Combination of an octapeptide and high molecular weight hyaluronic acid for topical application

Abstract

Described herein are compositions and methods comprising high molecular weight hyaluronic acid (high MW HA). In some embodiments, the high MW HA is formulated in combination with one or more peptides. In some embodiments, the peptide comprises 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 / 330,710, 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 is a non-sulfated glycosaminoglycan that is widely distributed throughout connective tissues such as skin, epithelial tissues, and nervous tissues. Skin is the most abundant source of hyaluronic acid (HA), accounting for 50% of total body HA. The HA content of the dermis is much greater than that of the epidermis, accounting for the majority of the 50% of total body HA present in the skin. HA-bound water in both the dermis and epidermis may be important for skin hydration. Hyaluronic acid has been used in topical compositions for the skin due to its ability to promote skin hydration. However, low molecular weight (MW) hyaluronic acid may promote the production of proinflammatory mediators that lead to undesirable effects.

[0004] Skin aging is associated with loss of skin moisture and reduced barrier function. Environmental influences such as excessive UV exposure may contribute to the visible progressive changes seen in aged skin. Aged or damaged skin may exhibit various signs of inflammation, which may be visible as increased skin redness, increased visibility of fine blood vessels, and certain skin conditions, including but not limited to rosacea. As such, improving skin hydration and reducing inflammation in the skin may provide benefits for treating damaged, inflamed, or aging skin. Application of topical compositions containing hyaluronic acid (HA) and HA derivatives has been used to treat damaged, inflamed, or aging skin. Typically, lower molecular weight (MW) HA, often characterized as HA less than 1 megadalton (MDa), may be used in the composition, either alone or in combination with high MW HA. However, such compositions containing low MW HA may have undesirable pro-inflammatory effects on the treated skin. The use of exclusively high MW HA in topical compositions may reduce or minimize the pro-inflammatory effects seen with lower MW HA while increasing the hydration capacity and regenerative activity within the extracellular matrix (ECM). Such compositions containing exclusively high MW HA may promote the endogenous production of high MW HA within epidermal and dermal cells without negative effects such as the production of pro-inflammatory mediators. Thus, topical compositions containing high MW HA would be highly advantageous in treating aged or damaged skin or dry skin that exhibits a loss of barrier function. Summary of the Invention

[0005] overview Described herein are compositions and methods that include topical formulations and applications of high MW HA. In some embodiments, the compositions and methods described herein include topical formulations of high MW HA used to reduce skin inflammation. In some embodiments, the compositions and methods described herein include a mixture of high MW HA and a peptide (e.g., an octapeptide) that provides a synergistic effect on markers of skin health, healing, and hydration.

[0006] In some cases, application of high MW HA can result in increased skin hydration and reduced visibility of fine lines, demonstrating the benefits of applying a composition that includes high MW HA. In some cases, additional active agents can be included in the composition to stimulate production of high MW HA in epidermal and dermal cell types.

[0007] In one aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a topical composition for reducing skin inflammation comprising an octapeptide and a high molecular weight (MW) hyaluronic acid (HA) or derivatives thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivatives thereof has a molecular weight of at least 1 MDa. In some embodiments, the high MW HA or derivatives thereof has a molecular weight of about 1 MDa to 4 MDa. In some embodiments, the high MW HA or derivatives thereof comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprises sodium hyaluronate crosspolymer.

[0008] 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), or GPGKNGDD (SEQ ID NO:7). In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO:5). In some embodiments, the octapeptide comprises octapeptide-45.

[0009] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0010] 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.

[0011] In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.

[0012] In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises a Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyldecanone.

[0013] In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof.

[0014] In some embodiments, the composition is aqueous.

[0015] In some embodiments, reduction in skin 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.

[0016] 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.

[0017] In some embodiments, the composition increases skin hydration in the individual's skin after application of the composition.

[0018] In some embodiments, the composition promotes HA synthesis. In some embodiments, the composition stimulates hyaluronan synthase (HAS) activity. In some embodiments, the HAS comprises HAS2.

[0019] In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0020] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a method for reducing inflammation in the skin of an individual, the method comprising applying to the skin a topical composition comprising an octapeptide and a high molecular weight (MW) hyaluronic acid (HA) or derivative thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivative thereof has a molecular weight of at least 1 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.

[0021] 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), or GPGKNGDD (SEQ ID NO:7). In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO:5). In some embodiments, the octapeptide comprises octapeptide-45.

[0022] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0023] 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.

[0024] In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.

[0025] 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 a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, a phosphatidylserine, a Tremella fuciformis extract, or a hydroxymethoxyphenyldecanone, or a combination thereof.

[0026] In some embodiments, the composition is aqueous.

[0027] In some embodiments, the reduction in skin 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.

[0028] 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.

[0029] 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.

[0030] In some embodiments, the composition stimulates hyaluronan synthase (HAS) activity. In some embodiments, the HAS comprises HAS2.

[0031] In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0032] In some embodiments, the composition reduces the appearance of birthmarks, aging spots, or wrinkles.

[0033] In some embodiments, the composition is applied 1, 2, 3, 4, 5, 6, 7, or 8 times daily.

[0034] In some embodiments, the individual is a human.

[0035] In another aspect, which may be combined with any other aspect or embodiment, the present disclosure relates to a topical composition for reducing skin inflammation comprising an octapeptide and hyaluronic acid (HA) or a derivative thereof, wherein the composition reduces skin inflammation. In some embodiments, the HA or derivative thereof has a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or 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.

[0036] 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), or GPGKNGDD (SEQ ID NO:7). In some embodiments, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO:5). In some embodiments, the octapeptide comprises octapeptide-45.

[0037] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate.

[0038] 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.

[0039] In some embodiments, the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.

[0040] 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 a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, a phosphatidylserine, a Tremella fuciformis extract, or a hydroxymethoxyphenyldecanone, or a combination thereof.

[0041] In some embodiments, the composition is aqueous.

[0042] In some embodiments, reduction in skin 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.

[0043] In some embodiments, the composition increases CD44 expression.

[0044] 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.

[0045] In some embodiments, the composition increases skin hydration in the individual's skin after application of the composition.

[0046] In some embodiments, the composition promotes HA synthesis. In some embodiments, the composition stimulates hyaluronan synthase (HAS) activity. In some embodiments, the HAS comprises HAS2.

[0047] In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the hyaluronidase comprises hyaluronidase 2 (HYAL2).

[0048] In some embodiments, the composition reduces or reverses the symptoms of solar elastosis.

[0049] In some embodiments, the composition stimulates the synthesis of high MW HA having a MW of at least 0.5 MDa. [Brief description of the drawings]

[0050] 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.

