C. Roberts and F. Vellaccio. The amino acids in the peptides of the present invention may be present in their natural L configuration, their non-natural D configuration, or as a racemic mixture.

[0031] As used herein, the term "peptide" shall also refer to the salts, deproteinized forms, acylated forms of the peptide, deacylated forms of the peptide, enantiomers, diastereomers, racemates, prodrugs, and hydrates of the peptide mentioned above, unless otherwise specified. 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 can be added to help prevent enzymatic degradation.

[0032] The term “prodrug” refers to any precursor compound that can generate or release the aforementioned 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 groups. Suitable protecting groups for the carboxylic acid group are esters such as benzyl esters or t-butyl esters.

[0033] Useful bioactive nut peptides according to this disclosure often include one or more amino acid residues selected from leucine, proline, or combinations thereof. In preferred embodiments, the one or more bioactive nut peptides include from 2 to about 20 amino acids, in which one or more of the amino acids are leucine, proline, or combinations thereof. Non-limiting examples of bioactive nut peptides having from 2 to about 20 amino acid residues, in which at least one amino acid residue is leucine or proline, include the peptides represented by SEQ ID NOS: 2-5, 7-9, and 11-46. The one or more bioactive nut peptides comprising at least one leucine or proline residue have from 2 to about 15 amino acids, preferably from 2 to about 12 amino acids, more preferably from 2 to about 12 amino acids, and even more preferably from 2 to about 10 amino acids.

[0034] Non-limiting examples of bioactive nut peptides having 2 amino acids (dipeptides) include the dipeptides represented by SEQ ID NOS: 1-12. Non-limiting examples of bioactive nut peptides having 2 amino acids (dipeptides) that have at least one leucine or proline residue include the dipeptides represented by SEQ ID NOS: 2-5 and 7-12. Accordingly, one or more nut peptides can be selected from peptides represented by SEQ ID NOS: 2-5 and 7-12. In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 2-5 and 7-12 and have from 3 to about 20 amino acids, from 3 to about 15 amino acids, from 3 to about 12 amino acids, from 3 to about 10 amino acids, from 3 to about 8 amino acids.In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 2-5 and 7-12 and have from 4 to about 20 amino acids, from 4 to about 15 amino acids, from 4 to about 12 amino acids, from 4 to about 10 amino acids, or from 4 to . about 8 amino acids, from 5 to about 20 amino acids, from 5 to about 18 amino acids, from 5 to about 15 amino acids, from 5 to about 12 amino acids, from 5 to about 10 amino acids, from 5 to about 8 amino acids, from 6 to about 20 amino acids, from 6 to about 18 amino acids, from 6 to about 15 amino acids, from 6 to about 12 amino acids, or from 6 to about 10 amino acids.

[0035] In various embodiments, one or more nut peptides are selected from peptides having an additional amino acid sequence represented by SEQ ID NOS: 1-12. In other embodiments, one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 1 to 12 and have from 3 to about 20 amino acids, from 3 to about 15 amino acids, from 3 to about 12 amino acids, from 3 to about 10 amino acids, from 3 to about 8 amino acids.In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 1 to 12 and have from 4 to about 20 amino acids, from 4 to about 15 amino acids, from 4 to about 12 amino acids, from 4 to about 10 amino acids, or from 4 to about 8 amino acids, from 5 to about 20 amino acids, from 5 to about 18 amino acids, from 5 to about 15 amino acids, from 5 to about 12 amino acids, from 5 to about 10 amino acids, from 5 to about 8 amino acids, from 6 to about 20 amino acids, from 6 to about 18 amino acids, from 6 to about 15 amino acids, from 6 to about 12 amino acids, or from 6 to about 10 amino acids.

[0036] The one or more bioactive nut peptides may include peptides having 3 amino acids. Non-limiting examples of bioactive nut peptides having 3 amino acids include the peptides represented by SEQ ID NOS: 13-23. Therefore, in various embodiments, the one or more nut peptides may be selected from the peptides represented by SEQ ID NOS: 13-23. In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 13 to 23 and have from 3 to about 20 amino acids, from 3 to about 15 amino acids, from 3 to about 12 amino acids, from 3 to about 10 amino acids, or from 3 to about 8 amino acids.In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 13-23 and have from 4 to about 20 amino acids, from 4 to about 15 amino acids, from 4 to about 12 amino acids, from 4 to about 10 amino acids, or from 4 to about 8 amino acids, from 5 to about 20 amino acids, from 5 to about 18 amino acids, from 5 to about 15 amino acids, from 5 to about 12 amino acids, from 5 to about 10 amino acids, from 5 to about 8 amino acids, from 6 to about 20 amino acids, from 6 to about 18 amino acids. amino acids, from 6 to about 15 amino acids, from 6 to about 12 amino acids, or from 6 to about 10 amino acids.

[0037] The one or more bioactive nut peptides may include peptides having 4 amino acids. Non-limiting examples of bioactive nut peptides having 4 amino acids include the peptides represented by SEQ ID NOS: 24-32. Therefore, in various embodiments, the one or more nut peptides are selected from the peptides represented by SEQ ID NOS: 24-32. In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 24 to 32 and having from 4 to about 20 amino acids, from 4 to about 15 amino acids, from 4 to about 12 amino acids, from 4 to about 10 amino acids, from 4 to about 8 amino acids.In other embodiments, the one or more bioactive nut peptides are selected from one or more amino acid sequences represented by SEQ ID NOS: 24 to 32 and have from 5 to about 20 amino acids, from 5 to about 18 amino acids, from 5 to about 15 amino acids, from 5 to about 12 amino acids, from 5 to about 10 amino acids, from 5 to about 8 amino acids, from 6 to about 20 amino acids, from 6 to about 18 amino acids, from 6 to about 15 amino acids, from 6 to about 12 amino acids, or from 6 to about 10 amino acids.

[0038] The one or more bioactive nut peptides may include peptides having 5 amino acids. Non-limiting examples of bioactive nut peptides having 5 amino acids include the peptides represented by SEQ ID NOS: 33-39. Therefore, in various embodiments, the one or more nut peptides are selected from the peptides represented by SEQ ID NOS: 33-39. In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 33 to 39 and have from 5 to about 20 amino acids, from 5 to about 15 amino acids, from 5 to about 12 amino acids, from 5 to about 10 amino acids, or from 5 to about 8 amino acids.In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 33 to 39 and have from 6 to about 20 amino acids, from 6 to about 18 amino acids, from 6 to about 15 amino acids, from 6 to about 12 amino acids, or from 6 to about 10 amino acids.

[0039] The one or more bioactive nut peptides may include peptides having 6 amino acids. Non-limiting examples of bioactive nut peptides having 6 amino acids include the peptides represented by SEQ ID NOS: 40-44. Therefore, in various embodiments, the one or more nut peptides are selected from the peptides represented by SEQ ID NOS: 40-44. In other embodiments For production, one or more bioactive nut peptides are chosen from peptides having one or more amino acid sequences represented by SEQ ID NOS: 40 to 44 and have from 6 to about 20 amino acids, from 6 to about 15 amino acids, from 6 to about 12 amino acids, from 6 to about 10 amino acids, from 6 to about 8 amino acids.

[0040] The one or more bioactive nut peptides may include peptides having 7 amino acids. A non-limiting example of a bioactive nut peptide having 7 amino acids is the peptide represented by SEQ ID NO: 45. Therefore, in various embodiments, the one or more nut peptides include the peptide represented by SEQ ID NO: 45. In other embodiments, the one or more bioactive nut peptides are selected from peptides having one or more amino acid sequences represented by SEQ ID NOS: 40 to 44 and have from 7 to about 20 amino acids, from 7 to about 15 amino acids, from 7 to about 12 amino acids, from 7 to about 10 amino acids, from 7 to about 8 amino acids.

[0041] The one or more bioactive nut peptides include peptides having 8 amino acids. A non-limiting example of a bioactive nut peptide having 8 amino acids is the peptide represented by SEQ ID NO: 46. Therefore, in various embodiments, the one or more nut peptides include the peptide represented by SEQ ID NO: 46. In other embodiments, the one or more bioactive nut peptides are selected from peptides having the amino acid sequences represented by SEQ ID NOS: 46 and have from 8 to about 20 amino acids, from 8 to about 15 amino acids, from 8 to about 12 amino acids, or from 8 to about 10 amino acids.

