Octapeptides for topical application

Abstract

Described herein are compositions and methods comprising one or more peptides, including an octapeptide, having amino acid sequences characteristic of those found in collagen and / or elastin.

Description

[Technical field]

[0001] cross reference This application claims the benefit of U.S. Provisional Patent Application No. 63 / 330,752, filed April 13, 2022, which is incorporated by reference herein in its entirety.

[0002] Incorporation by Reference 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. To the extent that the publications and patents or patent applications incorporated by reference conflict with the disclosure contained herein, the present specification is intended to supersede and / or take precedence over any such conflicting material. [Background technology]

[0003] background Hyaluronic acid (HA) plays an important role in cellular and extracellular matrix (ECM) homeostasis in the skin. HA is involved in regulating hydration, cell turnover, metabolism, tissue repair, nutrient exchange, and protection against free radical damage. HA-bound water in both the dermis and epidermis is important for skin hydration. Although HA decreases with aging, the HA content in skin actually remains constant with age. The difference is that HA becomes increasingly tissue-bound and more resistant to extraction. This HA encapsulated within tissue proteins may be limited in its hydration capacity, and the lower molecular weight (MW) HA present in these tissue-bound forms may be pro-inflammatory. Thus, topical compositions that accelerate the synthesis of high MW HA would be beneficial to improve skin health.

[0004] Elastin is an important extracellular matrix (ECM) protein that provides stretch, resilience, elasticity, and elasticity to tissues and organs, including skin. Normal levels of elastic fiber production, organization, and integration with other skin extracellular matrix proteins, proteoglycans, and glycosaminoglycans are key to maintaining healthy skin structure, function, and youthful appearance. Elastin production decreases with age, and elastin degrades upon exposure to environmental insults, especially sun exposure, causing loss of skin structural integrity and contributing to sagging or crepiness of the skin and associated undesirable changes in appearance. Disruption of the elastic fiber network also leads to suboptimal wound healing and impaired structure and appearance of scars and stretch marks. Thus, topical compositions that can enhance the production of elastic fibers would be highly advantageous to improve and / or restore the appearance, texture, elasticity, and wound healing capacity of skin. Summary of the Invention

[0005] overview Skin aging is associated with loss of skin moisture and reduced barrier function.Aged or damaged skin may show various signs of inflammation, which may be visible as certain skin conditions, including but not limited to increased skin redness, increased visibility of fine blood vessels, and rosacea.Environmental influences, such as excessive UV exposure, may contribute to the progressive visible changes seen in aged skin.Topical compositions and methods that promote the inherent high MW HA production in skin or increase elastin production may be useful for treating various skin conditions.

[0006] In many cases, improving skin hydration and reducing inflammation in the skin can help treat damaged, inflamed, or aging skin. Application of topical compositions containing peptides has been used to treat damaged, inflamed, or aging skin.

[0007] Described herein are compositions and methods that include topical formulations and applications of octapeptides. In some embodiments, the compositions and methods described herein reduce skin inflammation, the appearance of solar elastosis, and / or signs of skin aging. In some embodiments, the compositions and methods described herein improve skin health, appearance, and hydration. In some embodiments, the compositions and methods described herein increase HA production and / or increase elastin production.

[0008] There remains a need to promote proper skin hydration and healthy skin tone, texture and elasticity in individuals.Skin aging is associated with loss of skin moisture, and dry skin leads to reduced barrier function, as well as reduced elastin production and degradation of elastin fiber network.Environmental influences such as excessive UV exposure also contribute to the visible progressive changes seen in aging skin, such as accelerated development of fine lines and wrinkles, and reduced skin elasticity.There is a need for topical compositions and methods that promote the production of inherent high MW HA in epidermal and dermal cells to improve skin health, and promote elastin production to restore or improve the appearance of aged or damaged skin.

[0009] In one aspect, which may be combined with any other aspect or embodiment, the disclosure relates to a topical composition for stimulating synthesis of high molecular weight hyaluronic acid (high MW HA), the composition comprising a synthetic, isolated, or recombinant polypeptide comprising an amino acid sequence selected from the group consisting of GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), or GPGKNGDD (SEQ ID NO: 7), the polypeptide comprising 20 or fewer amino acids. In some embodiments, the amino acid sequence is GPHGVREA (SEQ ID NO: 1). In some embodiments, the polypeptide is an octapeptide. In some embodiments, the polypeptide comprises octapeptide-45.

[0010] In some embodiments, the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivative thereof, the high MW HA having a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivative thereof has a molecular weight of about 1 MDa to 4 MDa. In some embodiments, the high MW HA or derivative comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprises sodium hyaluronate crosspolymer.

[0011] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0012] In some embodiments, the composition comprises a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, the hexapeptide-11 is encapsulated in a liposome.

[0013] In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.

[0014] In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyldecanone. In some embodiments, the composition comprises high molecular weight hyaluronic acid, synthetic tripeptide, hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, the composition comprises hexapeptide-12. In some embodiments, the composition comprises tripeptide-1. In some embodiments, the composition comprises tripeptide-1 and hexapeptide-12.

[0015] In some embodiments, the composition is aqueous.

[0016] In some embodiments, the composition is effective to achieve a reduction in inflammation, as assessed by a reduction or lack of increase in the expression levels of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), or cluster of differentiation-80 (CD80), or a combination thereof.

[0017] In some embodiments, the composition increases CD44 expression.

[0018] In some embodiments, the composition reduces redness of the individual's skin after application of the composition, hi some embodiments, the redness is measured by photography.

[0019] In some embodiments, the composition increases skin hydration in the individual's skin after application of the composition, hi some embodiments, the composition reduces or reverses the signs of solar elastosis.

[0020] In some embodiments, the composition upregulates hyaluronic acid synthase (HAS) expression. In some embodiments, the HAS comprises HAS2.

[0021] In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0022] In another aspect, which may be combined with any other aspect or embodiment, the disclosure relates to a method for reducing inflammation in the skin of an individual, the method comprising applying a topical composition comprising a synthetic, isolated, or recombinant polypeptide comprising the amino acid sequence GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), or GPGKNGDD (SEQ ID NO: 7), wherein the polypeptide comprises 20 or fewer amino acids. In some embodiments, the amino acid sequence is GPHGVREA (SEQ ID NO: 1). In some embodiments, the polypeptide is an octapeptide. In some embodiments, the polypeptide comprises octapeptide-45.

[0023] In some embodiments, the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivative thereof, the high MW HA having a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivative thereof has a molecular weight of about 1 MDa to 4 MDa. In some embodiments, the high MW HA or derivative comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprises sodium hyaluronate crosspolymer.

[0024] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0025] In some embodiments, the composition comprises a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, the hexapeptide-11 is encapsulated in a liposome.

[0026] In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.

[0027] In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyldecanone.

[0028] In some embodiments, the composition comprises a synthetic tripeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, the composition comprises hexapeptide-12. In some embodiments, the composition comprises tripeptide-1. In some embodiments, the composition comprises tripeptide-1 and hexapeptide-12.

[0029] In some embodiments, the composition is aqueous.

[0030] In some embodiments, reduced inflammation is assessed by a reduction or lack of increase in the expression levels of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), or cluster of differentiation-80 (CD80), or a combination thereof.

[0031] In some embodiments, the composition increases CD44 expression.

[0032] In some embodiments, the composition reduces redness of the individual's skin after application of the composition, hi some embodiments, the redness is measured by photography.

[0033] In some embodiments, the composition increases skin hydration in the individual's skin after application of the composition, hi some embodiments, the composition promotes HA synthesis.

[0034] In some embodiments, the composition upregulates hyaluronic acid synthase (HAS) expression. In some embodiments, the HAS comprises HAS2.

[0035] In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0036] In some embodiments, the composition reduces the appearance of birthmarks, aging spots, or wrinkles.

[0037] In some embodiments, the composition is applied 1, 2, 3, 4, 5, 6, 7, or 8 times daily.

[0038] In some embodiments, the individual is a human.

[0039] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a topical composition for promoting skin repair, the topical composition comprising a synthetic, isolated, or recombinant octapeptide comprising the amino acid sequence Gly-X1-X2-X3-X4-X5-X6-X7. In some embodiments, X1 is Pro, Asp, or Leu. In some embodiments, X2 is His, Gly, Met, or Gln. In some embodiments, X3 is Gly, Asp, Pro, or Lys. In some embodiments, X4 is Val, Gly, Pro, Phe, or Asn. In some embodiments, X5 is Arg, Ala, Ser, Gln, or Gly. In some embodiments, X6 is Glu, Ser, Gly, Arg, or Asp. In some embodiments, X7 is Ala, Pro, or Asp. In some embodiments, the octapeptide comprises an amino acid sequence characteristic of collagen or elastin. In some embodiments, the amino acid sequence comprises GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). In some embodiments, the amino acid sequence is GPHGVREA (SEQ ID NO: 1). In some embodiments, the polypeptide comprises octapeptide-45.

[0040] In some embodiments, the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivative thereof, the high MW HA having a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivative comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprises sodium hyaluronate crosspolymer.

[0041] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0042] In some embodiments, the composition comprises a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, the hexapeptide-11 is encapsulated in a liposome.

[0043] In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.

[0044] In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyldecanone.

[0045] In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, the composition comprises hexapeptide-12. In some embodiments, the composition comprises tripeptide-1. In some embodiments, the composition comprises tripeptide-1 and hexapeptide-12.

[0046] In some embodiments, the composition is aqueous.

[0047] In some embodiments, reduced inflammation is assessed by a reduction or lack of increase in the expression levels of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), or cluster of differentiation-80 (CD80), or a combination thereof.

[0048] In some embodiments, the composition increases CD44 expression.

[0049] In some embodiments, the composition reduces redness of the individual's skin after application of the composition, hi some embodiments, the redness is measured by photography.

[0050] In some embodiments, the composition increases skin hydration in the skin of an individual after application of the composition. In some embodiments, the composition promotes HA synthesis. In some embodiments, the composition promotes elastin production.

[0051] In some embodiments, the composition upregulates hyaluronic acid synthase (HAS) expression. In some embodiments, the HAS comprises HAS2. In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0052] In some embodiments, the composition reduces or reverses the symptoms of solar elastosis.

[0053] In some embodiments, the composition stimulates the synthesis of high MW HA having a MW of at least 0.5 MDa.

[0054] In another aspect, which may be combined with any other aspect or embodiment, the disclosure relates to a topical composition for increasing elastin production in mammalian skin comprising a polypeptide comprising an amino acid sequence selected from the group consisting of GPHGVREA (SEQ ID NO:1), GDGDGASA (SEQ ID NO:2), GPMGPSGP (SEQ ID NO:3), GLGPGARA (SEQ ID NO:4), GPQGFQGP (SEQ ID NO:5), GPMGPRGP (SEQ ID NO:6), or GPGKNGDD (SEQ ID NO:7), wherein the polypeptide comprises 20 or fewer amino acids; tripeptide-1; and hexapeptide-12.

[0055] In some embodiments, the polypeptide is an octapeptide. In some embodiments, the amino acid sequence of the polypeptide comprises GPHGVREA (SEQ ID NO: 1). In some embodiments, the octapeptide is octapeptide-45.

[0056] In some embodiments, the composition further comprises hexapeptide-11, tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate, lactoferrin, phosphatidylserine, Tremella fuciformis extract, hydroxymethoxyphenyldecanone, or any combination thereof. In some embodiments, the composition comprises high molecular weight (MW) hyaluronic acid (HA) or a derivative thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa.

[0057] In some embodiments, the composition comprises hexapeptide-11.

[0058] In some embodiments, the composition comprises lactoferrin.

[0059] In some embodiments, the composition comprises phosphatidylserine.

[0060] In some embodiments, the composition comprises a Tremella fuciformis extract.

[0061] In some embodiments, the composition comprises tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0062] In some embodiments, the composition comprises hydroxymethoxyphenyldecanone.

[0063] In some embodiments, any one of hexapeptide-11, tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate, lactoferrin, phosphatidylserine, Tremella fuciformis extract, hydroxymethoxyphenyldecanone, or any combination thereof is encapsulated in a liposome.

[0064] In some embodiments, the composition is aqueous.

[0065] In some embodiments, the composition is effective to increase production of tropoelastin and microfibril associated protein 4 (MFAP4). In some embodiments, the composition is effective to upregulate expression of S100A2, LAMA3, MERTK, or ITGB4. In some embodiments, the composition upregulates one or more genes associated with a pathway associated with ECM regulation.

[0066] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of treating or restoring aging skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0067] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of upregulating MFAP4, S100A2, LAMA3, MERTK, or ITGB4 expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0068] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing MFAP4 expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0069] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing S100A2 expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0070] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing LAMA3 expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0071] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing MERTK expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0072] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing ITGB4 expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0073] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of upregulating genes associated with pathways associated with ECM regulation in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0074] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing MFAP4 expression in the skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0075] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method of increasing elastin production in human skin, the method comprising administering to the skin a composition according to any of the embodiments described herein.

[0076] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a composition according to any of the embodiments described herein for use in treating or restoring aging skin or increasing elastin production in human skin.

[0077] In some embodiments, the composition increases the expression of MFAP4, S100A2, LAMA3, MERTK, or ITGB4 in the skin.

[0078] In some embodiments, the composition upregulates expression of genes associated with pathways associated with ECM regulation in the skin.

[0079] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for promoting wound healing comprising administering to the skin a topical composition according to any of the embodiments described herein.

[0080] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for promoting wound healing or wound closure comprising administering to the skin a topical composition according to any of the embodiments described herein.