[0051] [Figure 1] 2 is a graph of the fold change in hyaluronan synthase (HAS2) mRNA gene expression in dermal fibroblasts following treatment with the compounds listed below the bars compared to untreated cells. [Figure 2A] FIG. 1 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] FIG. 1 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] FIG. 1 is a graph of the fold change in early growth response 3 (EGR3) mRNA gene expression in keratinocytes following treatment with the compounds listed below the bars 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] 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, and waiting a second 15 minute period after treatment before measuring hydration. [Figure 9] FIG. 1 is a table summarizing participant evaluations following cleansing of the skin and satisfaction survey results at various time points following the start of treatment. [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] Photographs and analysis of subjects at baseline and at various time points after initiation of treatment with regular skin care using a composition comprising high MW HA and an octapeptide, a sunscreen, and a gentle cleanser. Photographs taken at baseline and at week 4 of treatment are shown to demonstrate the reduction in redness and the reduction of fine lines and wrinkles in the third study. [Figure 14B] Photographs and analysis of subjects at baseline and at various time points after initiation of treatment with regular skin care using a composition comprising high MW HA and an octapeptide, a sunscreen, and a gentle cleanser. Photographs of the forehead, nose, and upper cheek taken at baseline and at week 4 of treatment are shown to demonstrate reduced redness, diminished fine lines and wrinkles, and evened out skin tone and texture. [Figure 15] Photographs and analysis of subjects are shown at baseline and at various time points after the start of treatment with regular skin care using a composition containing high MW HA and octapeptide, sunscreen, and a gentle cleanser. Photographs were taken at baseline and at week 8 of treatment (top panel) to demonstrate the reduction in redness and the reduction in fine lines and wrinkles. The bottom panel images include only the red channel to demonstrate the effect of redness reduction alone after treatment. [Figure 16]Photographs and analysis of subjects are shown at baseline and at various time points after the start of treatment with regular skin care using a composition containing high MW HA and octapeptide, sunscreen, and a gentle cleanser. Photographs were taken at baseline and at weeks 2 and 4 of treatment (top panel) to demonstrate the reduction in redness and the reduction of fine lines and wrinkles. The bottom panel images include only the red channel to demonstrate the effect of redness reduction alone after treatment. [Figure 17A] Photographs are shown from a subject 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, an SPF 30+ sunscreen, an optional ultra-light moisturizer after week 4, and a gentle cleanser. Photographs taken at baseline and 4 weeks after treatment are shown to demonstrate improvements to the skin of a female subject, age 39. [Figure 17B] 17A shows photographs from a subject 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, an SPF 30+ sunscreen, an optional ultra-light moisturizer after week 4, and a gentle cleanser. Analysis of the photographs from FIG. 17A is shown using a VISIA® skin analysis system to demonstrate the reduction in facial red areas after treatment. [Figure 18] Photographs are shown taken at baseline, 2 weeks, 4 weeks, and 8 weeks after treatment (from left to right) to demonstrate the improvement to progressive effect of skin improvement in a female subject, age 35. The subject was treated with usual skin care using a composition containing high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after week 4, and a gentle cleanser. [Figure 19A]Close-up photographs taken at baseline and 8 weeks after treatment are shown to demonstrate improvement in pore size, skin tone, and skin texture in a female subject, age 35. The subject was treated with usual skin care using a composition comprising high MW HA and Octapeptide-45, SPF 30+, optional Ultra Light Moisturizer after 4 weeks, and a Gentle Cleanser. [Figure 19B] Analysis of photographs from Figure 18 taken at baseline and 8 weeks after treatment using a VISIA® Skin Analysis System to demonstrate reduction in facial redness areas. Subjects were treated 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. [Figure 19C] To demonstrate the reduction in facial pore size, an analysis of photographs from Figure 18 taken at baseline and 8 weeks after treatment is shown using a VISIA® Skin Analysis System. Subjects were treated 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. [Figure 19D] Shown are close-up photographs of the eye and cheek area of ​​a female subject, age 35, from Figure 18 taken at baseline and 8 weeks after treatment to demonstrate improvement in fine lines, wrinkles, and redness reduction. The subject was treated with usual skin care using a composition comprising high MW HA and octapeptide-45, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser. [Figure 19E] Shown is an analysis of photographs of the eye and cheek area of ​​a female subject, age 35, from Figure 18 taken at baseline and 8 weeks after treatment using a VISIA® Skin Analysis System to demonstrate reduced wrinkle density, area, and distribution. The subject was treated 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. [Figure 20A] Photographs taken at baseline and 8 weeks after treatment are shown to demonstrate improvements to skin tone, texture, and reduced redness in a female subject, age 38. The subject was treated with usual skin care using a composition containing high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser. [Figure 20B] Analysis of the photograph from Figure 20A using a VISIA® Skin Analysis System is shown to demonstrate the improvements resulting in a more uniform skin texture in the facial area after treatment. The subject was treated with a composition containing high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and regular skin care using a gentle cleanser. [Figure 21A] Photographs are shown taken at baseline, 2 weeks, 4 weeks, and 8 weeks after treatment (from left to right) to demonstrate the improvement of progressive reduction in skin inflammation, reduction in skin redness, and reduction in visibility of fine blood vessels in a male subject, age 48. The subject was treated 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 21B] Shown are close-up photographs of the upper left face from the male subject, age 48, from Figure 21A taken at baseline and 8 weeks after treatment to demonstrate improved rosacea and inflammation. The subject was treated 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. [Figure 21C] Shown are close-up photographs of the right cheek from the male subject, age 48, from Figure 21A taken at baseline and 8 weeks after treatment to demonstrate skin improvements including reduced visibility of fine blood vessels. The subject was treated 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. [Figure 22A]Close-up photographs of the right cheek and mouth area of ​​a female subject, age 73, taken at baseline and 2 weeks after treatment to demonstrate improvement of fine lines, deep wrinkles, and skin crepiness. The subject was treated with usual skin care using a composition containing high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser. [Figure 22B] Photographs of the right side of the face of a female subject, age 73, from Figure 22A are shown taken at baseline and 4 weeks after treatment to demonstrate the progressive improvement of fine lines, deep wrinkles, and fine skin lines. The subject was treated with usual skin care using a composition comprising high MW HA and octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser. [Figure 22C] Photographs of the right side of the face of a female subject, age 73, from Figure 22A are shown taken at baseline and 8 weeks after treatment to demonstrate the progressive improvement of fine lines, deep wrinkles, and fine skin lines. The subject was treated with usual skin care using a composition containing high MW HA and octapeptide, SPF 30+, optional ultra-light moisturizer after 4 weeks, and a gentle cleanser. [Figure 23A] Figures 23A-C show tissue sections of periauricular biopsies of subjects at baseline and 8 weeks after 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. Subjects were treated with regular skin care using a composition comprising high MW HA and octapeptide, SPF 30+, and optional ultra-light moisturizer after 4 weeks, and a gentle cleanser. [Figure 23B] See legend to Figure 23A. [Figure 23C] See legend to Figure 23A. [Figure 24A]Figures 24A-C show tissue sections of periauricular biopsies of subjects at baseline and 8 weeks after treatment to demonstrate significant upregulation of CD44 expression in epidermal and dermal structures. Figure 24A shows paired sections from a first subject at 200x magnification. Figures 24B-C show paired sections from a second subject at 200x and 400x magnification, respectively. The subjects were treated with regular skin care using a formulation of a composition comprising high MW HA and an octapeptide, SPF 30+, optional ultra-light moisturizer after week 4, and a gentle cleanser. [Figure 24B] See legend to Figure 24A. [Figure 24C] See legend to Figure 24A. [Diagram 25] 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.

[0052] 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

[0053] 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.

[0054] 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 hyaluronic acid (HA) and HA derivatives has been used to treat damaged, inflamed, or aging skin.Usually, lower molecular weight (MW) HA, often characterized as HA less than 1 megadalton (MDa), can be used in the composition, either alone or in combination with high MW HA.However, such compositions containing low MW HA can have undesirable pro-inflammatory effects on treated skin.

[0055] Using exclusively high MW HA in a topical composition may reduce or minimize the pro-inflammatory effects seen with lower MW HA while increasing hydration capacity and regenerative activity within the extracellular matrix (ECM). Such compositions containing exclusively high MW HA may promote endogenous production of high MW HA within epidermal and dermal cells without negative effects such as production of pro-inflammatory mediators. In some cases, application of high MW HA may result in increased hydration of the skin and reduced visibility of fine lines, demonstrating the benefits of applying a composition containing high MW HA. In some cases, additional active agents may be included in the composition to stimulate production of high MW HA in epidermal and dermal cell types.

[0056] Compositions and methods comprising topical formulations and applications of high MW HA are described herein. In some embodiments, the compositions and methods described herein comprise topical formulations comprising high MW HA used to reduce skin inflammation. In some embodiments, the compositions and methods described herein comprise a mixture of high MW HA and a peptide (e.g., an octapeptide) that provides a synergistic effect on markers of skin health, healing, and hydration. In some embodiments, a composition comprising high MW HA and an 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 high MW HA in the composition reduces levels of inflammation and redness in treated skin.

