Senotherapeutic peptides for hair use

Abstract

Polypeptides which can provide a senotherapeutic effect are provided herein. The polypeptides can be formulated for hair use and can be applied to the scalp of a subject to provide a senotherapeutic effect in the subject or in the cells of the subject.

Description

WSGR Docket No. 56292-708.601SENOTHERAPEUTIC PEPTIDES FOR HAIR USECROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of US provisional application no. 63 / 759,916, filed February 18, 2025, which is incorporated by reference in its entirety.BACKGROUND

[0002] Hair loss, also known as alopecia, refers to a temporary or permanent loss of hair, typically from the scalp, and is a common cause of baldness. Hair loss can be caused by numerous factors. For example, hair loss can be caused by stress, hormone imbalance, disease, or aging. Accumulation of cellular senescence is also a known contributor to hair loss. Cellular senescence is not only a product of aging and disease, but it can also actively contribute to further senescence induction in a positive feedback cycle, impacting health and aging of other tissues. While there are multiple hair loss treatments available, altering one or more steps involved in cellular senescence may prevent or reserve hair loss. Altering cellular senescence may even promote hair growth.SUMMARY

[0003] Described herein are methods for treating hair, the methods comprising: applying to the hair of a subject a composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids. In another aspect, disclosed herein are methods for maintaining or increasing hair bulk, hair density, hair thickness, hair volume, or hair length in a subject, the methods comprising administering to a skin having hair a composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids. In another aspect, disclosed herein are methods for reducing hair loss in a subject, the methods comprising administering to a skin having hair a composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids.

[0004] In some embodiments, the polypeptide comprises at least 10 amino acids, 15 amino acids, or 20 amino acids. In some embodiments, the polypeptide is an isolated, synthetic, or recombinant polypeptide.

[0005] In some embodiments, the composition maintains or increases hair bulk, hair density, hair thickness, hair volume, or hair length. In some embodiments, the composition reduces hair loss. In some embodiments, the composition increases proliferation of one or more hair follicleWSGR Docket No. 56292-708.601cells. In some embodiments, the composition decreases hair follicle miniaturization of one or more hair follicle cells. In some embodiments, the composition decreases senescence of one or more hair follicle cells. In some embodiments, the one or more hair follicle cells comprises outer root sheath cells, inner root sheath cells, or dermal papilla cells, or a combination thereof. In some embodiments, the composition increases proliferation of one or more hair follicle cells.

[0006] In some embodiments, amount of the polypeptide in the composition is about 500 nM to about 500 pM. In some embodiments, amount of the polypeptide in the composition is about 0.001% (w / w) to about 5% (w / w). In some embodiments, amount of the polypeptide in the composition is about 0.01% (w / w) to about 1% (w / w).

[0007] In some embodiments, the polypeptide is formulated as a topical composition. In some embodiments, the topical composition comprises a shampoo, a conditioner, a serum, a cream, a lotion, a gel, a liquid, an oil, a powder, an emulsion, a moisturizer, a foam, a mousse, an aerosol, or a spray.

[0008] In some embodiments, the polypeptide is formulated with a second peptide. In some embodiments, the polypeptide is formulated with one or more of a plant extract, an amino acid, an emollient, a hydrating agent, a vitamin or a vitamin derivative, an antioxidant, and a viscosity modifying agent. In some embodiments, the plant extract comprises one or more of Arctium Majus root extract, Morus Nigra leaf extract, Panax ginseng root extract, Pisum Sativum (pea) sprout extract, Irish moss extract, soy extract, Helianthus annuus (sunflower) sprout extract, Rehmannia Chinensis root extract, and Oryza Sativa (rice) bran extract. In some embodiments, the amino acid comprises one or more of tyrosine, arginine, citrulline, alanine, glycine, phenylalanine, cysteine, methionine, serine, glutamic acid, ornithine, and derivatives thereof. In some embodiments, the hydrating agent comprises on or more of glycerin, squalane, sorbitol, hyaluronic acid, hyaluronic acid derivatives, sodium hyaluronate, sodium hyaluronate crosspolymer, niacinamide, glycoproteins, pyrrolidone carboxylic acid (PCA), lysine HC1, allantoin, and algae extract. In some embodiments, the emollient comprises one or more of plant oil, mineral oil, shea butter, cocoa butter, petrolatum, a fatty acid, a triglyceride, a benzoate, a myristate, a palmitate, a stearate, a glycolipid, a phospholipid, squalane, glycerin, ceramide, and algae extract. In some embodiments, the plant oil comprises one or more of rose hip oil, andiroba oil, grape seed oil, avocado oil, plum seed oil, pracaxi oil, Calycophyllum spruceanum oil, almond oil, and argan oil. In some embodiments, the vitamin comprises one or more of vitamin A, vitamin D, vitamin E, vitamin F, vitamin K, vitamin Bl (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B7 (biotin), vitamin B6, vitamin B 12 (cyanocobalamin), vitamin B9, folic acid, niacinamide, or derivatives thereof.WSGR Docket No. 56292-708.601

[0009] In some embodiments, senescence is assessed by one or more of senescence associated P-galactosidase activity level, a ratio of ATRX foci / cell, pl6 expression, IL-8 expression, and Ki-67 expression.

[0010] In some embodiments, the subject has hair loss. In some embodiments, the hair loss is due to androgenic alopecia, alopecia areata, telogen effluvium, anagen affluvium, or a combination thereof.

[0011] In some embodiments, the polypeptide comprises one or more of ETAKHWLKGI, ATAKAWLKGI, KLKGILRGAA, and WLKGILREAA.

[0012] In another aspect, disclosed herein are compositions for treating hair, the compositions comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids. In another aspect, disclosed herein are compositions for maintaining or increasing hair bulk, hair density, hair thickness, hair volume, or hair length in a subject, the compositions comprising an effective amount of a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids, and a cosmetic excipient. In another aspect, disclosed herein are compositions for reducing hair loss in a subject, the compositions comprising an effective amount of a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids, and a cosmetic excipient.INCORPORATION BY REFERENCE

[0013] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the featuresand advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:

[0015] FIG. 1 illustrates the effects of individual polypeptides on progeria fibroblast cell number and senescence level.

[0016] FIG.2 illustrates the senotherapeutic effects of four polypeptides, Peptide 14 (panel A), Peptide 13 (panel B), Peptide 15 (panel C), and Peptide 16 (panel D), on senescent fibroblasts. *p<0.05; **p<0.01; ***p<0.001.WSGR Docket No. 56292-708.601

[0017] FIG. 3 illustrates a senotherapeutic effect of a polypeptide which promotes a higher number of cells with less ATRX foci / nuclei (panel A), a lower average of ATRX foci / nuclei (panel B), and a higher number of cells with less than 10 ATRX foci / nuclei (panel C). *p<0.05; **p<0.01.

[0018] FIG. 4 illustrates the effect of a senotherapeutic polypeptide which can decrease the number of senescent fibroblasts in a cell population during a 3 -week long exposure keeping the senotherapeutic effect for at least one week after treatment (panel A) without inducing cell toxicity or significantly affecting cellular proliferation during this period (panel B). ***p<0.001; ****p<0.0001.

[0019] FIG. 5 illustrates that treatment with a polypeptide can promote a dose response decrease in cellular senescence, as measured by average ATRX foci accumulation per cell in cells derived from multiple donors.

[0020] FIG.6 illustrates that cellular senescence can be induced in fibroblasts with exposure to etoposide (panel A), treatment of etoposide-induced senescent cells with a polypeptide can result in reduced senescence (panel B), cellular senescence can be induced by UVB exposure (panel C), and treatment of a UVB treated sample with a polypeptide can result in reduced senescence (panel D). *p<0.05; **p<0.01; ***p<0.001.

[0021] FIG. 7 illustrates that human skin equivalents treated with a polypeptide can show altered gene expression in which p 16 can have a significantly lower expression in polypeptide treated epidermis and dermis, compared to untreated control; and IL-8 and MMP-1 were significantly less expressed in polypeptide-treated dermis, compared to untreated counterparts (panel C). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

[0022] FIG.8 illustrates pAkt S473 can be significantly decreasedin both epidermal and dermal samples treated with a polypeptide (panel A), polypeptide treatment can decrease SA-B gal staining in UVB exposed samples indicating protection from UVB-induced cellular senescence (panel B), and that a polypeptide can increase the expression of SIRT6 and BLM (panel C). *p<0.05; **p<0.01; ***p<0.001.

[0023] FIG.9 illustrates the predicted three dimensional structures of two polypeptides, Peptide 14 (panel A) and Peptide 13 (panel B), as well as the superposition of the two polypeptides (panel C).

[0024] FIG. 10 illustrates relative mRNA expression levels of pl6, BLIMP1, ZYGI IB, IL-8, Ki-67, ZIC1, MMP1, HAS2 of the epidermal and dermal layers of 3D skin equivalents treated with a control, Peptide 14, Peptide 13, or Retinoic Acid. Data are presented as 2-ddCt normalized to GAPDH and untreated control. *p<0.05.WSGR Docket No. 56292-708.601

[0025] FIG. 11 illustrates prolonged life and healthspan of Caenorhabditis elegcms (C. elegans) as acquired data indicating that treatment with either 1 pM or 2 pM polypeptide improved worm thrashing (panel A), pumping (panel B), and median lifespan (panel C). *p<0.05; **p<0.01.

[0026] FIG. 12 illustrates the effect of polypeptide sequence LKGIL (SEQ ID NO:6) (A, B, C), and polypeptide sequence WLKGI (SEQ ID NO:7) (D, E, F) to decrease cellular senescence without promoting cell death. In panels A and D, the y-axis indicates the relative senescence level normalized to untreated control. In panels B and D, y-axis indicates the relative cell number normalized to untreated control. In panels C and F, y-axis indicates the average ATRX foci accumulation per cell. *p<0.05; **p<0.01.

[0027] FIG. 13 illustrates relative senescence level (panel A), relative gene expression of CDKN1 A (panel B), and relative gene expression of IL-6 (panel C) of outer root sheath cells after treatment with corticotropin-releasing hormone (CRH) only or treatment with CRH and Peptide 14. *p<0.05; **p<0.01.

[0028] FIGs. 14-15 illustrates representative images of human scalpsbefore and after a 3 -month treatment of Peptide 14.

[0029] FIG. 16 illustrates a HairMetrix® analysis of a clinical study participant at 0 months and after 3 months of using Peptide 14 twice daily.DETAILED DESCRIPTION

[0030] Hair loss is a common condition that can have multiple causes including stress, hormone imbalance, or nutrient deficiency. Hairloss can also be a result of organismal aging or diseases and disorders, such as telogen effluvium and androgenetic alopecia. In telogen effluvium, hair follicles may enter a resting phase, causing hair to fall out due to lack of hair growth. Hair health and density can be associated with lower levels of inflammatory and age-related cytokines IL-ip and IL-6 compared to counterparts with hairloss. Increased levels of IL-ip and IL-6 have been observed in the serum of patients with several age-associated disorders, including cardiovascular disease (CVD), Alzheimer’s disease, and type II diabetes. In some cases, in older adults’ serum IL-6 can be associated with all-cause mortality, CVD, cancer, and liver-related mortality.

[0031] Current treatment options for hair loss include hair transplants, laser therapy, and medication, all of which have various drawbacks and risks. For example, minoxidil is a common treatment for hair loss. Minoxidil acts by opening potassium channels in hair follicle and vascular smooth muscle cells, therefore can have systemic effects on blood pressure. When applied topically, minoxidil has been shown to cause skin irritation. Finasteride is also frequently prescribed for hair loss. However, finasteride has been shown to have limited effectWSGR Docket No. 56292-708.601in older men (e.g., those over 60), and its side effects include increased risk of cancer. Due to the limited safety and effectiveness of current hair loss treatments, new treatments are needed.

[0032] One mechanism of hair loss that could be harnessed for therapeutic applications is cellular senescence. Cellular senescence of hair follicle cells is known to contribute to hair loss. Senescent cells can be identified by senescence -associated beta-galactosidase (SA-BGal) production, pl6 expression, presentation of SASP, and / or alpha thalassemia / mental retardation X-linked chromatin remodeling protein (ATRX) foci accumulation in the nuclei. Functional alterations that can also distinguish senescent cells, include, but are not limited to, decreased proliferation capacity and resistance to mitogenic stimuli. While cellular senescence can be affected by multiple factors, aging and disease are key contributors to cellular senescence and hair loss. In particular, aging can largely result from a functional decline in the ability to maintain tissue homeostasis and integrity, possibly coupled with diminished responses to physiological demands under conditions of stress.

[0033] In a mosaic model of aging, senescent cells can be induced by intrinsic and extrinsic stimuli, such as time / age, UV exposure and smoking, among others. According to this model, senescent cells accumulate in the tissue and actively promote tissue aging by altering the local microenvironment through a senescence-associated secretory phenotype (SASP) composed of proinflammatory cytokines among others. In some cases, senescent cells further promote tissue aging by compromising stem cell renewal and promoting senescence of otherwise normal cells. Therefore, senescent cells are not only a product of aging, but are also active players in the aging process.

[0034] Provided herein are polypeptides, compositions comprising polypeptides and other components, and methods of use thereof. Provided herein are methods for increasing or maintaining hair bulk or maintaining or stimulating hair growth in a subject, the method comprising administering to a skin having hair an effective amount of a polypeptide. Provided herein are methods for increasing or maintaining hair density, hair thickness, hair volume, or hair length in a subject, the method comprising administering to a skin having hair an effective amount of a polypeptide. Provided herein are methods for reducing hair loss in a subject, the method comprising administering to a skin having hair an effective amount of a polypeptide. The polypeptides and compositions comprising the polypeptides can provide an anti -senescent effect (e.g., on cells of a subject). Polypeptides can promote a decrease of senescence levels in cells and tissues by promoting apoptosis, promoting DNA repair, and / or inhibiting DNA-damage induced senescence. Anti-senescent effects can include, but are not limited to, increased cell renewal rates, decreased ATRX nuclear foci accumulation, decreased pl 6 expression, lower SASP production, decreased senescence-associated beta-galactosidase production, decreasedWSGR Docket No. 56292-708.601hair follicle miniaturization, increased hair thickness, increased hair growth, increased hair bulk, increased hair density, increased hair volume, increased hair length, and reduction in hair loss. The polypeptides and compositions comprising the polypeptides provided herein may inhibit, prevent, or slow hair loss, including aging-associated hair loss. The polypeptides and compositions comprising the polypeptides may increase or maintain hair bulk or may maintain or stimulate hair growth. The polypeptides and compositions comprising the polypeptides may increase or maintain hair density, hair thickness, hair volume, or hair length . Polypeptides have properties such as multifunctional behavior which can make them useful for cosmetic or therapeutic applications, including senotherapy.Definitions

[0035] It is to be understood that the methods and compositions described herein are not limited to the particular methodology, protocols, and reagents described herein and as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the methods and compositions described herein, which will be limited only by the appended claims.