[0042] Particularly useful bioactive nut peptides include peptides having the amino acid sequences represented by SEQ ID NOS: 45 and 46, referred to throughout this disclosure as peptide (I) (or nut peptide (I)) (SEQ ID NO: 45) and peptide (II) (or nut peptide (II)) (SEQ ID NO: 46).

[0043] Peptide I Thr-Trp-Leu-Pro-Leu-Pro-Arg (TWLPLPR) (SEQ ID NO: 45), and

[0044] Peptide II Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys (YVLLPSPK) (SEQ ID NO: 46).

[0045] Accordingly, in a preferred embodiment, the one or more bioactive nut peptides represented by SEQ ID NOS: 45 and 46 (Peptide (I) and Peptide (II)). In other embodiments, the one or more bioactive nut peptides are selected from peptides having the amino acid sequences represented by SEQ ID NOS: 45 and 46 and have from 7 to about 20 amino acids, from 7 to about 15 amino acids, from 7 to about 12 amino acids, from 7 to about 10 amino acids, from 7 to about 8 amino acids, from 8 to about 20 amino acids, from 8 to about 15 amino acids, from 8 to about 12 amino acids, or from 8 to about 10 amino acids.

[0046] In a particularly preferred embodiment, the one or more bioactive nut peptides include both the peptide represented by SEQ ID NO: 45 and the peptide represented by SEQ ID NO: 46.

[0047] In other embodiments, one or more bioactive nut peptides may be selected from the peptides of formula (I) and formula (II), shown below.

[0048] X-Thr-Trp-Leu-Pro-Leu-Pro-Arg-Z (I)

[0049] X-Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys-Z (II),

[0050] in which,

[0051] - X represents the primary amine function of the N-terminal amino acid, free or substituted by a protecting group selected from an acetyl group, a benzoyl group, a tosyl group or a benzyloxycarbonyl group, and

[0052] - Z represents the hydroxyl group of the carboxyl function of the amino acid C- terminal, free or substituted by a protecting group chosen from a Ci-C20, NH2, NHY or NYY, in which Y represents a C1-C4 alkyl.

[0053] Bioactive nut peptides can be obtained by hydrolyzing nut proteins into small molecular peptides with molecular weights between the molecular weights of individual amino acids and the molecular weights of proteins. This can be achieved using biological or chemical processes. For example, bioactive peptides can be prepared by enzymatic procedures, fermentation, and chemical processes.

[0054] Bioactive nut 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 by condensation of fragments. In principle, the seemingly simple formation of a peptide bond can be accomplished using any of the procedures available in organic chemistry for the synthesis of carboxylic acid amides.

[0055] 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 the reactive side chains are protected by a temporary protecting group. Once the amino acid is fixed to the resin, the resin is filtered and washed to remove excess byproducts and reagents. Next, the N-alpha protecting group is removed in a deprotection process, and the resin is washed again to remove excess byproducts and reagents. Then, the next amino acid is coupled to the fixed 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 complete. Then, typically, all protecting groups are removed, the peptide resin is washed, and the peptide is cleaved from the resin.

[0056] Enzymatic hydrolysis involves the use of commercial enzymes to obtain bioactive peptides. The enzymes are responsible for cleaving the peptide bonds established in the protein, thereby releasing the encrypted peptide. For the enzyme to exert its activity, it is important that the enzyme binds to the substrate and proceeds with enzymatic catalysis. The enzyme possesses 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 interactions, or Van der Waals interactions.Enzymatic hydrolysis can generally be carried out 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).

[0057] Microbial fermentation is a biotechnological process used to obtain bioactive peptides. This process involves the use of microorganisms capable of producing proteolytic enzymes, with the aim of having these enzymes hydrolyze proteins into shorter peptides. The microorganisms generally used are bacteria, fungi, or yeasts, which may be present in the substrate natively 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.

[0058] Immersion fermentation uses a culture of microorganisms in a liquid medium containing nutrients. This system is suitable for microorganisms with high water-level activity, 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 features the advantage of releasing nutrients in a controlled manner and is suitable for fungi and microorganisms with lower moisture requirements.

[0059] Bioactive nut peptides can be produced using any process known to those skilled in the art, such as those disclosed in Merrifield, RB, Solid Phase Peptide Synthesis I., J. AM. CHEM. SOC. 85:2149-2154 (1963); Carpino, LA 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, RB 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 Synthesizer OfNovel Design, IN: PEPTIDES 1984 (Ragnarsson U., ed.) Almqvist and Wiksell Int., Stockholm (Sweden), p. 185-188. Pharmaceutical or cosmetic compositions

[0060] One or more bioactive nut peptides are typically incorporated into a pharmaceutical or cosmetic composition for application to the scalp. Pharmaceutical and cosmetic compositions include one or more bioactive nut peptides and one or more physiologically acceptable carriers. 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, and the like.

[0061] The pharmaceutical or cosmetic composition includes an amount of one or more bioactive nut peptides sufficient to ensure that a therapeutically effective amount of one or more nut peptides is delivered to the scalp during use. The term "therapeutically effective amount" as used herein refers to an amount sufficient to treat or prevent hair loss, or to treat a scalp condition. Throughout the disclosure, reference to application or delivery to the scalp includes both topical administration and application to hair and hair follicles located within and extending from the scalp. In various embodiments, the pharmaceutical or cosmetic composition includes from approximately 0.001 to approximately 10% by weight of one or more bioactive nut peptides.In additional embodiments, the pharmaceutical or cosmetic compositions include from about 0.001 to about 8% by weight, from about 0.001 to about 5% by weight, from about 0.001 to about 3% by weight, from about 0.001 to about 2% by weight, from about 0.001 to about 1% by weight, from about 0.001 to about 0.5% by weight, 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 2% by weight, from about 0.01 to about 1% by weight, from about 0.01 to about 0.5. % by weight, from approximately 0.1 to approximately 8% by weight, from approximately 0.1 to approximately 5% by weight, from approximately 0.1 to approximately 3% by weight, from approximately 0.1 to approximately 2% by weight, from approximately 0.1 to approximately 1% by weight, from approximately 0.1 to approximately 0.5% by weight, of one or more bioactive nut peptides. Preferably, the pharmaceutical or cosmetic composition includes approximately 0.05 to approximately 8% by weight, more preferably approximately 0.1 to approximately 5% by weight, and even more preferably approximately 0.1 to approximately 3% by weight of one or more bioactive nut peptides.

[0062] The therapeutically effective amount of one or more bioactive nut peptides will vary depending on the specific bioactive nut peptide and the combination of bioactive nut peptides. In various embodiments, the therapeutically effective amount can range from approximately 1 pg to approximately 50 mg (50,000 pg) per cm² of scalp skin. In other embodiments, the therapeutically effective amount of one or more walnut peptides is approximately 1 pg to approximately 40 mg, approximately 1 pg to approximately 30 mg, approximately 1 to approximately 20 mg, approximately 1 pg to approximately 10 mg, approximately 1 pg to approximately 8,000 pg, approximately 1 pg to approximately 5,000 pg, approximately 1 pg to approximately 2,000 pg, approximately 1 pg to approximately 1,000 pg, approximately 10 pg to approximately 40 mg, approximately 10 pg to approximately 30 mg, approximately 10 to approximately 20 mg, approximately 10 pg to approximately 10 mg, approximately 10 pg to approximately 8,000 pg, approximately 10 pg to approximately 5,000 pg, from approximately 10 pg to approximately 2000 pg,from approximately 10 pg to approximately 1000 pg, from approximately 100 pg to approximately 50 mg, from approximately 100 pg to approximately 40 mg, from approximately 100 pg to approximately 30 mg, from approximately 100 pg to approximately 20 mg, from approximately 100 pg to approximately 10 mg, from approximately 100 pg to approximately 8000 pg, from approximately 100 pg to approximately 5000 pg, from approximately 100 pg to approximately 2000 pg, from approximately 100 pg to approximately 1000 pg, from approximately 500 pg to approximately 50 mg, from approximately 500 pg to approximately 40 mg, from approximately 500 pg to approximately 30 mg, from approximately 500 pg to approximately 30 mg, from approximately 500 pg to approximately 20 mg, from approximately 500 pg to approximately 10 mg, from approximately 500 to approximately 8,000 pg, from approximately 500 pg to approximately 5,000 pg, from approximately 500 pg to approximately 2,000 pg, or from approximately 500 pg to approximately 1,000 pg per cm² of scalp skin.