[0081] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for treating a skin wound comprising administering to the skin a topical composition according to any of the embodiments described herein. [Brief description of the drawings]

[0082] This patent application contains at least one drawing executed in color. Copies of this patent or patent application with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0083] [Figure 1] Shown is a graph of fold change in hyaluronan synthase (HAS2) mRNA gene expression in dermal fibroblasts following treatment with the compounds listed below the bar compared to untreated cells. [Figure 2A] Shown is a graph of fold change in hyaluronan synthase (HAS2) mRNA gene expression in keratinocytes following treatment with the compounds listed below the bars compared to untreated cells. [Figure 2B] Shown is a graph of the fold change in hyaluronidase 2 (HYAL2) mRNA gene expression in keratinocytes following treatment with the compounds listed below the bars compared to untreated cells. [Diagram 3] Shown is a graph of fold change in early growth response 3 (EGR3) mRNA gene expression in keratinocytes following treatment with the compounds listed below the bar compared to untreated cells. [Figure 4] 1 shows an SDS-PAGE gel with concentrated supernatant from cultured human dermal fibroblasts demonstrating the effect of treatment with various compounds. [Diagram 5] 1 shows an SDS-PAGE gel with concentrated supernatant from cultured human dermal fibroblasts demonstrating the effect of treatment with various compounds. [Figure 6] FIG. 1 shows a graph of dose-response stimulation of high molecular weight HA production by octapeptide treatment in dermal fibroblasts. [Figure 7] 1 is a line plot demonstrating the progressive increase in average hydration level for clean skin 15 minutes after cleansing of facial skin. [Figure 8] FIG. 1 is a bar graph demonstrating the progressive increase in average hydration levels for cleansed skin after cleansing facial skin, waiting a first 15 minute period after cleansing, applying a treatment with a composition comprising high MW HA and an octapeptide, and waiting a second 15 minute period after treatment before measuring hydration. [Figure 9] 1 is a table summarizing the results of participant evaluations and satisfaction surveys following cleansing of the skin at various time points following the initiation of treatment with a composition comprising high MW HA and an octapeptide. [Figure 10] 1 is a table summarizing the results of a Participant Assessment Survey of the Participant's Own Skin Condition beginning at the start of treatment (baseline) and at various time points after the start of treatment. [Figure 11] 1 is a table of participant satisfaction and statistical analysis of the differences between baseline ratings and evaluations at various time points after the start of treatment. [Figure 12] 1 is a chart comparing the mean investigator assessment scores for a given measure of skin condition at baseline and after 8 weeks of treatment. [Figure 13] 1 is a chart comparing the mean investigator-assessed scores for skin moisture / hydration at baseline and after 8 weeks of treatment. [Figure 14A] Figures 14-21 are photographs, analyses, and tissue sections from subjects at baseline and at various time points after initiation of treatment with usual skin care using a composition comprising high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after week 4, and a gentle cleanser. Figure 14A shows photographs taken at baseline and 4 weeks after treatment to demonstrate improvements to the skin in a female subject, age 39. [Figure 14B] Analysis of the photograph from FIG. 14A using a VISIA® Skin Analysis System is shown to demonstrate the reduction in facial red areas following treatment. [Figure 15] Photographs taken at baseline, 2 weeks, 4 weeks, and 8 weeks after treatment (from left to right) are shown to demonstrate the progressive effect of skin improvement in a female subject, age 35. [Figure 16A] 1 shows close-up photographs taken at baseline and 8 weeks after treatment to demonstrate improvement in pore size, skin tone, and skin texture in a female subject, age 35. [Figure 16B]Analysis of photographs from FIG. 15 taken at baseline and 8 weeks after treatment using the VISIA® Skin Analysis System to demonstrate the reduction in facial red areas is shown. [Figure 16C] To demonstrate the reduction in facial pore size, an analysis of photographs from FIG. 15 taken at baseline and 8 weeks after treatment is shown using a VISIA® Skin Analysis System. [Figure 16D] Shown are close-up photographs of the eye and cheek area of ​​a female subject, age 35, from FIG. 15 taken at baseline and 8 weeks post-treatment to demonstrate the improvement in fine lines, wrinkles, and redness reduction. [Figure 16E] Shown is an analysis of photographs of the eye and cheek area of ​​a female subject, age 35, from FIG. 15 taken at baseline and 8 weeks post-treatment using a VISIA® Skin Analysis System to demonstrate reduced wrinkle density, area, and distribution. [Figure 17A] Photographs taken at baseline and 8 weeks after treatment are shown to demonstrate improvements in skin tone, texture, and reduced redness in a female subject, age 38. [Figure 17B] Analysis of the photograph from FIG. 17A using a VISIA® Skin Analysis System is shown to demonstrate the improvements resulting in more uniform skin texture in the facial areas following treatment. [Figure 18A] Photographs taken at baseline, 2 weeks, 4 weeks, and 8 weeks after treatment (from left to right) are shown to demonstrate the improvement in a male subject, age 48, with progressive reduction in skin inflammation, reduction in skin redness, and reduction in visibility of small blood vessels. [Figure 18B] Shown are close-up photographs of the upper left face from the male subject, age 48, from FIG. 18A taken at baseline and 8 weeks post-treatment to demonstrate improved rosacea and inflammation. [Figure 18C] Shown are close-up photographs of the right cheek from the male subject, age 48, from FIG. 18A taken at baseline and 8 weeks post-treatment to demonstrate skin improvements including reduced visibility of fine blood vessels. [Figure 19A]Shown are close-up photographs of the right cheek and mouth area of ​​a female subject, age 73, taken at baseline and 2 weeks post-treatment to demonstrate improvement of fine lines, deep wrinkles, and fine skin lines. [Figure 19B] Photographs of the right side of the face of a female subject, age 73, from FIG. 19A are shown taken at baseline and 4 weeks after treatment to demonstrate the progressive improvement of fine lines, deep wrinkles, and fine skin lines. [Figure 19C] Photographs of the right side of the face of a female subject, age 73, from FIG. 19A are shown taken at baseline and 8 weeks after treatment to demonstrate the progressive improvement of fine lines, deep wrinkles, and fine skin lines. [Figure 20A] Figures 20A-C show histological sections of periauricular biopsies from subjects at baseline and 8 weeks post-treatment to demonstrate significant improvement in solar elastosis extracellular matrix (ECM). Paired sections from three separate subjects at a given time point are stained with hematoxylin and eosin. Magnification is 200x. [Figure 20B] See legend to Figure 20A. [Figure 20C] See legend to Figure 20A. [Figure 21A] Figures 21A-C show histological sections of periauricular biopsies from subjects at baseline and 8 weeks post-treatment to demonstrate significant upregulation of CD44 expression in epidermal and dermal structures in the third experiment. Figure 21A shows paired sections from the first subject, with a magnification of 200x. [Figure 21B] 21B-C are paired sections from a second subject, at 200x and 400x magnifications, respectively. [Figure 21C] See legend to Figure 21B. [Figure 22] FIG. 1 shows a diagram of the chemical structure of octapeptide-45, which contains the amino acid sequence Gly-Pro-His-Gly-Val-Arg-Glu-Ala. [Figure 23A]Figures 23A-23D are bar graphs showing results (percent change relative to untreated control) from in vitro gene expression studies on human dermal fibroblasts cultured and treated with tripeptide-1 + hexapeptide-12 (TriHex), octapeptide-45, and TriHex + octapeptide-45. Figure 23A shows results for S100A2 expression. Figure 23B shows results for LAMA3 expression. Figure 23C shows results for MERTK expression. Figure 23D shows results for ITGB4 expression. All treatments were at 20 μg / mL. [Figure 23B] See legend to Figure 23A. [Figure 23C] See legend to Figure 23A. [Figure 23D] See legend to Figure 23A. [Figure 24] FIG. 1 is a schematic diagram of an ex vivo experimental model used to test the effect on protein production for peptide compositions according to the present disclosure. [Diagram 25] FIG. 10 is a schematic diagram of a model showing tropoelastin and MFAP4 production in elastin fiber assembly. (Reproduced from P. Bartosz et al., 291 J. Biol. Chem. 1103-14 (Jan. 2016) (FIG. 10).) [Figure 26] FIG. 1 is a cross-sectional view of a human skin sample showing the location of the epidermis, papillary dermis, and reticular dermis. [Figure 27] Stained human skin sample cross sections are shown for untreated skin (left) and skin treated with tripeptide-1 + hexapeptide-12 (TriHex) (second from left), octapeptide-45 (second from right), and TriHex + octapeptide-45 (right). Blue staining represents DAPI for nuclei and cells. Red staining represents MFAP4. [Figure 28]Stained human skin sample cross sections are shown for untreated skin (left) and skin treated with tripeptide-1 + hexapeptide-12 (TriHex) (second from left), octapeptide-45 (second from right), and TriHex + octapeptide-45 (right). Blue stain represents DAPI for nuclei and cells. Red stain represents tropoelastin (TE).

[0084] The novel features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0085] Detailed Description Skin aging is associated with the loss of skin moisture and the reduction of barrier function.Aged or damaged skin may show various signs of inflammation, which may be visible as specific skin conditions, including but not limited to increased skin redness, increased visibility of fine blood vessels, and rosacea.Environmental influences, such as excessive exposure to ultraviolet light, may contribute to the progressive changes seen in aged skin.

[0086] In many cases, improving skin hydration and reducing inflammation in the skin can help treat damaged, inflamed, or aging skin. Application of topical compositions containing peptides has been used to treat damaged, inflamed, or aging skin.

[0087] Described herein are compositions and methods comprising topical formulations and applications of the octapeptide. In some embodiments, the compositions and methods described herein reduce skin inflammation, the appearance of solar elastosis, and / or signs of skin aging. In some embodiments, the compositions and methods described herein improve skin health, appearance, and hydration. In some embodiments, a composition comprising the octapeptide results in increased synthesis of high MW HA compared to skin not treated with the composition. In some embodiments, the octapeptide in the composition stimulates synthesis of high MW HA. In some embodiments, the composition increases elastin production.

[0088] As described herein, a group of active agents was identified that stimulated high MW HA production in both epidermal and dermal cell types. Members of one class of these active agents (e.g., octapeptides) contain amino acid sequence features present in both collagen and elastin. Upon application of a topical formulation containing octapeptides, endogenous signaling pathways may recognize these short peptide sequences that mimic extracellular matrix (ECM) degradation products and upregulate the innate response to produce high MW HA. These novel approaches to skin treatment described herein, including high MW HA+peptide topical application, aim to increase the production of exclusively high MW HA in both epidermal and dermal cells. These formulations of compositions and methods of using them were created to maximize hydration capacity in both the epidermis and dermis while promoting regenerative activity within the ECM. Application of the octapeptide formulation resulted in a quantifiable increase in skin hydration, a concomitant reduction in redness, inflammation, and the appearance of fine lines and wrinkles, as well as a notable reversal of solar elastosis ECM structure to a healthier, denser ECM network.

[0089] In some embodiments, the composition comprising an octapeptide increases the production of elastin in the skin. In some embodiments, the composition comprises octapeptide-45, tripeptide-1, and hexapeptide-12. In some embodiments, the composition upregulates expression of genes associated with ECM regulation in the skin. In some embodiments, the composition increases MFAP4 expression. In some embodiments, the composition increases tropoelastin (TE) production. In some embodiments, the composition upregulates SA1002 expression. In some embodiments, the composition upregulates LAMA3 expression. In some embodiments, the composition upregulates MERTK expression. In some embodiments, the composition upregulates ITGB4 expression. In some embodiments, the composition upregulates gene expression for one or more pathways associated with ECM regulation.

[0090] composition peptide Described herein are compositions comprising octapeptides, as well as methods of preparing and using such compositions. In some embodiments, the compositions and methods comprise one or more peptides for inducing new elastic fiber formation in human skin or stimulating hyaluronic acid (HA) production. In some embodiments, the compositions and methods comprise one or more peptides for stimulating the production of high molecular weight HA. In some embodiments, the octapeptide comprises octapeptide-45. In some embodiments, the octapeptide-45 is a synthetic octapeptide. In some embodiments, the octapeptide-45 is a synthetic octapeptide having the chemical structure shown in FIG. 22.

[0091] The compositions and methods described herein, in some embodiments, comprise one or more octapeptides. In some embodiments, the one or more octapeptides comprise the octapeptide amino acid sequence H-Gly-X 1 -X 2 -X 3 -X 4 -X5 -X 6 -X 7 In some embodiments, the octapeptide amino acid sequence comprises H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 1 is Pro, Asp, or Leu. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 2 is His, Gly, Met, or Gln. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 3 is Gly, Asp, Pro, or Lys. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 4 is Val, Gly, Pro, Phe, or Asn. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 5is Arg, Ala, Ser, Gln, or Gly. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 6 is Glu, Ser, Gly, Arg, or Asp. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 7 is Ala, Pro, or Asp. In some embodiments, the octapeptide amino acid sequence is H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 1 is Pro, Asp, or Leu, and X 2 is His, Gly, Met, or Gln, and X 3 is Gly, Asp, Pro, or Lys, and X 4 is Val, Gly, Pro, Phe, or Asn, and X 5 is Arg, Ala, Ser, Gln, or Gly; X 6 is Glu, Ser, Gly, Arg, or Asp; X 7 is Ala, Pro, or Asp. In some embodiments, the octapeptide amino acid sequence is H-Gly-Pro-Met-Gly-Pro-X 5 -Gly-Pro-OH, X 5is Ser or Arg. In some embodiments, the octapeptide amino acid sequence comprises one or more of GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide is GPHGVREA. In some embodiments, the octapeptide has the chemical structure of octapeptide-45. In some embodiments, the octapeptide is octapeptide-45. In some embodiments, the octapeptide-45 is GPHGVREA.

[0092] In some embodiments, the compositions and methods described herein include one or more octapeptides. In some embodiments, the one or more octapeptides include an octapeptide having sequence characteristics found in collagen and / or elastin. In some embodiments, the octapeptide having sequence characteristics found in collagen and / or elastin includes the amino acid sequence GXXXXXX. In some embodiments, the octapeptide having sequence characteristics found in collagen and / or elastin includes the amino acid sequence H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X 6 -X 7 -OH, X 1 is Pro, Asp, or Leu, and X 2 is His, Gly, Met, or Gln, and X 3 is Gly, Asp, Pro, or Lys, and X 4 is Val, Gly, Pro, Phe, or Asn, and X 5 is Arg, Ala, Ser, Gln, or Gly; X 6 is Glu, Ser, Gly, Arg, or Asp; X 7 is Ala, Pro, or Asp.

[0093] In some embodiments, the composition comprises an octapeptide, a hexapeptide, or a tripeptide, or a combination thereof. In some embodiments, the composition comprises a peptide having amino acid sequence characteristics found in collagen and / or elastin. In some embodiments, the composition comprises a peptide having similar or identical amino acid sequence characteristics to the chemical or enzymatic degradation products of collagen. In some embodiments, the composition comprises a peptide having similar or identical amino acid sequence characteristics to the chemical or enzymatic degradation products of elastin. In some embodiments, the composition and method comprises one or more peptides for inducing new elastic fiber formation or stimulating elastin production in human skin. In some embodiments, the composition and method comprises one or more peptides for inducing new elastic fiber formation or stimulating collagen production in human skin. In some embodiments, the composition and method comprises one or more peptides for inducing new elastic fiber formation or turnover of older, thinner collagen fibers with thicker, healthier collagen fibers in human skin. In some embodiments, the composition and method comprises one or more peptides for inducing new elastic fiber formation or stimulating elastin and collagen production in human skin. In some embodiments, one or more elastin-derived peptides are hexapeptides. In some embodiments, one or more elastin-derived peptides include the amino acid sequence XGVXXG. In some embodiments, one or more elastin-derived peptides include a sequence according to the amino acid sequence IGVAPG, VGVAPG, or VGVTAG. The elastin-derived peptides described herein (e.g., XGVXXG) are used in some embodiments to regenerate damaged or aged human skin. In some embodiments, the compositions and methods described herein include one or more tripeptides, one or more hexapeptides, one or more octapeptides, or a combination thereof. In some embodiments, one or more peptides (e.g., elastin-derived peptides) are synthetic.In some embodiments, the compositions and methods described herein include a mixture of peptides (e.g., peptides derived from elastin) that provide a synergistic effect (e.g., gene expression). In some embodiments, the compositions and methods that include one or more peptides having amino acid sequences characteristic of those found in collagen and / or elastin include an octapeptide.

[0094] In some embodiments, the peptide is an elastin-derived peptide. In some embodiments, the elastin-derived peptide comprises the amino acid sequence XGVXXG. In some embodiments, the elastin-derived peptide comprises a sequence according to the amino acid sequence IGVAPG, VGVAPG, or VGVTAG. In some embodiments, the compositions and methods described herein further comprise one or more tripeptides.