[0036] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly indicates otherwise.

[0037] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which the disclosures described herein belong. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure described herein, the preferred methods, devices, and materials are now described.

[0038] In a peptide or polypeptide, suitable conservative substitutions of amino acids are known to those of skill in this art and can be made generally without altering the biological activity of the resulting molecule. Watson et al. (1987, Molecular Biology of the Gene, 4th Edition, The Benjamin Cummings Pub. Co., p. 224) is incorporated herein by reference. The amino acids may be either in the L- or D-isomeric form. When an amino acid residue is part of a polypeptide chain, the D-isomeric form of the amino acid can be substituted for the L-amino acid residue, as long as the desired functional property is retained. The amino acids herein can be represented by their standard IUPAC 1 -letter code or 3 -letter code. An amino acid residue represented by “X” or “Xxx” refers to any one of the naturally occurring or non -naturally occurring amino acid residues known in the art or to a modification of a nearby residue. Amino acid substitutions are typically of single residues, such substitutions are preferably made with those set forth in Table 1 , but may be of multiple residues, either clustered or dispersed. An amino acid can be replacedWSGR Docket No. 56292-708.601with a different naturally occurring or a non-conventional amino acid residue. Such substitutions may be classified as “conservative,” in which case an amino acid residue contained in a polypeptide is replaced with another naturally occurring amino acid of similar character either in relation to polarity, side chain functionality, or size. Additions encompass the addition of one or more naturally occurring or non-conventional amino acid residues. Deletion encompasses the deletion of one or more amino acid residues.Table 1: Conservative and Non-Conservative Amino Acid Substitutions

[0039] Substitutions encompassedby the present disclosure may also be “non -conservative,” in which an amino acid residue which is present in a polypeptide is substituted with an amino acid having different properties, such as a naturally -occurring amino acid from a different group (e.g., substituting a charged or hydrophobic amino acid with alanine), or alternatively, in which a naturally-occurring amino acid is substituted with a non-conventional amino acid.WSGR Docket No. 56292-708.601

[0040] The term “analog(s)” as used herein refers to a composition that retains the same structure or function (e.g., binding to a receptor) as a polypeptide, such as the same protein from a different organism. Examples of analogs include mimetics or peptidomimetics, peptides, small and large organic or inorganic compounds, as well as derivatives and variants of a polypeptide herein. Such derivatives and variants refer to polypeptides that differ from the naturally occurring polypeptides by one or more amino acid deletions, additions, substitutions, or sidechain modifications. In some embodiments, a peptide analog is a peptide in which one or more of the amino acids has undergone side-chain modifications. Examples of side-chain modifications contemplated by the present disclosure include modifications of amino groups such as by reductive alkylation by reaction with an aldehyde followed by reduction with NaBH4; amidination with methylacetimidate; acylation with acetic anhydride; carbamoylation of amino groups with cyanate; trinitrobenzylation of amino groups with 2,4,6 -trinitrobenzene sulphonic acid (TNBS); acylation of amino groups with succinic anhydride and tetrahydrophthalic anhydride; and pyridoxylation of lysine with pyridoxal -5 -phosphate followed by reduction with NaBH4. In some embodiments, a peptide analog is one in which the guanidine group of arginine residue(s) is modified by the formation of heterocyclic condensation products with reagents such as 2,3 -butanedione, phenylglyoxal and glyoxal; carboxyl group(s) is modified by carbodiimide activation via O-acylisourea formation followed by subsequent derivatization, for example, to a corresponding amide; sulphydryl group(s) may be modified by methods such as carboxymethylation with iodoacetic acid or iodoacetamide; performic acid oxidation to cysteic acid; formation of mixed disulphides with other thiol compounds; reaction with maleimide, maleic anhydride or other substituted maleimide; formation of mercurial derivatives using 4-chloromercuribenzoate, 4-chloromercuriphenylsulphonic acid, phenylmercury chloride, 2-chloromercuri-4-nitrophenol and other mercurials; carbamoylation with cyanate at alkaline pH. In any of the analogs herein, any modification of cysteine residues preferably do not affect the ability of the peptide to form the necessary disulphide bonds. In some embodiments, a peptide analog comprises tryptophan residue(s) that are modified, for example, by oxidation with N-bromosuccinimide or alkylation of the indole ring with 2-hydroxy-5-nitrobenzyl bromide or sulphenyl halides; tyrosine residues altered by nitration with tetranitromethane to form a 3-nitrotyrosine derivative; imidazole ring(s) of a histidine residue modification accomplished by alkylation with iodoacetic acid derivatives or N-carb ethoxylation with diethylpyrocarbonate; proline residue(s) modified by, for example, hydroxylation in the 4 -position; glycosylation variants from a completely unglycosylated molecule to a modified glycosylated molecule; and altered glycosylation patterns as a result from expression of recombinant molecules in different host cells.WSGR Docket No. 56292-708.601

[0041] The term “isolated” means altered from its natural state; i.e., if it occurs in nature, it has been changed or removed from its original environment, or both. For example, a naturally occurring polynucleotide or a polypeptide naturally present in a living animal in its natural state is not “isolated,” but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is “isolated,” as the term is employed herein.

[0042] The terms “protein,” “peptide,” “oligopeptide,” or “polypeptide” as used herein refer to any composition that includes two or more amino acids joined together by a peptide bond. It will be appreciated that polypeptides often contain amino acids other than the 20 amino acids commonly referred to as the 20 naturally occurring amino acids, and that many amino acids, including the terminal amino acids, may be modified in a given polypeptide, either by natural processes such as glycosylation and other post -translational modifications, or by chemical modification techniques which are well known in the art. The known modifications, which may be present in polypeptides of the present disclosure, include, but are not limited to, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of a flavonoid or a heme moiety, covalent attachment of a polynucleotide or polynucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphatidylinositol, cross -linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cystine, formation of pyro glutamate, formylation, gamma-carboxylation, glycation, glycosylation, glycosylphosphatidyl inositol (GPI) membrane anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to polypeptides such as arginylation and ubiquiti nation. The term “protein” also includes “artificial proteins” which refers to linear or non-linear polypeptides, consisting of alternating repeats of a polypeptide (e.g., SEQIDNOs:l-7)and a spacer. A DNA construct encoding the polypeptide and spacer alternate repeats can be synthesized using methods known in the art see Rotzschke et al., 1997, Proc. Natl. Acad. Sci. USA 94:14642-14647).

[0043] The term “purified” as used herein to describe a polypeptide, polynucleotide, or other composition, refers to such polypeptide, polynucleotide, or other composition separated from one or more compounds which are usually associated with it in nature. Such other compositions can be, for example, other polypeptides or polynucleotides, carbohydrates, lipids, etc. The term “purified” can also be used to specify the separation of monomeric polypeptides of the disclosure from oligomeric forms such as homo- or hetero-dimers, trimers, etc. A substantially pure polypeptide typically comprises at least about 50%, 60%, 70%, 80%, or 90% weight / weight of a polypeptide sample, or more preferably at least about 95%, 96%, 97%, 98%, 99%, or 99.5% weight / weight of a polypeptide sample. As a preferred embodiment, a polypeptide of the presentWSGR Docket No. 56292-708.601disclosure is atleast 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% pure relative to heterologous polypeptides.

[0044] The term “subject,” or “patient” as used herein, encompasses animals. In some embodiments, the subject may be a mammal. Examples of mammals include, but are not limited to, any member of the Mammalian class: humans, non -human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, and guinea pigs, and the like. The mammal can be a human.

[0045] The terms “treat,” “treating,” or “treatment,” as used herein, include delaying the onset of, reducing the occurrence of, or ameliorating at least one symptom of a disease or condition, preventing additional symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition either prophylactically and / or therapeutically.

[0046] The term “therapeutically acceptable,” as used herein, refers to a material, including but not limited, to a salt, carrier, or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively nontoxic, i.e.,the material may be administered to a subject without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0047] The term “carrier,” as used herein, refers to relatively nontoxic chemical compounds or agents that facilitate the incorporation of a compound into cells or tissues.

[0048] The term “diluent,” as used herein, refers to chemical compounds that are used to dilute the compound of interest prior to delivery. Diluents can also be used to stabilize compounds because they can provide a more stable environment. Salts dissolved in buffered solutions (which also can provide pH control or maintenance) are utilized as diluents in the art, including, but not limited to, a phosphate buffered saline solution.

[0049] The terms “effective amount” or “therapeutically effective amount,” as used herein, refer to a sufficient amount of an agent or a compound being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and / or alleviation of the signs, symptoms, orcauses of a disease, or any other desired alteration of a biological system. For example, an “effective amount” for therapeutic uses is the amount of the composition comprising a compound as disclosed herein required to provide a clinically significant decrease in disease symptoms. An appropriate “effective” amount in any individual case may be determined using techniques, such as a dose escalation study.WSGR Docket No. 56292-708.601Hair Follicle

[0050] Usually, hair has two parts, a hair shaft and a hair root, with the hair shaft being the visible portion protruding from the skin. Hair is produced in the hair follicle. Hair follicles are located in the dermal layer of the skin. Typically, a hair follicle comprises a dermal papilla, a hair matrix, a root sheath, and a bulge. The papilla may be located at the bottom of the hair follicle and is made up of connective tissue. The dermal papilla is often surrounded by the hair matrix. Usually, the root sheath has an outer root sheath, a companion layer, and an inner root sheath. The bulge may be located in the outer root sheath and contains multiple types of stem cells which differentiate into the hair follicle subtypes. The hair follicle is surrounded by a connective tissue sheath and connected to nerves, glands, muscles, and blood vessels.

[0051] Usually, the dermal papilla cells (DPCs) are a highly active group of cells derived from the dermis mesenchyme. Located at the base of the hair follicles, the dermal papillae may play a crucial role in hair growth cycle by inducing follicle development from the epidermis to produce hair fiber. Often, hair growth is tightly regulated by the epithelial-mesenchymal interaction of hair follicular cells, which involves various molecular pathways including BMP and Wnt signaling. Dermal papilla cells have been shown to gradually lose their hair inductivity and proliferation capacity in long-term cultures. In some cases, dermal papilla cells may undergo premature aging in diseases such as androgenetic alopecia and senescent alopecia. In some cases, aging of dermal papilla cells has been linked to decreases Wnt and beta-catenin signaling. In some cases, increasing Wnt and beta-catenin signaling has been associate with increased hair follicle reconstruction in senescent dermal papilla cells.

[0052] Typically, the outer root sheath surrounds the hair fiber and inner root sheath. The ORS may be derived from the epidermis by downward growth. The ORS comprises basal, prickle cells and granular layers near the opening of the follicle onto the skin surface, and modified glycogen-rich prickle cells in the deeper parts of the follicle. The ORS may have several cell populations which play a role in certain functions of the hair follicle, such as serving as a sensory organ and an immunologic sentinel of the skin. Often, the ORS has been shown to contain nestin-expressing stem cells, with a variable location of the nestin -expressing stem cells based on the hair cycle. In some cases, the ORS represents an alternate source of keratinocytes, which are suitable for skin replacement.

[0053] The companion layer may be located between the outer root sheath and the inner root sheath.WSGR Docket No. 56292-708.601

[0054] The inner root sheath is located between the outer root sheath and the hair fiber. The inner root sheath comprises a Henle’s layer, a Huxley’s layer, and a cuticle of inner root sheath. The inner root sheath helps to shape new hair and is involved in keratinization.

[0055] Hair follicles may regulate hair growth though a multiphase process. During anagen, the growth phase, new cells may be formed, stuck together, hardened, and add to the hair shaft, extending the length of the hair. Healthy hair follicles generally stay in the growth phase for about 2-7 years. The catagen phase may occur at the end of the anagen phase. During the catagen phase, a bulb of keratin may be attached to the bottom of the hair shaft to keep it in place. During the telogen phase, hair growth typically does not occur. Entry into the telogen phase can be triggered by stressors such as stress or hormonal changes. Finally, hair follicles may enter the exogen phase during which shedding occurs.

[0056] Corticotropin-releasing hormone (CRH) can also play a role in hair loss. CRH is a peptide hormone secreted by the hypothalamus in response to stress or other stimuli. In some embodiments, CRH plays a key role in initiating the hypothalamic-pituitary-adrenal (HP A) axis. Upon release, CRH can stimulate the pituitary gland to secrete adrenocorticotropic hormone (ACTH). In some embodiments, the secretion of ACHT may trigger the adrenal glands to release cortisol, the body's primary stress hormone. In some embodiments, CRH indirectly affects hair follicles through cortisol production and inflammatory pathways. In some embodiments, chronic stress and elevated CRH levels may disrupt the hair cycle, leading to hair follicle miniaturization or a premature shift to the telogen (resting) phase, contributing to stress -related hair shedding or telogen effluvium. In some embodiments, CRH and its downstream signaling pathways can accelerate cellular senescence in skin cells and hair follicles. Chronic stress and prolonged exposure to elevated cortisol levels can induce oxidative stress, DNA damage, and inflammation, and / or driving senescence in keratinocytes and dermal fibroblasts. In hair follicles, this may result in structural and functional decline, contributing to hair thinning and hair loss over time.Polypeptides

[0057] Polypeptides and compositions comprising polypeptides as provided herein can provide a senotherapeutic effect, e.g., the polypeptide can reduce senescence, such as by halting senescence, preventing senescence, inhibiting senescence, reversing senescence, destroying senescent cells, killing senescent cells, removing senescent cells, or by any suitable mechanism of reducing the burden or effects of senescent cell accumulation. Such polypeptides can in some cases comprise the amino acid sequence LKGI (SEQ ID NO:5). Compositions comprising such polypeptides can be employed or used to provide a senotherapeutic effect.WSGR Docket No. 56292-708.601

[0058] The polypeptides (e.g., senotherapeutic polypeptides) can comprise at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21,22, 23,24, 25,26, 27,28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 or more amino acids. In some cases, the polypeptides may be no more than 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21,22, 23,24, 25,26, 27,28, 29,30, 31, 32, 33, 34, 35, 36, 37,38, 39,40, 45, 50, 55,60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acids long. In some cases, the polypeptides canbe from 4 to 25, from 4 to 15, or from 4 to 10 amino acids in length. In some embodiments, the polypeptides can comprise at least 30, 40, 50, 60, 70, 80, 90, 100, or more amino acids. In certain embodiments, the polypeptides may comprise less than 100, 90, 80, 70, 60, 50, 40, 30, or fewer amino acids.