[0063] In a preferred embodiment, the pharmaceutical or cosmetic composition includes two or more bioactive nut peptides. In other embodiments, the pharmaceutical or cosmetic composition includes two or more bioactive nut peptides provided that at least one of the bioactive nut peptides comprises or consists of the amino acid sequence represented by SEQ ID NO: 45. In another embodiment, the pharmaceutical or cosmetic composition includes two or more bioactive nut peptides provided that at least one of the bioactive nut peptides comprises or consists of the amino acid sequence represented by SEQ ID NO: 46. In a preferred embodiment, the pharmaceutical composition or cosmetic includes both a peptide comprising or consisting of the amino acid sequence represented by SEQ ID NO: 45 and a peptide comprising or consisting of the amino acid sequence represented by SEQ ID NO: 46, preferably the peptide represented by SEQ ID NO: 45 and the peptide represented by SEQ ID NO: 46 (i.e. peptides (I) and (II)).

[0064] For pharmaceutical or cosmetic compositions which include peptide (I), the amount of peptide (I) in the compositions will vary but is typically in an amount of about 0.01 to about 8% by weight, relative to the total weight of the pharmaceutical or cosmetic composition. Preferably, the pharmaceutical or cosmetic composition comprises about 0.01 to about 5% by weight, about 0.01 to about 3% by weight, about 0.01 to about 2% by weight, about 0.01 to about 1% by weight, about 0.01 to about 0.5% by weight, about 0.1 to about 8% by weight, about 0.1 to about 5% by weight, about 0.1 to about 3% by weight, about 0.1 to about 2% by weight, about 0.1 to about 1% by weight or about 0.1 to about 0.5% by weight of peptide (I), relative to the total weight of the pharmaceutical or cosmetic composition.

[0065] For pharmaceutical or cosmetic compositions that include peptide (II), the amount of peptide (II) in the compositions will vary but is typically in an amount of about 0.01 to about 8% by weight, relative to the total weight of the pharmaceutical or cosmetic composition. Preferably, the pharmaceutical or cosmetic composition comprises about 0.01 to about 5% by weight, about 0.01 to about 3% by weight, about 0.01 to about 2% by weight, about 0.01 to about 1% by weight, about 0.01 to about 0.5% by weight, about 0.1 to about 8% by weight, about 0.1 to about 5% by weight, about 0.1 to about 3% by weight, about 0.1 to about 2% by weight, about 0.1 to about 1% by weight or about 0.1 to about 0.5% by weight of peptide (II), relative to the total weight of the pharmaceutical or cosmetic composition.

[0066] The peptide (I) and the peptide (II) can be in a weight ratio of about 1:10 to about 10:1. Preferably, the peptide (I) and the peptide (II) are in a weight ratio of about 1:5 to about 5:1, more preferably about 1:2 to about 2:1, and even more preferably about 1:1.

[0067] Two or more bioactive nut peptides will often work synergistically to treat or prevent hair loss, or to treat scalp conditions. For example, two or more bioactive nut peptides may synergistically prevent (or slow) hair loss and treat scalp conditions. For example, two or more bioactive nut peptides may synergistically reduce the occurrence and / or severity of dandruff, scalp psoriasis, and acne. "Synergism," as used here, occurs when two substances or factors, or more of them, work together in such a way that their combined effect is greater than the sum of their individual effects. In other words, their interaction produces a more effective or beneficial result than if they worked alone. For example, the synergistic activity or result based on a combination of two or more bioactive nut peptides can be at least 5%, at least 10%, or at least 25% greater than the sum of the individual contributions of the peptides.

[0068] Pharmaceutical and cosmetic compositions include one or more bioactive nut peptides and a physiologically acceptable carrier. A "physiological carrier" as used here is a carrier that is suitable and safe for application to the human scalp. A particularly common physiologically acceptable carrier is water. However, physiologically acceptable carriers may be oils, fats, organic solvents, and the like, provided they are suitable and safe for application to the scalp. 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 in this disclosure may be lotions, creams, serums, sprays, emulsions, gels, powders, dispersions, ointments, sticks, pastes or foams.

[0069] Pharmaceutical and cosmetic compositions including 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 foams. In addition to a physiologically acceptable carrier, the pharmaceutical and cosmetic composition may optionally include one or more of the following:

[0070] Emollients

[0071] Suspending agents,

[0072] Emulsifying agents and

[0073] Thickeners. Emollients

[0074] 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 usually remain on the skin's surface or in the stratum corneum and act as a moisturizer or lubricant and reduce destratification. Non-limiting examples of emollients include acetylarginine, acetylated lanolin, seaweed extract, and polyethylene glycol-6 esters derived from oil. apricot kernel, 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, dimyristyle tartrate, 5-lauroyl glutamate distearate, etilavokadate, ethylhexyl myristate, glyceryl isostearate, glyceryl oleate, geksildetsilstearate, geksilizostearate, hydrogenated palm glycerides, hydrogenated soybean glycerides, hydrogenated fat glycerides isostearylneopentanoate, 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, commercial PAL 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. Suspenders

[0075] The pharmaceutical or cosmetic composition of the present invention may optionally include one or more suspending agents, preferably in a concentration effective for suspending the water-insoluble material in a dispersed form within the compositions or for modifying the viscosity of the composition. These concentrations will vary. However, in some embodiments, the pharmaceutical and cosmetic composition includes from approximately 0.1% to approximately 10%, more preferably from approximately 0.25% to approximately 5.0%, or even more preferably from approximately 0.5% to approximately 3% by weight, of one or more suspending agents, relative to the total weight of the composition.Non-limiting examples include vinyl polymers, such as crosslinked acrylic acid polymers known as carbomers; 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), algal colloids (algal 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, and acrylate polymers such as polyacrylate. sodium, polyacrylate, polyacrylamide, polyethyleneimine and water-soluble inorganic materials such as bentonite, aluminum magnesium silicate, laponite, hectorite and anhydrous silicic acid.

[0076] Other optional suspending agents include crystalline suspending agents that can be resolved into acyl derivatives, long-chain amine oxides, long-chain acyl derivatives, 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 monoethanolamide); and glyceryl esters (e.g., glyceryl distearate, trihydroxystearine, tribhengen).Other suitable suspending agents include primary amines containing a fatty alkyl fragment with at least about 16 carbon atoms, examples of which include palmitamine or stearamine, and secondary amines containing two fatty alkyl fragments, each with 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 maleic anhydride / methyl vinyl ether copolymer. Emulsifying agents

[0077] 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 alkylene polyoxide is esterified at one end with a fatty acid and esterified (i.e., via an ether bond) at the other end with a fatty alcohol].Non-limiting examples of nonionic surfactants derived from said alkylene oxide include ceteth-6, ceteth-10, ceteth-12, cetetareth-6, cetetareth-10, cetetareth-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 sulfate, PEG-10 glyceryl stearate, PEG-30 glyceryl cocoate, PEG-80 glyceryl cocoate, and PEG-30 glyceryl cocoate. PEG-200 glyceryl, PEG-8 dilaurate, PEG-10 distearate, and mixtures thereof. Other applicable nonionic surfactants include polyhydroxyamide fatty acid surfactants. One surfactant in particular... The preferred surfactant corresponding to the above structure is coco alkylamide N-methylglucoside. The preferred surfactants among nonionic surfactants are those 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 sugar esters and 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 these 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 useful nonionic surfactants here is a mixture of fatty acid esters based on a mixture of sorbitan or a fatty acid ester of sorbitol and a fatty acid ester of sucrose, where the fatty acid in each example is preferably C8-C24, more preferably C10-C20.

[0078] The one or more emulsifying agents are present in a sufficient quantity to emulsify the pharmaceutical or cosmetic compositions. For example, the one or more emulsifying agents may be present in an amount of approximately 0.1 to approximately 10% by weight, relative to the total weight of the pharmaceutical or cosmetic composition. In other embodiments, the one or more emulsifying agent(s) may be in an amount of about 0.1 to about 8% by weight, about 0.1 to about 5% by weight, about 0.1 to about 3% by weight, about 0.1 to about 2% by weight, about 0.5 to about 10% by weight, about 0.5 to about 8% by weight, about 0.5 to about 5% by weight, about 0.5 to about 2% by weight or about 0.5 to about 1% by weight, relative to a total weight of the cosmetic composition. Thickeners

[0079] Thickeners suitable for inclusion in pharmaceutical or cosmetic compositions are described herein. Non-limiting examples include acrylamide copolymer, agarose, amylopectin, bentonite, calcium alginate, the calcium carboxymethylcellulose, carbomer, carboxymethylchitin, cellulose gum, dextrin, gelatin, hydroxymethyl hydroxycellulose hydroxypropyl, hydroxyethyl 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.

