Compositions and methods for stimulating hyaluronic acid

Abstract

Compositions and methods for stimulating endogenous hyaluronic acid production are provided herein, which can improve skin hydration and elasticity. TIFF2023544824000002.tif76156

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 089,424, filed Oct. 8, 2020, and U.S. Provisional Patent Application No. 63 / 218,068, filed Jul. 2, 2021, each of which is incorporated herein by reference in its entirety.

Background Art

[0002] Background Hyaluronic acid is a non - sulfated glycosaminoglycan that is widely distributed throughout connective tissue, epithelial tissue, and nerve tissue such as the skin. Hyaluronic acid is used in topical compositions for the skin due to its ability to promote skin hydration. However, low - molecular - weight hyaluronic acid can promote the production of pro - inflammatory mediators. Therefore, there is a need to provide topical compositions that promote endogenous hyaluronic acid production without associated adverse effects such as the production of pro - inflammatory mediators.

Summary of the Invention

[0003] Brief Summary The skin contains 50% of the body's hyaluronic acid (HA). HA is a major component of the skin's extracellular matrix (ECM) and is present in the epidermis, dermis, and the basement membrane between them. HA is also observed intracellularly and plays an important role in metabolism, cell turnover, differentiation, cell migration, tissue repair, hydration, nutrient exchange, and protection against free - radical damage. The rapid turnover of HA suggests that HA may also be important as a conduit for the removal of toxic substances. Natural HA and modified cross - linked HA have been used to help the skin maintain and even regain elasticity, tone, and moisture. Topical compositions are needed to promote endogenous HA stimulation within the skin.

[0004] Embodiments described herein are topical compositions for stimulating hyaluronic acid, comprising a synthetic tripeptide, an octapeptide, and a hexapeptide, the topical compositions stimulating hyaluronic acid. In one feature, the synthetic tripeptide comprises tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate. In one feature, the octapeptide is encapsulated in liposomes. In one feature, the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). In one feature, the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1). In one feature, the octapeptide contains the amino acid sequence GPMGPSGP (SEQ ID NO: 2). In one feature, the octapeptide contains the amino acid sequence GLGPGARA (SEQ ID NO: 3). In one feature, the octapeptide contains the amino acid sequence GPQGFQGP (SEQ ID NO: 4). In one feature, the octapeptide contains the amino acid sequence GPHGVREA (SEQ ID NO: 5). In one feature, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 6). In one feature, the octapeptide contains the amino acid sequence GPGKNGDD (SEQ ID NO: 7). In one feature, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 8). In one feature, the hexapeptide is hexapeptide-11. In one feature, hexapeptide-11 is encapsulated in liposomes. In one feature, the topical composition further comprises lactoferrin. In one feature, lactoferrin is encapsulated in liposomes. In another feature, the topical composition further comprises phosphatidylserine. In yet another feature, the topical composition further comprises Tremella fuciformis extract. In yet another feature, the topical composition further comprises sodium hyaluronate crosspolymer.In one feature, the topical composition further comprises hydroxymethoxyphenyl decane. In one feature, the topical composition is aqueous. In one feature, the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide contains the amino acid sequence GPQGFQGP (SEQ ID NO: 4), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide contains the amino acid sequence GPHGVREA (SEQ ID NO: 5), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 6), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof.In one feature, the octapeptide contains the amino acid sequence GPGKNGDD (SEQ ID NO: 7), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 8), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof.

[0005] Embodiments described herein are methods for stimulating hyaluronic acid production, comprising the step of administering a topical composition comprising a synthetic tripeptide, an octapeptide, and a hexapeptide. In one feature, the synthetic tripeptide comprises tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate. In one feature, the octapeptide is encapsulated in liposomes. In one feature, the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). In one feature, the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1). In one feature, the octapeptide contains the amino acid sequence GPMGPSGP (SEQ ID NO: 2). In one feature, the octapeptide contains the amino acid sequence GLGPGARA (SEQ ID NO: 3). In one feature, the octapeptide contains the amino acid sequence GPQGFQGP (SEQ ID NO: 4). In one feature, the octapeptide contains the amino acid sequence GPHGVREA (SEQ ID NO: 5). In one feature, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 6). In one feature, the octapeptide contains the amino acid sequence GPGKNGDD (SEQ ID NO: 7). In one feature, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 8). In one feature, the hexapeptide is hexapeptide-11. In one feature, hexapeptide-11 is encapsulated in liposomes. In one feature, the topical composition further comprises lactoferrin. In one feature, lactoferrin is encapsulated in liposomes. In one feature, the topical composition further comprises phosphatidylserine. In one feature, the topical composition further comprises Trellis fusiformis extract. In one feature, the topical composition further comprises sodium hyaluronate crosspolymer. In one feature, the topical composition further comprises hydroxymethoxyphenyldecanone.In one feature, the octapeptide contains the amino acid sequence GPMGPSGP (SEQ ID NO: 2), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide contains the amino acid sequence GLGPGARA (SEQ ID NO: 3), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide contains the amino acid sequence GPQGFQGP (SEQ ID NO: 4), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one characteristic, the octapeptide contains the amino acid sequence GPGKNGDD (SEQ ID NO: 7), and the hexapeptide is hexapeptide-11.In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8), and the hexapeptide is hexapeptide-11. In one feature, the topical composition further comprises lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. In one feature, the topical composition is aqueous. In one feature, the topical composition improves skin moisture. In one feature, the topical composition reduces the appearance of bruises, age spots, or wrinkles. In one feature, the topical composition is administered 1, 2, 3, 4, 5, 6, 7, or 8 times a day. In one feature, the individual is human. [Brief explanation of the drawing]