[0057] composition High Molecular Weight Hyaluronic Acid Described herein are compositions comprising high molecular weight hyaluronic acid (high MW HA), as well as methods of preparing and using such compositions. In some embodiments, the high MW HA comprises derivatives of high MW HA. In some embodiments, the compositions comprising high MW HA are formulated for topical application. In some embodiments, the compositions comprising high MW HA are formulated for topical application to reduce skin inflammation. In some embodiments, the reduction in skin inflammation is present in the skin near the site of application. In some embodiments, the reduction in skin inflammation is evident in the epidermis near the site of application. In some embodiments, the reduction in skin inflammation is evident in the dermis near the site of application. In some embodiments, the compositions comprising high MW HA are formulated for topical application to impart an anti-inflammatory effect as part of a topical application treatment routine. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of macrophages near the site of application. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of eosinophils near the site of application. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of neutrophils near the site of application. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of lymphocytes near the site of application. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of αβ T lymphocytes near the application site. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of γδ T lymphocytes near the application site. In some embodiments, the anti-inflammatory effect comprises a reduction in the number of B lymphocytes near the application site.

[0058] In some embodiments, the compositions described herein comprise a high MW HA, the high MW HA being above a particular molecular weight measurement. In some embodiments, the compositions comprise a high MW HA, the high MW HA having a molecular weight of at least about 0.5 MDa, 1.0 MDa, 1.5 MDa, 2.0 MDa, 2.5 MDa, 3.0 MDa, 3.5 MDa, 4 MDa, or 5 MDa. In some embodiments, the compositions comprise a high MW HA, the high MW HA having a molecular weight of at most about 2.0 MDa, 2.5 MDa, 3.0 MDa, 3.5 MDa, 4 MDa, or 5 MDa. In some embodiments, the compositions comprise a high MW HA, the high MW HA having a molecular weight of about 0.5 MDa to about 5 MDa, about 1 MDa to about 5 MDa, about 1 MDa to about 3 MDa, or about 2 MDa to about 3 MDa. In some embodiments, the compositions comprise a high MW HA, the high MW HA or derivative thereof comprises sodium hyaluronate. In some embodiments, the composition further comprises a sodium hyaluronate crosspolymer, hi some embodiments, the composition comprises high MW HA in liposomes.

[0059] In some embodiments, the composition comprising high MW HA is formulated for topical application to reduce skin inflammation. In some embodiments, the composition comprising high MW HA reduces visible redness in the skin. In some embodiments, the reduction in visible redness in the skin comprises a reduction in inflammation. In some embodiments, the reduction in visible redness in the skin comprises a reduction in the visibility of small blood vessels. In some embodiments, the reduction in visible redness in the skin comprises a reduction in the visibility of small blood vessels near the surface of the individual's skin. In some embodiments, the composition comprising high MW HA reduces visible redness in the facial skin of an individual.

[0060] In some embodiments, the composition comprises a peptide and 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 4 MDa. In some embodiments, the formulation comprises an octapeptide and a high MW HA with a molecular weight of at least about 0.5 MDa. In some embodiments, the formulation comprises an octapeptide and a high MW HA with a molecular weight of at least about 1 MDa. In some embodiments, the formulation comprises an octapeptide and a high MW HA with a molecular weight of at least about 1.5 MDa. In some embodiments, the formulation comprises an octapeptide and a high MW HA with a molecular weight of at least about 2.0 MDa. In some embodiments, the formulation comprises an octapeptide and a high MW HA with a molecular weight of about 1 MDa to about 5 MDa.

[0061] In some embodiments, the high MW HA formulation is provided at at least or about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 milligrams per milliliter (mg / ml). In some embodiments, the high MW HA formulation is provided in the range of about 0.01 to about 100 mg / ml. In some embodiments, the high MW HA formulation is provided in the range of about 0.1 to about 100, about 1 to about 100, about 5 to about 50, or about 1 to about 10 mg / ml. In some embodiments, the high MW HA formulation is provided at at least or about 6 mg / ml. In some embodiments, the high MW HA formulation is provided at at least or about 18 mg / ml.

[0062] In some embodiments, a high MW formulation is provided at at least or about 0.00001%, 0.0003%, 0.0005%, 0.001%, 0.001%, 0.005%, 0.0055%, 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% by weight (wt.%). In some embodiments, a high MW HA formulation 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, a high MW HA formulation 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 ppm. In some embodiments, a high MW HA formulation 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, a high MW HA formulation is provided in the range of about 1 to about 10 ppm.

[0063] In some embodiments, high MW HA formulations are 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, high MW HA formulations 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. In some embodiments, high MW HA formulations are provided at at least about 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, or greater than 2000 micrograms per milliliter (μg / mL). In some embodiments, high MW HA formulations are provided in the range of about 30 to about 2000 μg / mL. In some embodiments, high MW HA formulations are provided in the range of about 40 to about 1000, about 50 to about 900, about 60 to about 800, about 70 to about 700, about 80 to about 600, or about 90 to about 500 μg / mL. In some embodiments, high MW HA formulations are provided at at least or about 150 μg / mL. In some embodiments, high MW HA formulations are provided at at least or about 450 μg / mL.

[0064] peptide Provided herein are compositions comprising one or more peptides. In some embodiments, the compositions comprise one or more peptides having amino acid sequence characteristics commonly found in one or more extracellular matrix proteins of skin. In some embodiments, the compositions comprise one or more peptides having amino acid sequence characteristics found in collagen and / or elastin. In some embodiments, the compositions comprise one or more peptides having amino acid sequence characteristics found in collagen. In some embodiments, the compositions comprise one or more peptides having amino acid sequence characteristics found in elastin. In some embodiments, the compositions and methods comprise a mixture of one or more peptides obtained after chemical or enzymatic degradation of collagen. In some embodiments, the compositions and methods comprise a mixture of one or more peptides obtained after chemical or enzymatic degradation of elastin (e.g., bovine elastin). In some embodiments, the compositions and methods comprise a mixture of one or more peptides obtained after chemical or enzymatic degradation of collagen and / or elastin. In some embodiments, the compositions and methods comprise one or more peptides for inducing new elastic fiber formation or stimulating elastin production in human skin. In some embodiments, the compositions and methods include one or more peptides for inducing new elastic fiber formation or stimulating collagen production in human skin. In some embodiments, the compositions and methods include 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 compositions and methods include one or more peptides for inducing new elastic fiber formation or stimulating elastin and collagen production in human skin. In some embodiments, the compositions and methods include one or more peptides for inducing new elastic fiber formation or stimulating hyaluronic acid (HA) production in human skin. In some embodiments, the compositions and methods include one or more peptides for stimulating the production of high molecular weight HA.In some embodiments, the one or more elastin-derived peptides are hexapeptides. In some embodiments, the one or more elastin-derived peptides include the amino acid sequence XGVXXG. In some embodiments, the 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 further include one or more tripeptides, one or more hexapeptides, one or more octapeptides, or combinations thereof. In some embodiments, the 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., elastin-derived peptides) that provide a synergistic effect (e.g., gene expression). In some embodiments, the compositions and methods including one or more peptides having amino acid sequence characteristics found in collagen and / or elastin further include an octapeptide.

[0065] In some embodiments, the peptide comprises an octapeptide, a hexapeptide, or a tripeptide, or any combination thereof. In some embodiments, the octapeptide comprises octapeptide-45. In some embodiments, octapeptide-45 is a synthetic octapeptide having the chemical structure shown in Figure 25.