[0080] 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 commercially available carboxylic acid polymers useful here include carbomers, which are homopolymers of acrylic acid crosslinked with allyl ethers of sucrose or pentaerythritol. Carbomers are available in BF Goodrich's Carbopol® 900 range (e.g., Carbopol® 954).In addition, other suitable carboxylic acid-based polymeric agents include C10-30 alkyl acrylate copolymers with one or more monomers of acrylic acid, methacrylic acid, or esters of one of its short chains (i.e., a Cl-4 alcohol), the crosslinking agent being an allyl ether of sucrose or pentaerythritol. These copolymers are known as C10-C30 alkyl acrylate / acrylate crosslinked polymers and are commercially available under the names 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, C10-30 acrylate / alkyl acrylate crosslinked polymers, or mixtures thereof.

[0081] Furthermore, according to certain 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 cellulose. Alkyl-substituted celluloses are also useful. In these polymers, the hydroxyl groups of the cellulosic 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 bond. Typically, these polymers are straight-chain or branched-chain esters of C10-30 alcohols and hydroxyalkyl celluloses. Examples of alkyl groups applicable in this application include groups selected from stearyl, isostearyl, lauryl, myristyle, cetyl, isocetyl, cocoyl (e.g., an alkyl group derived from coconut oil alcohols), palmityl, oleyl, 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 stated material 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 known cosmetic and pharmaceutical ingredients in the art. Other useful polysaccharides include scleroglucans, which are straight-chain (1-3) glucose units linked (1-6), where one in three glucose units is linked; a commercially available example is Clearogel™ CS 11 by Michel Mercier Products Inc. (Mountainside, New Jersey). .

[0082] Other useful thickeners include those derived from natural sources. Non-limiting examples include acacia gum, agar-agar, algin, alginic acid, ammonium alginate, amylopectin, calcium alginate, calcium carrageenan, carnitine, carrageenan, dextrin, gelatin, gellan gum, guar gum hydrochloride, gydrohydrochloride, 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, sodium carboxymethyl dextran, dextran, sodium carrageenan, tragacanth gum, xanthan gum, or mixtures thereof.

[0083] Furthermore, the compositions may optionally contain polyacrylamide polymers, in particular nonionic polyacrylamide polymers, including substituted branched or unbranched polymers. Other polyacrylamide polymers useful here include multiblock copolymers of acrylamides and acrylic acid-substituted acrylamides and substituted acrylic acids.

[0084] One or more thickeners are present in sufficient quantity to thicken the pharmaceutical or cosmetic compositions. For example, one or more thickeners may be present in an amount of approximately 0.05 to approximately 5% by weight, relative to the total weight of the pharmaceutical or cosmetic composition. In other embodiments, one or more thickeners may be in an amount of about 0.05 to about 3% by weight, about 0.05 to about 2% by weight, about 0.05 to about 1% by weight, about 0.1 to about 5% by weight, about 0.1 to about 3% by weight, about 0.1 to about 2% by weight, about 0.1 to about 1% by weight, about 0.5 to about 5% by weight, about 0.5 to about 3% by weight or about 0.5 to about 2% by weight, relative to the total weight of the pharmaceutical or cosmetic composition. Additional active agents

[0085] It may be useful to combine one or more bioactive nut peptides with one or more additional active agents to treat or prevent hair loss and to treat scalp conditions. For example, additional active agents include hair growth promoters and agents for treating scalp conditions. Non-limiting examples of hair growth promoters include androgen receptor inhibitors, androgen antagonists, and antiandrogens.More specific but not limiting examples of hair growth-promoting agents include episteride, finasteride, cyproterone acetate, alpharadiol, minoxidil, bimatoprost, bicalcutamide, spironolactone, flutamide, lantanoprost, dutasteride, ketoconazole, tofacitinib, ruxolitinib, tacrolimus, bimatoprost, latanoprost, spironolactone, aldactone, kenalog-10, kenalog-40, triamcinolone, azofidine, sulfasalazine and sulfazine.

[0086] Non-limiting examples of active agents for the treatment of scalp conditions include anti-dandruff agents, agents for the treatment of psoriasis and / or scalp inflammation, anti-acne agents, and combinations thereof. Non-limiting examples of anti-dandruff agents include zinc pyrithione, ketoxonazole, selenium sulfide, coal tar, and salicylic acid. Non-limiting examples of agents for the treatment of psoriasis and / or inflammation include tacrolimus, calcipotriene, corticosteroids, salicyclic acid, and coal tar. Non-limiting examples of anti-acne agents include salicylic acid, benzoyl peroxide, alpha-hydroxy acids (AHAs), tea tree oil, retinoids, and ceramides.

[0087] In a preferred embodiment, one or more of the additional active agents is biotin.

[0088] In a preferred embodiment, one or more of the additional active agents is baicalin or Scutellaria baicalensis extract.

[0089] In a preferred embodiment, one or more additional active agents are botanical extracts. Non-limiting examples of botanical extracts that are useful for treating or preventing hair loss include rosemary extract, peppermint extract, saw palmetto extract, ginseng extract, nettle extract, aloe vera extract, green tea extract, bamboo extract, horsetail extract, fenugreek extract, grape seed extracts (and / or proanthoxyanidins), skullcap, and eclipta.

[0090] In a preferred embodiment, the pharmaceutical or cosmetic composition includes one or more additional active ingredients selected from the plants and botanical extracts of Eclipta alba, Polygonum multiflorum, Pisum sativum, Momordica charanita, Astragalus membranaceus, Sophora flavescens, Angelica sinensis, caffeine, Malus domestica, Vitis viniferis, Rosmarinus officinalis, Polygonum multiflorum, Eclipta prostratum, Punica granatum, Scutellaria baicalensis, Ganoderma lucidum, Ocimum basilicum, Moringa olefiera, Chenopodium quinoa, and lupine protein. The one or more additional active ingredients of this disclosure preferably include those presented in US20240075089.

[0091] A list of preferred plant extracts that are useful as additional active agents is provided below.

[0092] [Tables 1] Botanical extract Approximate extract ratio Preferred standardization Preferred constituents Chenopodium quinoa Eclipta prostrata 10 5% lactones as wedelol actone Tannins, saponins, niotin, ecliptin Lupinus malus domesticus 30 50% polyphenols by UV Polyphenols, pectin, potassium Momordica charantia 10 5% active principles by UV Charantin, momordicosides, momordicin Moringa olifera Ocimum basilicum Pisum sativum Rosmarinus offic inalis Scutellaria baica lensis 12 30% of flavones in the form of baicalin e by UV Flavones (baicalin, baicalein, wogonin) terols Vitis viniferis 10-15 >=95% Proanthocyanadins in the form of catechins by HPLC 5-15% of catechin Monomers, >=80% of procyan idol oligomers A list of additional preferred active agents is provided below with

[0093]

[0094]

[0095] Preferred minimum and maximum concentrations (percentage by weight) for use in pharmaceutical or cosmetic compositions of this case. [Tables 2] Ingredients Min Max Biotin 0.005% 1.00% Caffeine 0.0001% 1.00% Chenopodium quinoa 0.1% 2.00% Eclipta prostrata 0.01% 4.00% Lupinus 0.01% 6.00% Matus domestica 0.01% 4.00% Methylsulfonylmethane 0.005% 2.00% Momordica charantia 0.01% 4.00% Moringa olifiera 0.01% 6.00% Ocimum basilicum 0.01% 6.00% Pisum sativum 0.01% 6.00% Rosmarinus officinalis 0.01% 4.00% Scutellaria baicalensis 0.01% 4.00% Vitamin E (alpha-tocopherol) 0.005 % 2.00% Vitis viniferis 0.01% 4.00% In a preferred embodiment, the pharmaceutical or cosmetic composition includes one or more additional active agents selected from Eclipta prostrata, Scutellaria baicalensis and proanthocyanidins, and optionally any of or a combination of Malus domestica, Rosmarinus officinalis, Moringa olifera and biotin.