[0095] Peptide concentration The compositions described herein, in some embodiments, include an octapeptide. In some embodiments, the octapeptide is present in an amount of at least 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05% by weight based on the total weight of the composition. %, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or greater than 10% by weight (wt.%). In some embodiments, the octapeptide is provided in the range of about 0.00001 wt.% to 1.0 wt.%, 0.0001 wt.% to 0.01 wt.%, about 0.0001 wt.% to 0.001 wt.%, about 0.0001 wt.% to 0.0005 wt.%, about 0.25 wt.% to about 10 wt.%, about 0.5 wt.% to about 8 wt.%, about 0.75 wt.% to about 6 wt.%, or about 1 wt.% to about 4 wt.%. In some embodiments, the octapeptide is provided at a concentration of at least about 0.10, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 parts per million (ppm). In some embodiments, the octapeptide is provided in a range of about 0.1 ppm to about 200 ppm, about 1 ppm to about 200 ppm, about 1 to about 150 ppm, about 1 to about 100 ppm, about 1 to about 100 ppm, about 1 to about 50 ppm, about 1 to about 25 ppm, about 1 to about 20 ppm, about 1 to about 10 ppm, or about 10 to about 100 ppm. In some embodiments, the octapeptide is provided at a concentration of about 10 ppm. In the composition, the octapeptide may be present in an amount of about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, e.g., 50 ppm to 150 ppm.In the compositions, the hexapeptide is typically present in an amount of about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, e.g., 50 ppm to 150 ppm. In some embodiments, the octapeptide is provided at a concentration of at least about 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 μg / mL. In some embodiments, the octapeptide is provided at a concentration of about 1 to about 200 μg / mL, about 1 to about 150 μg / mL, about 1 to about 100 μg / mL, about 1 to about 50 μg / mL, about 1 to about 10 μg / mL, about 10 to about 100 μg / mL, about 10 to about 50 μg / mL, about 20 to about 100 μg / mL, or about 30 to about 100 μg / mL. In some embodiments, the octapeptide is provided at a concentration of about 10 μg / mL. In some embodiments, the octapeptide is provided at a concentration of about 10 μg / mL to 20 μg / mL. In some embodiments, the octapeptide is provided at a concentration of about 20 μg / mL. In some embodiments, the octapeptide is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter.

[0096] The compositions described herein include varying concentrations of peptides. In some cases, the peptides are present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 ppm. In some cases, the peptides are present in the range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 ppm. In some cases, the peptide is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or greater than 1000 micrograms per milliliter (μg / mL). In some cases, the peptide is present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 micrograms per milliliter. In some cases, the peptide is present at about 0.01% to about 10% by weight, about 0.01% to about 0.02% by weight, about 0.01% to about 0.03% by weight, about 0.01% to about 0.04% by weight, about 0.01% to about 0.05% by weight, about 0.01% to about 0.1% by weight, about 1% to about 5% by weight, or about 1% to about 10% by weight (wt.%).

[0097] The compositions described herein, in some embodiments, comprise one or more peptides. In some cases, a peptide of the one or more peptides is present at about 50 ppm or less to 1000, 5000, 10000, 50000, 100000, 500000 ppm or more, such as 100 ppm of peptide, or any other suitable amount. In some cases, a peptide of the one or more peptides is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or more than 1000 ppm. In some cases, a peptide of the one or more peptides is present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 ppm. In some cases, a peptide of the one or more peptides is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or greater than 1000 micrograms per milliliter (μg / mL). In some cases, a peptide of the one or more peptides is present in the range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 micrograms per milliliter. In some cases, a peptide of the one or more peptides is present in about 0.01% to about 10% by weight, about 0.01% to about 0.02% by weight, about 0.01% to about 0.03% by weight, about 0.01% to about 0.04% by weight, about 0.01% to about 0.05% by weight, about 0.01% to about 0.1% by weight, about 1% to about 5% by weight, or about 1% to about 10% by weight (wt.%).In some embodiments, a peptide of the one or more peptides is at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.0 5%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or greater than 10% by weight (wt.%). In some embodiments, a peptide of the one or more peptides is provided in the range of about 0.25% to about 10%, about 0.5% to about 8%, about 0.75% to about 6%, or about 1% to about 4% by weight. In some embodiments, each peptide of the one or more peptides is provided in a range of about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.01% to about 3% by weight, or about 0.02% to about 2% by weight.

[0098] In some embodiments, the tripeptide is a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate. In the composition, the tripeptide is typically present in an amount of about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, for example, 50 ppm to 150 ppm. In the composition, the hexapeptide is typically present in an amount of about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, for example, 50 ppm to 150 ppm.

[0099] In some embodiments, the tripeptide is tripeptide-1. In some embodiments, tripeptide-1 is at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.0 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or greater than 10% by weight (wt.%). In some embodiments, tripeptide-1 is provided in the range of about 0.01 wt.% to about 2 wt.%, about 0.25 wt.% to about 10 wt.%, about 0.5 wt.% to about 8 wt.%, about 0.75 wt.% to about 6 wt.%, or about 1 wt.% to about 4 wt.%. In some embodiments, tripeptide-1 is provided at at least about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or greater than 25 ppm. In some embodiments, tripeptide-1 is provided in the range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 ppm. In some embodiments, tripeptide-1 is provided in the range of about 1 to about 10 ppm. In some embodiments, tripeptide-1 is provided at at least about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or greater than 25 micrograms per milliliter (μg / mL). In some embodiments, tripeptide-1 is provided in the range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter.

[0100] In some embodiments, the elastin-derived peptide is a synthetic peptide. In some embodiments, the elastin-derived peptide is a peptide that comprises elastin-derived VGVAPG, IGVAPG, VGVTAG (specified in IGF-1 binding protein-1 (IGFBP-1)), or derivatives thereof, that mimics the elastin-derived VGVAPG, IGVAPG, VGVTAG (specified in IGF-1 binding protein-1 (IGFBP-1)), or derivatives thereof. In some embodiments, the elastin-derived peptide is a synthetic peptide that comprises elastin-derived VGVAPG, IGVAPG, VGVTAG (specified in IGF-1 binding protein-1 (IGFBP-1)), or derivatives thereof, that mimics the elastin-derived VGVAPG, IGVAPG, VGVTAG (specified in IGF-1 binding protein-1 (IGFBP-1)), or derivatives thereof. In some embodiments, the elastin-derived peptide comprises the amino acid sequence XGVXXG. In some embodiments, the elastin-derived peptide comprises a sequence according to the amino acid sequence IGVAPG, VGVAPG, or VGVTAG. In some embodiments, the synthetic peptide is a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, the hexapeptide-11 is encapsulated in a liposome. In some embodiments, the tripeptide is encapsulated in a liposome. In some embodiments, the octapeptide is encapsulated in a liposome. In some embodiments, the octapeptide-45 is encapsulated in a liposome. In some embodiments, TriHex and octapeptide-45 are encapsulated in a liposome. In some embodiments, the synthetic peptide is lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome. In some embodiments, encapsulation in a liposome increases the efficiency of peptide delivery to cells. In some embodiments, encapsulation in a liposome increases the efficiency of peptide delivery to cells in deeper layers of the skin. In some embodiments, encapsulation in a liposome increases the efficiency of peptide delivery to cells in deeper epidermal layers of the skin. In some embodiments, encapsulation in a liposome increases the efficiency of peptide delivery to cells in deeper dermal layers of the skin. In some embodiments, encapsulation in a liposome increases the half-life of the peptide in the formulation.In some embodiments, liposomal encapsulation increases the half-life of the tripeptide in the formulation. In some embodiments, liposomal encapsulation increases the half-life of hexapeptide-11 in the formulation. In some embodiments, liposomal encapsulation increases the half-life of the octapeptide in the formulation. In some embodiments, liposomal encapsulation increases the half-life of the octapeptide-45 in the formulation. In some embodiments, liposomal encapsulation results in a formulation with sustained release of the peptide upon application. In some embodiments, liposomal encapsulation results in a formulation with sustained release of the peptide upon topical application. In some embodiments, liposomal encapsulation results in a formulation with sustained release of the peptide upon topical application to facial skin.

[0101] In some embodiments, the elastin derived peptides are at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05% by weight , 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10% or more by weight (wt.%). In some embodiments, the elastin-derived peptides are provided in the range of about 0.00001% to about 10% by weight, about 0.0001% to about 0.01% by weight, about 0.0003% to about 9% by weight, about 0.0005% to about 8% by weight, about 0.001% to about 4% by weight, or about 0.001% to 2% by weight (wt.%), or any range or value therebetween. In some embodiments, the elastin-derived peptides are provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or greater than 25 ppm. In some embodiments, the elastin-derived peptides are provided in the range of about 1 to about 10 ppm. In some embodiments, the elastin-derived peptides are provided in the range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 ppm. In some embodiments, the elastin-derived peptides are provided at least about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or greater than 25 micrograms per milliliter (μg / mL). In some embodiments, the elastin-derived peptides are provided in the range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter.

[0102] In some embodiments, hexapeptide-12 (VGVAPG) is at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05 %, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or greater than 10% by weight (wt.%). In some embodiments, hexapeptide-12 is provided in the range of about 0.00001% to about 10% by weight, about 0.0001% to about 0.01% by weight, about 0.0003% to about 9% by weight, about 0.0005% to about 8% by weight, or about 0.001% to about 4% by weight, about 0.001% to about 2% by weight (wt.%), or any range or value therebetween. In some embodiments, hexapeptide-12 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm. In some embodiments, hexapeptide-12 is provided in the range of about 1 to about 10 ppm. In some embodiments, hexapeptide-12 is provided in the range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 ppm. In some embodiments, hexapeptide-12 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or greater than 25 micrograms per milliliter (μg / mL). In some embodiments, hexapeptide-12 is provided in the range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter.

[0103] The compositions described herein, in some embodiments, include hexapeptide-11. In some embodiments, hexapeptide-11 is at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, It is provided at 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10% or greater than 10% by weight (wt.%). In some embodiments, hexapeptide-11 is provided in the range of about 0.00001% to about 10% by weight, about 0.0001% to about 0.01% by weight, about 0.0003% to about 8% by weight, about 0.0005% to about 6% by weight, about 0.001% to about 4% by weight, about 0.001% to about 2% by weight, about 0.005% to about 2% by weight, or about 0.01% to about 1% by weight (wt.%), or any range or value therebetween. In some embodiments, hexapeptide-11 is provided in the range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2%. In some embodiments, hexapeptide-11 is provided in the range of about 0.005% to about 0.02% by weight. In some embodiments, hexapeptide-11 is provided at at least about 0.1 ppm, 3 ppm, 5 ppm, 10 ppm, 50 ppm, 55 ppm, 500 ppm, 1,000 ppm, 2,500 ppm, 5,000 ppm, or greater than 5,000 ppm. In some embodiments, hexapeptide-11 is provided in the range of about 5 ppm to about 100 ppm, about 10 ppm to about 1000 ppm, about 50 ppm to about 1500 ppm, or about 500 ppm to about 5,000 ppm. In some embodiments, hexapeptide-11 is about 1000 ppm.In some embodiments, hexapeptide-11 is provided at at least about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, or greater than 500 micrograms per milliliter (μg / mL). In some embodiments, hexapeptide-11 is provided in the range of about 25 to about 250, about 50 to about 200, about 75 to about 150, about 200 to about 300, or about 200 to about 400 micrograms per milliliter.

[0104] Peptide Modifications The peptides can be functionalized. For example, the peptides can be functionalized with fatty acids such as myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, and the like. Examples include palmitoyl hexapeptide-12 (Pal-VGVAPG), palmitoyl tripeptide-1 (Pal-GHK), myristoyl hexapeptide-12 (Myr-VGVAPG), and myristoyl tripeptide-1 (Myr-GHK). Palmitoyl or myristoyl functionalization may be desirable in certain embodiments, as they exhibit enhanced penetration when compared to other fatty acids. In some embodiments, the peptide is functionalized with a chemical group. For example, the peptide is functionalized with acetyl. In some cases, the peptide is functionalized with a functional group containing 14 carbons or less. In some cases, the peptide is functionalized with a functional group containing 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or less carbons or more than 20 carbons. In some cases, the peptide is non-palmitoylated. Without being limited to a particular theory, in some embodiments, the incorporation of the peptide into a liposome increases the lipophilicity of the peptide, whether functionalized or not functionalized.

[0105] Some embodiments of the methods and compositions provided herein include glycine-histidine-lysine (GHK) as the first peptide. GHK is a peptide sequence that is rarely found in common protein classes, but is frequently found in extracellular matrix proteins. The small size of GHK allows it to approach membrane receptors much more easily than larger peptides. Furthermore, its unique copper-binding structure enhances copper transport in and out of cells, promoting wound healing through several different but related pathways. Due to its strong copper-binding structure, GHK can be provided in the form of GHK-Cu (copper-bound GHK form).

[0106] In some embodiments, the compositions described herein comprise an octapeptide. In some embodiments, the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO:1), GPMGPSGP (SEQ ID NO:2), GLGPGARA (SEQ ID NO:3), GPQGFQGP (SEQ ID NO:4), GPHGVREA (SEQ ID NO:5), GPMGPRGP (SEQ ID NO:6), GPGKNGDD (SEQ ID NO:7), or GPMGPRGP (SEQ ID NO:8). In some embodiments, the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO:1). In some embodiments, the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO:2). In some embodiments, the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO:3). In some embodiments, the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO:4). In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO:5). In some embodiments, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO:6). In some embodiments, the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO:7). In some embodiments, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8). In some embodiments, the octapeptide-45 is GPHGVREA (SEQ ID NO: 5).

[0107] In some embodiments, the octapeptide is a synthetic peptide. In some embodiments, the octapeptide is a peptide that stimulates HA synthesis. In some embodiments, the octapeptide is a peptide that stimulates high molecular weight hyaluronic acid (high MW HA) synthesis. In some embodiments, the octapeptide is a peptide that stimulates high MW HA synthesis in the dermis. In some embodiments, the octapeptide is a peptide that stimulates high MW HA synthesis in the skin. In some embodiments, the octapeptide comprises an octapeptide having the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide comprises GPHGVREA. In some embodiments, the octapeptide is GPHGVREA. In some embodiments, the octapeptide comprises the chemical structure of octapeptide-45 (Figure 22).

[0108] Liposomes In some embodiments, the compositions according to the present disclosure include liposomes for improved distribution, efficacy, bioavailability, and / or activity. The liposome composition may improve the distribution, efficacy, bioavailability, and / or activity of the active ingredient by improving delivery and tissue (e.g., skin) penetration. In some cases, the improved delivery and skin penetration is due to the active ingredient being incorporated (e.g., encapsulated) in the liposome. In some cases, the active ingredient is one or more peptides encapsulated in the liposome.

[0109] The liposome compositions described herein may include a peptide encapsulated in the liposome. In some embodiments, the peptide encapsulated in the liposome is selected from the group consisting of hexapeptide-12, tripeptide-1, one or more octapeptides (e.g., octapeptide-45), hexapeptide-11, hexapeptide-38, tetrapeptide-2, or any combination thereof. In some embodiments, the peptide encapsulated in the liposome is functionalized with an acetyl group, a palmitoyl group, a myristoyl group, or any combination thereof.

[0110] The liposome compositions described herein may include various components encapsulated in the liposomes. In some embodiments, the component is lactoferrin. In some embodiments, the component is phosphatidylserine. In some embodiments, the component is Ledum palustre extract. In some embodiments, the component is Arnica montana extract. In some embodiments, the component is sodium hyaluronate. In some embodiments, the component is larger than 50 kDa.

[0111] Lecithin and other phospholipids can be used to prepare liposomes containing the peptide compositions described herein. In some embodiments, liposomes are used to prepare one or more peptides. In some embodiments, the peptides are functionalized with acetyl groups. The formation of lipid vesicles occurs when phospholipids such as lecithin are placed in water, which results in the formation of a bilayer or a series of bilayers, each separated by water molecules, when sufficient energy is provided. Liposomes can be made by sonicating phospholipids in water. Low shear rates create multi-layered liposomes. Continued high shear sonication tends to form smaller unilamellar liposomes. Hydrophobic chemicals can dissolve in the phospholipid bilayer membrane. The lipid bilayer of the liposome delivers the peptide compositions described herein.

[0112] The phospholipids used to prepare the liposomal compositions described herein may comprise a transition phase temperature of about 10° C. to about 25° C. In some cases, the phospholipids comprise a transition phase temperature of about 10° C., 12° C., 14° C., 16° C., 18° C., 20° C., 22° C., 24° C., 26° C., 28° C., 30° C., 32° C., 34° C., 36° C., 38° C., 40° C., or greater than 40° C. In some cases, the phospholipids comprise a transition phase temperature in the range of about 10° C. to about 40° C., about 12° C. to about 36° C., about 14° C. to about 32° C., about 16° C. to about 20° C., or about 21° C. to about 25° C.