[0006] [Figure 1] The graph shows the expression of hyaluronic acid synthase 2 (HAS2) in fibroblasts. [Figure 2] The graph shows the expression of hyaluronic acid synthase 2 (HAS2) in keratinocytes. [Figure 3] The graph shows the expression of hyaluronidase 2 (HYAL2) in keratinocytes. [Figure 4] The SDS-PAGE gel is shown demonstrating the effects of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment in the first experiment. [Figure 5] This shows an SDS-PAGE gel demonstrating the effect of the octapeptide compound on hyaluronic acid production in human fibroblasts 72 hours after treatment in the first experiment. [Figure 6] The second experiment shows an SDS-PAGE gel demonstrating the effects of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment. [Figure 7] The second experiment shows an SDS-PAGE gel demonstrating the effects of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment. [Figure 8] Figure 8A shows the SDS-PAGE gel of octapeptide-induced HA production in fibroblasts, and Figure 8B shows a graph quantifying the results. [Figure 9] This graph demonstrates EGR3 gene expression after treatment with various compounds.

[0007] Embedding by reference All publications, patents, and patent applications referenced herein are incorporated by reference to the same extent as each individual publication, patent, or patent application is specifically and individually indicated as being incorporated by reference. [Modes for carrying out the invention]

[0008] Detailed explanation definition Throughout this disclosure, various embodiments are presented in range form. It should be understood that the range form description is for convenience and brevity only and should not be interpreted as an inalienable limitation on the scope of any embodiment. Therefore, unless the context clearly indicates otherwise, the range description should be considered to specifically disclose all possible subranges and individual numerical values ​​within a range up to one-tenth of the lower limit unit. For example, a range description such as 1-6 should be considered to specifically disclose subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, and individual values ​​within those ranges, e.g., 1.1, 2, 2.3, 5, and 5.9. This applies regardless of the width of the range. These intervening upper and lower limits may independently be included within smaller ranges and are incorporated within this disclosure, subject to any specifically excluded limitations within the described range. If the scope described includes one or both of the limitations, the scope excluding one or both of those limitations is also included in this disclosure, unless the context otherwise explicitly indicates.

[0009] The technical terms used herein are intended solely to describe specific embodiments and are not intended to limit any embodiments. Where used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well, unless the context otherwise explicitly indicates. The terms “comprises” and / or “comprising,” where used herein, specify the presence of the described features, integers, steps, actions, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, actions, elements, components, and / or groups thereof. Where used herein, the terms “and / or” include any combination of one or more of the items listed in relation.

[0010] Where used herein, unless otherwise specifically stated or evident from the context, the term “about” with respect to a number or range of numbers is understood to mean the stated number and its range plus or minus 10%, or, for any enumerated range of values, 10% below the enumerated lower limit and 10% above the enumerated upper limit.

[0011] composition Hyaluronic acid (HA) is a major component of the extracellular matrix (ECM) of the skin. HA plays a crucial role in metabolism, cell turnover, differentiation, cell migration, tissue repair, hydration, nutrient exchange, and protection against free radical damage. Topical compositions are needed to promote endogenous HA stimulation within the skin.

[0012] This specification describes compositions and methods for stimulating hyaluronic acid (HA). The compositions and methods described herein may include HA, such as high molecular weight HA. The compositions and methods described herein may further include one or more agents that stimulate cells in the skin to produce more HA. The compositions and methods described herein may promote endogenous hyaluronic acid (HA) stimulation. The compositions and methods described herein may promote the production of high molecular weight HA (HMW-HA). In some embodiments, the compositions described herein promote HA synthesis. In some embodiments, the compositions described herein promote the gene expression of genes involved in HA synthesis (e.g., HAS2, HYAL2, EGR3).

[0013] Hyaluronic acid This specification describes compositions comprising hyaluronic acid. In some embodiments, the hyaluronic acid is high molecular weight hyaluronic acid. In some embodiments, the hyaluronic acid is synthetic. In some embodiments, the hyaluronic acid has improved water-binding capacity. In some embodiments, the hyaluronic acid is crosslinked.

[0014] In general, the term "hyaluronic acid" can also encompass all variants and combinations of variants of hyaluronic acid, hyaluronic acid salts, or hyaluronans, including various chain lengths and charge states, as well as various chemical modifications, including crosslinking. In some cases, hyaluronic acid includes hyaluronic acid salts with various counterions, such as sodium hyaluronate. Various modifications of hyaluronic acid, such as oxidation (e.g., oxidation of the -CH2OH group to -CHO and / or -COOH, periodate oxidation of adjacent hydroxyl groups), optionally followed by reduction (e.g., reduction of -CHO to -CH2OH, or coupling with amines to form imines, followed by reduction to secondary amines), sulfation, deamidation (optionally followed by deamination or amide formation with new acids), esterification, crosslinking (e.g., substitution with various compounds using crosslinking agents or carbodiimide co-couplings), coupling (including coupling of different molecules such as proteins, peptides, and active drug components to hyaluronic acid), and deacetylation, are also encompassed by this term. Other examples of modifications include isourea, hydrazide, bromosyanide, monoepoxide, and monosulfone coupling.