[0096] In a preferred embodiment, one or more additional active agents are selected from among the proanthocyanidins, which include procyanidin, procyanadin, anthocyanidin, anthocyanadin, anthocyanin, celphinidin, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, and petunidin. Proanthocyanidins are part of a group of compounds called polyphenols and belong to a subclass of compounds called flavonoids, which are found in many plants, including: apples, pine bark, cinnamon, aronia, cocoa beans, grape seeds, grape skin, blueberry, cranberry, blackcurrant, green tea, black tea, cocoa beans, Quercus petraea, and Q. heartwood.Robur, agaf palm, green beans (Vicia faba), gallipoli rose, bilberry, cranberry, elderberry, chokecherry, blackcurrant, blueberry, strawberry, persimmon, banana, carob, Chinese quince, chokecherries, rose stems, medlar, blackberry, plum, apricot, walnut, silver oleaster, pomegranate, triticale, sorghum, red cabbage, birch and ginkgo biloba.

[0097] In various embodiments, the total amount of additional active agents in the pharmaceutical or cosmetic composition, other than one or more bioactive nut peptides, is from more than zero to about 9% by weight, from more than zero to about 8% by weight, from more than zero to about 7% by weight, from more than zero to about 6% by weight, from more than zero to about 5% by weight, from more than zero to about 4% by weight, from more than zero to about 3% by weight, from more than zero to about 2% by weight; from about 10 ppm to about 10% by weight (100,000 ppm), from about 10 ppm to about 5% by weight (50,000 ppm), from about 10 ppm to about 2.5% by weight (25,000 ppm), from about 10 ppm to about 1% by weight (10,000 ppm), from about 10 ppm to about 0.5% by weight (5,000 ppm), from about 10 ppm to about 0.3% by weight (3,000 ppm), from about 10 ppm to about 0.2% by weight (2,000 ppm), from about 10 ppm to about 0.1% by weight (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, relative to the total weight of the pharmaceutical or cosmetic composition. Optional exclusions

[0098] Bioactive nut peptides may be derived from nut proteins or may be synthesized. For example, nut peptides are obtained by enzymatic hydrolysis, fermentation hydrolysis, or chemical hydrolysis of nut proteins or produced synthetically, for example, by solid-phase synthesis. In various embodiments, one or more bioactive nut peptides are isolated from other constituents of the nut and walnut tree from which the bioactive nut peptide is derived. Nuts and walnut trees include constituents such as fats, proteins, fibers, vitamins, minerals, phytosterols, etc. The term "walnut tree" in the context of this disclosure includes all parts of the walnut tree, such as the roots, trunk, bark, branches, leaves, hulls, and shells.Consequently, the one or more nut peptides useful in the processes and compositions disclosed herein may be isolated from other constituents of the walnut or walnut tree. For example, the one or more nut peptides may be free or substantially free of nut fats, nut peptides and proteins (other than the one or more specified bioactive nut peptides), nut fibers, and the like. Another pharmaceutical or cosmetic composition comprising one or more nut peptides may be free or substantially free of nut fats, nut peptides and proteins (other than the one or more specified bioactive nut peptides), nut fibers, and the like.

[0099] Nuts, particularly their shells, are useful for producing a variety of dye compounds. The principal dye compounds of walnuts and walnut trees are juglone and tannins. Therefore, one or more bioactive nut peptides can be isolated from the dye compounds of walnuts and walnut trees, including juglone and tannins. Similarly, pharmaceutical or cosmetic compositions comprising one or more bioactive nut peptides are preferably free or substantially free of dye compounds from walnuts and walnut trees.For example, in various embodiments, the pharmaceutical or cosmetic composition does not comprise any tincture compound derived from walnuts and nuts or comprises less than 1% by weight, preferably less than 0.1% by weight, more preferably less than 0.01% by weight, more preferably less than 0.001% by weight, even more preferably less than 0.0001% by weight, and even more preferably less than 0.00001% by weight, relative to the total weight of the pharmaceutical or cosmetic composition.

[0100] In a preferred embodiment, bioactive walnut peptides are isolated from juglone, a compound naturally occurring mainly in the shells and leaves of walnut trees, particularly black walnut (Juglans nigra). Juglone is a type of naphthoquinone. The bioactive walnut peptides are preferably isolated Nut naphthoquinones, including juglone. Pharmaceutical or cosmetic compositions comprising one or more bioactive nut peptides are preferably free or substantially free of juglone. For example, in various embodiments, the pharmaceutical or cosmetic composition does not include juglone or includes less than 1% by weight, preferably less than 0.1% by weight, more preferably less than 0.01% by weight, more preferably less than 0.001% by weight, even more preferably less than 0.0001% by weight, and even more preferably less than 0.00001% by weight, relative to the total weight of the pharmaceutical or cosmetic composition.

[0101] Furthermore, pharmaceutical or cosmetic compositions comprising one or more bioactive nut peptides may be free or substantially free of nut naphthoquinones, including juglone. For example, in various embodiments, the pharmaceutical or cosmetic composition does not comprise nut naphthoquinones or comprises less than 1% by weight, preferably less than 0.1% by weight, more preferably less than 0.01% by weight, more preferably less than 0.001% by weight, even more preferably less than 0.0001% by weight, and even more preferably less than 0.00001% by weight, relative to the total weight of the pharmaceutical or cosmetic composition.

[0102] In a preferred embodiment, the bioactive walnut peptides are isolated from tannins. Tannins are polyphenolic compounds found in many plants and are known for their astringent properties. In walnuts, tannins are mainly present in the skin or shell of the fruit. These compounds contribute to the bitter taste of walnut skins and have antioxidant and antimicrobial properties. Although they are more concentrated in the shell and leaf of the walnut, some tannins are also present in the edible part of the walnut, albeit in smaller quantities. In a preferred embodiment, the pharmaceutical or cosmetic composition comprising one or more bioactive walnut peptides is free or substantially free of tannins.For example, in various embodiments, the pharmaceutical or cosmetic composition does not include nut tannins or includes less than 1% by weight, preferably less than 0.1% by weight, more preferably less than 0.01% by weight, more preferably less than 0.001% by weight, even more preferably less than 0.0001% by weight, and even more preferably less than 0.00001% by weight, relative to the total weight of the pharmaceutical or cosmetic composition.

[0103] In various embodiments, the one or more bioactive nut peptides do not include the peptide represented by SEQ ID NO: 47 (also referred to as peptide (III) or nut peptide (III) throughout the disclosure).

[0104] Peptide (III) KVPPLLY (Lys-Val-Pro-Pro-Leu-Leu-Tyr) (SEQ ID NO: 47)

[0105] For example, the one or more bioactive nut peptides may comprise less than 1% by weight of peptide (III) relative to the total weight of the one or more bioactive nut peptides. More preferably, the one or more bioactive nut peptides may include less than 0.1% by weight, more preferably less than 0.01% by weight, more preferably less than 0.001% by weight, even more preferably less than 0.0001% by weight, and even more preferably less than 0.00001% by weight of peptide (III), relative to the total weight of the one or more bioactive nut peptides. Similarly, pharmaceutical or cosmetic compositions comprising the one or more bioactive nut peptides may be free of or essentially free of peptide (III).For example, pharmaceutical or cosmetic compositions comprising one or more bioactive nut peptides may include less than 1% by weight of peptide(III), relative to the total weight of the pharmaceutical or cosmetic composition. Preferably, the pharmaceutical or cosmetic composition includes less than 0.01% by weight, more preferably less than 0.001% by weight, even more preferably less than 0.0001% by weight, and even more preferably less than 0.00001% by weight of peptide(III), relative to the total weight of the pharmaceutical or cosmetic composition. Processing methods

[0106] As already mentioned, bioactive nut peptides are particularly useful for topical application to the scalp in processes for treating or preventing hair loss, and for treating scalp conditions. More specifically, bioactive nut peptides are useful in processes for increasing hair density, increasing follicular density, increasing hair shaft thickness, increasing hair length, preventing hair loss, reducing hair loss, or any combination thereof, or, for example, in an individual who needs it. The processes include applying an effective amount of one or more bioactive nut peptides to the scalp of an individual, preferably an individual who needs it.In another embodiment, the individual in need is an individual with one or more disorders chosen from among alopecia grand, androgenic alopecia, alopecia areata, alopecia universalis, involutional alopecia, trichotillomania, telogen effluvium, anagen effluvium, cicatricial alopecia, alopecia with scarring, scalp thinning, hair shaft abnormalities, infectious hair disorders, genetic disorders, and hair loss due to chemotherapy, hormonal imbalance, fungal infection, medication use, chemical hair treatment, or aging.