[0113] The topical composition may contain micelles, or aggregates of surfactant molecules, dispersed in an aqueous solution. Micelles can be prepared by dispersing an oil solvent in an aqueous solution containing a surfactant, where the surfactant concentration exceeds the critical micelle concentration. The resulting composition contains micelles, i.e., spherical oil droplets.

[0114] The liposome composition may contain micelles or aggregates of surfactant molecules dispersed in an aqueous solution. Micelles can be prepared by dispersing an oil solvent in an aqueous solution containing a surfactant, where the surfactant concentration exceeds the critical micelle concentration. The resulting formulation contains micelles, i.e., spherical oil droplets surrounded by a membrane of polar surfactant molecules, dispersed in an aqueous solvent.

[0115] In some embodiments, methods are described herein for preparing compositions comprising peptides encapsulated in liposomes, comprising combining a peptide and a solvent to form a mixture and contacting the mixture with an aqueous solution comprising liposomes. In some cases, the contacting is performed at a temperature of about 10° C. to about 25° C. In some cases, the contacting is performed at about 10° C., 12° C., 14° C., 16° C., 18° C., 20° C., 22° C., 24° C., 26° C., 28° C., 30° C., 32° C., 34° C., 36° C., 38° C., 40° C., or greater than 40° C. In some cases, the contacting is performed at a temperature ranging from about 10° C. to about 40° C., from about 12° C. to about 36° C., from about 14° C. to about 32° C., from about 16° C. to about 20° C., or from about 21° C. to about 25° C.

[0116] The method for preparing a composition comprising a peptide encapsulated in liposomes may include the use of a solvent. In some cases, the solvent is water. In some cases, the solvent is an organic solvent. Exemplary organic solvents include, but are not limited to, petroleum ether, cyclohexane, toluene, carbon tetrachloride, dichloromethane, chloroform, diethyl ether, diisopropyl ether, ethyl acetate, butanol, n-propanol, ethanol, methanol, polyethylene glycol, propylene glycol, and pyridine. In some cases, the solvent is glycol. In some cases, the solvent is butylene glycol. In some cases, the solvent is caprylyl glycol. In some cases, the solvent is propanediol (propylene glycol).

[0117] Solvents can be used in various percentages. In some cases, the solvent is provided at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight. The solvent can be propanediol, butylene glycol, or caprylyl glycol.

[0118] The methods described herein, in some embodiments, include combining a peptide and a solvent to form a mixture and contacting the mixture with an aqueous solution containing liposomes, the aqueous solution comprising a percentage of water and a percentage of liposomes. In some cases, the aqueous solution comprises at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more than 90% water. In some cases, the aqueous solution comprises in the range of about 10% to about 95%, about 20% to about 90%, about 30% to about 85%, about 40% to about 80%, or about 50% to about 60% water. In some cases, the aqueous solution comprises at least or about 20%, 30%, 40%, 50%, 60%, or more than 60% liposomes. In some cases, the aqueous solution comprises liposomes in the range of about 10% to about 80%, about 20% to about 70%, or about 30% to about 60%. The ratio of liposomes to water can be in the range of about 1:9 to about 3:7. In some cases, the ratio of liposomes to water can be at least or about 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, or 1:2.

[0119] The methods for producing liposomal compositions described herein can result in an entrapment efficiency of 100% or less. In some cases, the entrapment efficiency is 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5% or less.

[0120] Described herein are liposome compositions in which the peptide constitutes a percentage of the composition. In some embodiments, the peptide is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% of the composition. In some embodiments, the peptide is provided at at least about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 24%, 26%, 28%, 30%, or more than 30% of the composition. In some embodiments, the peptide is provided in the range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 5%, or about 0.02% to about 2% by weight. In some embodiments, the peptide is provided at about 0.03% of the composition.

[0121] Described herein are liposome compositions in which liposomes constitute a percentage of the composition. In some embodiments, liposomes are provided at at least about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 24%, 26%, 28%, 30%, or greater than 30% of the composition. In some embodiments, liposomes are provided in the range of about 5% to about 90%, about 10% to about 80%, about 20% to about 70%, about 30% to about 60%, about 10% to about 30%, or about 20% to about 40%. In some embodiments, liposomes are provided at about 30%. In some embodiments, liposomes are provided at 27%.

[0122] In some embodiments, the liposome compositions described herein comprise an average particle size of up to 220 nanometers (nm). In some cases, the average particle size is up to 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, 230 nm, 240 nm, 250 nm, 260 nm, 270 nm, 280 nm, 290 nm, 300 nm, 320 nm, 340 nm, 360 nm, 380 nm, or 400 nm. In some cases, the average particle size is about 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, 230 nm, 240 nm, 250 nm, 260 nm, 270 nm, 280 nm, 290 nm, 300 nm, 320 nm, 340 nm, 360 nm, 380 nm, or 400 nm. In some cases, the average particle size ranges from about 50 nm to about 500 nm, from about 100 nm to about 400 nm, from about 150 nm to about 220 nm, from about 180 nm to about 220 nm, or from about 190 nm to about 210 nm.

[0123] In some cases, the liposome composition includes an active agent having a molecular weight of about 600 Daltons (Da) or less. In some cases, the active agent has a molecular weight of at least or about 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, or more than 1000 Daltons (Da). In some cases, the active agent has a molecular weight of at least about 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 4000, 5000, 6000, or more than 6000 Daltons (Da). In some cases, the active agent has a molecular weight in the range of about 50 to about 1000, about 100 to about 900, about 200 to about 800, about 300 to about 700, or about 400 to about 600 Daltons (Da). In some cases, the active agent is a peptide disclosed herein. In some cases, the active agent is a peptide encapsulated in a liposome.

[0124] The polydispersity index (PdI) of the liposome compositions described herein, in some embodiments, ranges from 0 to about 0.2. In some cases, the polydispersity index is about 0.01, 0.025, 0.05, 0.1, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, or 0.8. In some cases, the polydispersity index is in the range of about 0.01 to about 0.8, about 0.025 to about 0.75, about 0.05 to about 0.6, or about 0.1 to about 0.3.

[0125] In some cases, the intercept of the liposome compositions described herein ranges from about 0.85 to about 0.95. In some cases, the intercept is an amplitude. In some cases, the intercept is at least or about 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, or 0.95.

[0126] In some embodiments, the liposome comprises propanediol, lecithin, or a combination thereof, in some embodiments, the propanediol is provided at at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt.%). In some embodiments, propanediol is provided in the range of about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.01% to about 3% by weight, or about 0.02% to about 2% by weight. In some embodiments, lecithin is provided at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt.%) based on the total weight of the composition. In some embodiments, the lecithin is provided in the range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2% by weight based on the total weight of the composition. In some embodiments, the liposome comprises propanediol and lecithin.In some embodiments, propanediol and lecithin are provided at at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt.%) based on the total weight of the composition. In some embodiments, the propanediol and lecithin are provided in the range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2% by weight, based on the total weight of the composition. In some embodiments, the propanediol and lecithin are provided in about 0.90% by weight, based on the total weight of the composition.

[0127] Described herein are liposome compositions that include improved distribution, efficacy, bioavailability, and / or activity. The liposome compositions may include improved distribution, efficacy, bioavailability, and / or activity compared to compositions that do not contain liposomes. In some cases, the distribution is improved by at least or about 0.5-fold, 1.0-fold, 1.5-fold, 2.0-fold, 2.5-fold, 3.0-fold, 4.0-fold, 4.5-fold, 5-fold, or more than 5-fold compared to compositions that do not contain liposomes. In some cases, the efficacy is improved by at least or about 0.5-fold, 1.0-fold, 1.5-fold, 2.0-fold, 2.5-fold, 3.0-fold, 4.0-fold, 4.5-fold, 5-fold, or more than 5-fold compared to compositions that do not contain liposomes. In some cases, bioavailability is improved by at least or about 0.5-fold, 1.0-fold, 1.5-fold, 2.0-fold, 2.5-fold, 3.0-fold, 4.0-fold, 4.5-fold, 5-fold, or more than 5-fold compared to compositions that do not contain liposomes. In some cases, activity is improved by at least or about 0.5-fold, 1.0-fold, 1.5-fold, 2.0-fold, 2.5-fold, 3.0-fold, 4.0-fold, 4.5-fold, 5-fold, or more than 5-fold compared to compositions that do not contain liposomes. Distribution, efficacy, bioavailability, and / or activity may be improved by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90% compared to compositions that do not contain liposomes.

[0128] The liposome compositions and methods described herein are, in some embodiments, topical compositions. In some cases, the liposome compositions do not include oil. In some cases, the liposome compositions do not include preservatives. In some embodiments, the liposome formulation is an aqueous formulation. In some embodiments, the liposome formulation is an anhydrous formulation. In some cases, the liposome composition comprises a pH in the range of about 5 to about 8. In some cases, the liposome composition comprises a pH of at least or about 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0129] The methods and compositions described herein may result in improved follicular penetration. In some cases, follicular penetration is improved by at least or about 0.5 times, 1.0 times, 1.5 times, 2.0 times, 2.5 times, 3.0 times, 4.0 times, 4.5 times, 5 times, or more than 5 times. Follicular penetration may be improved by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90%. In some cases, the compositions result in follicular penetration to a depth of at least or about 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 7, 8, 9, 10, or more than 10 millimeters.

[0130] Gene expression The compositions described herein, in some embodiments, modify gene expression in skin cells. In some embodiments, exposure of dermal cells (e.g., dermal fibroblasts) to an octapeptide results in upregulation of HAS2 expression in the dermal cells. In some embodiments, the dermal cells comprise dermal fibroblasts. In some embodiments, the octapeptide comprises one or more octapeptides having the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide is octapeptide-45.

[0131] In some embodiments, exposure of dermal cells (e.g., dermal fibroblasts) or epidermal cells (e.g., keratinocytes) to a hexapeptide (e.g., hexapeptide-11) results in upregulation of HAS2 expression. In some embodiments, exposure of dermal cells (e.g., dermal fibroblasts) or epidermal cells (e.g., keratinocytes) to a tripeptide and a hexapeptide (TriHex-Hexapeptide-12+Tripeptide-1) results in upregulation of HAS2 expression. In some embodiments, exposure of epidermal cells (e.g., keratinocytes) to a hexapeptide (e.g., hexapeptide-11) results in upregulation of HAS2 expression. In some embodiments, exposure of epidermal keratinocytes to TriHex results in upregulation of HAS2 expression. In some embodiments, exposure of epidermal keratinocytes to hexapeptide-11 results in upregulation of HAS2 expression.

[0132] In some embodiments, exposure of epidermal cells (e.g., keratinocytes) to a hexapeptide (e.g., hexapeptide-11) results in the downregulation of HYAL2 expression. In some embodiments, exposure of epidermal keratinocytes to hexapeptide-11 results in the downregulation of HYAL2 expression.

[0133] In some embodiments, exposure of epidermal cells to a hexapeptide or an octapeptide results in upregulation of EGR3 expression. In some embodiments, exposure of epidermal (e.g., keratinocyte) cells to tripeptide-1 and hexapeptide-12 (TriHex) results in upregulation of EGR3 expression. In some embodiments, exposure of epidermal cells to hexapeptide-11 results in upregulation of EGR3 expression. In some embodiments, exposure of epidermal keratinocytes to a hexapeptide results in upregulation of EGR3 expression. In some embodiments, exposure of epidermal keratinocytes to hexapeptide-11 results in upregulation of EGR3 expression. In some embodiments, exposure of epidermal keratinocytes to an octapeptide (e.g., octapeptide-45) results in upregulation of EGR3 expression.

[0134] In some embodiments, exposure of human dermal fibroblasts to TriHex results in upregulation of LAMA3 expression. In some embodiments, exposure of human dermal fibroblasts to TriHex results in upregulation of MERTK expression. In some embodiments, exposure of human dermal fibroblasts to an octapeptide (e.g., octapeptide-45) results in upregulation of S100A2 expression. In some embodiments, exposure of human dermal fibroblasts to an octapeptide (e.g., octapeptide-45) results in upregulation of LAMA3 expression. In some embodiments, exposure of human dermal fibroblasts to an octapeptide (e.g., octapeptide-45) results in upregulation of MERTK expression. In some embodiments, exposure of human dermal fibroblasts to an octapeptide (e.g., octapeptide-45) results in upregulation of ITGB4 expression.

[0135] In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in upregulation of S100A2 expression. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in synergistic upregulation of S100A2 expression when compared to expression following exposure to TriHex alone or the octapeptide alone. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in upregulation of LAMA3 expression. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in synergistic upregulation of LAMA3 expression when compared to expression following exposure to TriHex alone or the octapeptide alone. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in upregulation of MERTK expression. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in synergistic upregulation of MERTK expression when compared to expression following exposure to TriHex alone or the octapeptide alone. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in upregulation of ITGB4 expression. In some embodiments, exposure of human dermal fibroblasts to TriHex + octapeptide (e.g., octapeptide-45) results in synergistic upregulation of ITGB4 expression when compared to expression following exposure to TriHex alone or the octapeptide alone.

[0136] Hyaluronic Acid Synthesis In some embodiments, a composition comprising the octapeptide results in increased synthesis of high MW HA compared to skin not treated with the composition. In some embodiments, the octapeptide in the composition stimulates synthesis of high MW HA. In some embodiments, the increase in HA synthesis is due to stimulation of hyaluronic acid synthase (HAS) activity. In some embodiments, the HAS comprises HAS2. In some embodiments, the increase in HA synthesis is due to downregulation of hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2). In some embodiments, the octapeptide stimulates HAS2 in fibroblasts, an enzyme that stimulates HA production. In some embodiments, the octapeptide is encapsulated in a phospholipid delivery system for increased absorption and activity. In some embodiments, the hexapeptide provides upregulation of HAS2 in keratinocytes and strong downregulation of HYAL2 in keratinocytes. In some embodiments, the compositions described herein stimulate synthesis of HA in dermal and / or epidermal cells. In some embodiments, the HA synthesized upon application of the composition comprises a higher MW HA than HA synthesized without the composition. In some embodiments, the high MW HA produced by stimulation has a MW of at least 0.5 MDa, 1.0 MDa, 1.5 MDa, or 2.0 MDa. In some embodiments, the high MW HA produced by stimulation has a MW of about 1 MDa to 4 MDa. In some embodiments, exposure of dermal cells to the octapeptide stimulates the production of HA. In some embodiments, exposure of dermal cells to octapeptide-45 stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to the octapeptide stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to the octapeptide at a concentration of at least about 100 μg / ml stimulates the production of HA. In some embodiments, exposure of epidermal cells to the octapeptide stimulates the production of HA. In some embodiments, exposure of epidermal cells to octapeptide stimulates the production of HA.In some embodiments, exposure of epidermal keratinocytes to the octapeptide stimulates the production of HA. In some embodiments, exposure of epidermal keratinocytes to octapeptide-45 stimulates the production of HA.

[0137] In some embodiments, exposure of dermal fibroblasts to lactoferrin at a concentration of at least about 500 μg / ml stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to phosphatidylserine at a concentration of at least about 500 μg / ml stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to Tremella at a concentration of at least about 500 μg / ml stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to hydroxymethoxphenyl decanone at a concentration of at least about 250 μg / ml stimulates the production of HA.