[0059] Examples of polypeptides which can provide a senotherapeutic effect are provided in Table 2 below.Table 2: Example Polypeptide Amino Acid Sequences

[0060] A polypeptide can be an isolated or recombinant polypeptide, which can comprise an amino acid sequence of XiX2X3X4X5X6X7X8X9Xio . Amino acids included in the polypeptide can comprise a natural amino acid, which can include alanine (Ala, A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine (Cys, C), glutamine (Glu, Q), glutamic acid (Glu, E), glycine (Gly, G), histidine (His, H), isoleucine (He, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y), or valine (Vai, V).

[0061] In some cases, the isolated or recombinant polypeptide comprising an amino acid sequence of XiX2X3X4X5X6X7X8X9Xio can comprise an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity to a first sequence, ETAKHWLKGI (SEQ ID NO:1), wherein Xi is E, X2is T, X4is K, X6is W, X7is L, X9is G, and Xi0is I; and wherein at least either (i) X3is not S; or (ii) if X5is any amino acid then X8is not G; or (iii) if X8is any amino acid then X5is not N; or (iv) any one of (i), (ii), or (iii), where the sequence can optionally comprise 1, 2, 3, or 4 conservative amino acid substitutions. In some cases, the isolated or recombinant polypeptide can include an amino acid sequence having at least 70%,WSGR Docket No. 56292-708.60175%, 80%, 85%, 90%, 95%, or 100% identity to a sequence of ETAKHWLKGI (SEQ ID NO:1). The present disclosure also contemplates analogs, such as peptidomimetics of the above.

[0062] In some cases, the isolated or recombinant polypeptide comprising an amino acid sequence of XiX2X3X4X5X6X7X8X9Xio can comprise an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity to a second sequence, ATAKAWLKGI (SEQ ID NO:2), wherein Xiis A, X2is T, X3is A, X4is K, X5is A, X6is W, X7is L, X8is K, X9is G, and Xio is I. Such a recombinant polypeptide can optionally comprise 1, 2, 3, or 4 conservative amino acid substitutions. In some cases, the isolated or recombinant polypeptide can include an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, or 100% identity to a sequence of ATAKAWLKGI (SEQ ID NO:2). The present disclosure also contemplates analogs, such as peptidomimetics of the above.

[0063] In some cases, the isolated or recombinant polypeptide comprising an amino acid sequence of XiX2X3X4X5X6X7X8X9Xio can comprise an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity to a third sequence, KLKGILRGAA (SEQ ID NO:3), wherein at least either (i) if X9is any amino acid then X3is notN; or (ii) if X3is any amino acid then X9is not S; or (iii) if X4is any amino acid then X7is not L; or (iv) if X7is any amino acid then X4is not S; or (v) any one of (i), (ii), (iii), or (iv), where the sequence can optionally comprise 1, 2, 3, or 4 conservative amino acid substitutions. In some cases, the isolated or recombinant polypeptide can include an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 95%, or 100% identity to a sequence of KLKGILRGAA (SEQ IDNO:3). The present disclosure also contemplates analogs, such as peptidomimetics of the above.

[0064] In some cases, the isolated or recombinant polypeptide comprising an amino acid sequence of XiX2X3X4X5X6X7X8X9Xio can comprise an amino acid sequence having at least 80%, 85%, 90%, or 95% identity to a fourth sequence, WLKGILREAA (SEQ ID NO:4), wherein Xi is W, X2is L, X3is K, X4is G, X5is I, X6is L, X7is R, X8is E, X9is A, and Xi0is A. Such a recombinant polypeptide can optionally comprise 1, 2, 3, or 4 conservative amino acid substitutions. In some cases, the isolated or recombinant polypeptide can include an amino acid sequence having at least 80%, 85%, 90%, 95%, or 100% identity to a sequence of WLKGILREAA (SEQ ID NO:4). The present disclosure also contemplates analogs, such as peptidomimetics of the above.

[0065] In some cases, a polypeptide can comprise the amino acid sequence LKGI (SEQ ID NO:5), LKGIL (SEQ ID NO:6), or WLKGI (SEQ ID NO:7) (see Table 3 below).Table 3: Example Polypeptide Amino Acid SequencesWSGR Docket No. 56292-708.601

[0066] A polypeptide comprising one of SEQ ID NOs:5-7 may comprise at least 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, or 50 amino acids. In some embodiments, a polypeptide comprising one of SEQ ID NOs:5-7 may comprise no more than 4, 5, 6, 7, 8, 9, 10,20, 30,40, 50, 60, 70, 80, 90, or 100 amino acids. SEQ ID NOs:5-7 can be at the N-terminus of a polypeptide, atthe C-terminus of a polypeptide, or between the N-terminus and C-terminus of a polypeptide. In some cases, a polypeptide can comprise more than one of SEQ ID NOs:5-7.

[0067] The polypeptide can be isolated, substantially pure, or purified. In some cases, an isolated polypeptide can be (i) synthesized chemically or (ii) expressed in a host cell and purified away from associated and contaminating proteins. In some cases, the polypeptide can be present in a host cell as the expression product of a portion of an expression vector, and can be linked to a protein moiety or linked to a chemical moiety.

[0068] Analogs, including peptidomimetics, of the disclosed polypeptides can provide a senotherapeutic effect. The peptides and polypeptides disclosed herein may include peptidomimetic equivalents.

[0069] In some cases, as discussed above, a polypeptide can have a sequence identity to a polypeptide described herein. Sequence identity of a polypeptide can refer to an exact amino acid-to-amino acid correspondence of two polypeptide sequences. In some cases, techniques for determining sequence identity can include determining the amino acid sequence and comparing the amino acid sequence to a second amino acid sequence. Two or more sequences can be compared by determining their percent identity, or the number of exact matches between two aligned sequences divided by the length of the longer sequence and multiplied by 100. Percent identity may also be determined, for example, by comparing sequence information using the advanced BLAST computer program, including, e.g., version 2.2.9, available from the National Institutes of Health. The BLAST program is based on the alignment method of Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 87:2264-2268 (1990) and as discussed in Altschul, et al., J. Mol. Biol., 215:403-410 (1990); Karlin and Altschul, Proc. Natl. Acad. Sci. USA, 90:5873-5877 (1993); and Altschul etal., Nucleic Acids Res. 25:3389-3402 (1997). The program may be used to determine percent identity over the entire length of the polypeptides being compared. Default parameters are provided to optimize searches with short query sequences in, for example, the blastp program. The program also allows the use of an SEG filter to mask -off segments of the query sequences as determined by the SEG program of Wootton and Federhen, Computers and Chemistry 17:149-163 (1993).WSGR Docket No. 56292-708.601Compositions

[0070] Disclosed herein are compositions comprising one or more of the polypeptides described herein. In some cases, a composition can be used to reduce hair loss, to increase or maintain hair bulk, or to maintain or stimulate hair growth. In some cases, a composition can be used to increase or maintain hair density, hair thickness, hair volume, or hair length. In some embodiments, the compositions may be senotherapeutic. In some cases, a composition can be used to treat an age-related disease or condition or age -associated disorder, e.g., to delay the onset of, reduce the occurrence of, or ameliorate the age-related disease or condition or age-associated disorder, including but not limited to hair loss.

[0071] In some cases, a composition may include, e.g., an effective amount of a polypeptide, alone or in combination, with one or more vehicles (e.g., therapeutically acceptable compositions or therapeutically acceptable carriers) and other therapeutically effective compounds. In some embodiments, the effective amount of the polypeptide refers to having a desired effect on a subject, including but not limited to a cell, a tissue, or an organism, treated with the composition. In some embodiments, the effective amount of the polypeptide has minimal or low effect systemically on the treated subject. In some embodiments, the effective amount of the polypeptide has maximal effect locally at or near the treated area.

[0072] In some embodiments, the formulation can be configured to penetrate topically from the scalp or the epidermis to the hair follicle or the dermis. In some embodiments, the formulation is configured to penetrate topically through the epidermis layers. In some embodiments, the formulation is configured to penetrate topically into the hair follicle. In some embodiments, the formulation is configured to penetrate to the hair follicle cell (e.g., the dermal papilla cells or the outer root sheath cells). In some embodiments, the effective amount of a polypeptide is at least 1 nM, 5 nM, 10 nM, 50 nM, 100 nM, 500 nM, 1 pM, at least 10 pM, at least 25 pM, at least 50 pM, at least75 pM, atleast 100 pM, at least 150 pM, at least 200 pM, at least 250 pM, at least 300 pM, at least 350 pM, at least 400 pM, at least 450 pM or at least 500 pM. In some instances, the effective amount of a polypeptide is between about 1 nMto about 1000 nM, about 5 nM to about 750 nM, about 25 nM to about 750 nM, or about 50 nM to about 500 nM. In some instances, the effective amount of a polypeptide is between about 50 nMto about 500 pM, about25 pMto about250 pM, about50 pMto about200 pM, orabout75 pMto about 125 pM In some instances, the effective amount of a polypeptide is at least 0.00001%, 0.00005%, 0.0001%, 0.0005%, 0.001%, at least 0.005%, at least 0.01%, at least 0.05%, at least 0.1%, atWSGR Docket No. 56292-708.601least 0.5%, at least 1%, at least 1.5%, at least 2%, at least 2.5%, at least 3%, at least 3.5%, at least 4%, at least 4.5%, or at least 5% (w / w) of the final composition. In some instances, the effective amount of a polypeptide between about 0.00001% to about 5%, 0.00001% to about 1%, 0.00001% to about 0.1%, about 0.001% to about 5%, about 0.005% to about 4%, about 0.005% to about3%, about0.005%to about 2%, about 0.005% to about 1%, about 0.005% to about 0.5% or about 0.001% to about 1% of the final composition. In some embodiments, the effective amount of the polypeptide for an in vivo application maybe at least 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, or 500 times than the amount used in an in vitro application. The effective amount of polypeptide results in some hair follicle or dermis penetration of the polypeptide, in some instances about 1% penetration, about 2% penetration, about 4% penetration, about 5% penetration, or about 10% penetration. In some instances, no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of the polypeptide in the composition applied onto the skin having hair penetrates into the hair follicle or the dermis. In some instances, no more than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of the polypeptide in the composition applied onto the skin having hair penetrates to the hair follicle cell (e.g., the dermal papilla cells or the outer root sheath cells). In some instances, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of the polypeptide in the composition applied onto the skin having hair penetrates into the hair follicle or the dermis. In some instances, at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% of the polypeptide in the composition applied onto the skin having hair penetrates to the hair follicle cell (e.g., the dermal papilla cells or the outer root sheath cells). In some instances, the amount used in an in vivo application is a factor of amount of hair follicle or dermal penetration in an in vitro penetration study. In some instances, the factor for the amountusedin an in vivo application is atleast2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 times the amount of in vitro hair follicle or dermal penetration.

[0073] In some cases, the compositions described herein can be administered with one or more additional nutraceutical, cosmetic, therapeutic, or pharmaceutical treatments (e.g., co-administered, sequentially administered, or co -formulated).

[0074] In some embodiments, the formulation may be used in conjunction with one or more treatments. In some embodiments, the formulation may be used with a sonic treatment, an ultrasonic treatment, a LED treatment, a light treatment, an electrical treatment, a radiofrequency treatment, or other dermatological treatments. In some embodiments, the composition is applied to the scalp or skin having hair before, after, or during the treatment.

[0075] A composition can be formulated for topical application. A composition, such as for topical application, can be formulated as a cosmetic composition. For example, the compositionWSGR Docket No. 56292-708.601may be formulated for application onto the scalp or skin having hair. In some embodiments, the composition is configured as a topical supplement. Formulations such as those for topical application can be a cream, an ointment, a gel, a liquid, a powder, a lotion, a serum, an emulsion, a moisturizer, a foam, a face mask, a mousse, an aerosol, a spray, a cleanser, a toner, a topical patch, a hydrogel patch, or a shampoo. Polypeptides applied topically can be applied to an affected area, to an area which may become affected in the future, a portion of the subject, or substantially the entire subject. In some cases, atopical treatment can be applied with a buffer, another topical treatment, a cream, or a moisturizer.

[0076] A composition as contemplated herein can also be edible, i.e., formulated as an edible supplement or beverage, such that the composition is formulated to be safely consumed by humans. In some cases, an edible composition can be therapeutically effective to treat hair loss, to increase or maintain hair bulk, or to maintain or stimulate hair growth. In some cases, an edible composition can be therapeutically effective to increase or maintain hair density, hair thickness, hair volume, or hair length. In some cases, an edible supplement can be configured as a tablet, capsule, chew, gummy, powder, food bar, meal replacement bar, or a food additive. In some cases, a beverage can be formulated to comprise water, a soda, a tea, coffee, milk, a juice, a shake, a drink, or other edible liquid.

[0077] The compositions, polypeptides, and methods provided herein may be formulated for a systemic administration. In some cases, the systemic administration comprises ophthalmic delivery, otologic delivery, nasal delivery, urogenital delivery, rectal delivery, dermal delivery, or delivery by injection. In some cases, the delivery by injection comprises intradermal, subcutaneous, intraarticular, intravitreal, intracerebral, intrathecal, epidural, intravenous, intracardiac, intramuscular, intraosseous, or intraperitoneal delivery. In some cases, the ophthalmic delivery includes but is not limited to nasal spray, ear drops, eye drops, ointment, hydrogel, insufflation, or mucoadhesive disc. In some embodiments, the systemic administration of the compositions, polypeptides, and methods provided herein maybe formulated as a fluid, a liquid, a hydrogel, an ointment, a solution, a suspension, or a dry -powder. In some embodiments, the systemic formulation comprises water, a saline, or a buffer. In some cases, the buffer comprises sodium bicarbonate, phosphate, sodium chloride, or sodium lactate. In some cases, the systemic delivery may be delivered as a bolus or an infusion.

[0078] The compositions, polypeptides, and methods provided herein may be formulated for an oral administration. In some cases, the oral administration comprises enteral administration, buccal administration, or administration by respiratory tract. Usually, the enteral administration is by the gastrointestinal tract. Such oral formulations may include carrier and dispersant components and may be in any suitable form, including aerosols (for oral or pulmonaryWSGR Docket No. 56292-708.601delivery), syrups, elixirs, tablets, including chewable tablets, hard or soft capsules, troches, lozenges, aqueous or oily suspensions, emulsions, cachets or pellets granulates, and dispersible powders. In some embodiments, the compositions and polypeptides described herein are formulated into solid dosage forms, including but not limited to tablets or capsules, which are suitable for simple oral administration of precise dosages. In some cases, oral administration provides an easy route for systemic administration, a higher patient compliance, a low-cost production, and a longer storage and shelf-life.

[0079] In some cases, the formulation for systemic delivery comprises components to improve bioavailability or stability of the compositions and polypeptides provided herein. Such components to improve stability may comprise polymers. Such polymers to improve stability may include but are not limited polyethylene glycol, polyacrylic acid, polyethylene acrylate, polygly colic acid, polylactic acid, polylactic-glycolic acid, sodium hyaluronate, chitosan, methylcellulose, alginate, cyclodextrin, or dextrin.