[0015] The compositions described herein, in some embodiments, include sodium hyaluronate crosspolymer. Sodium hyaluronate crosspolymer is a high molecular weight synthetic hyaluronic acid derived from non-animal sources that has high water-binding and moisturizing capabilities. Sodium hyaluronate crosspolymer is also a scavenger of harmful free radicals and has a unique gel structure with gel domains that hold tightly bound water.

[0016] In some embodiments, sodium hyaluronate cross polymer is provided at at least or about 0.0001 wt.%, 0.0005 wt.%, 0.001 wt.%, 0.005 wt.%, 0.01 wt.%, 0.02 wt.%, 0.05 wt.%, 0.10 wt.%, 0.20 wt.%, 0.25 wt.%, 0.50 wt.%, 0.75 wt.%, 1.0 wt.%, 1.5 wt.%, 2.0 wt.%, 2.5 wt.%, 3.0 wt.%, 3.5 wt.%, 4.0 wt.%, or more than 4.0 wt.%. In some embodiments, sodium hyaluronate cross polymer is provided at about 0.5 wt.%. In some embodiments, sodium hyaluronate cross polymer is provided in the range of about 0.0001 wt.% to about 4.0 wt.%, about 0.001 wt.% to about 4.0 wt.%, about 0.01 wt.% to about 3.0 wt.%, about 0.1 wt.% to about 2.5 wt.%, or about 0.50 wt.% to about 1.5 wt.%.

[0017] Peptide The peptides described herein promote endogenous hyaluronic acid (HA) stimulation in some embodiments. The peptides described herein promote HA synthesis in some embodiments. The peptides described herein (e.g., octapeptide) promote the production of high molecular weight HA (HMW-HA) in some embodiments. The peptides described herein promote the gene expression of genes involved in HA synthesis (e.g., HAS2, HYAL2, EGR3) in some embodiments.

[0018] The compositions described herein contain peptides at various concentrations. In some cases, the peptide is present at about 50 ppm or less to 1000, 5000, 10000, 50000, 100000, 500000 ppm or more, for example, 100 ppm of the peptide. 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 more than 1000 ppm. In some cases, the peptide 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 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 more than 1000 micrograms per milliliter (ug / mL). In some cases, the peptide 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, the peptide is present at about 0.01 wt.% to about 10%, about 0.01 wt.% to about 0.02 wt.%, about 0.01 wt.% to about 0.03 wt.%, about 0.01 wt.% to about 0.04 wt.%, about 0.01 wt.% to about 0.05 wt.%, about 0.01 wt.% to about 0.1 wt.%, about 1 wt.% to about 5 wt.%, or about 1 wt.% to about 10 wt.% (wt.%).

[0019] The compositions described herein, in some embodiments, comprise a plurality of peptides. In some cases, the peptides among the plurality are present in amounts ranging from about 50 ppm or less to 1000, 5000, 10000, 50000, 100000, 500000 ppm or more, for example, 100 ppm of peptide, or any other preferred amount. In some cases, the peptides among the plurality are present in amounts ranging from 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 ppm. In some cases, the peptides among the multiple peptides are present in concentrations ranging from approximately 1 to 100, 1 to 50, 1 to 40, 1 to 30, 1 to 20, 1 to 10, 5 to 90, 10 to 80, 20 to 60, or 30 to 50 ppm. In some cases, the peptides among the multiple peptides are present in concentrations of approximately 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 micrograms (ug / mL) per milliliter. In some cases, the peptides among the multiple peptides are present in amounts ranging from approximately 1 to approximately 100, approximately 1 to approximately 50, approximately 1 to approximately 40, approximately 1 to approximately 30, approximately 1 to approximately 20, approximately 1 to approximately 10, approximately 5 to approximately 90, approximately 10 to approximately 80, approximately 20 to approximately 60, or approximately 30 to approximately 50 micrograms per milliliter. In some cases, the peptides among the multiple peptides are present in amounts ranging from approximately 0.01% to approximately 10%, approximately 0.01% to approximately 0.02%, approximately 0.01% to approximately 0.03%, approximately 0.01% to approximately 0.04%, approximately 0.01% to approximately 0.05%, approximately 0.01% to approximately 0.1%, approximately 1% to approximately 5%, or approximately 1% to approximately 10% (wt.%).In some embodiments, the peptides among the plurality of peptides are provided in amounts of at least or about 0.00001% by weight, 0.0003% by weight, 0.0005% by weight, 0.001% by weight, 0.001% by weight, 0.005% by weight, 0.0055% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, the peptides among the multiple peptides are provided in amounts ranging from about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, each of the multiple peptides is provided in amounts ranging from 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.

[0020] In some embodiments, the compositions described herein include hexapeptide-11, octapeptide, synthetic peptide, or a combination thereof. In some embodiments, the synthetic peptide is tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate.

[0021] In some embodiments, hexapeptide-11 is present in amounts of at least or about 0.00001% by weight, 0.0003% by weight, 0.0005% by weight, 0.001% by weight, 0.001% by weight, 0.005% by weight, 0.0055% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1 0.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, 20% by weight, 30% by weight, 40% by weight, 50% by weight, 60% by weight, 70% by weight, 80% by weight, 90% by weight, or greater than 90% by weight (wt.%). In some embodiments, hexapeptide-11 is provided in the range of about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. 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% by weight to about 0.02% by weight. In some embodiments, hexapeptide-11 is provided in at least or more than about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or 250 ppm. In some embodiments, hexapeptide-11 is provided in the range of about 25 to about 250, about 50 to about 200, or about 75 to about 150 ppm. In some embodiments, hexapeptide-11 is provided in the range of about 10 to about 100 ppm. In some embodiments, hexapeptide-11 is provided in amounts of at least or more than 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or 250 micrograms (ug / mL) per milliliter. In some embodiments, hexapeptide-11 is provided in amounts ranging from about 25 to about 250, about 50 to about 200, or about 75 to about 150 micrograms per milliliter.