[0107] In a preferred embodiment, bioactive nut peptides are useful in processes for treating alopecia in an individual who needs them, wherein the individual who needs them is an individual suffering from adrenergic alopecia, telogen alopecia, patchy alopecia, traumatic alopecia, telogen effluvium, nutritional deficiencies, metabolic defects, marked weight loss, diabetes, hypervitaminosis, hypovitaminosis, zinc deficiency, alopecia vulgaris, pustular alopecia, erythrodermic alopecia, arthropathic alopecia, para-alopecia, palmoplantar pustulosis, ichthyoses, keratodermas and genodermatoses with pathological co-occurrence disorders. The processes include applying an effective amount of one or more bioactive nut peptides to the scalp of an individual who needs it.

[0108] In other embodiments, one or more bioactive nut peptides are useful in processes for stimulating or activating dermal papillary cells of the scalp and / or hair follicles, for potentiating fibroblast growth factor (FGF) and / or vascular endothelial growth factor (VEGF) in the scalp and / or hair follicles, for protecting or preserving hair follicles by regulating or managing oxidative stress (management of reactive oxygen species (ROS)) in the scalp and / or hair follicles, for downregulating cell cycle arrest and / or IL-1 (interleukin-1) signaling in hair follicles and / or scalp, for treating inflammation of hair follicles and / or scalp, or combinations thereof.

[0109] In a preferred embodiment, one or more bioactive nut peptides are useful in processes for treating scalp conditions. Non-limiting examples of scalp conditions include dandruff, scalp psoriasis, and acne.

[0110] For example, bioactive nut peptides are useful in processes for treating, preventing, or slowing hair loss. With regard to scalp conditions, bioactive nut peptides are particularly useful in processes for preventing and treating dandruff, scalp psoriasis, acne, or combinations thereof. Typically, one or more nut peptides are applied to the scalp in a pharmaceutical or cosmetic composition, which includes a physiologically acceptable carrier. Common and useful physiologically acceptable carriers include water, water-soluble solvents, and mixtures thereof.

[0111] It may be beneficial to use more than one (two or more) bioactive nut peptide in the processes and compositions described throughout the disclosure. Different bioactive nut peptides can act together to provide a variety of influences. beneficial to the scalp and hair. For example, one or more bioactive walnut peptides can treat hair loss by potentiating the secretion of growth factors and signaling molecules that interact with epithelial cells in the hair follicle, leading to cell proliferation and differentiation. One or more other bioactive walnut peptides can combat or prevent chemical changes or natural effects of aging that can shorten the hair growth phase (anagen phase), leading to thinner hair and ultimately hair loss. Due to their different mechanisms of action, two additional bioactive walnut peptides can be used together to synergistically treat, prevent (or slow) hair loss, and address scalp disorders.

[0112] In various embodiments, the use of two or more bioactive nut peptides is preferred. These two or more bioactive nut peptides may have similar activities or may offer different activities that treat or prevent hair loss and treat scalp conditions. In other embodiments, the use of three or more bioactive nut peptides is preferred. The use of multiple bioactive peptides allows for the modification of more than one physiological mechanism in skin treatment. Combinations of bioactive nut peptides can interact synergistically and provide benefits that exceed 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.

[0113] Various changes can be made to the compositions and processes described above without departing from the scope of the invention. Therefore, it is intended that any disclosure contained in the above description and in the examples given below shall be interpreted in an illustrative and not limiting manner. EXAMPLES Example 1

[0114] Inductivity of dermal papilla cells (DPCs)

[0115] Dermal papilla cell (DPC) inductivity refers to a state in which DPCs retain the ability to be stimulated or activated to promote hair follicle development. Growth factors, such as fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF), have been shown to stimulate DPCs and enhance their inductive properties, promoting hair growth (Ito et al., Decapeptide with fibroblast growth factor (FGF)-5 partial sequence inhibits hair growth suppressing activity ofFGF-5. J Cell Physio., 2003 Nov; 197(2):272-83).

[0116] Dermal papilla cells (DPCs) were cultured in a basal follicular dermal papilla cell medium (Promocell product number C-26501) supplemented with 4% fetal calf serum (FCS), 0.004 mL / mL bovine pituitary extract, 5 pg / mL insulin, and 1 nf / mL fibroblast growth factor (FGF). The dermal papilla cells were treated with bioactive walnut peptides for 24 hours using concentrations of 100 pM to 250 pM of each peptide. At the end of the culture period, RNA extraction and bulk RNA sequencing were performed. RNA was extracted from each sample according to standard protocols, quantified, and subjected to high-throughput RNA sequencing using DNA Nanoballs technology.

[0117] Bioinformatic analysis of transcriptomic data was performed using gene ontology-driven and gene panel-driven approaches (Examples 3 and 4). In the gene ontology (GO) terminology analysis, all significantly modulated genes were analyzed to see which predefined pathways they tend to enter (an unsupervised approach) (Example 3). In the gene panel approach, DP cell-specific marker genes were selected and separated into biological functions of interest. Then, a relative gene-panel score based on the integration of the p-value and the change factor was calculated for each gene panel relative to the negative control (a curative approach) (Example 4). Example 2 Autophagy inductance

[0118] The body requires a significant amount of energy for hair growth. During the active growth phase of hair follicles (anagen), metabolic demand is highest as hair cells divide and rapidly synthesize new proteins. Alpha-ketoglutarate (α-KG) is a key metabolite in cellular metabolism and plays an important role in autophagy, the process by which cells break down and recycle their own components. It has effects on cellular metabolism, oxidative stress, and signaling pathways, which are relevant to healthy hair growth. α-KG exhibits antioxidant properties that regulate oxidative stress in cells. Reducing oxidative stress can protect hair follicle cells from damage and promote their longevity and overall health.Furthermore, by managing reactive oxygen species (ROS), it helps prevent damage to hair follicle cells. Alpha-KG promotes hair growth through its roles in metabolism. Oxidative stress management, signaling pathways, and stem cell regulation. Its impact on these processes may contribute to healthier hair follicles and potentially improve hair growth.

[0119] Bioactive walnut peptides can be studied in mice, for example C57BL / 6 mice, or genetically modified strains exhibiting hair loss, with compositions containing one or more bioactive walnut peptides, such as the composition shown in Example 5. Various concentrations of one or more bioactive walnut peptides can be tested. Minoxidil, a vasodilator used to treat hair loss, can be used as a positive control (Messenger and Rundegren, Minoxidil: Mechanisms of Action on Hair Growth, Br. J. Dermatol. 150:186-194 (2004)).

[0120] Mice are divided into two groups: a treatment group and a control group. The treatment group receives one or two daily topical applications of the peptide compositions, while the control group receives a vehicle (placebo). The dorsal skin of the mice is shaved or plucked to facilitate observation of hair growth. The peptide compositions are applied once or twice daily for a specified period, for example, four or eight weeks. The mice are monitored throughout the treatment period for overall health and signs of hair growth. Changes in hair density, color, and texture are also noted.

[0121] At the end of the treatment period, the mice are euthanized in accordance with ethical guidelines, and skin samples are collected from both the treated and control areas for analysis. For histological and biochemical analysis, the collected skin samples are treated by impregnation with paraffin, sectioning, and staining for histological examination. Immunohistochemical or immunofluorescence techniques are used to detect autophagy markers, such as LC3 and p62, in the skin tissues. In addition, Western blotting of tissue extracts can be performed to quantify autophagy-related protein levels, and quantitative PCR can be used to assess autophagy-related gene expression.

[0122] Bioactive nut peptides can have their autophagy inductivity evaluated in mice as highlighted, for example, in Chai et al., Stimulation of Hair Growth by Small Molecules that Activate Autophagy, Cell Reports, 25:3143-3421 (2019) and Sun et al., Autophagy Induces Hair Follicle Stem Cell Activation and Hair Follicle Regeneration by Regulating Glycolysis, Cell and Bioscience, 14(6) (2024). Example 3 Gene ontology-driven analysis

[0123] Gene ontology-driven (GO) analysis is a method used in bioinformatics and computational biology to interpret and organize large sets of gene or protein data based on the gene ontology. It provides a structured and standardized mechanism for describing gene functions, processes, and cellular locations. To establish which biological processes, if any, are influenced by walnut peptide (I) (SEQ ID NO: 45), walnut peptide (II) (SEQ ID NO: 46), and walnut peptide (III) (SEQ ID NO: 47), an initial analysis of transcriptomic data was performed using GO-driven analysis, in which all genes that are significantly modulated are analyzed to see if they impact any predefined biological pathways. All genes that were statistically significantly upregulated or downregulated were analyzed for their association with biological processes.Multivariate results are presented as the ratio of differentially expressed genes in the dataset that match genes in public databases for any given biological process (rich ratio) and the significance of this association (Q value).