[0080] In some cases, a composition can comprise a skin or hair conditioning agent (e.g., a humectant, exfoliant, emollient, or hydrator). A humectant can be for moisturizing, reducing scaling, or stimulating removal of built-up scale from the skin. An exfoliant can be for the removal of old skin cells from the surface, and can be a physical exfoliant or a chemical exfoliant. An emollient can be a preparation or ingredient which can soften dry, rough, or flakey skin. A hydrator can be for moisturizing, reducing scaling, or stimulating removal of built-up scale from the skin. In some cases, emollient is an agent that prevents water loss and has a softening and soothing effect on skin. In some embodiments, emollients may comprise at least one of plant oils, mineral oil, shea butter, cocoa butter, petrolatum, fatty acids (animal oils, including emu, mink, and lanolin), triglycerides, benzoates, myristates, palmitates, stearates, glycolipids, phospholipids, squalene, glycerin, rosehip oil, andiroba oil, grape seed oil, avocado oil, plum seed oil, pracaxi oil, Calycophyllum spruceanum oil, almond oil, argan oil, caprylic / capric triglyceride, jojoba butter, jojoba oil, Spectrastat G2, ceramide, and algae extract. In some cases, the composition comprises a skin or hair hydrating agent, also referred to as a skin or hair hydrator. In some cases, the skin or hair hydrating agent include but are not limited to glycerin, squalene, sorbitol, hyaluronic acid, hyaluronic acid derivatives, sodium hyaluronate, sodium hyaluronate crosspolymer, niacinamide, glycoproteins, pyrrolidone carboxylic acid (PCA), lysine HC1, allantoin and algae extract. In some embodiments, the composition comprises at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% skin or hair conditioning agent. In some embodiments, the composition comprises about 1% to about 70%, about 1% to about 60%, about l%to about50%, about5%to about50%, about 5% to 45%, or about 5% to 40% skin or hair conditioning agent.WSGR Docket No. 56292-708.601

[0081] A composition can comprise a shine control agent, which can improve or regulate the shiny appearance of skin or hair. Shine control agents can be porous in nature. Such agents can provide a reservoir to absorb excess moisture to reduce the appearance of shine. Shine control agents can be silicas, magnesium aluminum silicates, talc, sericite and various organic copolymers. Particularly effective shine control agents can include silicates or carbonates that are formed by reaction of a carbonate or silicate with the alkali (IA) metals, alkaline earth (IA) metals, or transition metals, and silicas (silicon dioxide). Preferred shine control agents are selected from the group consisting of calcium silicates, amorphous silicas, calcium carbonates, magnesium carbonates, zinc carbonates, bentonite clay, and combinations thereof.

[0082] A composition can comprise a film forming agent, which can aid film substantivity and adhesion to the skin or hair. A film forming agent can improve long wear and non -transfer performance of a composition. Film forming agents can be water soluble, water insoluble, or water dispersing. Film forming agents can be 1) organic silicone resins, fluorinated silicone resins, copolymers of organic silicone resins, trimethylsiloxysilicate, GE’s copolymers of silicone resins, SF1318 (silicone resin and an organic ester of isostearic acid copolymer) and CF1301 (silicone resin and alpha methyl styrene copolymer), Dow Coming’s pressure sensitive adhesives copolymers of silicone resins and various PDMS’s (BIO -PSA series); and 2) acrylic and methacrylic polymers and resins, silicone-acrylate type copolymers and fluorinated versions of, including silicones plus polymer from 3M, KP545 from Shin-Etsu, alkyl-acrylate copolymers, KP 561 and 562 from Shin-Etsu; 3) decene / butene copolymer from Collaborative Labs; 4) polyvinyl based materials, PVP, PVP / VA, including Antaron / Ganex from ISP (PVP / Triacontene copolymer), Luviskol materials from BASF; polyurethanes, the Polyderm series from Alzo including but not limited to Poly derm PE / PA, Poly derm PPI-SLWS, Poly derm PPI-GH, LuvisetP.U.R. from BASF; 6) poly quaternium materials, Luviquat series from BASF; 7) acrylates copolymers and acrylates / acrylamide copolymers, Luvimer and Ultrahold series, both available from BASF; 8) styrene based materials; and 9) chitosan and chitosan based materials including cellulose and cellulose-based materials.

[0083] A composition can comprise a thickening agent or an emulsifying agent. A thickening agent may be used to increase the viscosity of liquid base materials to be used in a cosmetic composition. The selection of a particular thickening agent can depend on a type of composition desired (e.g., gel, cream, lotion, or wax based), the desired rheology, the liquid base material used, and other materials to be used in the composition. Examples of thickening agent or an emulsifying agent can include waxy materials such as candelilla, carnauba waxes, beeswax, spermaceti, carnauba, b ay sberry, montan, ozokerite, ceresin, paraffin, synthetic waxes such as Fisher-Tropsch waxes, silicone waxes (DC 2503 from Dow Corning), microcrystalline waxesWSGR Docket No. 56292-708.601and the like; soaps, such as the sodium and potassium salts of higher fatty acids, acids having from 12 to 22 carbon atoms; amides of higher fatty acids; higher fatty acid amides of alkylolamines; dibenzaldehyde-monosorbitol acetals; alkali metal and alkaline earth metal salts of the acetates, propionates and lactates; and mixtures thereof. Also useful are polymeric materials such as, locust bean gum, sodium alginate, sodium caseinate, egg albumin, gelatin agar, carrageenin gum sodium alginate, xanthan gum, quince seed extract, tragacanth gum, starch, chemically modified starches and the like, semi -synthetic polymeric materials such as cellulose, cellulose derivatives, cellulose ethers hydroxy ethyl cellulose, methyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, hydroxy propylmethyl cellulose, polyvinylpyrrolidone, polyvinylalcohol, guar gum, hydroxypropyl guar gum, soluble starch, cationic celluloses, cationic guars and the like and synthetic polymeric materials such as carboxyvinyl polymers, polyvinylpyrrolidone, polyvinyl alcohol polyacrylic acid polymers, poly(acrylic acid), carbomers, polymethacrylic acid polymers, polyvinyl acetate polymers, polyvinyl chloride polymers, polyvinylidene chloride polymers and the like. Inorganic thickeners may also be used such as aluminum silicates, such as, for example, bentonites, or a mixture of polyethylene glycol and polyethylene glycol stearate or distearate. An emulsifier may be used to help keep hydrophilic and hydrophobic ingredients from separating in an emulsion. In some cases, emulsifiers include but are not limited to Olivem, Oliwax LC, polysorbates, laureth-4, and potassium cetyl sulfate.

[0084] A cosmetic composition can provide a temporary change in an appearance or can provide a long-term change in an appearance. In some cases, a cosmetic composition can be formulated to provide a short-term change in an appearance as well as a long-term change in appearance.

[0085] A composition can comprise an additive that has an additive or synergistic effect when applied with a polypeptide as disclosed herein. For example, a composition comprising a polypeptide and an additive can have a greater effect on senescence, hair loss, hair bulk, hair density, hair thickness, hair volume, hair length, or hair growth than the individual effect of the additive, the polypeptide, or the sum of the individual effects of the additive and the polypeptide. Additivescan be an additional polypeptide, a glycosaminoglycan, a carbohydrate, a polyphenol, a protein, a lipid, a plant aqueous or oil extract, a nucleic acid, an antibody, a small molecule, a vitamin, a humectant, an emollient, or another suitable additive. In some embodiments, the composition comprises a UV blocker. In some embodiments, the UV blocker may include but is not limited to aminobenzoic acid, avobenzone, cinoxate, dioxybenzone, homosalate, meradimate, octocrylene, octinoxate, octisalate, oxybenzone, padimate O, ensulizole, sulisobenzone, titanium dioxide, trolamine salicylate, and zinc oxide.WSGR Docket No. 56292-708.601

[0086] Often the methods, systems, and compositions provided herein comprise a vitamin. In some instances, the vitamin provides skin or hair soothing, skin or hair restoring, skin or hair replenishing, and / or hydrating effects. In some instances, the vitamin provides antioxidant effects. In some instances, the vitamin acts as an emollient. In some instances, the vitamin is vitamin A, vitamin D, vitamin E, vitamin F, vitamin K, vitamin Bl (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B7 (biotin), vitamin B6, vitamin B 12 (cyanocobalamin), vitamin B9, folic acid, niacinamide, and mixtures thereof. In some instances, the composition comprises a derivative of a vitamin. In some instances, a derivative of a vitamin is used to improve stability of the vitamin in the composition and / or compatibility of the vitamin derivative with other ingredients in the composition. In some instances, the composition comprises vitamin B3 or its derivative and vitamin E or its derivative. In some instances, the composition comprises niacinamide and vitamin E or its derivative. In some instances, the composition comprises vitamin C or its derivative, vitamin B3 or its derivative, and vitamin E or its derivative. In some embodiments, the composition comprises at least 0.01%, 0.05%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% vitamin. In some embodiments, the composition comprises about 0.1% to about 10%, about 0.1% to about 5%, about 0.5% to about 10%, about 0.5% to about 5%, about 1% to 10%, or about 1% to 5% vitamin.

[0087] Compositions for topical administration can further comprise a carrier. The carrier may be a solution, an emulsion, an ointment, an oil, or a gel base. The gel base, for example, may comprise one or more of the following: petrolatum, lanolin, PEG(s), beeswax, mineral oil, diluents such as water and alcohol, emulsifiers, and / or stabilizers. Thickening agents may be present in a therapeutic composition for topical administration. If intended for transdermal administration, the composition may include a transdermal patch or iontophoresis device. In some cases, biodegradable microspheres (e.g., polylactic acid) may also be employed as carriers for a composition. In some cases, the transdermal patch is prepared to deliver the formulation to the epidermal layer of the skin having hair. In some cases, the transdermal patch is prepared to deliver the formulation to the epidermal and dermal layers of the skin having hair. In some cases, the formulation is prepared as to be minimally delivered systemically in the subject or is not intended to be delivered directly into the bloodstream of the subject.

[0088] A composition may also contain one or more diluents such as buffers, or one or more antioxidants such as ascorbic acid, low molecular weight polypeptides, polypeptides, amino acids, carbohydrates including glucose, sucrose or dextrins, chelating agents such as EDTA, glutathione and other stabilizers. Neutral buffered saline or saline mixed with nonspecific serumWSGR Docket No. 56292-708.601albumin are exemplary diluents. A product may be formulated as a lyophilizate using suitable excipient solutions (e.g., sucrose) as diluents.

[0089] A composition can also comprise one or more excipients, such as a therapeutic, nutraceutical, or cosmetic excipient. Examples of excipients can comprise antiadherents, binders, coatings, colors, disintegrants, flavors, glidants, lubricants, preservatives, sorbents, sweeteners, or vehicles.

[0090] Suitable excipients or stabilizers can be nontoxic to recipients at the dosages and concentrations employed, and can comprise buffers such as phosphate, citrate, and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid, vitamin E, and methionine; preservatives (such as, for example, octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; glucoonolactone and sodium benzoate; phenol, butyl or benzyl alcohol; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin or gelatin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA or EDTA alternatives (e.g. Biopure GLDA, Spectrastat G2); sugars such as sucrose, mannitol, trehalose, or sorbitol; salt -forming counter-ions such as sodium; metal complexes (e.g., Zn -protein complexes); and / or surfactants. In some instances, a surfactant includes, but is not limited to, polysorbate 20, polysorbate 80, alginate, a poloxamer, TRITON (t-octylphcnoxypolyethoxyethanol); nonionic detergent; sodium dodecyl sulfate (SDS); sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfohetame; lauryl-, yristyl-, linoleyl- or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl -betaine; lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-, myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine (e.g., lauroarnidopropyl); myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-dimethylamin sodium methyl cocoyl-, or disodium methyl oleyl-taurate; sorbitan monopalmitate; and the MONAQUAT series (Mona Industries. Inc., Paterson, NJ); polyethylene glycol (PEG), polypropylene glycol (PPG), copolymers of poloxy ethylene and poloxypropylene glycol (e.g., Pluronies / Poloxamer, PLURONIC® F68, etc.); or another suitable surfactant. In some cases, the composition can comprise squalene, natural oils, plant extracts, hyaluronic acid, or clay. In some cases, the plant extract comprises, Arctium Majus root extract, Morus Nigra leaf extract, Panax ginseng root extract, Pisum Sativum (pea) sprout extract, Irish moss extract, soy extract, Rehmannia Chinensis root extract, Oryza Sativa (rice) bran extract, Helianthus annuus (sunflower) sprout extract, or combinations thereof. In some cases, the composition comprises a skin penetrating enhancer to enhance the penetration of active ingredients into the skin. In some cases, the skin penetrating enhancer may include but are notWSGR Docket No. 56292-708.601limited to fatty acids, essential oils, urea, liposomes, microsphere, DMSO, azone, sodium PC A, squalane, and squalene.

[0091] In some embodiments, the composition comprises an amino acid. In some cases, the amino acid comprises tyrosine, arginine, citrulline, alanine, glycine, phenylalanine, cysteine, methionine, serine, glutamic acid, ornithine, or derivatives thereof.

[0092] In some embodiments, the formulation comprises a carrier, a microsphere, a liposome, or a micelle in order to carry the polypeptide and control the release time and / or penetration depth of the polypeptide in the through the skin.

[0093] In some embodiments, the polypeptide is functionalized. In some embodiments, the polypeptide is functionalized with a chemical group. In some embodiments, the polypeptide is functionalized with a functional group comprising no more than 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more than 20 carbons. In some embodiments, the polypeptide is functionalized with acetyl or palmitoyl.

[0094] A polypeptide or composition to be applied to a subject can be sterilized. This may be accomplished by, for example, filtration through sterile filtration membranes, or any other art-recognized method for sterilization.

[0095] A composition can comprise a therapeutically effective amount of a polypeptide or peptidomimetic in an amount which can delay the onset of, reduce the occurrence of, or ameliorate one or more symptoms, such as a symptom of hair loss, of an age-related disease or condition, or age-associated disorder. In some cases, a therapeutically effective amount can be an amount of a therapeutic agent (e.g., a polypeptide) that can provoke a therapeutic (e.g., senotherapeutic) or desired response in a subject. A therapeutically effective amount can be sufficient to cause a therapeutic benefitto the subject. The therapeutically effective amount can vary depending on a variety of factors including the active agent selected for use, and the age, weight, height, and / or general health of a subject to be treated.