[0022] In some embodiments, the octapeptide is provided in at least or about 0.00001% by weight, 0.0003% by weight, 0.0005% by weight, 0.001% by weight, 0.001% by weight, 0.005% by weight, 0.0055% by weight, 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, the octapeptide is provided in a range of about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, the octapeptide is provided in at least or greater than 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 25 ppm. In some embodiments, the octapeptide is provided in a range of about 1 to about 10 ppm. 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 ppm. In some embodiments, the octapeptide is provided in an amount of at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 micrograms (ug / mL) per milliliter. In some embodiments, the octapeptide is provided in an amount of at least or about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or more than 200 micrograms (ug / mL) per milliliter. In some embodiments, the octapeptide 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.In some embodiments, the octapeptide is provided in the range of about 25 to about 200, about 25 to about 150, about 50 to about 150, about 50 to about 125, about 10 to about 60, or about 20 to about 40 micrograms per milliliter.

[0023] In some embodiments, the synthetic peptide is present in at least or about 0.00001% by weight, 0.0003% by weight, 0.0005% by weight, 0.001% by weight, 0.001% by weight, 0.005% by weight, 0.0055% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, and 1.5% by weight. The synthetic peptides are provided in amounts of wt%, 2.0 wt%, 2.5 wt%, 3.0 wt%, 3.5 wt%, 4.0 wt%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, 20 wt%, 30 wt%, 40 wt%, 50 wt%, 60 wt%, 70 wt%, 80 wt%, 90 wt%, or greater than 90 wt% (wt.%). In some embodiments, the synthetic peptides are provided in amounts ranging from 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 synthetic peptide 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, the synthetic peptide is provided in the range of about 0.005% by weight to about 0.02% by weight. In some embodiments, the synthetic peptide is provided in at least or more than about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or 250 ppm. In some embodiments, the synthetic peptide is provided in the range of about 25 to about 250, about 50 to about 200, or about 75 to about 150 ppm. In some embodiments, the synthetic peptide is provided in amounts of at least or about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or more than 250 micrograms (ug / mL) per milliliter. In some embodiments, the synthetic peptide is provided in amounts of at least or about 5250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or more than 1000 micrograms (ug / mL) per milliliter.In some embodiments, the synthetic peptide is provided in the range of about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, about 100 to about 800, or about 100 to about 500 micrograms (ug / mL) per milliliter. In some embodiments, the synthetic peptide is provided in the range of about 25 to about 250, about 50 to about 200, or about 75 to about 150 micrograms (ug / mL) per milliliter. In some embodiments, the synthetic peptide is a synthetic tripeptide. In some embodiments, the synthetic peptide is tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate.

[0024] In some embodiments, tripeptide-1 is provided in at least or about 0.00001% by weight, 0.0003% by weight, 0.0005% by weight, 0.001% by weight, 0.001% by weight, 0.005% by weight, 0.0055% by weight, 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, tripeptide-1 is provided in the range of about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, tripeptide-1 is provided in at least or greater than 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 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 in amounts of at least or more than 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 25 micrograms (ug / mL) per milliliter. In some embodiments, tripeptide-1 is provided in amounts ranging from 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.

[0025] In some embodiments, hexapeptide-12 is provided in at least or about 0.00001% by weight, 0.0003% by weight, 0.0005% by weight, 0.001% by weight, 0.001% by weight, 0.005% by weight, 0.0055% by weight, 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, hexapeptide-12 is provided in concentrations ranging from about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, hexapeptide-12 is provided in concentrations of at least or greater than 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 25 ppm. In some embodiments, hexapeptide-12 is provided in concentrations ranging from about 1 to about 10 ppm. In some embodiments, hexapeptide-12 is provided in concentrations ranging from 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 in amounts of at least or more than 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 25 micrograms (ug / mL) per milliliter. In some embodiments, hexapeptide-12 is provided in amounts ranging from 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.

[0026] In exemplary embodiments, the weight ratio of the first peptide to the second peptide in the topical composition is 1 part of the first peptide to 0.2 to 10 parts of the second peptide, 1 to 10 parts of the second peptide, 1 to 8 parts of the second peptide, or 1 to 5.5 parts of the second peptide. The following nomenclature is used herein to refer to various amino acids: alanine (also referred to herein as "Ala" or "A"), arginine (also referred to herein as "Arg" or "R"), asparagine (also referred to herein as "Asn" or "N"), aspartic acid (also referred to herein as "Asp" or "D"), cysteine ​​(also referred to herein as "Cys" or "C"), glutamic acid (also referred to herein as "Glu" or "E"), glutamine (also referred to herein as "Gln" or "Q"), glycine (also referred to herein as "Gly" or "G"), histidine (also referred to herein as "His" or "H"), isoleucine (as specified herein). Fluorine (also referred to as "Ile" or "I" in this specification), leucine (also referred to as "Leu" or "L" in this specification), lysine (also referred to as "Lys" or "K" in this specification), methionine (also referred to as "Met" or "M" in this specification), phenylalanine (also referred to as "Phe" or "F" in this specification), proline (also referred to as "Pro" or "P" in this specification), serine (also referred to as "Ser" or "S" in this specification), threonine (also referred to as "Thr" or "T" in this specification), tryptophan (also referred to as "Trp" or "W" in this specification), tyrosine (also referred to as "Tyr" or "Y" in this specification), valine (also referred to as "Val" or "V" in this specification).