[0124] The literature-based panels used in the GO-driven analysis are shown below.

[0125] [Tables3] Literature-based panels: WNT PMO FGF SHH NR5A2 PM06 FGFR1 PTCH1 FZD2 NOG SPRY4 PTCH2 WNT5A PM02 FGF5 GLU SOX11 GREM11 FGF1 GLIS2 FRZB PM04 FGF7 DYRK1B FZD6 PM04 FGF14 NDP PM07 FGF20 DKK1 FST PIK3R1 JUN PM05 SPRY1 WIF1 SOSTDC1 MAPK12 SOX8 JUN FGFR3 SOX9 PITX2 FGF 18 SOX2 TLE4 BAMBI LEF1 —

[0126] Nut peptide (I)

[0127] Walnut peptide (I) (SEQ ID NO: 45) was tested on PD cells at concentrations of 100 pM and 250 pM and was found to influence biological pathways involved in hair growth. In particular, walnut peptide (I) (SEQ ID NO: 45) showed significant upregulation of genes involved in hair follicle development, angiogenesis, and hypoxia. In addition, walnut peptide (I) (SEQ ID NO: 45) significantly downregulated genes associated with cell cycle arrest and IL-1 (interleukin-1) signaling, which is associated with inflammation, the immune response, and apoptosis. The data therefore suggest that walnut peptide (I) (SEQ ID NO: 45) has a positive influence on biological pathways that treat or prevent hair loss, while simultaneously downregulating biological pathways that are detrimental to hair growth.

[0128] Nut peptide (II)

[0129] Walnut (II) peptide (SEQ ID NO: 46) was tested on PD cells at concentrations of 100 pM and 250 pM and was found to influence biological pathways involved in hair growth. Walnut (II) peptide (SEQ ID NO: 46) was found to positively influence biological pathways associated with mitochondrial activity and adenosine triphosphate (ATP) production. Surprisingly, walnut (II) peptide (SEQ ID NO: 46) downregulated biological pathways that are detrimental to hair growth; for example, it downregulated inflammatory pathways.

[0130] Nut peptide (III)

[0131] Walnut Peptide (III) (SEQ ID NO: 47) was tested on PD cells at concentrations of 100 pM and 250 pM and was found not to influence biological pathways involved in hair growth. Unlike Walnut Peptide (I) and Walnut Peptide (II), Walnut Peptide (III) did not show any benefits associated with the treatment or prevention of hair loss.

[0132] In summary, for the genes that were downregulated by Walnut Peptide (I), the associated pathways were more diverse. In particular, cell cycle arrest, IL1 signaling, IL4 response, and apoptosis regulation were downregulated, suggesting that Walnut Peptide (I) reduces negative signals causing adverse effects on CPD activity, potentially creating a healthier microenvironment. With walnut peptide (II) (at both concentrations), a different set of pathways was associated with the upregulated genes. Most of these genes were associated with mitochondrial and ATP / energy production pathways. This suggests that walnut peptide (II) has a different mechanism of action than walnut peptide (I) and acts on CPD activation to provide the energy needed for rapid growth. For the genes that were downregulated by walnut peptide (II), the associated pathways were similar to those of walnut peptide (I), with a strong association with antigen presentation (which could trigger inflammation) and cell cycle arrest. Furthermore, the genes responsible for the downregulation of ATP production were also downregulated with walnut peptide (II).

[0133] With Walnut Peptide (III), very few significant associations were observed for both upregulated and downregulated genes. Genes associated with ATP production and the hypoxia response were observed with the lowest concentration of Walnut Peptide (III), but overall, the effects were smaller compared with Walnut Peptides (I) and (II). Example 4 Gene panel-driven analysis

[0134] In addition to the GO-driven analysis described above, a gene panel (GP)-driven analysis was performed. A GP-driven analysis is used like a GO-driven analysis in genomics and clinical research to investigate specific sets of genes related to particular diseases, conditions, or biological processes. However, the GP-driven approach involves analyzing a predefined panel of genes associated with a specific condition or trait of interest. This is a type of "curative" approach. Genes are selected based on their known or suspected association with specific conditions, their role in relevant biological pathways, or their potential impact on biological outcomes. Often, panels are customized for a specific research project or clinical need.

[0135] GP-driven analysis allows for targeted analysis using gene panels belonging to pathways of interest that may not be found in the public domain. CPD-specific genes (identified from published literature) were isolated to determine the precise transcriptomic changes in CPD. Importantly, a panel of genes relevant to CPD inductivity was evaluated, along with autophagy and hypoxia genes. The aggregate score of each gene (whether positive or negative) was used to determine an effect relative to the negative control. The unique panels used for the tests are shown below.

[0136] [Tables4] Unique Panels Inductivity Autophagy Hypoxie LRP4 ATG9A SLC2A1 RGS2 ULK1 VEGFA TRPS1 WIPI1 HIF1A WNT5A ULK2 CA9 ALPL BCL2 EGLN3 SEMA4C PINK1 BNIP3 NOG NBR1 NDO WDR45 RBP1 SQSTM1 MEF2C ATG16L1 SOX2 GABARAP SPRY4 ATG3 PM04 ATG14 BAMBI MAP1LC3B LEF1 GABARAPL1 ATG10 OPTN UVRAG

[0137] With Walnut Peptide (I), a relative increase in inductivity and hypoxia was readily observed, while autophagy and WNT signaling were observed to a lesser extent. These results are consistent with the gene ontology-based (GO) analysis (Example 3) while also providing further insight into other positive regulators of CPDs relevant to anagen entry. Surprisingly, a strong downregulation of PMO signaling was observed, which is known to act against WNT signaling. In the targeted analysis, Walnut Peptide (I) induces major positive signals and reduces negative signals for CPD activity compared to the negative control.

[0138] Walnut peptides (II) and (III) had no direct effect on genes relevant to CPD activity in the context of anagen entry. Therefore, no large increase in relative scores compared to the negative control was observed. The data are illustrated by a pie chart showing relative gene scores in [Fig. 1]. [Fig. 2] shows walnut peptide (I) (SEQ ID NO: 45) upregulating growth signals and the anagen phase of follicular hair growth. It also shows walnut peptide (II) promoting mitochondrial function and adenosine triphosphate production, thus helping to meet the energy requirements for active hair growth.

[0139] Example 5 Example of a hair and scalp treatment composition

[0140] [Tables 5] INGREDIENTS % by weight Nitrogen peptide(s) PEPTIDE I, II OR THEIR COMBINATIONS 0.01 - 10% Thickeners AMMONIUM POLYACRYLOYLDIMETHYL TAURATE 0.8 XANTHAN GUM 0.2 Preservatives SODIUM BENZOATE 0.3 DISODIUM EDTA 0.1 2-PHENOXYETHANOL 0.7 Water WATER QS100

[0141] The preceding description illustrates and describes preferred embodiments of the invention. However, it should be understood that the inventions are suitable for use in various other combinations, modifications, and environments, and that they are suitable for changes or modifications in the scope of the inventive concepts as expressed herein, in accordance with the teachings above and / or the skills or knowledge of the relevant art. The embodiments described herein are further intended to explain the best known means 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 particular applications or uses thereof. Accordingly, the description is not intended to limit the inventions to the forms disclosed herein.Similarly, the appended claims are to be interpreted in accordance with the patent memorandum and include alternative embodiments.

[0142] As used herein, the terms "comprising", "having" and "including" are used in their broad and non-limiting sense.

[0143] The terms "a," "an," "the," and "the" are understood to encompass both the plural and the singular. Thus, the phrase "one of their mixtures" also refers to "their mixtures." Throughout the disclosure, the phrase "one of their mixtures" is used, following a list of items as shown in the following example where the letters A through F represent the items: "one or more items selected from the group consisting of A, B, C, D, E, F, and one of their mixtures." The phrase "one of their mixtures" does not require that the mixture include all of the items A, B, C, D, E, and F (although all of the items 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 chosen 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".

[0144] Similarly, the expression "one of their / its salts" also refers to "their / its salts". Thus, when the disclosure refers to "an item selected from the group consisting of A, B, C, D, E, F, one of their salts, and one of their mixtures", 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.