[0138] In some embodiments, exposure of skin to a composition comprising one or more octapeptides increases expression of CD44. In some embodiments, exposure of skin to a composition comprising octapeptide-45 increases expression of CD44. In some embodiments, exposure of epidermal keratinocytes and / or dermal fibroblasts to a composition increases expression of CD44 in the exposed cells. In some embodiments, increased expression of CD44 contributes to remodeling of the dermal ECM. In some embodiments, increased expression of CD44 contributes to remodeling of the dermal ECM reversing the appearance of solar elastosis. In some embodiments, increased expression of CD44 contributes to reduced skin redness. In some embodiments, increased expression of CD44 contributes to reduced skin inflammation. In some embodiments, increased expression of CD44 contributes to reduced appearance of wrinkles. In some embodiments, increased expression of CD44 contributes to reduced appearance of fine lines. In some embodiments, increased expression of CD44 contributes to reduced appearance of deep lines. In some embodiments, increased expression of CD44 contributes to reducing the appearance of deep lines in facial skin. In some embodiments, increased expression of CD44 contributes to reducing the appearance of fine lines. In some embodiments, increased expression of CD44 contributes to reducing the appearance of pore size. In some embodiments, increased expression of CD44 contributes to improving the appearance of skin texture. In some embodiments, increased expression of CD44 contributes to reducing skin dryness. In some embodiments, increased expression of CD44 contributes to improving skin hydration.

[0139] Hyaluronic acid In some embodiments described herein, the formulations for use on the skin comprise hyaluronic acid or a derivative thereof. In some embodiments, the hyaluronic acid comprises high molecular weight hyaluronic acid (high MW HA), or a derivative thereof. In some embodiments, the high MW HA comprises a high MW HA having an average molecular weight of at least about 0.5 MDa, 1.0 MDa, 1.5 MDa, 2.0 MDa, 3.0 MDa, 4.0 MDa, or 5.0 MDa. In some embodiments, the high MW HA comprises a high MW HA with a molecular weight of about 1 MDa to about 5 MDa. In some embodiments, the high MW HA comprises sodium hyaluronate. In some embodiments, the high MW HA comprises sodium hyaluronate crosspolymer.

[0140] Formulation components In some embodiments described herein, the composition comprises phosphatidylserine. In some embodiments described herein, the composition comprises Tremella fuciformis extract. In some embodiments described herein, the composition comprises hydroxymethoxyphenyldecanone. In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, a phosphatidylserine, a Tremella fuciformis extract, or a combination thereof. In some embodiments, the composition provided herein increases the production of high MW HA in skin cells. In some embodiments, the composition increases the production of high MW HA in epidermal cells. In some embodiments, the composition increases the production of high MW HA in dermal cells. In some embodiments, the composition provided herein increases barrier integrity below the basal layer. In some embodiments, the composition increases barrier thickness below the basal layer. In some embodiments, the composition increases expression of genes that regulate barrier function of the basal layer. In some embodiments, the composition increases barrier function of the skin. In some embodiments, the composition increases barrier function of the skin to prevent excessive water loss. In some embodiments, the composition increases the barrier function of the skin to maintain a higher moisture content. In some embodiments, the composition increases the barrier function of the skin to increase skin hydration. In some embodiments, the composition increases the barrier function of the skin to reduce skin dryness. In some embodiments, the composition increases the barrier function of the skin to reduce redness. In some embodiments, the composition increases the barrier function of the skin to reduce inflammation. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of wrinkles. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of fine lines.In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of deep lines in the skin. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of deep lines in facial skin. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of fine lines. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of pore size. In some embodiments, the composition increases the barrier function of the skin to improve the appearance of skin texture. In some embodiments, the composition is aqueous. In some embodiments, the composition reduces inflammation as assessed by a reduction or lack of increase in expression levels of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), or cluster of differentiation-80 (CD80), or a combination thereof. In some embodiments, the composition increases CD44 expression. In some embodiments, the composition reduces redness of the skin of an individual after applying the composition. In some embodiments, the composition reduces redness of the skin of an individual after applying the composition, and the reduction in redness is measured by photography. In some embodiments, the composition increases skin hydration in the skin of an individual after application of the composition. In some embodiments, the composition increases skin hydration in the skin of an individual after application of the composition. In some embodiments, the composition promotes HA synthesis. In some embodiments, the composition upregulates hyaluronic acid synthase (HAS) expression. In some embodiments, the composition upregulates hyaluronic acid synthase (HAS) expression, where the HAS comprises HAS2. In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the composition downregulates hyaluronidase expression, where the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0141] In some embodiments, the composition comprises tripeptide-1, hexapeptide-12, and octapeptide-45. In some embodiments, the composition increases elastin production. In some embodiments, the composition increases the production of tropoelastin. In some embodiments, the composition increases the production of microfibril associated protein 4 (MFAP4). In some embodiments, the composition upregulates SA1002 expression. In some embodiments, the composition upregulates LAMA3 expression. In some embodiments, the composition upregulates MERTK expression. In some embodiments, the composition upregulates ITGB4 expression. In some embodiments, the composition upregulates gene expression for one or more pathways associated with ECM regulation.

[0142] In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, a phosphatidylserine, a Tremella fuciformis extract, or a hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, the composition provided herein increases the production of high MW HA in skin cells. In some embodiments, the composition increases the production of high MW HA in epidermal cells. In some embodiments, the composition increases the production of high MW HA in dermal cells. In some embodiments, the composition provided herein reduces one or more of the following: the appearance of redness, skin inflammation, the appearance of rosacea, the appearance of erythema, the appearance of fine blood vessels in the skin, the appearance of wrinkles, the appearance of fine lines, the appearance of deep lines, the appearance of fine lines in the skin, the appearance of pore size, excess water loss in the skin, dry skin, or the appearance of solar elastosis. In some embodiments, the composition provided herein improves the appearance of skin texture and / or increases skin hydration. In some embodiments, the composition provided herein increases the rate of collagen turnover. In some embodiments, collagen turnover involves the replacement of old, thin collagen fibers in the dermal ECM with denser, healthier collagen fibers in the dermal ECM.

[0143] In some embodiments described herein, a topical composition for reducing inflammation includes an octapeptide and hyaluronic acid (HA) or a derivative thereof, wherein the composition reduces inflammation. In some embodiments, the HA or a derivative thereof has a molecular weight of at least 0.5 MDa. In some embodiments, the HA or a derivative thereof has a molecular weight of at least 1 MDa. In some embodiments, the high MW HA or derivative comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprises sodium hyaluronate crosspolymer. In some embodiments, the octapeptide comprises at least one of the amino acid sequences GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). In some embodiments, the octapeptide is the amino acid sequence GPHGVREA (SEQ ID NO: 5). In some embodiments, the octapeptide comprises octapeptide-45. In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate. In some embodiments, the composition comprises a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, the hexapeptide-11 is encapsulated in a liposome. In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome. In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises a Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyldecanone.In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, the composition is aqueous.

[0144] In some embodiments, the reduction in inflammation is assessed by a reduction or lack of increase in the expression levels of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), or cluster of differentiation-80 (CD80), or a combination thereof. In some embodiments, the composition increases CD44 expression. In some embodiments, the composition reduces redness in the skin of the individual after application of the composition. In some embodiments, the redness is measured by photography. In some embodiments, the composition increases skin hydration in the skin of the individual after application of the composition. In some embodiments, the composition promotes HA synthesis. In some embodiments, the composition upregulates hyaluronan synthase (HAS) expression. In some embodiments, the composition upregulates hyaluronan synthase (HAS) expression, where the HAS comprises HAS2. In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the composition downregulates hyaluronidase expression, where the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0145] In some embodiments, the composition reduces the appearance of fine lines. In some embodiments, the composition reduces the appearance of deep lines. In some embodiments, the composition reduces the appearance of deep lines in facial skin. In some embodiments, the composition reduces the appearance of wrinkles. In some embodiments, the composition reduces the appearance of age spots. In some embodiments, the composition reduces the appearance of fine lines. In some embodiments, the composition reduces the appearance of total area pore size. In some embodiments, the composition reduces skin dryness. In some embodiments, the composition improves the appearance of skin texture. In some embodiments, the composition improves the appearance of solar elastosis. In some embodiments, the composition reduces or reverses the signs of solar elastosis.

[0146] In some embodiments, the compositions provided herein act to increase the production of high MW HA in the skin. In some embodiments, the compositions provided herein act to increase the amount of high MW HA in the skin. In some embodiments, the compositions provided herein act to increase the production of high MW HA by one or more of epidermal cells, fibroblasts, keratinocytes, or other skin cells. In some embodiments, the compositions provided herein act to increase the deposition of high MW HA in the skin ECM. In some embodiments, application of the compositions provided herein results in an increase in the ratio of high MW HA to low MW HA in the skin. In some embodiments, application of the compositions provided herein results in an increase in the ratio of high MW HA to low MW HA in the epidermis, dermis, or combinations thereof. In some embodiments, the increase in the ratio of high MW HA to low MW HA in the skin results in reduced inflammation. In some embodiments, the increase in the ratio of high MW HA to low MW HA in the skin results in reduced visible redness. In some embodiments, the increase in the ratio of high MW HA to low MW HA in the skin results in reduced visible fine lines. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reduction in the visible deep lines. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reduction in the visible deep lines on the face of an individual. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reduction in fine skin lines. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in improved skin elasticity. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reduction in the total area of ​​pore size. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in an improvement in the visible skin texture. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reduction in skin dryness.In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in increased skin hydration. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reduction in the visible signs of solar elastosis. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a reversal of the visible signs of solar elastosis. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a remodeling of the dermal ECM to produce thicker collagen fibers. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a remodeling of the dermal ECM to produce denser collagen fibers. In some embodiments, increasing the ratio of high MW HA to low MW HA in the skin results in a remodeling of the dermal ECM to a healthier collagen fiber network. In some embodiments, increasing high MW HA in the skin results in a reduction in inflammation in the epidermis and an increase in high MW HA production in the dermis.

[0147] Stability testing Stability testing of compositions can be performed as follows: High temperature testing is now commonly used as a predictor of long term stability. High temperature testing can be performed at 37°C (98°F) and 45°C (113°F). If the product is stored at 45°C for 3 months (and shows acceptable stability), it should be stable at room temperature for 2 years. A good controlled temperature is 4°C (39°F), with most products showing excellent stability. In some cases, the product is subjected to -10°C (14°F) for 3 months.

[0148] In some cases, the stability of the product is evaluated by undergoing three cycles of temperature testing from -10°C (14°F) to 25°C (77°F). In such cases, the product is placed at -10°C for 24 hours, then placed at room temperature (25°C) for 24 hours. This completes one cycle. An even more stringent test is a five cycle test from -10°C to 45°C, which places a great deal of stress on the emulsion.

[0149] In some cases, the dispersed phase (of an oil-in-water emulsion) may separate and rise to the top of the emulsion, forming a layer of oil droplets. This phenomenon is called creaming. Creaming is one of the first signs of approaching emulsion instability. In some cases, one test method to predict creaming is centrifugation. In some cases, the emulsion is heated to 50° C. (122° F.) and centrifuged at 3000 rpm for 30 minutes. In some cases, the emulsion is examined for signs of creaming.

[0150] In some embodiments, the formulation and packaging may be sensitive to UV light. In some cases, the product is placed in a glass and the actual packaging is placed in a light box with broad spectrum output. In some cases, a separate glass bottle completely covered in aluminum foil acts as a control. In some cases, discoloration of the product may be observed.

[0151] For all the above tests, color, odor / scent, viscosity, pH value, and, if available, particle size uniformity and / or particle agglomeration under a microscope may be observed.

[0152] method Methods of Using Peptide Formulations Described herein are methods of preparing and using compositions comprising one or more peptides that comprise amino acid sequence characteristics found in collagen and / or elastin. In some embodiments, the peptide comprises an amino acid sequence characteristic found in elastin. In some embodiments, the peptide comprises an amino acid sequence characteristic found in collagen. In some embodiments, the method comprises topical application of the composition. In some embodiments, the one or more peptides comprise an octapeptide. In some embodiments, the octapeptide comprises the amino acid sequence H-Gly-X 1 -X 2 -X 3 -X 4 -X 5 -X6 -X 7 -OH, X 1 is Pro, Asp, or Leu, and X 2 is His, Gly, Met, or Gln, and X 3 is Gly, Asp, Pro, or Lys, and X 4 is Val, Gly, Pro, Phe, or Asn, and X 5 is Arg, Ala, Ser, Gln, or Gly; X 6 is Glu, Ser, Gly, Arg, or Asp; X 7 is Ala, Pro, or Asp. In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD, or any combination thereof. In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA. In some embodiments, the octapeptide comprises the chemical structure of octapeptide-45 (Figure 22).

[0153] Described herein are methods for reducing inflammation in the skin of an individual, comprising topical application of one or more peptides having amino acid sequence characteristics found in collagen or elastin. In some embodiments, the method for reducing inflammation in the skin of an individual comprises applying a topical composition comprising an octapeptide. In some embodiments, the octapeptide comprises at least one of the amino acid sequences GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). In some embodiments, the octapeptide comprises octapeptide-45.

[0154] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate. In some embodiments, the composition comprises a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, the hexapeptide-11 is encapsulated in a liposome. In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome. In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises a Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyldecanone. In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, a phosphatidylserine, a Tremella fuciformis extract, or a hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, the composition is aqueous. In some embodiments, the reduction in inflammation is assessed by a reduction or lack of increase in the expression levels of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), or cluster of differentiation-80 (CD80), or a combination thereof. In some embodiments, the composition increases CD44 expression. In some embodiments, the composition reduces redness of the individual's skin after application of the composition. In some embodiments, redness is measured by photography.

[0155] How to Increase Elastin Production In some embodiments, the present disclosure relates to a method of treating or restoring aging skin or increasing elastin production in the skin, such method comprising administering to the skin a topical composition disclosed herein. In some embodiments, the composition comprises tripeptide-1, hexapeptide-12, and octapeptide-45. In some embodiments, the composition increases elastin production. In some embodiments, the composition increases the production of tropoelastin. In some embodiments, the composition increases the production of microfibril associated protein 4 (MFAP4). In some embodiments, the composition upregulates SA1002 expression. In some embodiments, the composition upregulates LAMA3 expression. In some embodiments, the composition upregulates MERTK expression. In some embodiments, the composition upregulates ITGB4 expression. In some embodiments, the composition upregulates gene expression for one or more pathways related to ECM regulation.

[0156] How to Increase Skin Hydration In some embodiments, the composition increases skin hydration on the individual's skin after application of the composition. In some embodiments, the skin hydration on the individual's skin is measured at baseline. In some embodiments, the skin hydration on the individual's skin is measured 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the initiation of a skin treatment routine that includes application of the topical composition. In some embodiments, the individual completes a self-assessment of skin hydration at baseline. In some embodiments, the individual completes a self-assessment of skin hydration at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the initiation of a skin treatment routine that includes application of the topical composition. In some embodiments, the responsible physician completes an assessment of the individual's skin hydration at baseline. In some embodiments, the responsible physician completes an assessment of the individual's skin hydration at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the initiation of a skin treatment routine that includes application of the topical composition. In some embodiments, the individual is measured on the side of the face using a skin hydration sensor measurement system by Wearifi. In some embodiments, the individual is measured at baseline. In some embodiments, the individual is measured 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the start of a skin treatment routine that includes application of a topical composition. In some embodiments, the individual is measured for skin hydration 15 minutes after cleansing the individual's facial skin. In some embodiments, the individual is measured for skin hydration the first 15 minutes after cleansing the individual's facial skin and the second 15 minutes after application of the composition. In some embodiments, the method is described in which the individual is measured for skin hydration three times on a side of the face. In some embodiments, the individual is measured for skin hydration three times on a side of the face in the same facial area. In some embodiments, the individual is measured for skin hydration on the right and left sides of the face. In some embodiments, the individual is measured for skin hydration three times on each side of the right and left sides of the face. In some embodiments, the individual is measured for skin hydration three times on each side of the right and left sides of the face in the same facial area. In some embodiments, the individual has a skin hydration measurement that captures the water content across the stratum corneum and upper epidermis.In some embodiments, the individual has a skin hydration measurement, the measurement being a quantitative result of the volumetric ratio of water in skin tissue. In some embodiments, the individual has a skin hydration measurement, the measurement representing an absolute value that has a direct relationship to the effective hydration of the skin. In some embodiments, the individual has a skin hydration measurement taken at baseline and after the start of a skin treatment routine that includes application of a topical composition.