[0096] As is understood in the clinical context, an effective therapeutic amount of an active agent may or may not be achieved in conjunction with an other drug, compound, therapeutic, or pharmaceutical composition. Thus, an effective therapeutic amount may be considered in the context of administering one or more active agents, and a single active agent may be considered to be given in an effective amount if, in conjunction with one or more other active agents, a desirable result may be or is achieved. Accordingly, in some instances, one or more active agents may be administered to the subject. In other instances, treatment with an active agent described herein is conducted prior to, or after, one or more other treatment modalities described herein.WSGR Docket No. 56292-708.601

[0097] In some embodiments, the composition comprises Aqua (Water), 1,2 -Hexanediol, Acetyl Tyrosine, Alpha-Glucan Oligosaccharide, Arginine, Arctium Majus Root Extract, Biotin, Calcium Pantothenate, Cellulose Gum, Chondrus Crispus, Citric Acid, Citrulline, Decapeptide-52 (Peptide 14), Erythritol, Fisetin, Glycerin, Glucosamine HC1, Hydrolyzed Soy Protein, Hydroxyacetophenone, Inulin, Morus Nigra Leaf Extract, Niacinamide, Ornithine HC1, Oryza Sativa (Rice) Bran Water, Panax Ginseng Root Extract, Panthenol, PEG-12 Dimethicone, Phenoxyethanol, Pisum Sativum (Pea) Sprout Extract, Polyquatemium-11, Potassium Sorbate, Propanediol, Raspberry Ketone, Rehmannia Chinensis Root Extract, Sodium Benzoate, Sodium PCA, Tetrasodium Glutamate Diacetate, Xanthan Gum, Helianthus annuus (sunflower) sprout extract, or combinations thereof. In some embodiments, Panax ginseng root extract may stimulate blood flow and boost circulation to the scalp, which may provide hair follicles with better access to nutrients. In some embodiment, Fisetin may protects hair follicles from oxidative stress and inflammation. In some embodiments, Oryza sativa (rice) bran water may strengthen, smooth, and repair hair and / or act as a mild scalp exfoliator to help promote an optimal environment for hair growth. In some embodiments, Helianthus annuus (sunflower) sprout extract may protect against UV-induced oxidative stress, which may help maintain hair density, vitality, and overall hair health. In some embodiments, biotin may support keratin production, strengthen the hair shaft, and / or reduce breakage, which may promote fuller, thicker hair. In some embodiments, inulin and / or alpha-glucan oligosaccharide may promote a balanced scalp microbiome, which may be essential for scalp health and resilient hair growth.Polypeptide Synthesis

[0098] Also disclosed are isolated polynucleotides encoding one or more of the presently disclosed polypeptides. The isolated polynucleotides may be present in an expression vector comprising the isolated polynucleotides operably linked to a promoter. The expression vector may be present in an isolated cell (i.e., a recombinant cell transfected or transformed with the expression vector).

[0099] Suitable expression vectors may include bacterial, plant, fungal, insect, or animal host cell replication, and / or expression vectors that express the disclosed peptides, polypeptides, and variants thereof. Expression vectors may be used to transform appropriate host cells (e.g., E. colt). The transformed host cell may be cultivated or fermented such that the peptide or polypeptide is expressed constitutively or after adding a reagent that induces expression (e.g., via an inducible promoter). Expression vectors as contemplated herein may include control sequences that modulate expression of the encoded polypeptide. Expression control sequencesWSGR Docket No. 56292-708.601may include constitutive or inducible promoters (e.g., T3, T7, Lac, trp, or phoA), ribosome binding sites, or transcription terminators.

[0100] An expression vector can be utilized to transform host cells. Suitable host cells include bacterial, plant, fungal, insect, or animal host cells. Suitable bacteria include, but are not limited to: Gram-negative bacteria such as Escherichia species (e.g., E. colt), other Gramnegative bacteria, (e.g., Pseudomonas sp., such as Pseudomonas aeruginosa, or Caulobacter sp., such as Caulobacter crescentus), or Gram-positive bacteria (e.g., Bacillus sp., such as Bacillus subtilis). Suitable fungal cells may include yeast (e.g., Saccharomyces cerevisiae),

[0101] An expression vector can, for example, provide a mechanism for synthesis of a polypeptide. Synthesis can take place in a cell, for example an animal cell, a plant cell, a bacterial cell, or a yeast cell. An expression vector can comprise nucleic acids, e.g., DNA derived from a plasmid, cosmid, phasmid, or bacteriophage or synthesized by chemical or enzymatic means, into which one or more fragments of nucleic acid may be inserted or cloned which can encode one or more polypeptides described herein. An expression vector may be capable of autonomous replication in a defined host or organism such that the cloned sequence is reproduced. An expression vector can have a linear, circular, or supercoiled configuration and may be complexed with other vectors or other materials for certain purposes. The components of an expression vector can comprise, but are not limited to, a DNA molecule incorporating: (1) DNA; (2) a sequence encoding a therapeutic or desired product; or (3) regulatory elements for transcription, translation, RNA stability, and replication.

[0102] A polypeptide can be produced using an expression vector. In some cases, such production can comprise cultivating or fermenting a transformed host cell (e.g., a bacterial host cell as contemplated herein) which comprises an expression vector (as contemplated herein) which in turn comprises a nucleic acid molecule encoding the disclosed peptides, polypeptides, or variants thereof (as contemplated herein), wherein cultivation occurs under conditions which cause expression of the peptides, polypeptides, or variants; and isolating, separating, or purifying the peptides, polypeptides, or variants. The transformed bacteria may be cultivated or fermented using methods known in the art in order to express the peptides, polypeptides, or variants. An exemplary isolation, separation, or purification method may include one or more of the following steps: a cell disruption step, a clarification step (e.g., via centrifugation or filtration), a chromatographic separation step, a dialysis step, and a precipitation step.

[0103] In some other embodiments, the polypeptide can be chemically synthesized. Synthesis of a polypeptide can be performed using solution -phase techniques, solid-phase methods, or other suitable methods of polypeptide synthesis.WSGR Docket No. 56292-708.601Methods

[0104] Provided are methods for the use of the polypeptides and compositions disclosed herein. Such methods can comprise application of one or more of the polypeptides described herein to a subject. Methods described herein may increase or maintain hair bulk or may maintain or stimulate hair growth in a subject. Methods described herein may increase or maintain hair density, hair thickness, hair volume, or hair length in a subject. Methods described herein may delay the onset of, reduce the occurrence of, or ameliorate hair loss. Methods described herein may result in improvements in hair quality (e.g., hair health).

[0105] A polypeptide or composition canbe applied or administered to a subject to treat a condition directly or indirectly causing hair loss. A polypeptide or composition can be applied or administered to a subject as a treatment to increase or maintain hair bulk or to maintain or stimulate hair growth. A polypeptide or composition can be applied or administered to a subject as a treatment to increase or maintain hair density, hair thickness, hair volume, or hair length. A polypeptide or composition can be applied or administered to a subject as a treatmentto improve hair quality. Such methods can comprise administering to the subject a compound that promotes increased hair bulk or hair growth or applying topical treatment to a skin having hair.

[0106] In some embodiments, the compositions described herein are administered once per day, twice per day, three times per day or more. In some embodiments, the compositions described herein are administered twice daily administration, e.g., morning and evening. In some embodiments, the compositions described herein are administered daily, every day, every alternate day, five days a week, once a week, every other week, two weeks per month, three weeks per month, once a month, twice a month, three times per month, or more. In some embodiments, the compositions described herein are administered for at least 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 18 months, 2 years, 3 years, 4 years, 5 years, or longer. In some embodiments, the compositions described herein are administered daily to once a week. In some embodiments, the composition is directed to be applied directly to a dry or towel dried scalp or skin having hair in the morning and the evening. In some embodiments, formulation is a daily essential topical supplement scientifically formulated to improve or maintain hair bulk or hair growth. In some embodiments, formulation is a daily essential topical supplement scientifically formulated to improve or maintain hair density, hair thickness, hair volume, or hair length. In some embodiments, the user applies the compositions described herein comprising at least one of the polypeptides directly the scalp or skin having hair. In some embodiments, the compositions described herein is used in conjunction with other topical compositions, such as a UV blocker. In some embodiments, the compositions described herein isWSGR Docket No. 56292-708.601applied before, together with, or after the application of the other topical composition. In some embodiments, the compositions described herein are applied before or after use of a dermaroller, also referred herein as microneedling. In some embodiments, the dermaroller is rolled at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times in all directions. In some embodiments, the dermaroller is lifted between passes. In some embodiments, the compositions described herein are applied 1, 2, 3, 4, 5, 6, or 7 days before the use of a dermaroller. In some embodiments, the compositions described herein are applied 1, 2, 3, 4, 5, 6, or 7 days after the use of a dermaroller. In some embodiments, the compositions described herein are applied before or after use of another treatment. In some embodiments, the compositions described herein are applied 1, 2, 3, 4, 5, 6, or 7 days before another treatment. In some embodiments, the compositions described herein are applied 1, 2, 3, 4, 5, 6, or 7 days after another treatment. In some embodiments, the compositions described herein are massaged into the scalp using fingertips following administration.

[0107] The methods provide herein can comprise topical application of a polypeptide or composition. In some embodiments, topical application can comprise rubbing, spraying, dipping, dabbing, or otherwise applying a polypeptide or composition to skin having hair or scalp, or a combination thereof.

[0108] In some instances, the treatment may increase or maintain hair bulk, maintain or stimulate hair growth, or may reduce hair loss. In some instances, the treatment may increase or maintain hair density, hair thickness, hair volume, or hair length. In some instances, the treatment may increase proliferation, decrease miniaturization, or decrease senescence of hair follicle cells (e.g., outer root sheath cells, inner root sheath cells, or dermal papilla cells).

[0109] In some cases, the methods, systems, and compositions provided herein may decrease senescence of the outer root sheath (ORS) of the hair follicle. In some instances, decreasing senescence of the outer root sheath results in increased or maintained hair bulk or hair growth. In some instances, decreasing senescence of the outer root sheath results in increased or maintained hair density, hair thickness, hair volume, or hair length. In some instances, decreasing senescence of the outer root sheath results in decreased or maintained hair loss.

[0110] In some cases, the methods, systems, and compositions provided herein may decrease senescence of the dermal papilla. In some instances, decreasing senescence of the dermal papilla cells results in increased or maintained hair bulk or hair growth. In some instances, decreasing senescence of the dermal papilla cells results in increased or maintained hair density, hair thickness, hair volume, or hair length. In some instances, decreasing senescence of the dermal papilla cells results in decreased or maintained hair loss.WSGR Docket No. 56292-708.601

[0111] In some instances, the treatment results in the increase in hair bulk or hair growth or decreased hair loss by, for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 99%. In some instances, the treatment results in the increase hair density, hair thickness, hair volume, or hair length by, for example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 99%.

[0112] In some instances, hair growth, hair loss, or hair quality is assessed by a measurement of hair density, hair quantity, or hair length. In some instance, hair bulk is assessed by measurement of hair density, hair thickness, hair volume, or hair length. In some instances, the measurement of hair density, hair thickness, hair volume, hair quantity, or hair length is assessed through a phototrichogram, HairMetrix® analysis, visual inspection, ponytail circumference, daily hair count, dermoscopy, hair weight, contrasting felt examination, surface electron microscopy, trichogram, pull and tug examination, or scalp biopsy. A phototricogram may measure hair density and the growth phase of each hair. A HairMetrix® analysis may measure the number of healthy hairs (e.g., in anagen phase), the number of miniaturized hairs (e.g., vellus hairs), the ratio of healthy hairs to vellus hairs, hair density, and / or inter follicular mean distance. In some instances, root strength or hair shedding is measured by a tensile strain / stress test or a pull and tug examination. In some instances, hair density may be the number of hairs per unit area on the scalp or skin having hair.

[0113] In some instances, hair loss or hair growth is assessed by a measurement of a blood component level or gene or protein expression level. In some instances, the blood component may be prolactin, testosterone, DHEA, luteinizing hormone or follicular stimulating hormone. In some instances, the measured gene or protein may be P-galactosidase, CDKN1A (p21), IL-6, ATRX, pl6, IL-8, Ki-67, pRB, Bmi-1, androgen receptor, 5-alpha reductase, or prostaglandin synthase. In some instances, the gene or protein expression level is accessed in a biopsied hair sample.

[0114] In some instances, hair loss, hair growth, or hair quality is assessed by DNA methylation. In some instances, DNA methylation patterns may be used to assess cellular senescence or cell aging of hair follicle cells.

[0115] In some instances, a measurement shows an improvement in at least one of hair density, hair quantity, hair length, hair shedding, root strength, overall appearance, blood component level, gene or protein expression level or other metric of hair quality by at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% after the use of the composition as compared to before the use of the composition. In some instances, the improvement is presented as a mean percentage improvement (MPI) compared to baselinebefore the use of the composition. In some instances, MPI is at least 1%,WSGR Docket No. 56292-708.6012%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% after the use of the composition as compared to before the use of the composition. In some instances, the measurements are taken at 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 5 months, 6 months, or one year after the use of the composition. In some instances, the effect of the treatment is assessed by an expert in hair conditions, disorders, or diseases who analyzes one or more of the hair measurements. In some instances, the effect of the treatmentis assessedby the user themselves. In some instances, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the users may report an improvement in at least one of hair density, hair quantity, hair length, hair shedding, root strength, overall appearance, blood component level, gene or protein expression level or other metric of hair quality after using the methods, systems, and compositions disclosed herein.

[0116] In some cases, the methods, systems, and compositions provided herein may increase hair bulk or new hair growth. In some cases, the methods, systems, and compositions provided herein may increase or maintain hair density, hair thickness, hair volume, or hair length. In some cases, the methods, systems, and compositions provided herein may increase growth rate of existing hair.

[0117] In some cases, the methods, systems, and compositions provided herein may increase the number of hairs in anagen phase on a subject. In some cases, the methods, systems, and compositions provided herein may decrease the number of hair follicles in telogen phase on a subject. In some cases, the methods, systems, and compositions provided herein may decrease the number of hair follicles in exogen phase on a subject. In some cases, the methods, systems, and compositions provided herein may decrease the number of vellus hairs on a subject. In some cases, the methods, systems, and compositions provided herein may increase the ratio of healthy hairs (e.g., in anagen phase) to miniaturized hairs (e.g., vellus hairs) on a subject. In some cases, the methods, systems, and compositions provided herein may decrease the inter follicular mean distance of hair follicles on a subject.

[0118] In some cases, the methods, systems, and compositions provided herein may increase the length of time that a hair follicle is in anagen phase. In some cases, the methods, systems, and compositions provided herein may decrease the length of time that a hair follicle is in telogen phase. In some cases, the methods, systems, and compositions provided herein may decrease the length of time that a hair follicle is in exogen phase.

[0119] In some cases, the methods, systems, and compositions provided herein may promote growth, increase number, or reduce death of hairs in anagen phase. In some cases, theWSGR Docket No. 56292-708.601methods, systems, and compositions provided herein may promote growth, increase number, or reduce death of hairs in anagen phase more than vellus hairs.