[0145] The salts referred to throughout this 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 exhaustive. The appropriate counterions for the components described herein are known in art. This disclosure may not expressly identify all compounds mentioned throughout that may exist as salts or in ionized form, for example, when formulated in a pharmaceutical or cosmetic composition. Nevertheless, to the extent that salt or ionized forms of the compound exist and are known, they are understood to be encompassed within the scope of this disclosure, even if not expressly mentioned. For example, many surfactants may exist as salts or in ionized form in pharmaceutical or cosmetic compositions.Similarly, peptides, including bioactive nut peptides, can be in salt or ionized form, which can improve their stability or compatibility. Non-limiting examples of beneficial compounds include sodium, potassium, calcium, magnesium, chloride, sulfate, phosphate, acetate, citrate, TFA (trifluoroacetate), bicarbonate, and nitrate. Throughout this disclosure, when referring to a bioactive nut peptide, salts of bioactive nut peptides are understood to be included. if terms such as "one of their salts" or "their salts" are not expressly associated with peptides or each time a peptide is mentioned.

[0146] The expression "one or more" means "at least one" and therefore includes individual components as well as mixtures / combinations.

[0147] The term “plurality” means “more than one” or “two or more”.

[0148] Except in operational examples, or unless otherwise indicated, all numbers Indications of quantities of ingredients and / or reaction conditions can be modified in all cases by the term "approximately," meaning within + / - 5% of the stated amount. For example, a quantity "approximately 10% by weight" can include quantities as low as 9.5% by weight and as high as 10.5% by weight. Similarly, a quantity "approximately 50% by weight" includes quantities as low as 47.5% by weight and as high as 52.5% by weight.

[0149] All percentages, parts and ratios here are based on weight unless otherwise stated.

[0150] Some of the various component categories mentioned throughout the disclosure may overlap. However, a single overlapping component cannot simultaneously serve as two different components despite the overlap. Some alcohols function both as a preservative and as a water-soluble solvent, which can be useful in a physiologically acceptable carrier. If an embodiment, claim, or other quotation throughout the disclosure communicates that both a preservative and a water-soluble solvent are present, necessary, or required, a single alcohol such as ethanol cannot simultaneously serve as both a preservative and a water-soluble solvent. In this case, the ethanol will serve as either a preservative or a water-soluble solvent, but simultaneously as both.

[0151] As used herein, all provided ranges are intended to include each specific range within the given ranges, as well as a 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 can serve as a minimum or maximum value for inferring a subrange, etc.

[0152] The term “substantially free” or “essentially free” as used here means that up to 2% by weight of the element may be present. In other embodiments, however, up to 1.5% by weight, up to 1% by weight, up to 0.5% by weight, up to 0.1% by weight, or up to 0.01% by weight of the element may be present. For example, in the case where a pharmaceutical or cosmetic composition is substantially free or essentially free of a component, The pharmaceutical or cosmetic composition may include up to 2% by weight of the component. However, the pharmaceutical or cosmetic composition preferably includes 1.5% by weight or less, 1% by weight or less, 0.5% by weight or less, 0.1% by weight or less, 0.01% by weight or less, or no amount of the specified component.

[0153] As used herein, a "subject" may be a human primate, a non-human primate, a mammal, a rat, a mouse, a cow, a horse, a pig, a sheep, a goat, a dog, a cat, and the like. "Mammal" includes humans and domestic animals such as laboratory animals (e.g., mice, rats, monkeys, dogs, etc.) and domestic animals (e.g., cats, dogs, pigs, cattle, sheep, goats, horses, rabbits) and non-domestic animals such as wildlife and the like.

[0154] All components that are presented positively throughout the disclosure may be excluded negatively from the claims, for example, a claimed composition may be "free", "essentially free" (or "substantially free") of one or more components that are presented positively in this disclosure.

Claims

Demands

1. A bioactive nut-derived peptide, or composition comprising at least one bioactive nut-derived peptide, for use in the treatment or prevention of hair loss or dandruff, wherein one or more nut-derived peptides comprise from 2 to 20 amino acid residues and one or more amino acid sequences selected from: SEQ ID NO: 1 (Thr-Trp), SEQ ID NO: 2 (Trp-Leu), SEQ ID NO: 3 (Leu-Pro), SEQ ID NO: 4 (Pro-Leu), SEQ ID NO: 5 (Pro-Arg), SEQ ID NO: 6 (Tyr-Val), SEQ ID NO: 7 (Val-Leu), SEQ ID NO: 8 (Leu-Leu), SEQ ID NO: 9 (Leu-Pro), SEQ ID NO: 10 (Pro-Ser), SEQ ID NO: 11 (Ser-Pro), and SEQ ID NO: 12 (Pro-Lys).

2. Peptide or composition for use therein according to claim 1, wherein the one or more nut-derived peptides comprise from 3 to 20 amino acid residues and one or more amino acid sequences selected from: SEQ ID NO: 13 (Thr-Trp-Leu), SEQ ID NO: 14 (Trp-Leu-Pro), SEQ ID NO: 15 (Leu-Pro-Leu), SEQ ID NO: 16 (Pro-Leu-Pro), SEQ ID NO: 17 (Leu-Pro-Arg), SEQ ID NO: 18 (Tyr-Val-Leu), SEQ ID NO: 19 (Val-Leu-Leu), SEQ ID NO: 20 (Leu-Leu-Pro), SEQ ID NO: 21 (Leu-Pro-Ser), SEQ ID NO: 22 (Pro-Ser-Pro), and SEQ ID NO: 23 (Ser-Pro-Lys).

3. Peptide or composition for its use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise from 4 to 20 acid residues amino acid and one or more amino acid sequences chosen from: SEQ ID NO: 24 (Thr-Trp-Leu-Pro) SEQ ID NO: 25 (Trp-Leu-Pro-Leu), SEQ ID NO: 26 (Leu-Pro-Leu-Pro), SEQ ID NO: 27 (Pro-Leu-Pro-Arg), SEQ ID NO: 28 (Tyr-Val-Leu-Leu), SEQ ID NO: 29 (Val-Leu-Leu-Pro), SEQ ID NO: 30 (Leu-Leu-Pro-Ser), SEQ ID NO: 31 (Leu-Pro-Ser-Pro), and SEQ ID NO: 32 (Pro-Ser-Pro-Lys).

4. Peptide or composition for use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise from 5 to 18 amino acid residues.

5. Peptide or composition for use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise one or more amino acid sequences selected from: SEQ ID NO: 33 (Thr-Trp-Leu-Pro-Leu), SEQ ID NO: 34 (Trp-Leu-Pro-Leu-Pro), SEQ ID NO: 35 (Leu-Pro-Leu-Pro-Arg), SEQ ID NO: 36 (Tyr-Val-Leu-Leu-Pro), SEQ ID NO: 37 (Val-Leu-Leu-Pro-Ser), SEQ ID NO: 38 (Leu-Leu-Pro-Ser-Pro), and SEQ ID NO: 39 (Leu-Pro-Ser-Pro-Lys).

6. Peptide or composition for use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise from 6 to 18 amino acid residues.

7. Peptide or composition for use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise one or more amino acid residues selected from: SEQ ID NO: 40 (Thr-Trp-Leu-Pro-Leu-Pro), SEQ ID NO: 41 (Trp-Leu-Pro-Leu-Pro-Arg), SEQ ID NO: 42 (Tyr-Val-Leu-Leu-Pro-Ser), SEQ ID NO: 43 (Val-Leu-Leu-Pro-Ser-Pro), and SEQ ID NO: 44 (Leu-Leu-Pro-Ser-Pro-Lys).

8. Peptide or composition for use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise from 4 to 15 amino acid residues, preferably from 5 to 10 amino acid residues.

9. Peptide or composition for use according to any one of the preceding claims, wherein the one or more nut-derived peptides comprise one or more amino acid sequences selected from: SEQ ID NO: 45 (Thr-Trp-Leu-Pro-Leu-Pro-Arg), and SEQ ID NO: 46 (Tyr-Val-Leu-Leu-Pro-Ser-Pro-Lys).

10. Peptide or composition for use according to any one of the preceding claims, wherein one or more nut-derived peptides are selected from the compounds of Formula (I) and Formula (II): X-Thr-Trp-Leu-Pro-Leu-Pro-Arg-Z (I) XT yr-V al-Leu-Leu-Pro-Ser-Pro-Ly sZ (II), in which X represents the primary amine function of the N-terminal amino acid, free or substituted by a protecting group selected from an acetyl group, a benzoyl group, a tosyl group or a benzyloxycarbonyl group, and Z represents the hydroxyl group of the carboxyl function of the C-terminal amino acid, free or substituted by a protecting group selected from a Ci-C20, NH2, NHY, or NYY, in which Y represents a C1-C4 alkyl.

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