[0066] 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-X1-X2-X 3- X 4- X 5- X 6- X 7- In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X3- X 4- X 5- X 6- X 7- OH, and X is Pro, Asp, or Leu. In some embodiments, the octapeptide amino acid sequence is H-Gly-X-X-X 3- X 4- X 5- X 6- X 7- OH, and X2 is His, Gly, Met, or Gln. In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X 3- X 4- X 5- X 6- X 7- OH, and X3 is Gly, Asp, Pro, or Lys. In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X 3- X 4- X 5- X 6- X 7- OH, and X4 is Val, Gly, Pro, Phe, or Asn. In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X 3- X 4- X 5- X 6- X 7- OH, and X5 is Arg, Ala, Ser, Gln, or Gly. In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X 3- X 4- X 5- X 6- X 7- OH, and X6 is Glu, Ser, Gly, Arg, or Asp. In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X 3- X 4- X 5- X 6- X 7- OH, and X7 is Ala, Pro, or Asp. In some embodiments, the octapeptide amino acid sequence is H-Gly-X1-X2-X3- X 4- X 5- X 6- X 7- OH, X1 is Pro, Asp, or Leu, X2 is His, GIy, Met, or Gln, X3 is GIy, Asp, Pro, or Lys, X4 is Val, GIy, Pro, Phe, or Asn, X5 is Arg, Ala, Ser, Gln, or GIy, X6 is Glu, Ser, GIy, Arg, or Asp, and X7 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, and X5 is 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.

[0067] In some embodiments, the compositions and methods described herein include one or more octapeptides. In some embodiments, the octapeptide includes octapeptide-45. 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-X1-X2-X 3- X 4-X 5- X 6- X 7- OH, X1 is Pro, Asp, or Leu, X2 is His, Gly, Met, or Gln, X3 is Gly, Asp, Pro, or Lys, X4 is Val, Gly, Pro, Phe, or Asn, X5 is Arg, Ala, Ser, Gln, or Gly, X6 is Glu, Ser, Gly, Arg, or Asp, and X7 is Ala, Pro, or Asp. In some embodiments, the octapeptide having sequence characteristics found in collagen and / or elastin comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide having a sequence characteristic of collagen and / or elastin comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, GPGKNGDD, or any combination thereof.

[0068] 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), or GPGKNGDD (SEQ ID NO:7). 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).

[0069] In some embodiments, the octapeptide is a synthetic peptide having amino acid sequences characteristic of those found in collagen or elastin. In some embodiments, the octapeptide 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 that stimulates high MW HA synthesis in the dermis comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide that stimulates high MW HA synthesis in the dermis comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, GPGKNGDD, or any combination thereof. In some embodiments, the octapeptide that stimulates high MW HA synthesis in the dermis comprises the amino acid sequence GPHGVREA. In some embodiments, the octapeptide that stimulates the synthesis of high MW HA in the dermis comprises the chemical structure of octapeptide-45. In some embodiments, the octapeptide that stimulates the synthesis of high MW HA in the dermis comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide that stimulates the synthesis of high MW HA in the dermis comprises the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, GPGKNGDD, or any combination thereof. In some embodiments, the octapeptide that stimulates the synthesis of high MW HA in the dermis comprises the amino acid sequence GPHGVREA. In some embodiments, the octapeptide that stimulates the synthesis of high MW HA in the dermis comprises the chemical structure of octapeptide-45.

[0070] In some embodiments, the octapeptide 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, based on the total weight of the composition. It is provided at 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, 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.

[0071] 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.%).

[0072] 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.

[0073] 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. 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%, 0.005% or 0.005% 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 greater than 10% by weight (wt.%). In some embodiments, tripeptide-1 is provided in the range of about 0.25% to about 10%, about 0.001% to about 2%, about 0.5% to about 8%, about 0.75% to about 6%, or about 1% to about 4% by weight. 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 more 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.

[0074] 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.

[0075] 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.

[0076] 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 greater than 10% 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 0.0003% 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 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, 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 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.

[0077] 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%, 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-12 is provided in the range of about 0.00001% to about 10% 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, about 0.001% to about 2% by weight (wt.%), or any range or value therebetween. In some embodiments, hexapeptide-12 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, 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 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, 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.

[0078] 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 0.0003% to about 8% by weight, about 0.0005% to about 6% by weight, or about 0.001% to about 4% by weight, about 0.005% to about 2% by weight, about 0.001% 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.

[0079] 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.

[0080] 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).

[0081] 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.

[0082] The liposome composition 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 one or more octapeptides (e.g., octapeptide-45), one or more hexapeptides (e.g., hexapeptide-11, hexapeptide-38, hexapeptide-12), one or more tripeptides, one or more tetrapeptides (e.g., 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] 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).

[0090] 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.

[0091] 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.

[0092] 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.

[0093] 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.

[0094] 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%.

[0095] 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.

[0096] 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.

[0097] 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.

[0098] 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.

[0099] 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.

[0100] 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.

[0101] 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.

[0102] 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.

[0103] Gene expression The compositions described herein, in some embodiments, modify gene expression in skin cells. In some embodiments, exposure of dermal cells to the 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.

[0104] 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.

[0105] 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.

[0106] 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.

[0107] Hyaluronic Acid Synthesis The compositions described herein, in some embodiments, stimulate the production of HA in dermal and epidermal cells. In some embodiments, exposure of dermal cells to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of HA. In some embodiments, exposure of dermal cells to an 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 SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to an octapeptide stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to an octapeptide stimulates the production of HA. In some embodiments, exposure of epidermal cells to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of HA. In some embodiments, exposure of epidermal cells to octapeptide-45 stimulates the production of HA. In some embodiments, exposure of epidermal keratinocytes to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to Syn®-Hycan at a concentration of at least about 500 μg / ml stimulates the production of HA. 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 Hyalasome™ 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 hydroxymethoxyphenyl at a concentration of at least about 250 μg / ml stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to Aquaxyl at a concentration of at least about 500 μg / ml stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to octapeptide at a concentration of at least about 100 μg / ml stimulates the production of HA.

[0108] The compositions described herein, in some embodiments, stimulate the production of high MW HA in dermal and epidermal cells. In some embodiments, exposure of dermal cells to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of high MW HA. In some embodiments, exposure of dermal cells to octapeptide stimulates the production of high MW HA. In some embodiments, exposure of dermal cells to octapeptide-45 stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to octapeptide stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to octapeptide stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to octapeptide stimulates the production of high MW HA. In some embodiments, exposure of epidermal cells to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of high MW HA. In some embodiments, exposure of epidermal keratinocytes to SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™ stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to SynHycan at a concentration of at least about 500 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to lactoferrin at a concentration of at least about 500 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to phosphatidylserine at a concentration of at least about 500 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to Hyalasomes at a concentration of at least about 500 μg / ml stimulates the production of high MW HA.In some embodiments, exposure of dermal fibroblasts to Tremella at a concentration of at least about 500 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to hydroxymethoxyphenyl at a concentration of at least about 250 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to Aquaxyl at a concentration of at least about 500 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to Octapeptide at a concentration of at least about 10 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to Octapeptide-45 at a concentration of at least about 100 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to Octapeptide-45 at a concentration of at least about 1000 μg / ml stimulates the production of high MW HA. In some embodiments, exposure of dermal fibroblasts to octapeptide-45 at a concentration of at least about 100 μg / ml stimulates greater production of high MW HA than exposure of dermal fibroblasts to octapeptide-45 at a concentration of about 10 μg / ml. In some embodiments, exposure of dermal fibroblasts to octapeptide-45 at a concentration of at least about 1000 μg / ml stimulates greater production of high MW HA than exposure of dermal fibroblasts to octapeptide-45 at a concentration of about 100 μg / ml. In some embodiments, exposure of dermal fibroblasts to octapeptide-45 at a concentration of at least about 80 μg / ml stimulates greater production of high MW HA than exposure of dermal fibroblasts to octapeptide-45 at a concentration of about 10 μg / ml. In some embodiments, exposure of dermal fibroblasts to octapeptide-45 at a concentration of at least about 60 μg / ml stimulates greater production of high MW HA than exposure of dermal fibroblasts to octapeptide-45 at a concentration of about 10 μg / ml. In some embodiments, exposure of dermal fibroblasts to octapeptide-45 at a concentration of at least 40 μg / ml stimulates greater production of high MW HA than exposure of dermal fibroblasts to octapeptide-45 at a concentration of about 10 μg / ml.In some embodiments, exposure of dermal fibroblasts to octapeptide-45 at a concentration of at least 20 μg / ml stimulates the production of greater high MW HA than exposure of dermal fibroblasts to octapeptide-45 at a concentration of about 10 μg / ml. 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.