[0157] Methods for assessing the efficacy of topical applications In some embodiments, the individual completes a self-assessment survey on the characteristics of their facial skin. In some embodiments, the survey is completed at baseline, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, the survey is completed after cleansing the skin. In some embodiments, the survey is completed about 15 minutes after cleansing the skin. In some embodiments, the individual completes the survey after beginning a skin treatment routine that includes application of the topical composition. In some embodiments, the individual completes the survey using a scale to rate their assessment of their skin. In some embodiments, the individual completes the survey using a 5-point scale to rate their assessment of their skin. In some embodiments, the individual completes the survey at baseline and after beginning a skin treatment routine that includes application of the composition.

[0158] In some embodiments, the individual completes a self-assessment survey regarding overall satisfaction with the composition. In some embodiments, the individual completes a self-assessment survey regarding overall satisfaction with the composition. In some embodiments, the individual completes a self-assessment survey regarding overall satisfaction with the composition at baseline and at other time points. In some embodiments, the individual completes a self-assessment survey regarding overall satisfaction with the composition at baseline and at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, the overall satisfaction survey uses a scale that is compared to baseline. In some embodiments, the overall satisfaction survey uses a 7-point scale that is compared to baseline.

[0159] In some embodiments, the responsible physician completes an assessment of the individual's facial skin. In some embodiments, the individual begins a skin treatment routine that includes application of a topical composition provided herein. In some embodiments, the responsible physician completes an assessment of the individual's facial skin at baseline and at a later time point. In some embodiments, the later time point is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the first application. In some embodiments, the responsible physician completes an assessment of the individual's facial skin after cleansing the skin. In some embodiments, the responsible physician completes an assessment of the individual's facial skin about 15 minutes after cleansing the skin. In some embodiments, the responsible physician completes an assessment of the individual's facial skin to assess fine lines / wrinkles, fine lines, texture, erythema, dryness, or moisture / hydration. In some embodiments, the responsible physician completes an assessment of the individual's facial skin to assess fine lines / wrinkles, fine lines, texture, erythema, dryness, and moisture / hydration. In some embodiments, a scale is used to assess fine lines / wrinkles, fine lines, texture, erythema, dryness, and / or moisture / hydration. In some embodiments, a 10-point scale is used to assess fine lines / wrinkles, fine lines, texture, erythema, dryness, and / or moisture / hydration. In some embodiments, the scale parameters are 0 absent, 1-3 mild, 4-6 moderate, 7-9 severe. In some embodiments, methods are described in which the results of the investigator assessment are subjected to statistical analysis.

[0160] In some embodiments, the method of taking a photograph of an individual who has begun a skin treatment routine including application of a composition provided herein is as described herein. In some embodiments, photographs are taken showing the individual's facial skin at baseline and at 2 weeks, 4 weeks, and / or 8 weeks after beginning a skin treatment routine including application of the composition. In some embodiments, a portrait lens (85mm f / 1.4) is used as part of the imaging system to capture the individual's facial skin characteristics. In some embodiments, the photograph is taken after cleansing the skin. In some embodiments, the photograph is taken about 15 minutes after cleansing the skin. In some embodiments, the photograph is taken after cleansing the skin and after application of a topical test composition. In some embodiments, the photograph is taken after a first period of about 15 minutes after cleansing the skin, followed by a second period of about 15 minutes after application of the topical test composition. In some embodiments, a VISIA® skin analysis system is used to take the photograph. In some embodiments, a VISIA® skin analysis system is used to perform an analysis of the skin characteristics on the photograph taken. In some embodiments, a LifeViz® Infinity System is used to take the photograph. In some embodiments, a LifeViz® Infinity System is used to perform an analysis of skin features on the photograph taken. In some embodiments, a LifeViz® Micro System is used to take the photograph. In some embodiments, a LifeViz® Micro System is used to perform an analysis of skin features on the photograph taken. In some embodiments, visible skin redness is evaluated on the photograph of the individual. In some embodiments, visible skin inflammation is evaluated on the photograph of the individual. In some embodiments, visible skin wrinkles are evaluated on the photograph of the individual. In some embodiments, visible fine lines are evaluated on the photograph of the individual.In some embodiments, deep lines visible in a photograph of an individual are evaluated. In some embodiments, deep lines in facial skin visible in a photograph of an individual are evaluated. In some embodiments, fine lines in facial skin visible in a photograph of an individual are evaluated. In some embodiments, texture of facial skin visible in a photograph of an individual is evaluated. In some embodiments, pore size area of ​​facial skin visible in a photograph of an individual is evaluated. In some embodiments, rosacea of ​​facial skin visible in a photograph of an individual is evaluated. In some embodiments, fine blood vessels of facial skin visible in a photograph of an individual are evaluated. In some embodiments, solar elastosis of facial skin visible in a photograph of an individual is evaluated. In some embodiments, age spots on facial skin visible in a photograph of an individual are evaluated.

[0161] In some embodiments, punch biopsies may be taken around the ear at baseline and at 2, 4, and 8 weeks after application. In some embodiments, individuals may be instructed to apply the topical serum to the ear twice daily for the duration of the study. In some embodiments, an independent dermatopathologist evaluates the tissue before and after application of the test product. In some embodiments, methods are described in which biopsy samples are processed for histological analysis. In some embodiments, histological analysis includes CD44 immunohistochemistry, hemotoxylin and eosin staining. In some embodiments, hemotoxylin and eosin staining reveals the structure of the dermal extracellular matrix and the thickness and density of collagen fibers. In some embodiments, CD44 immunohistochemistry reveals the cellular location of HA production.

[0162] In some embodiments, topical application of the composition reduces inflammation in an individual. In some embodiments, methods are described in which topical application of the composition reduces skin inflammation in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition reduces skin redness in an individual. In some embodiments, methods are described in which topical application of the composition reduces skin redness in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition reduces the appearance of deep lines in an individual. In some embodiments, methods are described in which topical application of the composition reduces the appearance of deep lines in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition reduces the appearance of wrinkles in an individual. In some embodiments, methods are described in which topical application of the composition reduces the appearance of wrinkles in an individual when comparing baseline or another time point with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition reduces the appearance of fine lines in an individual. In some embodiments, methods are described in which topical application of the composition reduces the appearance of fine lines in an individual when comparing baseline or another time point with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition reduces skin dryness in an individual. In some embodiments, methods are described in which topical application of the composition reduces skin dryness in an individual when comparing baseline or another time point with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks.

[0163] In some embodiments, topical application of the composition increases skin hydration in an individual. In some embodiments, methods are described in which topical application of the composition increases skin hydration in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition improves the appearance of skin texture in an individual. In some embodiments, methods are described in which topical application of the composition improves the appearance of skin texture in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition improves the appearance of solar elastosis in an individual. In some embodiments, methods are described in which topical application of the composition improves the appearance of solar elastosis in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition reverses the appearance of solar elastosis in an individual. In some embodiments, methods are described wherein topical application of the composition reverses the appearance of solar elastosis in an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks.

[0164] In some embodiments, topical application of the composition stimulates HA synthesis in the skin of the individual. In some embodiments, methods are described in which topical application of the composition stimulates HA synthesis in the skin of the individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition upregulates hyaluronan synthase (HAS) expression in the skin of the individual. In some embodiments, methods are described in which topical application of the composition upregulates hyaluronan synthase (HAS) expression in the skin of the individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, stimulating hyaluronan synthase (HAS) activity in the skin of the individual comprises increasing expression of HAS2. In some embodiments, topical application of the composition downregulates hyaluronidase expression in the skin of the individual. In some embodiments, methods are described wherein topical application of the composition downregulates hyaluronidase expression in the skin of the individual when comparing baseline or another time point to weeks 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the composition downregulates hyaluronidase expression in the skin of the individual, including a decrease in expression of HYAL2.

[0165] In some embodiments, topical application of the composition reduces the appearance of bruising, age spots, or wrinkles on the skin of an individual. In some embodiments, methods are described in which topical application of the composition reduces the appearance of bruising, age spots, or wrinkles on the skin of an individual when comparing baseline or another time point with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, the reduction in the appearance of bruising, age spots, or wrinkles persists after the period of topical application of the composition has ended.

[0166] In some embodiments, topical application of the composition increases elastin production in the skin of an individual. In some embodiments, methods are described in which topical application of the composition stimulates elastin production in the skin of an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition upregulates MFAP4 expression. In some embodiments, topical application of the composition increases tropoelastin production. In some embodiments, methods are described in which topical application of the composition upregulates MFAP4 expression and / or tropoelastin production in the skin of an individual when comparing baseline or another time point to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, the composition upregulates expression of S100A2, LAMA3, MERTK, or ITGB4. In some embodiments, methods are described wherein topical application of the composition upregulates expression of S100A2, LAMA3, MERTK, or ITGB4 in the skin of an individual when compared to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks from baseline or another time point. In some embodiments, the increased elastin production, increased tropoelastin production, or upregulation of MFAP4, S100A2, LAMA3, MERTK, or ITGB4 persists after the period of topical application of the composition has ended.

[0167] In some embodiments, the topical application of the composition is applied 1, 2, 3, 4, 5, 6, 7, or 8 times a day as described herein. In some embodiments, the topical application is applied in the morning. In some embodiments, the topical application is applied in the afternoon. In some embodiments, the topical application is applied in the morning and in the afternoon. In some embodiments, the topical application is applied in the morning or in the afternoon. In some embodiments, the topical application is applied after cleansing the skin. In some embodiments, the topical application is applied a certain period of time after cleansing the skin. In some embodiments, the topical application is applied about 15 minutes after cleansing the skin.

[0168] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for promoting wound healing comprising administering a topical composition according to the present disclosure to the skin of a subject.

[0169] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for promoting wound healing or wound closure comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0170] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for treating a skin wound comprising administering a topical composition according to the present disclosure to the skin of a subject.

[0171] In some embodiments, wounds (e.g., skin wounds) are associated with the use of the device for anti-aging indications, which may include combating skin laxity, combating loss of skin texture and tone, reducing redness, reducing skin blemishes, reducing inflammation, reducing swelling, reducing skin itching, or any combination thereof.

[0172] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for promoting elastic fiber formation comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0173] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing scar formation comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0174] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing the effects of aging on the skin comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0175] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing or reversing skin aging comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0176] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing skin laxity comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0177] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for improving skin texture or improving skin tone comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0178] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for improving pigmentation comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0179] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing skin redness comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0180] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing skin problems or improving the appearance of skin problems comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0181] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing inflammation of the skin comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0182] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing skin swelling comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0183] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing skin itch comprising administering to the skin of a subject a topical composition according to the present disclosure.

[0184] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for promoting extracellular matrix (ECM) remodeling comprising administering a topical composition according to the present disclosure to the skin of a subject.

[0185] In any of the aspects or embodiments disclosed herein, the skin affected by the itch is not suffering from one or more of wounds, skin damage, signs of aging, bruising (e.g., suffering from bruising associated with a bodily procedure such as a filler injection), inflammation, redness, soreness, sensitivity, dryness, looseness, fine lines, poor contouring, discoloration, swelling, pigmentation disorders (e.g., hyperpigmentation), age spots, wrinkles, pain, discomfort, or dermatoporosis.

[0186] In some embodiments, the individual receiving the topical application is a human.

[0187] kit In some embodiments, a kit is provided herein that includes a composition comprising the peptides described herein. In some embodiments, the kit can be provided to a physician, other medical personnel, patient, or caregiver to administer. In some embodiments, the kit includes a container containing the composition in a suitable topical composition and instructions for administering the composition to a subject. The kit can include one or more additional therapeutic or other agents. For example, a kit containing a topical form of the composition can be provided with other skin care agents, such as cleansers, occlusive moisturizers, penetrating moisturizers, sunscreens, sunblocks, and the like. The kit can include the composition in bulk form, or can include separate doses of the composition for continuous or sequential administration. The kit can optionally include one or more diagnostic tools, administration tools, and / or instructions for use. The kit can include a suitable delivery device, such as a syringe, pump dispenser, single dose packet, and the like, along with instructions for administering the peptide composition and any other therapeutic or beneficial agent. The kit may optionally contain instructions for storage, reconstitution (if applicable), and administration of any or all of the therapeutic or beneficial agents included. The kit may contain multiple containers reflecting the number of doses to be given to a subject, or different products to be administered to a subject.

[0188] In some embodiments, the composition stimulates the skin renewal process and helps improve the appearance of the skin and the feeling of skin tightness. In some embodiments, the composition is suitable for all skin types and post-procedure skin. In some embodiments, the composition can be provided to the patient in bulk form to allow a suitable amount of peptide to be self-administered by the patient. In some embodiments, the patient can apply a sufficient amount of the composition to evenly cover the affected area or as otherwise directed by a physician. In some embodiments, cleansers, sunscreens, sunscreens, penetrating moisturizers, and / or occlusive moisturizers can be provided for administration before or after the topical composition of the embodiments.

[0189] In some embodiments, various examples of creams, ointments, lotions, solutions, gels, sprays, and patches may incorporate the compositions described herein in combination with penetration enhancers and other agents to improve skin health.

[0190] definition Throughout this disclosure, various embodiments are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of any embodiment. Thus, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values ​​within the range to the tenth of the unit of the lower limit, unless the context clearly dictates otherwise. For example, the description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual values ​​within that range, for example, 1.1, 2, 2.3, 5, and 5.9. This applies regardless of the breadth of the range. The upper and lower limits of these intervening ranges may independently be included in the smaller ranges and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where a stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the disclosure, unless the context clearly dictates otherwise.

[0191] The use of absolute or sequential terms, such as "will," "will not," "shall," "shall not," "must," "must not," "first," "initially," "next," "consequently," "before," "after," "lastly," and "finally" are not intended to limit the scope of the embodiments disclosed herein, but are exemplary.

[0192] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms unless the context clearly indicates otherwise. Furthermore, to the extent the terms "including," "includes," "having," "has," "with," or variations thereof are used in any of the detailed description and / or claims, such terms are intended to be inclusive in a similar manner as the term "comprising."

[0193] As used herein, the phrases "at least one," "one or more," and "and / or" are open-ended expressions that are both conjunctive and disjunctive in operation. For example, each of the phrases "at least one of A, B, and C," "at least one of A, B, or C," "one or more of A, B, and C," "one or more of A, B, or C," and "A, B, and / or C" means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B, and C together.

[0194] As used herein, "or" can refer to "and," "or," or "and / or," and can be used both exclusively and inclusively. For example, the term "A or B" can refer to "A or B," "A but not B," "A but not B," and "A and B." In some cases, the context may dictate a particular meaning.

[0195] The term "about" when referring to a number or numerical range means that the referenced number or numerical range is approximate within experimental variability (or within statistical experimental error) and that the number or numerical range may vary, for example, from 1% to 15% of the stated number or numerical range. In examples, the term "about" refers to ±10% of the stated number or value.

[0196] The terms "increased", "increase", or "increase" are generally used herein to mean an increase by a statically significant amount. In some embodiments, the term "increased" or "increase" means an increase of at least 10% compared to a reference level, e.g., an increase of at least about 10%, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to 100% increase, including a 100% increase, or any increase between 10-100%. Other examples of "increase" include an increase of at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, at least 1000-fold or more compared to a reference level.