[0120] In some cases, the methods, systems, and compositions provided herein may counteract the effect of CRH. In some embodiments, the peptides provided herein decrease cellular senescence. In some embodiments, the peptides provided herein partially or fully counteract the effect of CRH on cellular senescence. In some embodiments, the peptides provided herein decrease cellular senescence more than CRH increased cellular senescence. In some embodiments, the peptides provided herein decrease hair follicle miniaturization. In some embodiments, the peptides provided herein partially or fully counteract the effect of CRH on hair follicle miniaturization. In some embodiments, the peptides provided herein decreases hair follicle miniaturization more than CRH increases hair follicle miniaturization. In some embodiments, the peptides provided herein increase proliferation of hair follicle cells. In some embodiments, the peptides provided herein partially or fully counteract the effect of CRH on proliferation of hair follicle cells. In some embodiments, the peptides provided herein increase proliferation of the hair follicle cells more than CRH decreases proliferation. In some embodiments, the peptides provided herein increase hair bulk or hair growth. In some embodiments, the peptides provided herein partially or fully counteract the effect of CRH on hair bulk or hair growth. In some embodiments, the peptides provided herein increase hair bulk or hair growth more than CRH decreases hair bulk or hair growth. In some embodiments, the peptides provided herein partially or fully counteract the effect of CRH on hair density, hair thickness, hair volume, or hair length. In some embodiments, the peptides provided herein increase hair density, hair thickness, hair volume, or hair length more than CRH decreases hair density, hair thickness, hair volume, or hair length.

[0121] In some cases, the methods, systems, and compositions provided herein may reduce inflammation. In some cases, the methods, systems, and compositions provided herein may reduce expression levels of proteins involved in inflammation, interferon -gamma (IFN-y) and interleukin 10 (IL- 10), by the cells in the treated skin having hair.

[0122] In some cases, the methods, systems, and compositions provided herein may improve hair quality. In some cases, the methods, systems, and compositions provided herein may improve overall hair health. In some cases, the methods, systems, and compositions provided herein may increase hair fullness. In some cases, the methods, systems, and compositions provided herein may increase nourishment of the scalp.

[0123] In some cases, the methods described herein can delay the onset of, reduce the occurrence of, reduce the appearance of, or ameliorate a disease, a disorder, or a condition associated with the accumulation of senescent cells. A disease or disorder associated with theWSGR Docket No. 56292-708.601accumulation of senescent cells can be age related. In some cases, the disease or disorder can worsen overtime if untreated. An age-associated disorder can comprise a biological progression of events that occurs during a disease process that can affect the body, which can mimic or substantially mimic all or part of the aging events which occur in a normal subject. In some cases, this biological progression of events can occur over an accelerated time frame. An age-related disease or condition or age-associated disorder can relate to regular processes in the body, such as movement and eating capacity. In some cases, the age-related disease or condition or age-associated disorder can be a disease, condition, or disorder affecting hair growth, such as androgenic alopecia, alopecia areata, telogen effluvium, anagen affluvium, pattern hair loss, tinea capitis, trichotillomania, anagen effluvium, thyroid disease, lupus, or scalp aging. In some embodiments, the age-related disease or condition or age-associated disorder is male pattern baldness. In some embodiments, the male pattern baldness results in high temples, thinning at the crown of the head, and / or balding at the crown of the head.

[0124] An age-related disease or condition or age-associated disorder can be caused by UV damage, DNA damage, ATRX foci accumulation in cell nuclei, increased pl6 expression, increased senescence-associated P -galactosidase activity, accumulation of senescent cells in the tissue, increased SASP production, chemically induced senescence, chronological aging, decreased hyaluronic acid production, decreased expression of sirtuin 6, altered insulin -like growth factor- 1 (IGF-I) pathway signaling, increased production of matrix metallopeptidase 1 (MMP1), or genetic variants. In some instances, the age-related disease or condition or age-associated disorder is initiated or exacerbated by a therapeutic regimen, for example, a side effect of a therapeutic drug. An age-related disease or condition or age-associated disorder can affect the health or appearance of hair directly or indirectly. Topical application of a polypeptide or composition herein can improve the health or appearance of hair in some such cases.

[0125] An age-related disease or condition or age-associated disorder can comprise a cell proliferative disorder. A cell-proliferative disorder can affect the health or appearance of the hair. In some cases, a treatment administered for a cell -proliferative disorder, such as chemotherapy or radiation can affect the health or appearance of the hair. Topical application of a polypeptide or composition herein can improve the health or appearance of hair in some such cases.

[0126] Also provided herein are methods for treating the skin having hair of a subject comprising administering to a subject a composition that can promote a decrease in a number of senescent cells in a tissue or organism, inducing a pro-apoptotic state in the treated cells, inducing SIRT6 expression, preventing DNA-induced senescence, and / or enhancing DNA repair capacity. In some cases, a disease or condition can comprise hair loss, accumulation of senescentWSGR Docket No. 56292-708.601cells in the tissue, decreased hair density, decreased collagen production, increased MMP-1 production, decreased DNA repair capacity, decreased SIRT6 expression, inflammation, a senescence-associated secretory phenotype, or stem cell exhaustion of the hair follicle.

[0127] Methods can comprise administering to the subject a composition comprising a polypeptide that can promote a significant decrease in the number of senescent cells in the tissue or organism. A decrease in the number of senescent cells can comprise a pro-apoptotic state in the treated cells, inducing SIRT6 expression, preventing DNA-induced senescence, or enhancing DNA repair capacity. In some cases, the number of senescent cells in a sample, a portion of a subject (e.g., the scalp of a subject), and / or a subject can be reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In some cases, the senescence is assessed by a senescence associated P -galactosidase activity level, a ratio of ATRX foci / cell, pl 6 expression, IL-8 expression, or Ki -67 expression.

[0128] In cases where the subject is a human, the subject may be of any age. In some cases, the subject has an age-related disease or condition or age-associated disorder, is at risk for an age-related disease or condition or age-associated disorder, or is healthy. In some cases, the subject has hair loss. A subject can be male or female.

[0129] A polypeptide or compositions can be applied or administered to a cell, tissue, or subject. In some cases, application or administration of a polypeptide can result in a senotherapeutic effect in a cell, tissue, or subject. In some cases, a polypeptide can be administered to a subject, applied topically to a subject, or incubated with cultured cells to provide a senotherapeutic effect.

[0130] A cell can be a cultured cell or a cell isolated from a subject or from a cell line. Some examples of cultured cells can comprise a keratinocyte or a fibroblast or a melanocyte. A cell can be wild type or can be genetically modified. Some genetic modifications can promote senescence, such as genetic modifications in the p53 / p21 pathway, the pl6 / RB pathway, the, a mRNA or miR gene, among other RNA classes. In some cases, application of a polypeptide or composition to a cell can reduce senescence in the cell. In some cases, cells may comprise cells in vivo or in situ in an organism, including but not limited to animals, C. elegans, and humans.

[0131] A tissue can be a tissue which is a tissue of a subject or a tissue which has been isolated from a subject, i.e., ex vivo. In some cases, a tissue can be artificially grown. A tissue can comprise skin having hair, and examples of tissues can comprise healthy, diseased, or aged skin having hair, or healthy, diseased, or aged scalp. In some cases, application of a polypeptide or composition to a tissue can reduce senescence in one or more cells of the tissue or the entire tissue. In some cases, tissue may comprise tissue in vivo or in situ in an organism, including but not limited to animals, C. elegans, and humans.WSGR Docket No. 56292-708.601EXAMPLESExample 1

[0132] Primary fibroblasts isolated from Progeria patients can constitute a genetic model of early aging and cellular senescence in humans. Primary fibroblasts from Progeria patients were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96-well plates (1,000 cells per well) and, 6 hours after plating, were incubated with individual polypeptides from a proprietary library at 50 pM for 48 hours. Negative control comprised untreated cells, which received vehicle only; positive control group comprised cells incubated with 10 pM ABT-263, a senolytic compound for the same period. After incubation, relative cellular senescence (assessed by the activity of senescence associated B -galactosidase staining relative to untreated control) was analyzed as shown in FIG. 1, in which the Y axis indicates the total number of cells in the well (normalized to the untreated control), and the X axis represents senescence associated B-galactosidase staining intensity / nuclei (i.e., senescence level), also normalized to negative control. Three independent experiments including three technical replicates were performed. Polypeptides which promoted a significant decrease of cellular senescence to below 75%that of the untreated control sample were considered positive hits.

[0133] A total of 764 polypeptides were tested, among which 56 promoted a decrease in cellular senescence to below 75% that of the untreated control sample. Therefore, they were considered as positive hits, and putative senotherapeutic compounds. ABT-263, which was considered a positive control in the experiment, also promoted significant reduction of cellular senescence, but also cellular toxicity. This observation confirmed the senolytic characteristic of ABT-263, as well as the senotherapeutic potential of some of the tested polypeptides (FIG. 1). Example 2

[0134] Primary fibroblasts isolated from 3 healthy chronologically aged patients were used. Cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells were keptin a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96-well plates (4,000 cells per well) and, 6 hours after plating the cells were treated with one of 4 senotherapeutic polypeptides (Peptide 14, Peptide 13, Peptide 15, and Peptide 16) and incubated for 48 hours. Each polypeptide was tested at 6 different concentrations: 50 pM, 25 pM, 12.5 pM, 6.25 pM, 3.12 pM, and 1.56 pM, except for PeptideWSGR Docket No. 56292-708.60116, which was tested at 5 different concentrations: 25 pM, 12.5 pM, 6.25 pM, 3.12 pM, and 1.56 pM. A negative control comprised untreated cells, which received vehicle only. After incubation, relative cellular senescence (assessed by the activity of senescence associated B-galactosidase staining relative to untreated control) was analyzed (FIG.2, panels A-D), in which the Y axis indicates the relative senescence levels normalized to untreated control. Each column corresponds to a different concentration of a polypeptide. Three independent experiments (biological replicates) including three technical replicates were performed. Data was analyzed using Analysis of Variance (ANOVA) and a Bonferroni post -hoc tests. Statistical significance was determined as p values equal or lower than 0.05.

[0135] All polypeptides presented senotherapeutic potential in at least one of the tested concentrations, evidenced by the significant reduction of cellular senescence compared to untreated control. *p<0.05; **p<0.01; ***p<0.001 compared to untreated control (ctrl) (FIG. 2, panels A-D).Example 3

[0136] ATRX is a chromatin remodeling enzyme, which contributes to the formation of senescence associated heterochromatic foci. It increasingly accumulates in nuclear foci during senescence. Therefore, it constitutes a marker of cellular senescence. To investigate whether Peptide 14 decreased levels of cellular senescence, ATRX foci were analyzed in Peptide 14 treated (1 pM, 500 nM, 100 nM, and 10 nM) and untreated cells. To do so, primary fibroblasts isolated from 3 healthy chronologically aged (elder) donors were used. These cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS) and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96-well plates (4,000 cells per well) and, 6 hours after plating, were incubated for 48 hours with the Peptide 14 polypeptide in the aforementioned concentrations. A negative control comprised untreated cells, which received vehicle only. After incubation, relative cellular senescence was assessed. Briefly, immunostaining was performed by fixing, permeabilizing, and incubating the cells with anti-ATRX antibody, followed by secondary antibody. The number of nuclei and stained ATRX foci were counted. Panel A of FIG. 3 shows a representative graph showing the number of cells (Y axis) presenting a specific amount of ATRX foci / cell, represented as columns (X axis). The upper graph depicts untreated cells, while the lower graph depicts cells treated with Peptide 14 at 500 nM. Panel B of FIG. 3 shows the average number of ATRX foci / nuclei of fibroblasts treated with different conditions of Peptide 14 (columns). Panel C of FIG. 3 shows the percentage of cells presenting less than 10 ATRX foci / nuclei among fibroblasts treated with different conditions of Peptide 14 (columns). Three independentWSGR Docket No. 56292-708.601experiments (biological replicates) including three technical replicates were performed. Data in panels B and C of FIG. 3 were analyzed using ANOVA and Bonferroni post-hoc tests.Statistical significance was determined as p values equal or lower than 0.05.

[0137] Peptide 14 treatment significantly decreased ATRX foci / nuclei, when used at 500 nM and 50 nM, compared to untreated cells. In the same concentrations, Peptide 14 also increased the number of cells which presented less than 10 foci / nuclei. *p<0.05; **p<0.01 compared to untreated control (ctrl) (FIG. 3).Example 4

[0138] Human primary fibroblasts isolated from 3 healthy chronologically aged (elder) donors were used. These cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in T-75 flasks (250,000 cells per flask) and, 6 hours after plating, were incubated for 3 weeks (21 days) with Peptide 14 at 3.12 pM. A negative control comprised untreated cells, which received vehicle only. Day 0 was defined as the day cells started to be treated with Peptide 14. No Peptide 14 treatment was performed between days 21 and 28. Weekly, cellular senescence was assessed according to senescence associated P-galactosidase staining levels. Data was normalized to untreated group and plotted (FIG. 4, panel A). Cellular proliferation was also determined weekly. At days 7, 14, 21, and 28, cells were trypsinized and counted (FIG. 4, panel B). After counting, 250,000 cells were plated in new T-75 flasks. Three independent experiments (biological replicates) including three technical replicates were performed. Data was analyzed using T-test. Statistical significance was determined as p values equal or lower than 0.05.

[0139] Peptide 14 promoted a significant decrease (***p<0.001; ****p<0.0001) of cellular senescence beginning at the second week. After 21 days of treatment, the senotherapeutic effect of Peptide 14 was maintained for at least 7 days after polypeptide removal (between experimental days 21 and 28). No significant differences were observed regarding cell proliferation comparing Peptide 14 treated group and untreated control (FIG. 4).Example 5

[0140] Primary fibroblasts isolated from 7 healthy chronologically aged patients were used (patients were aleatory identified as patient 2, 3, 4, 5, 6, 7, or 8). These cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96-well plates (4,000 cells per well) and, 6 hours after plating, were incubated for 48 hours withWSGR Docket No. 56292-708.601Peptide 14 at 5 different concentrations: 25 pM (concentration 5), 12.5 pM (concentration 4), 6.25 pM (concentration 3), 3.12 pM (concentration 2), and 1.56 pM (concentration 1). A negative control comprised untreated cells (concentration 0), which received vehicle only. After incubation, relative cellular senescence was determined according to the average number of ATRX foci / nuclei quantified f oilowing ATRX immunostaining. Seven independent experiments (biological replicates) including three technical replicates were performed. Data was analyzed using a covariance test. Statistical significance was determined as p -values equal or lower than 0.05 (FIG. 5).