[0027] In some embodiments, the first peptide is a dipeptide. Preferred dipeptides include, but are not limited to, those having the following amino acid sequences: KK, KP, CK, KC, KT, DF, NF, VW, YR, or TT. In some embodiments, the dipeptide has the following amino acid sequence: KV. In other embodiments, the first peptide is a tripeptide. Preferred tripeptides include, but are not limited to, those having the following amino acid sequences: HGG, RKR, GHK, GKH, GGH, GHG, KFK, or KPK. In some embodiments, the tripeptide has the following amino acid sequence: KVK. In some embodiments, the first peptide is a tetrapeptide. Preferred tetrapeptides include, but are not limited to, those having the following amino acid sequences: GQPR, KTFK, AQTR, or RSRK. In some embodiments, the tetrapeptide has the following amino acid sequence: KDVY. In some embodiments, the second peptide is a pentapeptide. Suitable pentapeptides include, but are not limited to, those having the following amino acid sequences: KTTKS, YGGFX, or KLAAK. In some embodiments, the second peptide is a hexapeptide. Suitable hexapeptides include, but are not limited to, those having the following amino acid sequences: VGVAPG or GKTTKS. In some embodiments, the hexapeptide has the following amino acid sequence: FVAPFP. In some embodiments, the second peptide is a heptapeptide. Suitable heptapeptides include, but are not limited to, those having the amino acid sequence RGYYLLE, or heptapeptide-6 (prosirtuin peptide). The composition may contain two or more peptides, such as two dipeptides and one pentapeptide, one tripeptide and one hexapeptide, one dipeptide, one tripeptide, and one heptapeptide, provided that the composition contains at least one dipeptide, tripeptide, or tetrapeptide and at least one pentapeptide, hexapeptide, or heptapeptide.In some embodiments, the compositions described herein include hexapeptides, octapeptides, synthetic peptides, or combinations thereof. In some embodiments, one or more hexapeptides are hexapeptide-11. In some embodiments, one or more peptides are tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate.

[0028] Peptides can be functionalized. For example, peptides can be functionalized with fatty acids, such as myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoleidic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, and serotic acid. 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 because it exhibits enhanced penetration compared to other fatty acids. In some embodiments, peptides are functionalized with chemical groups. For example, peptides are functionalized with acetyl. Examples include acetyl hexapeptide-38 and acetyl tetrapeptide-2. In some cases, peptides are functionalized with functional groups containing 14 or fewer carbon atoms. In some cases, peptides are functionalized with functional groups containing 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 20 or more carbon atoms. In some cases, peptides are depalmitoylated. In some embodiments, though not limited to any particular theory, incorporating peptides into liposomes increases the lipophilicity of functionalized or unfunctionalized peptides.

[0029] 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 the general class of proteins 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 into and out of cells, promoting wound healing through several different but related pathways. Due to its strong copper-binding structure, GHK may be provided in the form of GHK-Cu (copper-bound GHK form).

[0030] In some embodiments, the compositions described herein contain an octapeptide. In some embodiments, the octapeptide contains the amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). In some embodiments, the octapeptide contains the amino acid sequence GDGDGASA (SEQ ID NO: 1). In some embodiments, the octapeptide contains the amino acid sequence GPMGPSGP (SEQ ID NO: 2). In some embodiments, the octapeptide contains the amino acid sequence GLGPGARA (SEQ ID NO: 3). In some embodiments, the octapeptide contains the amino acid sequence GPQGFQGP (SEQ ID NO: 4). In some embodiments, the octapeptide contains the amino acid sequence GPHGVREA (SEQ ID NO: 5). In some embodiments, the octapeptide contains the amino acid sequence GPMGPRGP (SEQ ID NO: 6). In some embodiments, the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7). In some embodiments, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8).

[0031] Peptides may be advantageously provided in a substrate suitable for combination with other components of a liposome composition. The substrate may contain one or more components such as a thickener / binder (e.g., pentaerythrityl tetraisostearate), a softener / dispersant (e.g., caprylic / capric triglyceride), a solvent (e.g., propylene carbonate), and / or a rheological modifier / antiprecipitating agent (e.g., disteadimonium hectorite).

[0032] Liposomes This specification describes liposome compositions for improved distribution, efficacy, bioavailability, and / or activity. Liposome compositions can improve the distribution, efficacy, bioavailability, and / or activity of active ingredients by improving delivery and tissue (e.g., skin) penetration. In some cases, the improved delivery and skin penetration are due to the active ingredient incorporated (e.g., encapsulated) within the liposome. In some cases, the active ingredient is a peptide encapsulated within the liposome.

[0033] The liposome compositions described herein may include peptides encapsulated in liposomes. In some embodiments, the peptide is hexapeptide-11. In some embodiments, the peptide is functionalized with palmitoyl groups. In some embodiments, the peptide is functionalized with acetyl groups.

[0034] The liposome compositions described herein may contain various components encapsulated in 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 greater than 50 kDa.