[0197] The terms "reduced", "reduced", or "reduction" are generally used herein to mean a reduction by a statistically significant amount. In some embodiments, "reduced" or "reduction" means a reduction of at least 10% compared to a reference level, e.g., a reduction of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% compared to a reference level, or up to a 100% reduction, including a 100% reduction (e.g., an absent or undetectable level compared to a reference level), or any reduction between 10 and 100%. In the context of a marker or symptom, these terms mean a statistically significant reduction in such levels. The reduction can be, for example, at least 10%, at least 20%, at least 30%, at least 40% or more, and is preferably reduced to a level that is accepted as within the normal range for individuals without a given disease. EXAMPLES

[0198] The following illustrative examples are representative of embodiments of the stimuli, systems, and methods described herein and are not meant to be limiting in any way.

[0199] Example 1. Gene Expression Assay An example is provided herein showing the effect of different active agents on gene expression in fibroblasts and keratinocytes.

[0200] method Cell Lines. Human adult dermal fibroblast and keratinocyte cell lines were treated with 11 different compound treatments (+ DMSO control as treatment number 12). Primary cells were cultured on 1 cm 2 Cells were seeded in cell specific medium at 5K or 10K cells per well. Cells were seeded in triplicate in 48-well plates (Greiner), one cell line per plate. Medium volume was 500ul per well. The outer two columns of the well were not used. All rows were used. Cells were incubated at 37°C, 5% CO 2 The cells were cultured in a 37° C. incubator for 2 days. After 48 hours, all cell cultures appeared homogenous and healthy, with no appreciable numbers of floating dead cells or evidence of cellular vacuolization that might indicate apoptotic or dying cells.

[0201] Compounds. Stocks of compounds 1-8 listed below were prepared in PBS. A 50 mg / ml stock of compound 9 ("Phos", phosphatidylserine) was prepared in DMSO, but did not completely dissolve in solution with extensive vortexing and warming. This stock was at 100x the final concentration. A 20 mg / ml stock of compound 10 (CBD) was prepared in DMSO. No undissolved particulates were observed, but the stock solution was slightly cloudy and not completely clear. This stock was at 200x the final concentration.

[0202] Cell treatment: 1. Lactoferrin (Lacto) 2.TCVRRAF (LCV) 3. Tripeptide-1 (Tri) 4. Hexapeptide-12 (Hex12) 5. Tripeptide-1 and Hexapeptide-12 (TriHex) 6. Hexapeptide-11 (Hex11) 7. Tranexamic acid 5% (Tran acid) 8. Octapeptide (Octa) 9. Phosphatidylserine (Phos) 10. CBD

[0203] Dosing. After 48 hours of adherent culture, fibroblasts and keratinocytes were treated with test compounds. Compounds were resuspended in the appropriate cell culture medium at final concentrations, adherent culture medium was removed, and compound-containing medium was added. Cells were exposed to the following compounds for 24 hours: Lactoferrin (500 μg / ml), TCVRRAF (100 μg / ml) (amino acid sequence extracted from lactoferrin), tripeptide-1-100 ppm (2.9 μg / ml), hexapeptide 12, 100 ppm (2.9 μg / ml), TriHex combination 200 ppm (2.9 μg / ml each), hexapeptide 11 (100 μg / ml), tranexamic acid 5% (500 μg / ml), octapeptide (100 μg / ml)-a proprietary peptide designed by R&D at Alastin (Alastin Skincare, Inc., a Galderma Company, Carlsbad CA), phosphatidylserine 500 μg / ml, or cannabidiol (CBD) 100 μg / ml. Control cells were left untreated.

[0204] RNA lysate preparation. After 24 hours of compound exposure, media was removed and cells were washed once with PBS. 100ul of RNA lysis buffer (Takara Bio Cat Num 635013, "10x RNA lysis buffer", diluted 1x) was added to wells, mixed thoroughly by trituration, combined in an RNAse-free microfuge tube, and immediately frozen at -30°C. Samples were prepared from one cell line (one plate) at a time. Plate array is 12x4, with each treatment in a row of three wells. Triplicate wells were lysed and combined into tubes in PCR tube strips. All samples were shipped frozen on dry ice for RNA extraction, library construction, and sequencing to 25M paired-end 100bp reads per sample.

[0205] All RNA samples were shipped frozen on dry ice for RNA extraction, library construction, and sequencing to 25M paired-end 100 bp reads per sample. Differentially expressed genes were identified and pathway enrichment was assessed using Reactome Pathway.

[0206] result Figure 1 shows data for hyaluronan synthase 2 (HAS2), the primary stimulator of HA in fibroblasts, in fibroblasts treated with various compounds. Data are presented as fold change in gene expression compared to untreated cells. The octapeptide showed superior stimulation of hyaluronan synthase 2 (HAS2) (Figure 1).

[0207] Figure 2A shows data for HAS2 in keratinocytes treated with various compounds. Data is presented as fold change in gene expression compared to untreated cells.

[0208] Figure 2B shows data for hyaluronidase 2 (HYAL2), a HA reductase, in keratinocytes treated with various compounds. Hexapeptide-11 showed a strong upregulation of HAS2 in keratinocytes and a strong downregulation of HYAL2 in keratinocytes (Figure 2A-B). Octapeptide showed a moderate downregulation of HYAL2 in keratinocytes. Data are presented as fold change in gene expression compared to untreated cells.

[0209] FIG. 3 shows data on early growth response 3 (EGR3) in keratinocytes treated with various compounds. HEX11 induced a significant upregulation of EGR3 by approximately 7-fold. EGR3 is a late epidermal regulator of differentiation highly expressed in the granular layer that may function to regulate the expression of skin barrier genes and strengthen the skin barrier. Data are presented as fold change in gene expression compared to untreated cells.

[0210] This example demonstrates that the peptides described herein are involved in regulating gene expression of genes involved in hyaluronic acid stimulation, hyaluronic acid turnover and maintenance, and the integrity and strength of the epidermal skin barrier.

[0211] Example 2. Selection of agents for in vitro HA production and size assessment Provided herein is an example of how octapeptide, SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™, and the complete formulation stimulate secretion of high molecular weight (high MW) hyaluronic acid (HA) from dermal fibroblasts (and keratinocytes). Provided herein is an assay for HA synthesis.

[0212] Agents identified by RNA-seq as novel HA production stimulators in fibroblasts were used together with other active agents previously determined to stimulate HA production to confirm the production of HA by fibroblasts and to determine the molecular weight (MW) / size of the HA produced. The components selected and treatment concentrations used are listed in Table 1. The rationale for the selection of these agents is described here.

[0213] Selection of agents for in vitro HA production and size assessment. Generally, the octapeptide (a unique peptide) upregulates hyaluronan synthase-2 (HAS2) gene expression in fibroblasts (based on the RNA-Seq data described in Example 1).

[0214] Lactoferrin may provide wound healing attributes, promote fibroblast proliferation, and increase HA secretion.

[0215] Syn-Hycan is a synthetic tripeptide that stimulates HA. Published evidence shows that the synthetic trifluoroacetate tripeptide tetradecylaminobutyroylvalylaminobutyric acid urea restores facial skin volume by stimulating HA synthesis. Syn-Hycan increases HA and CD44 in skin in vitro and ex vivo.

[0216] Phosphatidylserine stimulates hyaluronan synthesis. Sodium hyaluronate crosspolymer (Hyalasome) is an extremely high MW synthetic HA with exceptionally high water binding capacity resulting in excellent moisturizing properties.

[0217] Tremella fuciformis extract is derived from an edible mushroom and may provide high levels of moisture as a natural HA stimulator and has antioxidant properties.

[0218] Hydroxymethoxyphenyldecanone is an HA booster, antioxidant, and anti-irritant, which can stimulate dermal and epidermal hyaluronic acid levels by 259% and 198%, respectively, versus placebo in an ex vivo human skin model.

[0219] Hexapeptide-11 can upregulate HAS2 and downregulate the HA reductase hyaluronidase 2 (HYAL2) in keratinocytes (based on the RNA-seq data in Example 1).

[0220] Table 1. Compounds used to treat fibroblasts for HA production assessment by PAGE TIFF2025512393000002.tif69159

[0221] In vitro HA production and size assessment Dermal fibroblasts were cultured in growth medium to near confluence in 6-well plates. When they reached confluence, the growth medium was replaced with serum-free medium for 24 hours. The cells were then treated with the compounds shown in Table 1. After 72 hours, 100 μl of medium was collected from each treatment condition. The medium was concentrated in a SpeedVac concentrator to a final volume of approximately 10 μl. All 10 μl from each condition was loaded onto an acrylamide gel (NuPAGE 4-12% Bis-Tris protein gel, Invitrogen, Waltham, MA). Sodium hyaluronate, research grade, (HA2M) from Lifecore Biomedical (Chaska, MN) was reconstituted as recommended by the manufacturer and served as a MW reference representing high MW HA. The gel was run at 200 V for 3 hours to separate MW sizes from proteins and ECM components isolated from the supernatant with the HA2M reference as a comparator. To avoid overheating, the gel buffer was replaced with fresh room temperature buffer every 20 min. Gels were stained using Stains-All (Millipore Sigma, Burlington, MA) and destained according to the manufacturer's protocol. Bands migrating at the same size as the HA2M band indicated production of high MW HA.

[0222] Data showing the effect of several compounds on hyaluronic acid production in human dermal fibroblasts after 72 hours of treatment can be seen in Figure 4. Numbers 1-10 are the names of the compounds listed above in Table 1. Number 11 is the concentrated supernatant of untreated cells. Number 12 is the HA2M control (2 MDa / high MW HA) shown as a reference point. Figure 4 demonstrates that the fibroblasts produce HA in the range of 2 MDa / high MW HA, none of which produce low molecular weight (LMW) HA.

[0223] Verifying HA Fibroblasts were cultured as described above to near confluence in 6-well plates. The cells were then treated with compounds shown in Table 1. After 72 hours, 100 μl of medium was collected from each treatment condition. The medium was then subjected to hyaluronidase enzyme treatment (1 mg / ml) for 2 hours at 37° C. One condition was left undigested (100 μg / ml of the octapeptide). Samples were run on an SDS-PAGE gel, stained, and destained as described above. The results are shown in FIG. 5. Lanes 1-8 correspond to treatments using sample numbers 1-8 listed in Table 1. Lane 9 corresponds to the octapeptide treatment at 100 μg / ml that was left undigested. Lane 10 is the HA2M control (2 MDa / high MW HA) shown as a reference point. The absence of a band after digestion indicated that the band identified in the previous step was HA with a high molecular weight (of about 2 MDa).

[0224] The octapeptide stimulates high MW HA in a dose-responsive manner in dermal fibroblasts. Human dermal fibroblasts were treated with the octapeptide (1x and 10x, corresponding to 100 μg / ml and 1000 μg / ml, respectively) or left untreated. After 72 hours, supernatants were collected, concentrated, run on SDS-PAGE, stained and imaged. Experiments were performed in quadruplicate and the density of bands from each supernatant was quantified (in arbitrary units (AU)) using ImageJ. Data are graphed in FIG. 6 and represent the mean ± standard deviation of band density for each treatment condition. The large increase in signal density in the 10x octapeptide treatment indicated a dose-responsive increase in the production of high MW HA in dermal fibroblasts.

[0225] Example 3. Multicenter evaluation of topical hyaluronic acid and octapeptide formulations on facial skin This example includes a study on the progressive effectiveness of treatment using a topical HA and octapeptide formulation as part of a facial skin care routine in a third experiment. The formulations tested included topical application of normal skin care using a composition containing high MW HA and octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser in the third experiment. This study evaluated the efficacy and safety of a composition containing high MW HA and octapeptide on facial skin. Photographic and histological analysis of biopsies using a treatment formulation, a composition containing high MW HA and octapeptide, SPF 30+, optional topical application of ultra-light moisturizer after 4 weeks also constituted an aspect of the third experiment.

[0226] Materials and Methods This open-label clinical trial was conducted over a 4-month period from November 2021 to March 2022 and was approved by the Institutional Review Board, Advarra, Inc. (Columbia, MD). Eligible participants were healthy men and women aged 25 to 70 years who did not have any clinically significant unstable medical conditions, were willing to use only topical test products, and refrain from prolonged sun exposure, topical treatments, and procedures during the course of study participation. Exclusions for study participation included previous hypersensitivity or known allergy to any of the ingredients in the test products, use of retinol or topical agents containing active substances within 30 days, injectable toxins or resurfacing procedures within 3 months, injectable fillers and oral isotretinoin within 6 months. In addition, participants who were pregnant, breastfeeding, and planning to become pregnant during the study period were excluded.

[0227] Eligible enrolled participants were given a topical serum to apply twice daily and returned to the office for follow-up visits at weeks 2, 4, and 8. All participants were provided with a gentle cleanser for AM and PM use, as well as SPF30+ for AM use. At each visit, participants cleansed their facial skin and allowed it to acclimate for 15 minutes before assessment, photography, and measurements. Participants then applied the topical serum and allowed it to acclimate for 15 minutes before a second set of assessments, photography, and measurements.

[0228] Skin Hydration Sensor Measurements. At each visit, participants took three measurements on the right and left sides of the face using a skin hydration sensor measurement system by Wearifi (Chicago, IL). 15 minutes after cleansing the facial skin and 15 minutes after applying a topical test product. Measurements were performed in the same area at each visit. The measurement depth of the sensor was optimized to 50 μm to capture the water content across the stratum corneum and upper epidermis. The measurements are a quantitative result of the volumetric ratio of water in skin tissue, which represents an absolute value with a direct relationship to effective hydration.

[0229] Participant Assessment and Satisfaction. At all follow-up visits, 15 minutes after cleansing the skin, participants completed an assessment of their facial skin with the topical test product compared to baseline (using a 5-point scale) and overall satisfaction (using a 7-point scale). At each visit, 15 minutes after applying the topical serum, participants completed an assessment of their facial skin (using a 5-point scale).

[0230] Investigator Assessment. At baseline and week 8, the investigator completed a facial skin assessment 15 minutes after cleansing. A 10-point scale was used to assess fine lines / wrinkles, fine lines, texture, erythema, dryness, and moisture / hydration. Scale: 0 absent, 1-3 mild, 4-6 moderate, 7-9 severe.

[0231] Photography. At each visit, photographs were taken 15 minutes after cleansing the skin and after application of the test products. There were three different camera systems used in this study: VISIA® Skin Analysis System, (Canfield Scientific, Inc., Parsippany, NJ), LifeViz® Infinity and LifeViz® Micro (Quantificare, Inc. US, Cumming, GA).

[0232] Biopsies. 3 mm punch biopsies were performed around the ear at baseline and at weeks 2, 4, and 8 after application. Participants selected for biopsy were instructed to apply the topical serum around the ear twice daily for the duration of the study. An independent dermatopathologist evaluated the tissues before and after application of the study product.

[0233] Histology. Biopsies were fixed and sectioned for histological analysis according to standard protocols. Sectioned biopsies were mounted on slides and stained with hematoxylin and eosin to reveal epidermal and dermal cellular structures, including structures of the dermal ECM. Sectioned biopsies were mounted on slides and labeled for CD44 protein expression using anti-CD44 antibodies via immunohistochemistry according to standard protocols. CD44 labeling is shown in sections in a darker brown stain.

[0234] result An independent statistician completed the analyses using descriptive statistics, parametric and non-parametric tests.

[0235] The average hydration level in cleansed skin increases progressively with continued treatment. Analyses were completed for clean skin between baseline and follow-up visits, and between clean skin and after application of the test product at each visit. Pairwise comparisons between each time point were also calculated. 15 minutes after cleansing facial skin: At each follow-up visit, there was an increase in hydration measurements compared to baseline. At weeks 4 and 8, there was a statistically significant increase in hydration compared to baseline and the previous visit (Figure 7).