[0109] In some embodiments, exposure of the skin to a formulation comprising one or more octapeptides increases the expression of CD44. In some embodiments, exposure of the skin to a formulation comprising octapeptide-45 increases the expression of CD44. In some embodiments, exposure of epidermal keratinocytes to a formulation comprising octapeptide-45 increases the expression of CD44. In some embodiments, exposure of dermal fibroblasts to a formulation comprising octapeptide-45 increases the expression of CD44. 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 a reduction in skin redness. In some embodiments, increased expression of CD44 contributes to a reduction in skin inflammation. In some embodiments, increased expression of CD44 contributes to a reduction in the appearance of wrinkles. In some embodiments, increased expression of CD44 contributes to a reduction in the appearance of fine lines. In some embodiments, increased expression of CD44 contributes to a reduction in the 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.

[0110] Other Ingredients 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 skin 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 skin 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 stimulates hyaluronan synthase (HAS) activity. In some embodiments, the composition stimulates hyaluronan synthase (HAS) activity, 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).

[0111] In some embodiments, the composition increases high MW HA production in the skin. In some embodiments, the production of high MW HA results in a reduction in the appearance of redness. In some embodiments, the production of high MW HA reduces skin inflammation. In some embodiments, the production of high MW HA results in a reduction in the appearance of rosacea. In some embodiments, the production of high MW HA results in a reduction in the appearance of erythema. In some embodiments, the production of high MW HA results in a reduction in the appearance of fine blood vessels in the skin. In some embodiments, the production of high MW HA results in a reduction in the appearance of wrinkles. In some embodiments, the production of high MW HA results in a reduction in the appearance of fine lines. In some embodiments, the production of high MW HA results in a reduction in the appearance of deep lines. In some embodiments, the production of high MW HA results in a reduction in the appearance of deep lines in facial skin. In some embodiments, the production of high MW HA results in a reduction in the appearance of fine lines in the skin. In some embodiments, the production of high MW HA results in a reduction in the appearance of pore size. In some embodiments, the composition improves the appearance of skin texture. In some embodiments, the composition increases skin hydration. In some embodiments, the composition reduces excess water loss in the skin. In some embodiments, the composition increases high MW HA production in the skin to reduce dryness of the skin. In some embodiments, the composition reduces the appearance of solar elastosis. In some embodiments, the composition increases high MW HA production in the skin to increase the recycling of old, thin collagen fibers in the dermal ECM into denser, healthier collagen fibers in the dermal ECM.

[0112] In some embodiments described herein, the composition reduces skin inflammation. In some embodiments, the skin inflammation is caused by a skin condition. In some embodiments, the skin condition comprises rosacea, systemic lupus erythematosus, eczema, seborrheic eczema, dermatitis, seborrheic dermatitis, psoriasis, lichen pilaris, erythema nodosum, acne vulgaris, ichthyosis, carbuncles, cellulitis, or bacterial infection, or a combination thereof.

[0113] In some embodiments, described herein is a topical composition for reducing inflammation comprising a high MW HA. In some embodiments, the composition comprises an octapeptide. In some embodiments, the composition comprises hexapeptide-11. In some embodiments, the composition comprises lactoferrin. In some embodiments, the composition further comprises a Tremella fusiformis fruiting body extract. In some embodiments, the composition further comprises polyacrylate-13. In some embodiments, the composition further comprises glycerin. In some embodiments, the composition further comprises water. In some embodiments, the high MW HA comprises sodium hyaluronate. In some embodiments, the high MW HA comprises sodium hyaluronate crosspolymer.

[0114] In some embodiments, the composition acts on the surface of the skin. In some embodiments, ingredients acting on the surface of the skin include, but are not limited to, high MW HA, Tremella fuciformis extract, glycerin, xylitol, or combinations thereof. In some embodiments, ingredients acting on the surface of the skin promote skin hydration, reduction of skin inflammation and redness. In some embodiments, glycerin and / or xylitol act as humectants to limit water loss, stimulate HA and NMF production, promote water channeling in the skin, synergistically provide both immediate and long-term hydration, and reinforce barrier function by stimulating ceramide synthesis. In some embodiments, glycerin and / or xylitol enhance water circulation within the epidermis through their action on aquaporins and tight junctions. In some embodiments, Tremella fuciformis extract provides moisture to the skin. In some embodiments, HA provides moisture to the skin through its water binding capacity.

[0115] In some embodiments, the composition acts by penetrating the skin. In some embodiments, ingredients that act by penetrating the skin include, but are not limited to, octapeptides, hexapeptides, Tremella fusiformis extract, lactoferrin, hydroxymethoxyphenyldecanone, phosphatidylerine, or combinations thereof. 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.

[0116] In some embodiments described herein, a topical composition for reducing skin inflammation includes an octapeptide and hyaluronic acid (HA) or a derivative thereof, wherein the composition reduces skin 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), or GPGKNGDD (SEQ ID NO: 7). In some embodiments, the octapeptide comprises 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, a Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof.In some embodiments, the composition is aqueous.

[0117] In some embodiments, the reduction in skin 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 stimulates hyaluronan synthase (HAS) activity. In some embodiments, the composition stimulates hyaluronan synthase (HAS) activity, 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).

[0118] 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.

[0119] In some embodiments, a composition comprising an octapeptide and a high MW HA increases high MW HA comprising skin ECM. In some embodiments, a composition comprising an octapeptide and a high MW HA increases high MW HA comprising epidermal ECM. In some embodiments, a composition provided herein increases high MW HA comprising dermal ECM. In some embodiments, a composition increases high MW HA comprising epidermal or dermal ECM. In some embodiments, a composition increases high MW HA comprising epidermal and dermal ECM. In some embodiments, an increase in high MW HA synthesis is due to a synergistic effect of the high MW HA and the octapeptide in the composition. In some embodiments, a composition contributes to deposition of high MW HA in the epidermal ECM. In some embodiments, a composition stimulates deposition of high MW HA in the dermal ECM. In some embodiments, a composition contributes to deposition of high MW HA in the epidermal ECM and stimulates deposition of high MW HA in the dermal ECM. In some embodiments, a composition acts synergistically to increase the ratio of high MW HA to low MW HA in the skin. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the epidermis. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the dermis. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the epidermis and dermis. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and skin inflammation is reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and visible redness is reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and visible fine lines are reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and visible deep lines are reduced.In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the visible deep lines on the individual's face are reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the fine lines of the skin are reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the skin elasticity is improved. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the total area of ​​pore size is reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the visible skin texture is improved. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the skin dryness is reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and skin hydration is increased. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the visible signs of solar elastosis are reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the visible signs of solar elastosis are reduced. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the visible signs of solar elastosis are reversed. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the dermal ECM is remodeled to contain thicker collagen fibers. In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the dermal ECM is remodeled to contain denser collagen fibers, In some embodiments, the compositions act synergistically to increase the ratio of high MW HA to low MW HA in the skin and the dermal ECM is remodeled to a healthier collagen fiber network.In some embodiments, the composition comprises an octapeptide and high MW HA, where the high MW HA reduces skin inflammation in the epidermis and the octapeptide increases high MW HA production in the dermis.

[0120] 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), where most products will show excellent stability. In some cases, the product is subjected to -10°C (14°F) for 3 months.

[0121] 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.

[0122] 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.

[0123] 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.

[0124] 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.