[0141] Covariance analysis shows that the number of ATRX foci / nuclei was significantly reduced following Peptide 14 treatment. Peptide 14 efficacy followed a doseresponse pattern, with concentration and ATRX foci / nuclei being significantly correlated (p<0.0004) (FIG. 5).Example 6

[0142] Cellular senescence may be caused by several different stimuli. In order to assess whether Peptide 14 was effective against UVB -induced and chemically -induced senescence, human primary fibroblasts isolated from 3 healthy donors were used. These cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96-well plates (4,000 cells per well) and, 6 hours after plating, were submitted to either etoposide (20 pM) treatment for 24 hours, or exposed twice to 0.05 J / cm2of UVB radiation. Each UVB exposure corresponds to approximately 1 to 3 hours of daily sun exposure in April of major cities around the world (e.g. Auckland, NZ; Los Angeles, US; and Brasilia, BR). After the different senescence induction protocols, etoposide treated cells were incubated with Peptide 14 at 5 pM, 2.5 pM, or 1 pM for 48 hours. UVB exposed cells were treated with Peptide 14 at 5 pM for 48 hours. A negative control comprised untreated cells, which were submitted to stress, but received vehicle only. After incubation, relative cellular senescence (assessed by the activity of senescence associated P -galactosidase staining relative to untreated control) was analyzed and plotted in column graphs. ATRX foci were also assessed following ATRX immunofluorescence staining. Graphs were built using the average ATRX foci detected per nuclei. Three independent experiments (biological replicates) including three technical replicates were performed. Data was analyzed using with either t-test or ANOVA followed by a Bonferroni post -hoc test. Statistical significance was determined as p-values equal or lower than 0.05 (FIG. 6).

[0143] Etoposide treatment promoted a significant increase in cellular senescence level (p<0.001), and also a significant increase of ATRX foci nuclear accumulation (represented asWSGR Docket No. 56292-708.601the average number of ATRX foci / cell; p<0.05) (FIG. 6, panel A). When etoposide-stressed cells were treated with 2.5 pM or 5 pM of Peptide 14, senescence associated P -galactosidase staining was significantly reduced (*p<0.05) (FIG. 6, panel B, left graph). Average ATRX foci / cell was also significantly reduced when etoposide-exposed cells were treated with 2.5 pM Peptide 14, as shown in the right graph of panel B of FIG. 6. UVB exposure also promoted significant increase in cellular senescence, as assessedby senescence associated P-Galactosidase staining, 5 pM Peptide 14 treatment being able to significantly prevent cellular senescence (*p<0.05), as shown in the left graph of panel C of FIG. 6. Treatment with Peptide 14 did not significantly alter cell number, as shown in the right graph of panel C of FIG.6. UVB exposure promoted significant increase in the average number of ATRX foci per nuclei, and 5 pM Peptide 14 treatment significantly prevented cellular senescence, leading to significantly reduced ATRX foci / nuclei, compared to UVB treated samples which did not receive Peptide 14 (*p<0.01) (FIG.6, panel D).Example 7

[0144] Human primary fibroblasts and keratinocytes isolated from healthy elder donors were used to build human skin equivalents. Those skin equivalents were treated with 0.01% w.v. Peptide 14 for 5 days. Skin equivalents were characterized according to the expression of specific genes by reverse transcription-quantitative polymerase chain Reaction (RT-qPCR). Following treatment, epidermis and dermis were processed for RT-qPCR separately. For epidermis samples, glyceraldehyde 3 -phosphate dehydrogenase (GAPDH; ubiquitously expressed); pl 6 (associated to senescence), IL-8 (linked to irritation), and Ki -67 (associated to cell proliferation) were analyzed. For dermis samples, glyceraldehyde 3 -phosphate dehydrogenase (GAPDH; ubiquitously expressed); p 16 (associated to senescence), IL-8 (linked to irritation), Ki-67 (associated to cell proliferation); hyaluronic synthase 2 (HAS-2; associated with hyaluronic acid production), and matrix metalloprotease 1 (MMP1; associated to extracellular matrix protein degradation) were analyzed. CT values were analyzed using the 2’ AAcimehod. Average mRNA expression was normalized to GAPDH (ACt) and to the negative control group (AACt). Negative controls received formulation only. Three independent experiments were performed with three technical replicates. Data was analyzed using T-test. Statistical significance was determined as p-values equal or lower than 0.05 (FIG. 7)

[0145] Peptide 14 treatment led to significant decrease of pl 6 in epidermis (****p<0.0001) and dermis (**p<0.01); decreased expression of IL-8 in the dermis (*p<0.5); and decreased expression MMP-1 in the dermis (**p<0.01). The data corroborates the senotherapeutic potential of Peptide 14, safety and tolerability, as well as the beneficial effects of the polypeptide to skin gene expression, as shown in FIG. 7.WSGR Docket No. 56292-708.601Example 8

[0146] In order to shed light into the mechanism of action of Peptide 14 and similar Peptide 13, Akt S473 phosphorylation (FIG.8, panel A), senescence associated P -galactosidase staining (FIG. 8, panel B), as well as mRNA expression (FIG. 8, panel C) were investigated. For Akt S473 phosphorylation analysis using western blotting, human primary fibroblasts and keratinocytes were used. These cells were used to build human skin equivalents, which were kept in a 5% CO2, 37 °C and 95% humidity atmosphere air liquid interface. Then, skin equivalents were treated with IpM of either Peptide 14 or Peptide 13 for 5 days, and protein analysis was performed on the skin equivalents. Protein was isolated and quantified. Equal amounts of protein were loaded in polyacrylamide gels and transferred into a nitrocellulose membrane. GAPDH (loading control) and pAkt S473 antibodies were incubated with the membrane and staining was revealed by chemiluminescence. Relative pAkt S473 / GAPDH signal was compared between treated and non-treated samples. For senescence associated B-galactosidase staining experiments, fibroblasts were used. These cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) culture supplemented with 10% v.v. fetalbovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96 -well plates (4,000 cells per well) and incubated with basal medium for 6 hours to allow cell attachment. After, cells were exposed twice to 0.05 J / cm2. This was followed immediately by a second incubation, wherein Peptide 14 or Peptide 13 were added to the medium and left for 48 hours, when medium was changed, and cells were stained for senescence associated B-galactosidase. Untreated cells were incubated with vehicle only as negative controls (-). Relative staining was obtained after normalizing untreated control senescence associated B -galactosidase levels to 100%. For mRNA analysis, fibroblasts were used. These cells were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in either 6 -well plates (50,000 cells per well) and incubated with basal medium for 6 hours to allow cell attachment. After, cells were incubated for 48 hours with either Peptide 14 or Peptide 13. A negative control comprised untreated cells, which received vehicle only. Total RNA was isolated, samples were reverse transcribed, and mRNA expression of GAPDH, sirtuin 6 (SIRT6), BLM, and exonuclease 1 (EXO1) genes were determined using qPCR. Negative controls received vehicle only. CT values were analyzed using the 2'AAClmethod. Average mRNA expression was normalized to GAPDH (ACt) and to the negative control group (AACt). For all analysis, three independent experiments were performed with three technical replicates.WSGR Docket No. 56292-708.601Data was analyzed using a T-test. Statistical significance was determined as p values equal or lower than 0.05 (FIG. 8).

[0147] pAkt S473 was significantly decreased in both epidermal, as well as dermal samples treated with Peptide 14 (*p<0.05, and **p<0.01, respectively). Peptide 13 decreased pAkt S473 in dermal samples only (***p<0.001) (FIG. 8, panel A). For UVB and senescence associated B-galactosidase staining, it was observed that staining always increased following UVB exposure. Additionally, both Peptide 14 and Peptide 13 decreased staining in UVB-exposed samples (***p<0.001). Peptide 14 specifically led to increased SIRT6 and BLM expression in treated samples (*p<0.05) (FIG. 8).Example 9

[0148] Predicted three-dimensional structures of polypeptides having the amino acid sequence ETAKHWLKGI (SEQ ID NO:1) and ATAKAWLKGI (SEQ ID NO:2) were determined in water. The structure predictions are shown in FIG.9, panel A (SEQ ID NO: 1) and FIG. 9, panel B (SEQ ID NO:2). The structures were superimposed (FIG. 9, panel C) to illustrate the resemblance of the structures.Example 10A topically -applied formulation of Peptide 14 is made including niacin, vitamin E, at least one preservative, at least one emulsifier and between 50-150 pM Peptide 14. The topical formulation is applied to human scalp, resulting in increased hair growth.Example 11

[0149] A topically-applied formulation of Peptide 14 is made including niacin, vitamin E, at least one preservative, at least one emulsifier and between 75-100 pM Peptide 14. The topical formulation is applied to human scalp, resulting in increased hair growth.Example 12

[0150] An exemplary topically-applied formulation is shown below in Table 4. The topical formulation is applied to human scalp, resulting in increased hair density, hair quantity, hair length, root strength, and overall appearance as well as decreased hair shedding. The polypeptides in the formulation comprises at least one of the polypeptides disclosed herein.WSGR Docket No. 56292-708.601Example 13

[0151] An exemplary topically-applied formulation comprising Peptide 14 is shown below in Table 5. The topical formulation is applied to human scalp, resulting in the reduction of at least one of increased hair density, hair quantity, hair length, root strength, and overall appearance as well as decreased hair shedding. The polypeptides in the formulation comprises Peptide 14.Example 14.

[0152] To study the level of penetration by Peptide 14 through the depth of the scalp having hair, a diffusion study is performed using Franz cells and on ex vivo scalp culture sample. Fresh human scalp tissue of a female donor (79 years old) is cutin small pieces ~2.5cm x 2.5cm.

[0153] In the Franz cell study, the scalp tissue is treated with 10 pL of a formulated cream comprising 0.01% Peptide 14 and placed in a Franz cell with a contact area of 5 mm diameter (0.2cm2). The receptor chamber has 2 mL of PBS, pH 7.4. The Franz cells is kept for 24 hours at 32°C under agitation.

[0154] In the ex vivo skin culture study, the scalp tissue is treated with 2 pL of a formulated cream comprising 0.01% of Peptide 14 (for a total of 200 ng of Peptide 14). The scalp tissue is then placed in an air-liquid interface with DMEM media in the bottom and were kept for 24 hours at 37°C.

[0155] After 24 hours, the excess formulation on the scalp tissue is removed with a tissue paper, and the scalp tissue is washed 4 times in PBS. All the surrounding scalp tissue is also removed. The PBS in the receptor chamber (2 mL) and the scalp tissue are collected and frozen at -80°C until further analysis. The scalp tissue is analyzed using mass spectrometry for the Peptide 14 to determine amount of Peptide 14 that penetrates into the dermis.WSGR Docket No. 56292-708.601

[0156] In the Franz cell study, about 0.1 to 10% of the applied Peptide 14 was found in the scalp tissue. In the ex vivo scalp tissue culture study, about 0.1 to 10% of the applied Peptide 14 was found in the scalp tissue.Example 15.

[0157] 3D skin equivalent sections of samples built with cells from elder donors (71, 84 or 90 years old) were treated with IpM Peptide 14 or Peptide 13, or 20 pM Retinoic Acid (RA) and assessed for the effect of the various treatments of various markers of senescence, aging, and health.

[0158] FIG. 10 shows the relative mRNA expression levels of pl6, BLIMP1, ZYGI IB, IL-8, Ki-67, ZIC1, MMP1, HAS2 of the epidermal (Epi) and dermal (Der) layers of 3D skin equivalents treated with a control, Peptide 14, Peptide 13, or Retinoic Acid. Data are presented as 2-ddCt normalized to GAPDH and untreated control. *p<0.05. Peptide 13 and Peptide 14-treated samples generally had similar relative mRNA expression levels for pl 6, BLIMP1, ZYGI IB, IL-8, and Ki-67 in the epidermal layer, and for pl 6, MMP1, HAS2, IL-8, and Ki-67 in the dermal layer. Peptide 13 and Peptide 14-treated samples generally had lower relative mRNA expression levels than RA-treated samples for pl6, BLIMP1, ZYGI IB, IL-8, in the epidermal layer and for pl6, MMP1, IL-8, and Ki-67 in the dermal layer. Peptide 13 and Peptide 14-treated samples generally had higher relative mRNA expression levels than RA-treated samples for Ki-67 in the epidermal lay er and for HAS2 for the dermal lay er. Peptide 13 and Peptide 14-treated samples had similar relative mRNA expression levels as RA-treated samples for ZIC1 and Ki-67 for dermal layer.Example 16.

[0159] Senotherapeutic strategies can be linked to healthspan and lifespan extension in vivo. In order to assess whether Peptide 14 promotes extended healthspan and lifespan, Caenorhabditis elegans worms were used. Peptide 14 was added in the worm media (M9 buffer media) at different concentrations of 1 pM or 2 pM. Negative control worms received vehicle only. Two healthspan parameters were assessed i) pharyngeal pumping and ii) worm movement. Lifespan was also determined. For pharyngeal pumping analysis, 15 worms had their pharynx movement (pumping) observed daily and counted for 20 seconds. This experiment was repeated 3 times, by 2 differentblind observers, employing different populations of animals at different days. Statistical difference was detected by analyzing each individual time point by One-Way ANOVA and Dunnet’s post hoc test. All groups were compared with the H2O group. For worm movement analysis, the basic movement of C. elegans, also called thrashing, was measured daily for 15 worms. Observation duration was 30 seconds. The experiment was repeated 3 times, by 2 differentblind observers, employing different populations of worms at different days.WSGR Docket No. 56292-708.601Statistical difference was detected by analyzing each individual time point by One-Way ANOVA and Dunnef s post hoc, and all groups were compared with the H2O group. For lifespan analysis, 15 worms had their lifespan observed daily until the last worm died. This experiment was repeated 3 times, by 2 different blind observers, employing different populations of worms at different days. The average lifespan was measured. Statistical difference was detected by analyzing each individual time point by One-Way ANOVA and Dunnet’s post hoc, and all groups were compared with the H2O group (FIG. 11).

[0160] Treatment with either 1 pM or 2 pM of Peptide 14 improved worm thrashing (FIG. 11, panel A), pumping (FIG. 11, panel B), and lifespan (FIG. 11, panel C). While thrashing was significantly improved in day 1 (*p<0.05), pumping was significantly decreased at this day (*p<0.5). Taken together, the data suggest safety and efficacy of the tested peptides with regards to promoting healthspan and lifespan. For instance, on day 8, both thrashing (*p<0.05) and pumping (*p<0.05) were increased in the groups treated with Peptide 14 compared to control. Pumping decreases with aging in C. elegans, mainly as a result of muscular integrity loss. Even though not directly measured, defects in the macroscopic anatomy of C. elegans’’ pharynx (bent or swollen pharynx, both common features of this nematode aging) were also reduced in the 1 pM Peptide 14 group when compared to the H2O control group. No decrease in worm movement was detected in either time point or sample, suggesting that the peptide was not toxic to C. elegans at the concentrations tested. When used at 1 pM and 2 pM, Peptide 14 promoted a statistically significant increase in worm average lifespan (1 pM Peptide 14 **p<0.01; 2 pMpeptidel4 *p<0.05).Example 17.