[0236] At each visit, hydration measurements were statistically significantly higher as measured post-application compared to clean skin (Figure 8). At 4 and 8 weeks post-application, measurements were also significantly higher than baseline and post-application measurements at week 2.

[0237] Compilation of participant ratings and satisfaction after cleansing and application At weeks 2, 4, and 8, 15 minutes after cleansing the skin, participants who gave a favorable rating of either strongly agree or agree were statistically significantly greater compared to week 2 for the following ratings (Figure 9):

[0238] Fifteen minutes after application of the test product at all visits, the percentage of participants who rated favorably continued to increase with each visit (Figure 10).

[0239] As can be seen in Figure 11, participants' overall satisfaction was rated favorably on a scale of 4-6 and increased statistically significant: 90.0% at week 2, 97.8% at week 4, and 97.9% at week 8 (p=.0061).

[0240] Results of the investigator's evaluation The mean investigator assessment scores were compared across five categories of features related to facial skin health and appearance (Figure 12). Compared to baseline, all assessment grades were statistically significant at the 8 week mark, p<.0001. Grades for reduction in fine lines / wrinkles, reduction in fine lines, texture, erythema, and dryness, and increase in moisture / hydration.

[0241] The mean investigator assessment scores were compared across the study at baseline and 8 weeks after initiation of treatment. As demonstrated in Figure 13, there was a wide range of increases in the mean assessment of facial skin moisture / hydration.

[0242] Photos and Analysis: Treatment with HA Immerse Serum (high MW HA + octapeptide) formulation, SPF 30+, and optional Ultra Light Moisturizer after 4 weeks. Figures 14-21 are photographs, analyses, and tissue sections from subjects at baseline as part of the third experiment and at various time points after initiation of treatment with usual skin care using a composition comprising high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser.

[0243] FIG. 14A demonstrates improved appearance and reduced redness after 4 weeks in a female subject, age 39.

[0244] FIG. 14B shows the results of a VISIA® Skin Analysis System for the subject from FIG. 17A demonstrating a quantifiable reduction in erythema.

[0245] Figure 15 shows photographs taken at baseline, week 2, week 4, and week 8 (from left to right) to demonstrate the progressive effect of skin improvement in a female subject, age 35. When comparing each time point, visible progressive changes are evident improving the appearance of fine lines, skin texture, and redness, indicating that continued treatment provides progressively enhanced results.

[0246] Figures 16A-16C demonstrate the degree of improvement in quantifiable features of appearance in the subject from Figure 15 comparing baseline and week 8 measurements. Figure 16B shows the results of a VISIA® Skin Analysis System to show a measurable reduction in red areas on the face. Figure 16C quantifies the effect of treatment using the VISIA® Skin Analysis System to result in a reduction in the total extent of pore size on the subject's face.

[0247] Figures 16D-16E demonstrate the extent of reduction in the appearance of fine lines and wrinkles after 8 weeks of treatment. The reduction in the appearance of fine lines and wrinkles in the skin around the eyes is particularly noticeable. Figure 16E quantifies the effect of treatment using the VISIA® Skin Analysis System to result in a reduction in the visibility of fine lines and wrinkles on the subject's face.

[0248] Figures 17A-17B show photographs and analysis at baseline and 8 weeks post-treatment to demonstrate improvements to the appearance of skin texture in a female subject, age 38. Figure 17B quantifies the effect of treatment using the VISIA® Skin Analysis System to result in an improvement in the appearance of skin texture on the subject's face.

[0249] 18A-18C show photographs demonstrating the efficacy of treatment to show a significant reduction in the appearance of erythema and rosacea in a male subject, age 48.

[0250] FIG. 18A shows the progressive effect of treatment when comparing baseline to weeks 2, 4, and 8 in reducing visible redness in a male subject, age 48.

[0251] Figures 18B-18C show close-up views to demonstrate the degree of visible reduction in erythema and rosacea in both the left and right cheek areas in the subject from Figure 18A at week 8. The reduction in visibility of fine blood vessels after this treatment period is particularly evident.

[0252] 19A-19C show the progressive effect of reducing the visibility of deep wrinkles near the mouth, fine lines across the face, and reducing the appearance of fine skin lines resulting in the appearance of smoother, firmer skin when comparing baseline, week 2, week 4, and week 8 in a female subject, age 73.

[0253] Figures 20A-20C show tissue analysis within periauricular biopsies of three subjects at baseline and 8 weeks after treatment with a composition containing HA Immerse Serum (high MW HA + octapeptide). Figures 20A-20C show hematoxylin and eosin stained sections from biopsies from the same subjects compared at baseline and 8 weeks, respectively (3 subjects analyzed in total). The extent of dermal solar elastosis, visible as old, thin collagen fibers in the ECM, is evident in each baseline sample. After 8 weeks of use of HA Immerse, the old, thin collagen fibers in the dermis were replaced with thicker, healthier collagen fibers, producing a much denser, healthier dermis and ECM. This dramatic reversal of the cellular, extracellular, and structural features of solar elastosis was an unexpected and significant finding after only 8 weeks of treatment in all three subjects analyzed. The extent of recycling of old ECM and its replacement with denser, healthier ECM indicates the therapeutic benefit of treatment in both the superficial and deeper regions of the dermis.

[0254] Figures 21A-21C show histological analysis of CD44 staining within periauricular biopsies of two subjects at baseline and 8 weeks after treatment with HA Immerse Serum (high MW HA + octapeptide). Figure 21A shows a section from the first subject with CD44 expression labeled in brown via immunohistochemistry. As CD44 is a receptor for HA and has been shown to be upregulated upon increased extracellular HA, the increase in CD44 labeling in both the epidermis and dermis at 8 weeks indicates increased stimulation and secretion of HA. Figures 21B-21C show a section from the second subject with CD44 expression labeled in brown via immunohistochemistry. Similarly, the increase in CD44 expression at 8 weeks indicates increased stimulation and secretion of HA. Increased CD44 expression is seen throughout the epidermis and dermis.

[0255] Figure 22 shows the chemical structure of Octapeptide-45, which comprises the amino acid sequence Gly-Pro-His-Gly-Val-Arg-Glu-Ala. Octapeptide-45 is an octapeptide contained in the formulation of HA Immerse Serum.

[0256] Example 4. Exemplary Formulations Exemplary embodiments of the compositions described herein are provided in Tables 2 and 3 of weight concentration of the solutions expressed as % (w / w).

[0257] Table 2: Exemplary compositions TIFF2025512393000003.tif197161

[0258] Table 3: Exemplary compositions TIFF2025512393000004.tif51128

[0259] Example 5. Gene Expression Studies for Compositions Containing TriHex+Octapeptide method Human adult dermal fibroblast cell lines were cultured in medium from ZenBio (Durham, NC) in 48-well plates at 37 °C in 5% CO 2The cells were maintained at 37°C in an incubator. After 48 hours, the cells were treated with the compounds listed in Table 4 below. One group was left untreated for comparison purposes. After 24 hours, the cell medium was removed and the cells were washed once with PBS. The cells were lysed and generated using 100 μl of RNA lysis buffer ("10x RNA lysis buffer", Takara Bio, Catalog No. 635013) and diluted 1x in an RNAse-free microcentrifuge tube. RNA extraction, library construction, and sequencing were then performed on 25M paired-end 100bp reads per sample. Differentially expressed genes were identified and pathway enrichment was assessed using the Reactome Pathway database.

[0260] Table 4. Compositions for in vivo gene expression testing TIFF2025512393000005.tif33166

[0261] result As shown in Figure 23A-D, the combination of TriHex + Octapeptide-45 showed gene expression synergism for S100A2, LAMA3, MERTK, and ITGB4 genes. S100A2 is S100 calcium-binding protein A2. It is a secreted matrisome-associated protein expressed in the dermis. S100A2 activates protein phosphatase 5 (PP5) in a calcium-dependent manner. PP5 is a serine / threonine phosphatase involved in oxidative stress response. LAMA3 is laminin, alpha 3. LAMA3 is the gene encoding the alpha 3 chain of laminin 5, which is the main link between the epidermal basal cells and the papillary dermis. It initiates hemidesmosome formation and provides stable attachment of the epidermis to the dermis. Laminin 5 may also accelerate the assembly of the basement membrane and enhance the repair of damaged skin. MERTK is a tyrosine-protein kinase MER. MERTK transmits signals from the extracellular matrix to the cytoplasm by binding to several ligands, including LGALS3, TUB, TULP1, or GAS6. After activation by ligands, MERTK plays a role in various processes such as macrophage clearance of apoptotic cells, platelet aggregation, cytoskeletal rearrangements, and engulfment. ITGB4 is integrin beta 4. Partnered with integrin alpha 6, ITGB4 is a receptor for laminin. The α6β4 integrin protein is particularly important in strengthening and stabilizing the skin, as it is a component of hemidesmosomes in the epidermis.

[0262] Table 5 lists several pathways related to ECM regulation whose gene expression is upregulated by Octapeptide-45 or the combination of TriHex+Octapeptide-45, indicating therapeutic benefit in ECM regulation, including elastin synthesis. The p-values ​​are the results of over-representation analysis. Over-representation analysis is a statistical (hypergeometric distribution) test that determines whether a particular Reactome pathway is over-represented (enriched) in the data. This test produces a probability score, which is corrected for false discovery rate using the Benjamani-Hochberg method.

[0263] (Table 5) Genes enriched for pathways related to ECM regulation. TIFF2025512393000006.tif73166

[0264] Example 6. Ex vivo testing of MFAP4 and tropoelastin (TE) expression on skin samples treated with TriHex + octapeptide method With reference to FIG. 24, human skin tissue was degreased, cut into pieces with a diameter of approximately 7 mm, and cultured in cell culture medium (1.88 μM CaCl 2 The tissue samples were placed in a transwell insert with DMEM / F12 containing 2% fetal bovine serum, 50 μM adenine, 0.02 nM triiodothyronine, 1% ITS-X (insulin, transferrin, selenium mix), 1% glutagro (glutamine equivalent), 1% pen / strep, and 1% gentamicin. The medium was changed daily. The human skin tissue samples were equilibrated for 3 days and then treated with the compounds listed in Table 6 once a day for 7 days. After 7 days of treatment, the skin samples were fixed in formalin and embedded in paraffin according to industry standard procedures. The resulting blocks were prepared and sectioned for immunofluorescence staining.

[0265] Antibodies directed against tropoelastin (TE) (available from Elastin, Owensville, MO) and against microfibril associated protein 4 (MFAP4) (available from ThermoFisher Waltham, MA) were used to detect new elastin production in treated cells. Appropriate fluorescent secondary antibodies were used for detection by fluorescence microscopy. Images were acquired and analyzed using ImageJ software.

[0266] Data were obtained from a minimum of three donor skin samples, with replicates or triplicates performed for each compound treated.

[0267] Table 6. Compositions for ex vivo testing TIFF2025512393000007.tif33166

[0268] result Microfibril-associated protein 4 (MFAP4) is an extracellular glycoprotein found in elastic fibers. MFAP4 is a ligand for microfibrils and tropoelastin that actively promotes tropoelastin (TE) self-assembly and is involved in proper elastic fiber organization. See P. Bartosz et al., 291 J. Biol. Chem. 1103-14 (Jan. 2016) (Figure 10) (reproduced as Figure 25).

[0269] Figure 27 shows images of immunofluorescence staining of cells with an antibody directed against MFAP4. MFAP4 expression is increased with TriHex (second from left), octapeptide-45 (second from right), and TriHex + octapeptide-45 treatment (right). The increase in MFAP4 expression with TriHex + octapeptide-45 indicates greater infiltration of new fibrils into the papillary dermis (see Figure 26) than that observed after treatment with TriHex and octapeptide-45 alone. Expression of MFAP4 indicates an opportunity for the formation of new elastin fibers.

[0270] Figure 28 is an image of immunofluorescence staining of cells with an antibody directed against TE. TE expression is increased with TriHex, Octapeptide-45, and TriHex + Octapeptide-45. The increase in TE production with TriHex + Octapeptide-45 indicates greater infiltration of new elastin fibers into the papillary dermis than that observed after treatment with TriHex and Octapeptide-45 alone. In addition, fiber alignment is more organized with the combined treatment, indicating that the newly formed elastin fibers are more stable.

[0271] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. The present invention is not intended to be limited by the specific examples provided herein. Although the present invention has been described with reference to the foregoing specification, the description and illustration of the embodiments herein are not intended to be construed in a limiting sense. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the present invention. Furthermore, it should be understood that all aspects of the present invention are not limited to the specific depictions, configurations, or relative proportions described herein which depend upon a variety of conditions and variables. It should be understood that various alternatives to the embodiments of the present invention described herein may be used in practicing the present invention. It is therefore contemplated that the present invention also covers any such alternatives, modifications, variations, or equivalents. The following claims define the scope of the present invention, and it is intended that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

1. A synthesized, isolated, or recombinant polypeptide comprising the amino acid sequence GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), or GPGKNGDD (SEQ ID NO: 7), comprising 20 or fewer amino acids, and the polypeptide, Tripeptide-1 and, Hexapeptide-12 and A topical composition containing the above.

2. The composition according to claim 1, wherein the amino acid sequence is GPHGVREA (Sequence ID 1).

3. The composition according to claim 1, wherein the polypeptide is an octapeptide.

4. The composition according to claim 1, wherein the composition comprises high molecular weight (MW) hyaluronic acid (HA) or a derivative thereof, and the high MW HA has a molecular weight of at least 0.5 MDa.

5. The composition according to claim 4, wherein the high MW HA or its derivative has a molecular weight of about 1 MDa to 4 MDa.

6. The composition according to claim 4, wherein the high MW HA or its derivative comprises sodium hyaluronate.

7. The composition according to claim 1, further comprising a synthetic tripeptide.

8. The composition according to claim 7, wherein the synthetic tripeptide is urea trifluoroacetate tetradecylaminobutyroylvalylaminobutyric acid.

9. The composition according to claim 1, further comprising hexapeptide-11, lactoferrin, or phosphatidylserine, wherein the hexapeptide-11, lactoferrin, or phosphatidylserine are encapsulated in liposomes.

10. The composition according to claim 9, wherein the hexapeptide-11 is present in an amount of 0.1 ppm to 500 ppm.

11. The composition according to claim 1, further comprising Tremella fuciformis extract, hydroxymethoxyphenyldecanone, or a combination thereof.

12. The tripeptide-1 is present in an amount of 0.1 ppm to 200 ppm, The hexapeptide-12 is present in an amount of 0.1 ppm to 200 ppm, and The polypeptide is present in an amount of 0.1 ppm to 200 ppm. The composition according to claim 1.

13. A composition according to any one of claims 1 to 12 for reducing inflammation in the skin of an individual, promoting skin repair, treating or restoring aging skin, treating skin wounds, or promoting wound healing or wound closure.

14. By administering the composition to the skin, Decreased expression of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL-12b), differentiation antigen group 80 (CD80), or combinations thereof. Increased CD44 expression, Increased production of tropoelastin or microfibril-associated protein 4 (MFAP4), Upregulation of the expression of S100A2, LAMA3, MERTK, or ITGB4. Upregulation of gene expression in one or more pathways related to extracellular matrix (ECM) regulation, Reduction of redness of an individual's skin after applying the composition to the skin. Increase in skin hydration in an individual's skin after applying the composition to the skin, Reduction or reversal of the symptoms of photoelastic fibrosis, Reduction of the appearance of birthmarks, age spots, or wrinkles. Upregulation of hyaluronic acid synthase (HAS) expression, Promotion of hyaluronic acid synthesis, Increased elastin production in the skin, or Downregulation of hyaluronidase expression The composition according to claim 13, wherein at least one of the following is achieved.

15. The composition according to claim 13, wherein the composition is administered to the skin once, two, three, four, five, six, seven, or eight times a day.

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