[0125] method Methods of preparing and using high MW HA formulations Described herein are methods of preparing and using compositions comprising high MW HA. In some embodiments, a method for reducing inflammation in the skin of an individual comprises topical application of high molecular weight (MW) hyaluronic acid (HA) or a derivative thereof. In some embodiments, methods are described in which the high MW HA comprises a molecular weight of at least about 0.5 MDa, 1 MDa, 1.5 MDa, 2 MDa, 3 MDa, 4 MDa, or 5 MDa. In some embodiments, methods are described in which the high MW HA comprises a molecular weight of about 1 MDa to about 5 MDa, about 1 MDa to about 4 MDa, about 1 MDa to about 3 MDa, 1.5 MDa to 3 MDa, or about 2 MDa to about 3 MDa. In some embodiments, methods are described in which the high MW HA or a derivative thereof comprises sodium hyaluronate. In some embodiments, methods are described in which the sodium hyaluronate comprises a sodium hyaluronate crosspolymer. In some embodiments, a method for reducing inflammation in the skin of an individual comprises topical application of high molecular weight (MW) hyaluronic acid (HA) or a derivative thereof and an excipient.

[0126] The methods for reducing inflammation in the skin of an individual described herein include topical application of a high molecular weight (MW) hyaluronic acid (HA) or derivatives thereof. In some embodiments, the methods include topical application of a high MW HA formulation that results in a visible reduction in inflammation in the skin of the individual. In some embodiments, the visible reduction in inflammation in the skin of the individual is photographically documented. In some embodiments, the visible reduction in inflammation in the skin of the individual is photographically analyzed. In some embodiments, the methods include a visible reduction in inflammation in the skin of the individual, where the redness of the skin is reduced. In some embodiments, the methods include a photographic analysis of the redness of the skin, including Canfield redness quantification. In some embodiments, the methods include a visible reduction in inflammation in the skin of the individual, where the reduced inflammation persists after the individual has discontinued a treatment schedule of topical application of the composition. In some embodiments, the methods include a topical application of a high MW HA formulation that results in a visible reduction in inflammation in the skin of the individual, where the ratio of high MW HA to low MW HA in the epidermis is increased. In some embodiments, the methods include a reduction in immune cell density near the site of topical application of the high MW HA formulation.

[0127] Methods of Using High MW HA and Peptide Formulations Methods are described herein that include one or more peptides having amino acid sequence characteristics found in collagen. Methods are also described herein that include one or more peptides having amino acid sequence characteristics found in elastin. Methods are described herein that include one or more peptides having amino acid sequence characteristics found in collagen and elastin. In some embodiments, the method includes topical application of a composition that includes one or more peptides having amino acid sequence characteristics found in collagen. In some embodiments, the method includes topical application of a composition that includes one or more peptides having amino acid sequence characteristics found in elastin. In some embodiments, the method includes topical application of a composition that includes one or more peptides having amino acid sequence characteristics found in collagen or elastin. In some embodiments, the method includes topical application of a composition that includes one or more peptides having amino acid sequence characteristics found in collagen and elastin. In some embodiments, the one or more peptides include an octapeptide. In some embodiments, the octapeptide has the amino acid sequence H-Gly-X1-X2-X 3- X 4- X 5- X 6- X 7-OH, X1 is Pro, Asp, or Leu, X2 is His, GIy, Met, or Gln, X3 is GIy, Asp, Pro, or Lys, X4 is Val, GIy, Pro, Phe, or Asn, X5 is Arg, Ala, Ser, Gln, or GIy, X6 is Glu, Ser, GIy, Arg, or Asp, and X7 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 25).

[0128] 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 and high molecular weight (MW) hyaluronic acid (HA) or a derivative thereof. In some embodiments, methods are described in which 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), or GPGKNGDD (SEQ ID NO: 7). In some embodiments, methods are described in which the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). In some embodiments, methods are described in which the octapeptide comprises octapeptide-45. In some embodiments, methods are described in which the composition comprises a synthetic tripeptide. In some embodiments, a method is described in which the synthetic tripeptide is tetradecylaminobutyroylvalylaminobutyric acid urea trifluoroacetate. In some embodiments, a method is described in which the composition comprises a hexapeptide. In some embodiments, a method is described in which the hexapeptide is hexapeptide-11. In some embodiments, a method is described in which the hexapeptide-11 is encapsulated in a liposome. In some embodiments, a method is described in which the composition comprises lactoferrin. In some embodiments, a method is described in which the lactoferrin is encapsulated in a liposome. In some embodiments, a method is described in which the composition comprises phosphatidylserine. In some embodiments, a method is described in which the composition comprises a Tremella fuciformis extract. In some embodiments, a method is described in which the composition comprises hydroxymethoxyphenyldecanone.In some embodiments, methods are described in which the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyldecanone, or a combination thereof. In some embodiments, methods are described in which the composition is aqueous. In some embodiments, methods are described in which 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, methods are described in which the composition increases CD44 expression. In some embodiments, methods are described in which the composition reduces redness of the individual's skin after applying the composition. In some embodiments, methods are described in which the redness is measured by photography.

[0129] In some embodiments, the method of increasing skin hydration on the skin of an individual after the composition is applied is as described herein. In some embodiments, the skin hydration on the skin of an individual is measured at baseline. In some embodiments, the skin hydration on the skin of an 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 the application of a 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 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the start of a skin treatment routine that includes the application of a 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 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks after the start of a skin treatment routine that includes the application of a 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 beginning a skin treatment routine that includes application of a topical composition.

[0130] In some embodiments, the method of an individual completing a self-assessment survey on their facial skin characteristics is as described herein. In some embodiments, the method is described in which the survey is completed at baseline, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, the method is described in which the survey is completed after cleansing the skin. In some embodiments, the method is described in which the survey is completed about 15 minutes after cleansing the skin. In some embodiments, the method is described in which an individual completes the survey after the start of a skin treatment routine that includes the application of a topical composition. In some embodiments, the method is described in which an individual completes the survey using a scale to rate their skin evaluation. In some embodiments, the method is described in which an individual completes the survey using a 5-point scale to rate their skin evaluation. In some embodiments, the method is described in which an individual completes the survey on their facial skin characteristics at baseline and after the start of a skin treatment routine that includes the application of a topical composition.

[0131] In some embodiments, the method of the responsible physician completing the assessment of the individual's facial skin is as described herein. In some embodiments, the individual begins a skin treatment routine that includes application of a topical composition. 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, the responsible physician completes an assessment of the individual's facial skin to assess fine lines / wrinkles, fine lines, texture, erythema, dryness, and / or 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 include 0 absent, 1-3 mild, 4-6 moderate, and 7-9 severe.

[0132] In some embodiments, the method is as described herein, in which a photograph of an individual who has begun a skin treatment routine including application of a topical composition is taken. In some embodiments, a portrait lens (85mm f / 1.4) is used as part of an imaging system to capture the individual's facial skin characteristics. In some embodiments, a method is described in which 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 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 the skin characteristics on the photograph taken. In some embodiments, the LifeViz® Micro System is used to take the photograph. In some embodiments, the LifeViz® Micro System is used to perform an analysis of skin features on the photograph taken. In some embodiments, skin redness visible in the photograph of the individual is evaluated. In some embodiments, skin inflammation visible in the photograph of the individual is evaluated. In some embodiments, skin wrinkles visible in the photograph of the individual is evaluated. In some embodiments, fine lines visible in the photograph of the individual are evaluated. In some embodiments, deep lines visible in the photograph of the individual are evaluated. In some embodiments, deep lines in facial skin visible in the photograph of the individual are evaluated.In some embodiments, fine lines on the facial skin visible in the individual's photograph are evaluated. In some embodiments, texture of the facial skin visible in the individual's photograph is evaluated. In some embodiments, pore size area of ​​the facial skin visible in the individual's photograph is evaluated. In some embodiments, rosacea of ​​the facial skin visible in the individual's photograph is evaluated. In some embodiments, fine blood vessels of the facial skin visible in the individual's photograph are evaluated. In some embodiments, solar elastosis of the facial skin visible in the individual's photograph is evaluated. In some embodiments, age spots on the facial skin visible in the individual's photograph are evaluated.