[0161] Skin cells from Progeria patients were used as a model of aging because of the high levels of cellular senescence. Primary fibroblasts from Progeria patients were cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 10%v.v. fetalbovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U.mL -1). Cells cultured at 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, the cells were seeded in 96 -well plates (4,000 cells per well) and, 6 hours after plating, were incubated with 500 nM, 5 pM or 50 pM of polypeptide sequence LKGIL (SEQ ID NO: 6) or WLKGI (SEQ ID NO: 7). Negative control comprised untreated cells, which received vehicle only. After incubation, relative cellular senescence, which was assessed by the activity of senescence associated P -galactosidase staining relative to untreated control and by the quantification of ATRX foci / nuclei, was analyzed. Three independent experiments (biological replicates) including three technical replicates were performed. Data was analyzed using ANOVA and a Bonferroni post -hoc test.WSGR Docket No. 56292-708.601Statistical significance was determined as p values equal or lower than 0.05, where *p<0.05 and ** p<0.01.FIG. 12 shows the effect of polypeptide sequences LKGIL (SEQ ID NO: 6) (A, B, C), and WLKGI (SEQ ID NO: 7) (D, E, F) to decrease cellular senescence without promoting cell death. In panels A and D, the y-axis indicates the relative senescence level normalized to untreated control. In panels B and D, y-axis indicates the relative cell number normalized to untreated control. In panels C and F, y-axis indicates the average ATRX foci accumulation per cell. Treatment of cells with LKGIL (SEQ ID NO: 6) significantly decreased senescence associated P-galactosidase staining and the average number of ATRX foci / nuclei, when used at 5 pM and 50 pM, compared to untreated cells (p<0.01 for P-galactosidase staining and p<0.05 for ATRX foci / nuclei). Treatment of cells with WLKGI (SEQ ID NO: 7) decreased senescence associated P-galactosidase staining levels when used at 5 pM and 50 pM as compared to untreated cells (p<0.05 for 5 pM and p<0.01 for 50 pM). No cellular toxicity was observed in the tested concentrations.Example 18.

[0162] Additional polypeptides were tested for suitability as an anti -senescent agent. Cells were incubated with one of the polypeptides. Negative control comprised untreated cells, which received vehicle only. After incubation, relative cellular senescence (assessed by the activity of senescence associated P-galactosidase staining relative to negative control) and relative cell proliferation relative to negative control were analyzed. A number of polypeptides showed relative cellular senescence lower than 1, having decreased cellular senescence than the untreated negative control, and maintained cell proliferation at or above that of the untreated negative control. Examples of such polypeptides shown in Table 6.

[0163] Further, a dose-dependent effects of polypeptides of Table 6 on cell senescence were studied. The cells were incubated various doses of the polypeptides ranging from 1.26 pM to 50 pM (1.26 pM, 3.12 pM, 6.25 pM, 12.5 pM, 25 pM, 50 pM). There was noticeable decrease in cell senescence even at lower doses for the polypeptides.WSGR Docket No. 56292-708.601

[0164] While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the disclosure described herein maybe employed in practicing the disclosure. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby.Example 19.

[0165] Primary ORS cells were cultured in Mesenchymal Stem Cell Medium (ScienCell) supplemented with 10% v.v. fetal bovine serum (FBS), and 1% v.v. of penicillin / streptomycin solution (1,000 U / mL). Cells were kept in a 5% CO2, 37 °C and 95% humidity atmosphere. After expansion, these cells were seeded in 96 -well plates (5,000 cells per well) and overnight after plating, were serum starved in serum free Mesenchymal Stem Cell Medium. After 24 hours, cells were incubated with a combination of 0.4 pM CRH and 12.5 pM Peptide 14 or Peptide 14 alone for 72 hours. Negative control comprised untreated cells, which received vehicle only. After incubation, relative cellular senescence (assessed by the activity of senescence associated B-galactosidase staining, CDKN1 A (p21) gene expression, and IL-6 gene expression relative to untreated control) was analyzed. Six technical replicates were performed. Data was analyzed using unpaired t-test. Statistical significance was determined as p values equal or lower than 0.05 (FIG. 13).

[0166] Treatment of cells with CRH significantly increased senescence associated B-galactosidase staining (FIG. 13, panel A), increased gene expression of the senescence marker CDKN1A (p21) (FIG. 13, panel B), and significantly increased gene expression of the inflammatory marker IL-6 (FIG. 13, panel C) compared to untreated cells. Treatment of cells with Pep 14 formula and CRH significantly decreased senescence associated B-galactosidaseWSGR Docket No. 56292-708.601staining (FIG. 13, panel A), significantly decreased gene expression of the senescence marker CDKN1A (p21) (FIG. 13, panel B), and significantly decreased gene expression of the inflammatory marker IL-6 (FIG. 13, panel C) compared to cells treated with CRH alone. No cellular toxicity was observed.Example 20.

[0167] A beta test was performed using human participants. Participants were asked to use a Peptide 14 formulation for 3 months. The Peptide 14 formulation included Aqua (Water), 1,2-Hexanediol, Acetyl Tyrosine, Alpha-Glucan Oligosaccharide, Arginine, Arctium Maj us Root Extract, Biotin, Calcium Pantothenate, Cellulose Gum, Chondrus Crispus, Citric Acid, Citrulline, Decapeptide-52 (Peptide 14), Erythritol, Fisetin, Glycerin, Glucosamine HC1, Hydrolyzed Soy Protein, Hydroxyacetophenone, Inulin, Morus Nigra Leaf Extract, Niacinamide, Ornithine HC1, Oryza Sativa (Rice) Bran Water, Panax Ginseng Root Extract, Panthenol, PEG-12 Dimethicone, Phenoxyethanol, Pisum Sativum (Pea) Sprout Extract, Polyquatemium-11, Potassium Sorbate, Propanediol, Raspberry Ketone, Rehmannia Chinensis Root Extract, Sodium Benzoate, Sodium PC A, Tetrasodium Glutamate Diacetate, and Xanthan Gum. Participants were instructed to discontinue use of all other leave-in hair growth products that are applied to the scalp during the beta testing period. As they used the Peptide 14 formulation, participants were asked to take note of how their hair and scalp felt, and any improvements in hair density, volume, length and thickness. Participants took a survey at approximately 1 month, 2 months, and 3 months of use. Participant also took pictures of their scalp before use and after 3 months of use (FIGs. 14-15).

[0168] In the survey at 3 months of use, 82% of participants indicated they noticed new hair growth, 73% indicated that their hair was visibly denser, and 64% indicated that their hair was visibly healthier (Table 7).Example 23.

[0169] A double-blinded clinical study was performed. 30 participants were enrolled (7 male and 23 female). All men presented with male pattern baldness (high temples, thinning at the crown of the head, and / or bald at the crown of the head). Participants were asked to use a Peptide 14 formulation and a dermaroller twice a day for 3 months. The Peptide 14 formulation included Aqua (Water), 1,2-Hexanediol, Acetyl Tyrosine, Alpha-Glucan Oligosaccharide, Arginine, Arctium Majus Root Extract, Biotin, Calcium Pantothenate, Cellulose Gum, ChondrusWSGR Docket No. 56292-708.601Crispus, Citric Acid, Citrulline, Decapeptide-52 (Peptide 14), Erythritol, Fisetin, Glycerin, Glucosamine HC1, Hydrolyzed Soy Protein, Hydroxyacetophenone, Inulin, Morus Nigra Leaf Extract, Niacinamide, Ornithine HC1, Oryza Sativa (Rice) Bran Water, Panax Ginseng Root Extract, Panthenol, PEG-12 Dimethicone, Phenoxyethanol, Pisum Sativum (Pea) Sprout Extract, Polyquatemium-11, Potassium Sorbate, Propanediol, Raspberry Ketone, Rehmannia Chinensis Root Extract, Sodium Benzoate, Sodium PC A, Tetrasodium Glutamate Diacetate, and Xanthan Gum.

[0170] Participants took a survey after 3 months of use. In the survey at 3 months of use, 80% of participants indicated that they observed improvements in their hair, 77% indicated that they observed new hair growth, and 77% indicated that they observed an improvement in the overall health of their hair (Table 8).

[0171] Participants also underwent HairMetrix® analysis at 0 months and 3 months. In the HairMetrix® analysis, an imaging wand is used to capture images of the scalp, then an AI-powered assessment technique is used to determine the number of hairs and the health of each hair. Green hairs usually represent healthy, active, or “terminal” hairs, which are in the anagen phase of growth, and strong follicles with no significant signs of thinning, damage, or loss. Yellow hairs represent miniaturized or “vellus” hairs, which are tinner, weaker, and in a more compromised state, and follicles in the telogen or resting phase, often associated with hair shedding or early signs of thinning. By analyzing the proportions of green and yellow hairs, HairMetrix® can provide insights into hair density, scalp health, and the progression of hair thinning or loss. An exemplary HairMetrix® analysis is shown in FIG. 17. The HairMetrix® analysis showed that 70% of users experienced an increase in hair density within 3 months, 6 out of 7 men experienced an increase in hair thickness within 3 months, and 6 out of 7 men experienced a significant increase in anagen (growth -phase) hairs within 3 months (Table 9).WSGR Docket No. 56292-708.601

Claims

WSGR Docket No. 56292-708.601CLAIMS WHAT IS CLAIMED IS:

1. A method for treating hair, the method comprising: applying to the hair of a subject a composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids.

2. A method for maintaining or increasing hair bulk, hair density, hair thickness, hair volume, or hair length in a subject, the method comprising administering to a skin having hair a composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids.

3. A method for reducing hair loss in a subject, the method comprising administering to a skin having hair a composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids.

4. The method of any one of preceding claims, wherein the polypeptide comprises at least 10 amino acids, 15 amino acids, or 20 amino acids.

5. The method of any one of preceding claims, wherein the polypeptide is an isolated, synthetic, or recombinant polypeptide.

6. The method of any one of preceding claims, wherein the composition maintains or increases hair bulk, hair density, hair thickness, hair volume, or hair length.

7. The method of any one of preceding claims, wherein the composition reduces hair loss.

8. The method of any one of preceding claims, wherein the composition increases proliferation of one or more hair follicle cells.

9. The method of any one of preceding claims, wherein the composition decreases hair follicle miniaturization of one or more hair follicle cells.

10. The method of any one of preceding claims, wherein the composition decreases senescence of one or more hair follicle cells.WSGR Docket No. 56292-708.60111. The method of one of claims 7 to 9, wherein the one or more hair follicle cells comprises outer root sheath cells, inner root sheath cells, or dermal papilla cells, or a combination thereof.

12. The method of any one of preceding claims, wherein the composition increases proliferation of one or more hair follicle cells.

13. The method of any one of preceding claims, wherein amount of the polypeptide in the composition is about 500 nM to about 500 pM.

14. The method of any one of preceding claims, wherein amount of the polypeptide in the composition is about 0.001% (w / w) to about 5% (w / w).

15. The method of any one of preceding claims, wherein amount of the polypeptide in the composition is about 0.01% (w / w) to about 1% (w / w).

16. The method of any one of preceding claims, wherein the polypeptide is formulated as a topical composition.

17. The method of claim 12, wherein the topical composition comprises a shampoo, a conditioner, a serum, a cream, a lotion, a gel, a liquid, an oil, a powder, an emulsion, a moisturizer, a foam, a mousse, an aerosol, or a spray.

18. The method of any one of preceding claims, wherein the polypeptide is formulated with a second peptide.

19. The method of any one of preceding claims, wherein the polypeptide is formulated with one or more of a plant extract, an amino acid, an emollient, a hydrating agent, a vitamin or a vitamin derivative, an antioxidant, and a viscosity modifying agent.

20. The method of any one of preceding claims, wherein the plant extract comprises one or more of Arctium Majus root extract, Morus Nigra leaf extract, Panax ginseng root extract, Pisum Sativum (pea) sprout extract, Irish moss extract, soy extract, Helianthus annuus (sunflower) sprout extract, Rehmannia Chinensis root extract, and Oryza Sativa (rice) bran extract.

21. The method of any one of preceding claims, wherein the amino acid comprises one or more of tyrosine, arginine, citrulline, alanine, glycine, phenylalanine, cysteine, methionine, serine, glutamic acid, ornithine, and derivatives thereof.WSGR Docket No. 56292-708.60122. The method of any one of preceding claims, wherein the hydrating agent comprises on or more of glycerin, squalane, sorbitol, hyaluronic acid, hyaluronic acid derivatives, sodium hyaluronate, sodium hyaluronate crosspolymer, niacinamide, glycoproteins, pyrrolidone carboxylic acid (PCA), lysine HC1, allantoin, and algae extract.

23. The method of any one of preceding claims, wherein the emollient comprises one or more of plant oil, mineral oil, shea butter, cocoa butter, petrolatum, a fatty acid, a triglyceride, a benzoate, a myristate, a palmitate, a stearate, a glycolipid, a phospholipid, squalane, glycerin, ceramide, and algae extract.

24. The method of any one of preceding claims, wherein the plant oil comprises one or more of rose hip oil, andiroba oil, grape seed oil, avocado oil, plum seed oil, pracaxi oil, Calycophyllum spruceanum oil, almond oil, and argan oil.

25. The method of any one of preceding claims, wherein the vitamin comprises one or more of vitamin A, vitamin D, vitamin E, vitamin F, vitamin K, vitamin Bl (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B7 (biotin), vitamin B6, vitamin B12 (cyanocobalamin), vitamin B9, folic acid, niacinamide, or derivatives thereof.

26. The method of any one of preceding claims, wherein senescence is assessed by one or more of senescence associated P -galactosidase activity level, a ratio of ATRX foci / cell, pl6 expression, IL-8 expression, and Ki-67 expression.

27. The method of any one of preceding claims, wherein the subject has hair loss.

28. The method of claim 26, wherein the hair loss is due to androgenic alopecia, alopecia areata, telogen effluvium, anagen affluvium, or a combination thereof.

29. The method of any one of preceding claims, wherein the polypeptide comprises one or more of ETAKHWLKGI, ATAKAWLKGI, KLKGILRGAA, and WLKGILREAA.

30. A composition for treating hair, the composition comprising a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids.

31. A composition for maintaining or increasing hair bulk, hair density, hair thickness, hair volume, or hair length in a subject, the composition comprising an effectiveWSGR Docket No. 56292-708.601amount of a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids, and a cosmetic excipient.

32. A composition for reducing hair loss in a subject, the composition comprising an effective amount of a polypeptide comprising an amino acid sequence of WLKGI or an analog thereof, wherein the polypeptide comprises no more than 100 amino acids, and a cosmetic excipient.

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