[0035] Liposomes containing the peptide compositions described herein may be prepared using lecithin and other phospholipids. In some embodiments, liposomes are used to prepare one or more peptides. In some embodiments, the peptides are functionalized with acetyl groups. Lipid vesicle formation occurs when phospholipids such as lecithin are placed in water and, as a result, when sufficient energy is supplied, form one bilayer or a series of bilayers, each separated by water molecules. Liposomes can be prepared by sonicating phospholipids in water. Low shear rates produce multilayer liposomes. Continuous high-shear sonication tends to form smaller monolayer liposomes. Hydrophobic chemicals can be dissolved in the phospholipid bilayer membrane. The lipid bilayer of the liposome delivers the peptide compositions described herein.

[0036] The phospholipids used to prepare the liposome compositions described herein may have transition phase temperatures ranging from about 10°C to about 25°C. In some cases, the phospholipids have transition phase temperatures ranging from 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 above 40°C. In some cases, the phospholipids have transition phase temperatures ranging from 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.

[0037] Topical compositions may contain micelles dispersed in an aqueous solution, or aggregates of surfactant molecules. Micelles can be prepared by dispersing an oil solvent in an aqueous solution containing a surfactant, where the concentration of the surfactant exceeds the critical micelle concentration. The resulting composition contains micelles, i.e., spherical oil droplets.

[0038] Liposome compositions 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 concentration of the surfactant exceeds the critical micelle concentration. The resulting formulation contains micelles, i.e., spherical oil droplets surrounded by a film of polar surfactant molecules dispersed in an aqueous solvent.

[0039] In some embodiments, methods for preparing compositions comprising liposome-encapsulated peptides are described herein, comprising the steps of combining the peptides and a solvent to form a mixture, and contacting the mixture with an aqueous solution containing liposomes. In some cases, the contact occurs at a temperature of about 10°C to about 25°C. In some cases, the contact occurs at temperatures 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 above 40°C. In some cases, the contact occurs at temperatures 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.

[0040] A method for preparing a composition containing a peptide encapsulated in liposomes may involve 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).

[0041] The solvent can be used in various proportions. In some cases, the solvent is provided in amounts of 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%. The solvent may be propanediol, butylene glycol, or caprylyl glycol.

[0042] In some embodiments, the methods described herein include the steps of combining a peptide and a solvent to form a mixture, and contacting the mixture with an aqueous solution containing liposomes, wherein the aqueous solution contains a certain proportion of water and a certain proportion of liposomes. In some cases, the aqueous solution contains at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more than 90% water. In some cases, the aqueous solution contains water 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%. In some cases, the aqueous solution contains at least or about 20%, 30%, 40%, 50%, 60%, or more than 60% liposomes. In some cases, the aqueous solution contains 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 may be in the range of approximately 1:9 to approximately 3:7. In some cases, the ratio of liposomes to water may be at least approximately 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, or 1:2.

[0043] The methods for producing liposome compositions described herein may result in capture efficacy of 100% or less. In some cases, the capture efficacy is 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5% or less.

[0044] Liposome compositions are described herein, and peptides constitute a certain proportion of the composition. In some embodiments, peptides are provided in at least or about 0.0001%, 0.0005%, 0.00055%, 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% of the composition. In some embodiments, the peptide is provided in an amount of at least or 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 an amount ranging from about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.01% to about 5% by weight, or about 0.02% to about 2% by weight. In some embodiments, the peptide is provided in an amount of about 0.03% of the composition.

[0045] Liposome compositions are described herein, and liposomes constitute a certain proportion of the composition. In some embodiments, liposomes are provided in at least or 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, 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 in about 30%. In some embodiments, liposomes are provided in 27%.

[0046] The liposome compositions described herein, in some embodiments, consist of an average particle size of at most 220 nanometers (nm). In some cases, the average particle size is at most 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 approximately 100nm, 105nm, 110nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm, 150nm, 155nm, 160nm, 165nm, 170nm, 175nm, 180nm, 185nm, 190nm, 195nm, 200nm, 205nm, 210nm, 215nm, 220nm, 230nm, 240nm, 250nm, 260nm, 270nm, 280nm, 290nm, 300nm, 320nm, 340nm, 360nm, 380nm, or 400nm. In some cases, the average particle size is in the range of approximately 50 nm to 500 nm, approximately 100 nm to 400 nm, approximately 150 nm to 220 nm, approximately 180 nm to 220 nm, or approximately 190 nm to 210 nm.

[0047] In some cases, the liposome composition contains an activator having a molecular weight of about 600 daltons (Da) or less. In some cases, the activator has a molecular weight of at least 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 greater than 1000 daltons (Da). In some cases, the activator has a molecular weight of at least 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 activator 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 activator is a peptide. In some cases, the activator is a peptide encapsulated in liposomes.

[0048] The polydispersity index (PdI) of the liposome compositions described herein is in the range of 0 to about 0.2 in some embodiments. 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.

[0049] In some cases, the section of the liposome composition described herein is in the range of about 0.85 to about 0.95. In some cases, the section is amplitude. In some cases, the section is at least or about 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, or 0.95.

[0050] In some embodiments, the liposomes comprise propanediol, lecithin, or a combination thereof. In some embodiments, the propanediol is provided in at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, the 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 in an amount of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, lecithin is provided in amounts ranging from 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, the liposomes contain propanediol and lecithin.In some embodiments, propanediol and lecithin are provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, propanediol and lecithin are provided in amounts ranging from 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, propanediol and lecithin are provided in about 0.90% by weight.