[0133] 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 around 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 the biopsy samples are processed for histological analysis. In some embodiments, the histological analysis includes CD44 immunohistochemistry, hemotoxylin and eosin staining. In some embodiments, hemotoxylin and eosin staining reveals the structure of the dermal extracellular matrix, as well as the thickness and density of collagen fibers. In some embodiments, CD44 immunohistochemistry reveals the cellular location of HA production.

[0134] 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 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 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 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 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 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 to 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 to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks.

[0135] 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 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 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 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 to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks.

[0136] 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 to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 weeks. In some embodiments, topical application of the composition stimulates hyaluronan synthase (HAS) activity in the skin of the individual. In some embodiments, methods are described in which topical application of the composition stimulates hyaluronan synthase (HAS) activity in the skin of the individual when comparing baseline 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 to weeks 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, downregulation of hyaluronidase expression in the skin of the individual comprises a decrease in expression of HYAL2.

[0137] 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 to 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.

[0138] 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.

[0139] In some embodiments, the individual receiving the topical application, wherein the individual is a human.

[0140] kit Some embodiments of the methods and compositions provided herein include kits that include the peptides provided herein. In some embodiments, the kits may be provided to a physician, other medical personnel, a patient, or a caregiver to administer. In some embodiments, the kits include a container containing the peptide composition in a suitable topical composition and instructions for administering the peptide composition to a subject. The kits may also optionally contain one or more additional therapeutic or other agents. For example, a kit containing a peptide composition in topical form may be provided with other skin care agents, such as cleansers, occlusive moisturizers, penetrating moisturizers, sunscreens, sunblocks, and the like. The kits may contain the peptide composition in bulk form, or may contain separate doses of the peptide composition for continuous or sequential administration. The kits may optionally contain one or more diagnostic tools, administration tools, and / or instructions for use. The kits may contain 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 agents. 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.

[0141] In some embodiments, the composition also works with the skin's own natural renewal process, helping to improve the appearance of the skin and the firmness of the skin. The topical composition is suitable for all skin types and post-procedure skin. The topical composition can be provided to the patient in bulk form to allow a suitable amount of peptide to be self-administered by the patient. For example, the patient can apply a sufficient amount of the composition to evenly cover the affected area, or as otherwise directed by a physician. In certain embodiments, it may be desirable to incorporate additional therapeutic or active agents into the topical composition. Alternatively, supplemental therapies or agents can be administered separately. For example, cleansers, sunscreens, sunscreens, penetrating moisturizers, and / or occlusive moisturizers can be provided for administration before or after the topical composition of the embodiments.

[0142] Various examples of creams, ointments, lotions, solutions, gels, sprays, and patches may incorporate the peptide compositions described herein as active ingredients in combination with penetration enhancers and other active agents that act synergistically on the skin to promote wound healing or wound closure, or to treat chronic skin wounds.

[0143] 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.

[0144] 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.

[0145] 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."

[0146] 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.

[0147] 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.

[0148] 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.

[0149] 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.

[0150] 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.

[0151] 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. EXAMPLES

[0152] 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.

[0153] Example 1. Gene Expression Assay This example shows the effect of different active agents on gene expression in fibroblasts and keratinocytes.

[0154] 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 2Cells were seeded in cell specific medium at 5K or 10K cells per well. Cells were seeded in triplicate in 48-well plates, one cell line per plate. Medium volume was 500ul per well. The outer two columns of the wells were not used. Cells were cultured for 2 days in a 37°C, 5% CO2 incubator. After 48 hours, all cell cultures were homogenous and appeared healthy, with no appreciable numbers of floating dead cells or evidence of cellular vacuolation that could indicate apoptotic or dying cells.

[0155] Compounds. Stocks of compounds 1-8 listed below were prepared in PBS at 100x the concentration indicated by the red type. A 50mg / 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 20mg / 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.

[0156] 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

[0157] 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 (500μg / ml), TCVRRAF (100μg / ml) (amino acid sequence extracted from lactoferrin), Tri-peptide-1-100ppm (2.9μg / ml), Hexapeptide 12, 100ppm (2.9μg / ml), TriHex combination 200ppm (2.9μg / ml each), Hexapeptide 11 (100μg / ml), Tranexamic acid 5% (500μg / ml), Octapeptide (100μg / ml) - a proprietary peptide designed by Alastin, Phosphatidylserine 500ug / ml, or Cannabidiol (CBD) 100μg / ml. Control cells were left untreated.

[0158] RNA lysate preparation. After 24 hours of compound exposure, media was removed and cells were washed once with PBS. 100ul of RNA lysis buffer was added to wells, mixed thoroughly by trituration, combined in an RNAse-free microfuge tube, and immediately frozen at -30°C. Triplicate wells were thawed and combined in 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.

[0159] 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.

[0160] 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).

[0161] 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.

[0162] 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.

[0163] 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.

[0164] 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.

[0165] Example 2. Selection of agents for in vitro HA production and size assessment This example assays whether the octapeptide, SymDecanox™, Tremella, lactoferrin, phosphatidylserine, Hylasome™, Aquaxyl™, and complete formulations stimulate the secretion of high molecular weight (high MW) hyaluronic acid (HA) from dermal fibroblasts (and keratinocytes). This example also assays for HA synthesis.

[0166] 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 assess HA production by fibroblasts and 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.

[0167] Selection of agents for in vitro HA production and size assessment. The octapeptide (a unique peptide) upregulates hyaluronan synthase-2 (HAS2) gene expression in fibroblasts (based on the RNA-Seq data described in Example 1).

[0168] Lactoferrin may provide wound healing attributes, promote fibroblast proliferation, and increase HA secretion.

[0169] Syn-Hycan is a synthetic tripeptide that stimulates HA. The synthetic trifluoroacetate tripeptide tetradecylaminobutyroylvalylaminobutyric acid urea may restore facial skin volume by stimulating HA synthesis. Syn-Hycan increases HA and CD44 in skin in vitro and ex vivo.

[0170] Phosphatidylserine stimulates hyaluronan synthesis.

[0171] Sodium hyaluronate crosspolymer (Hyalasome) is an extremely high MW synthetic HA with exceptionally high water binding capacity resulting in excellent moisturizing properties.

[0172] Tremella fuciformis extract is derived from an edible mushroom and provides high levels of moisture as a natural HA stimulator and has antioxidant properties.

[0173] Hydroxymethoxyphenyldecanone is an HA booster, antioxidant, and anti-irritant, and can stimulate dermal and epidermal hyaluronic acid levels in an ex vivo human skin model.

[0174] Hexapeptide-11 can upregulate HAS2 in keratinocytes (based on the RNA-seq data in Example 1) and strongly downregulate the HA reductase hyaluronidase 2 (HYAL2).

[0175] Table 1. Compounds used to treat fibroblasts for HA production assessment by PAGE TIFF2025512392000002.tif67170

[0176] 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.

[0177] 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 megadaltons / 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, and none of them produce low molecular weight (LMW) HA.

[0178] Verifying HA Fibroblasts were cultured as described above to near confluence in 6-well plates. Cells were then treated with compounds shown in Table 1. After 72 hours, 100 μl of medium was collected from each treatment condition. Medium was subjected to hyaluronidase enzyme treatment (1 mg / ml) for 2 hours at 37° C. One condition was left undigested (100 μg / ml of octapeptide). Samples were run on SDS-PAGE gels, stained, and destained as described. 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 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).

[0179] 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 10 μg / ml and 100 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.