[0051] This specification describes liposome compositions having improved distribution, efficacy, bioavailability, and / or activity. Liposome compositions may have improved distribution, efficacy, bioavailability, and / or activity compared to compositions without liposomes. In some cases, the distribution is improved by at least or about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.5, 5, or more than 5 times compared to compositions without liposomes. In some cases, the efficacy is improved by at least or about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.5, 5, or more than 5 times compared to compositions without liposomes. In some cases, bioavailability is improved by at least 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.5, 5, or more than 5 times compared to compositions without liposomes. In some cases, activity is improved by at least 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.5, 5, or more than 5 times compared to compositions without liposomes. Distribution, efficacy, bioavailability, and / or activity may be improved by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90% compared to compositions without liposomes.

[0052] The liposome compositions and methods described herein are, in some embodiments, topical compositions. In some cases, the liposome compositions are oil-free. In some cases, the liposome compositions are preservative-free. In some embodiments, the liposome formulations are aqueous formulations. In some embodiments, the liposome formulations are anhydrous formulations. In some embodiments, the liposome compositions have a pH in the range of about 5 to about 8. In some embodiments, the liposome compositions have a pH of at least or about 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0053] 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, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.5, 5, 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 depths 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.

[0054] Phosphatidylserine The compositions described herein, in some embodiments, contain phosphatidylserine. Exposure of phosphatidylserine from the inner cell membrane of erythrocytes may induce phagocytosis of erythrocytes. See Chang CF, Goods BA, Askenase MH, et al. Erythrocyte efferocytosis modulates macrophages towards recovery after intracerebral hemorrhage. The Journal of Clinical Investigation. 2018;128(2):607-624.

[0055] In some embodiments, phosphatidylserine is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, phosphatidylserine is provided in amounts ranging from about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, phosphatidylserine is provided in the range of about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.005% to about 0.1% by weight, about 0.01% to about 3% by weight, or about 0.02% to about 2% by weight. In some embodiments, phosphatidylserine is provided in the range of about 0.005% to about 0.02% by weight. In some embodiments, phosphatidylserine is provided in about 0.05% by weight. In some embodiments, phosphatidylserine is provided in about 0.25% by weight. In some embodiments, phosphatidylserine is provided in about 1% by weight. In some embodiments, phosphatidylserine is provided in amounts of at least or more than 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or more than 1000 micrograms (ug / mL) per milliliter. In some embodiments, phosphatidylserine is provided in amounts ranging from about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, about 100 to about 600, or about 100 to about 500 micrograms (ug / mL) per milliliter.

[0056] Lactoferrin In some embodiments, the compositions described herein include transferrin. In some embodiments, the transferrin is lactoferrin. In some embodiments, the lactoferrin is encapsulated in liposomes. Lactoferrin has wound-healing properties, promotes fibroblast proliferation, and increases hyaluronan secretion. See Saito S, Takayama Y, Mizumachi K, Suzuki C. Lactoferrin promotes hyaluronan synthesis in human dermal fibroblasts. Biotechnology Letters. 2011;33(1):33-39, and Takayama Y. Effects of Lactoferrin on Skin Wound Healing. In: Lactoferrin and its Role in Wound Healing. 2012:87-100.

[0057] In some embodiments, lactoferrin is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, lactoferrin is provided in amounts ranging from about 0.005% by weight to about 0.1% by weight, about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, lactoferrin is provided in amounts ranging from about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.01% to about 2.5% by weight, or about 0.02% to about 2% by weight. In some embodiments, lactoferrin is provided in about 0.025%. In some embodiments, lactoferrin is provided in about 0.05%. In some embodiments, lactoferrin is provided in about 0.10%. In some embodiments, lactoferrin is provided in at least or more than about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or 1000 micrograms (ug / mL) per milliliter. In some embodiments, lactoferrin is provided in the range of about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, or about 100 to about 500 micrograms (ug / mL) per milliliter.

[0058] Hydroxymethoxyphenyldecanone The compositions described herein, in some embodiments, contain hydroxymethoxyphenyldecanone. In some embodiments, hydroxymethoxyphenyldecanone is a potent endogenous hyaluronic acid enhancer, antioxidant, anti-irritant, or a combination thereof.

[0059] In some embodiments, hydroxymethoxyphenyldecanone is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, hydroxymethoxyphenyldecanone is provided in amounts ranging from about 0.25% by weight to about 10% by weight, about 0.1% by weight to about 2.5% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, hydroxymethoxyphenyldecanone is provided in amounts ranging from 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.

[0060] Tremella fuciformis The compositions described herein, in some embodiments, include Trellis fusiformis extract. In some embodiments, the Trellis fusiformis extract is derived from an edible mushroom. In some embodiments, the Trellis fusiformis extract provides moisture and antioxidant properties. In some embodiments, the Trellis fusiformis extract provides moisture from naturally occurring hyaluronic acid.

[0061] In some embodiments, Trellera fusiformis extract is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, Trellera fusiformis extract is provided in amounts ranging from about 0.25% by weight to about 10% by weight, about 0.1% by weight to about 2.5% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, Trellera fusiformis extract is provided in amounts ranging from 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, Trellera fusiformis extract is provided in amounts of at least or more than about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or 1000 micrograms (ug / mL) per milliliter. In some embodiments, Trellera fusiformis extract is provided in the range of about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, about 100 to about 600, or about 100 to about 500 micrograms (ug / mL) per milliliter.