[0180] Example 3. Multicenter evaluation of topical hyaluronic acid formulations on facial skin This example includes a study on the progressive effect of treatments using different topical formulations as part of a facial skin care routine, comparing results across all treatments using different formulations. The formulations tested included topical application of a composition containing high MW HA and octapeptide in combination with either SilkSHIELD™ (available at www.alastin.com) or HydraTint™ SPF (available at www.alastin.com) combined with a facial care routine using a gentle cleanser. A composition containing high MW HA and octapeptide in combination with SPF30+, optional ultra-light moisturizer after 4 weeks, and regular skin care using a gentle cleanser was also tested. This study evaluated the efficacy and safety of topical compositions containing high MW HA and octapeptide on facial skin. A photo analysis was performed using the first treatment formulation (a composition containing high MW HA and octapeptide in combination with either SilkSHIELD™ or HydraTint™ SPF sunscreen). Photographic and histological analysis of biopsies using a second treatment formulation, a composition comprising high MW HA and an octapeptide, SPF 30+, and optional topical application of an ultra-light moisturizer after 4 weeks was performed.

[0181] 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 topicals 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.

[0182] 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 morning and afternoon use, and SPF30+ for morning use. At each visit, participants cleansed their facial skin and allowed it to acclimate for 15 minutes before assessment, photography, and measurement. Participants then applied the topical composition and allowed it to acclimate for 15 minutes before a second set of assessment, photography, and measurement. The HA immerse composition with formulated percentages of ingredients is listed in Table 2 with the weight concentration of the solution expressed as % w / w.

[0183] Table 2: Exemplary compositions TIFF2025512392000003.tif197161

[0184] Table 3: Exemplary compositions TIFF2025512392000004.tif51128

[0185] Various embodiments of the compositions described herein include the ranges of components listed in Tables 2 and 3 in weight concentration of the solution expressed as % w / w.

[0186] 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.

[0187] Participant Assessment and Satisfaction. At all follow-up visits, 15 minutes after cleansing the skin, participants completed an assessment of their facial skin (using a 5-point scale) and overall satisfaction (using a 7-point scale) with the topical test product compared to baseline. In addition, at each visit, 15 minutes after applying the topical composition, participants completed an assessment of their facial skin (using a 5-point scale).

[0188] 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 parameters: 0 absent, 1-3 mild, 4-6 moderate, 7-9 severe.

[0189] 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).

[0190] 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.

[0191] 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.

[0192] Results: Two participants were terminated early due to excessive sun exposure. An independent statistician completed the analyses using descriptive statistics, parametric and nonparametric tests.

[0193] The average hydration level in cleansed skin increases progressively with continued treatment. Analysis was completed for clean skin between baseline and follow-up visits, and between clean skin and after application of the composition 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 previous visit (Figure 7).

[0194] At each visit, hydration measurements were statistically significantly higher as measured after application compared to clean skin (Figure 8). At 4 and 8 weeks after application, measurements were also significantly higher than baseline and week 2.

[0195] 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):

[0196] 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).

[0197] 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).

[0198] 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<0.0001. Grades for reduction in fine lines / wrinkles, reduction in fine lines, texture, erythema, and dryness, and increase in moisture / hydration.

[0199] 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.

[0200] Photos and Analysis Figures 14-16 are photographs and analyses of subjects at baseline and at various time points after initiation of treatment with a formulation containing high MW HA and an octapeptide, either SilkSHIELD™ or HydraTint™ SPF sunscreen, and usual skin care using a gentle cleanser.

[0201] FIG. 14A demonstrates a significant improvement in skin appearance in a male subject, comparing baseline (left photo) and week 4 (right photo). Facial redness was reduced and fine lines were minimized. It is noteworthy that these significant effects were recorded after only 4 weeks of treatment. FIG. 14B more clearly shows the improvement in minimizing the appearance of fine lines and wrinkles in this close-up view of the subject's forehead from FIG. 14A.

[0202] Figure 15 demonstrates a significant improvement in skin appearance in a female subject, comparing baseline (left photo) and week 8 (right photo). There was a reduction in redness in the subject after the treatment period. The bottom panel shows only the red channel of the digital photo to specifically demonstrate the reduction in redness after treatment, apart from any other alterations in skin tone. Examples include Canfield redness quantification.

[0203] FIG. 16 demonstrates a significant improvement in skin appearance in a female subject, comparing baseline (left photo) to weeks 2 (middle photo) and 4 (right photo). Of note was the reduction in redness in the subject evident at the earliest time point assessed (week 2), and continued reduction in redness through week 4. The bottom panel displays only the red channel of the digital photo to specifically demonstrate the post-treatment redness reduction apart from any other alterations in skin tone. Examples include Canfield redness quantification.

[0204] Photos and Analysis: HA Immerse Serum formulation, SPF 30+, and optional Ultra Light Moisturizer treatment after 4 weeks Figures 17-24 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 4 weeks, and a gentle cleanser in the third experiment.

[0205] FIG. 17A demonstrates improved appearance and reduced redness after 4 weeks in a female subject, age 39.

[0206] FIG. 17B shows the results of a VISIA® Skin Analysis System for the subject from FIG. 17A demonstrating a quantifiable reduction in erythema.

[0207] Figure 18 shows photographs taken at baseline, week 2, week 4, and week 8 (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.

[0208] Figures 19A-19C demonstrate the degree of improvement in quantifiable features of appearance in the subject from Figure 18 comparing baseline and week 8 measurements. Figure 19B shows the results of a VISIA® Skin Analysis System to show a measurable reduction in red areas on the face. Figure 19C 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.

[0209] Figures 19D-19E 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 19E 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.

[0210] Figures 20A-20B 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 20B 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.

[0211] 21A-21C 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.

[0212] FIG. 21A 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.

[0213] Figures 21B-21C 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 21A at week 8. The reduction in visibility of fine blood vessels after this treatment period is particularly evident.

[0214] 22A-22C 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 lines on the skin resulting in the appearance of smoother, firmer skin when comparing baseline, week 2, week 4, and week 8 in a female subject, age 73.

[0215] Figures 23A-23C show tissue analysis within periauricular biopsies of three subjects at baseline and 8 weeks after treatment with a composition comprising high MW HA+octapeptide. Figures 23A-23C 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 the composition, 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 ones indicates the therapeutic benefit of the treatment in both the superficial and deeper regions of the dermis.

[0216] Figures 24A-24C show histological analysis of CD44 staining within periauricular biopsies of two subjects at baseline and 8 weeks after treatment with a composition comprising high MW HA + octapeptide. Figure 24A 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 24B-24C 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.

[0217] FIG. 25 shows a diagram of the chemical structure of octapeptide-45, which comprises the amino acid sequence Glu-Pro-His-Gly-Val-Arg-Glu-Ala.

[0218] Although the foregoing disclosure has been described in some detail for purposes of clarity and understanding, it will be apparent to those skilled in the art from reading this disclosure that various changes in form and detail can be made without departing from the true scope of the present disclosure. For example, all of the techniques and apparatus described above can be used in various combinations. All publications, patents, patent applications, and / or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and / or other document was individually and separately indicated to be incorporated by reference for all purposes.

Claims

1. Octapeptide and High molecular weight (MW) hyaluronic acid (HA) or its derivatives and A topical composition comprising, The aforementioned high MW HA has a molecular weight of at least 0.5 MDa. The aforementioned topical composition.

2. The composition according to claim 1, wherein the octapeptide comprises at least one of 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), or GPGKNGDD (SEQ ID NO: 7).

3. The composition according to claim 1, wherein the octapeptide is GPHGVREA (SEQ ID NO: 5).

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

5. The composition according to claim 1, wherein the high MW HA or its derivative comprises sodium hyaluronate.

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

7. The composition according to claim 6, wherein the synthetic tripeptide is urea trifluoroacetate tetradecylaminobutyroylvalylaminobutyric acid.

8. The composition according to claim 1, comprising a hexapeptide.

9. The composition according to claim 8, wherein the hexapeptide is hexapeptide-11.

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

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

12. The composition according to claim 1, comprising tripeptide-1 and hexapeptide-12.

13. The composition according to any one of claims 1 to 12 for reducing inflammation in the skin of an individual, treating or restoring aging skin, treating a skin wound, 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, Reduction of skin redness in an individual after application of the composition to the skin. The increase in skin hydration in the skin of an individual 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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