[0062] Chemically cross-linked hyaluronic acid The compositions described herein include, in some embodiments, chemically crosslinked hyaluronic acid (e.g., Hylasome®). In some embodiments, the chemically crosslinked hyaluronic acid is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, chemically crosslinked hyaluronic acid is provided in the range of about 0.005% to about 0.1% by weight, about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, chemically crosslinked hyaluronic acid 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 2.5% by weight, or about 0.02% to about 2% by weight. In some embodiments, chemically crosslinked hyaluronic acid is provided in about 0.025%. In some embodiments, chemically crosslinked hyaluronic acid is provided in about 0.05%. In some embodiments, chemically crosslinked hyaluronic acid is provided in about 0.10%. In some embodiments, chemically crosslinked hyaluronic acid is provided in amounts of at least or more than 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or more than 1000 micrograms (ug / mL) per milliliter. In some embodiments, chemically crosslinked hyaluronic acid is provided in amounts ranging from about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, or about 100 to about 500 micrograms (ug / mL) per milliliter.

[0063] Other components The compositions described herein include SymDecanox® in some embodiments. In some embodiments, SymDecanox® is provided in at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, SymDecanox® is provided in the range of about 0.005% by weight to about 0.1% by weight, about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, SymDecanox® is offered in amounts ranging from about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.01% to about 2.5% by weight, or about 0.02% to about 2% by weight. In some embodiments, SymDecanox® is offered in amounts ranging from about 0.025%. In some embodiments, SymDecanox® is offered in amounts ranging from about 0.05%. In some embodiments, SymDecanox® is offered in amounts ranging from about 0.10%. In some embodiments, SymDecanox® is offered in concentrations of at least or more than 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or more than 1000 micrograms (ug / mL) per milliliter. In some embodiments, SymDecanox® is offered in concentrations ranging from about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, or about 100 to about 500 micrograms (ug / mL) per milliliter.

[0064] The compositions described herein include Aquaxyl® in some embodiments. In some embodiments, Aquaxyl® is provided in at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, Aquaxyl® is provided in the range of about 0.005% by weight to about 0.1% by weight, about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, Aquaxyl® is offered in amounts ranging from about 0.001% to about 6% by weight, about 0.002% to about 4% by weight, about 0.01% to about 2.5% by weight, or about 0.02% to about 2% by weight. In some embodiments, Aquaxyl® is offered in an amount of about 0.025%. In some embodiments, Aquaxyl® is offered in an amount of about 0.05%. In some embodiments, Aquaxyl® is offered in an amount of about 0.10%. In some embodiments, Aquaxyl® is offered in amounts of at least or more than 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or more than 1000 micrograms (ug / mL) per milliliter. In some embodiments, Aquaxyl® is offered in amounts ranging from about 5 to about 1000, about 10 to about 900, about 30 to about 800, about 50 to about 700, about 60 to about 600, or about 100 to about 500 micrograms (ug / mL) per milliliter.

[0065] Other components may include anti-inflammatory agents, antioxidants, and solubility enhancers. Exemplary anti-irritants include, but are not limited to, panthenyl triacetate and naringenin. Panthenyl triacetate and naringenin are natural plant extracts that reduce skin redness and water loss. Typical amounts of anti-irritants used in compositions are 1% to 4% by weight (wt.%).

[0066] Exemplary antioxidants include, but are not limited to, Dunaliella salina extract and squalane. Dunaliella salina extract contains components such as beta-carotene, which may exhibit antioxidant activity. Typical amounts of anti-inflammatory agents used in compositions are 0.1% to 2.5% by weight (wt.%). In some embodiments, Dunaliella salina extract is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight. In some embodiments, Dunaliella salina extract is provided in the range of about 0.001% to about 4.0%, about 0.01% to about 3.0%, about 0.1% to about 2.5%, or about 0.50% to about 1.5%. In some embodiments, squalane is provided in at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight. In some embodiments, squalane is provided in amounts ranging from about 0.001% to about 4.0%, about 0.01% to about 3.0%, about 0.1% to about 2.5%, or about 0.50% to about 1.5%. In some embodiments, Dunaliella salina extract and squalane are provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight. In some embodiments, Dunaliella salina extract and squalane are provided in concentrations ranging from about 0.001% to about 4.0%, about 0.01% to about 3.0%, about 0.1% to about 2.5%, or about 0.50% to about 1.5%.

[0067] In some embodiments, the composition comprises a siloxane polymer. In some embodiments, the siloxane polymer is caprylyl methicone. In some embodiments, caprylyl methicone is provided in at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4.0% by weight (wt.%). In some embodiments, caprylyl methicone is provided in about 0.5% by weight. In some embodiments, caprylylmethicone is provided in amounts ranging from about 0.001% to about 4.0% by weight, about 0.01% to about 3.0% by weight, about 0.1% to about 2.5% by weight, or about 0.50% to about 1.5% by weight. In some embodiments, caprylylmethicone is provided in about 0.25% by weight. In some embodiments, caprylylmethicone is provided in about 1% by weight.

[0068] Bentonite clay can be used in combination with peptides to impart penetration and adsorption properties to compositions and assist in emulsion stabilization. Other clays such as hectorite and magnesium aluminum silicate can also be used. Organically modified clay compounds can be obtained by modifying bentonite or other clays. Salts of fatty acids (e.g., hydrogenated fatty acids) (e.g., quaternary ammonium salts) can be reacted with hectorite or other clays. As provided herein, fatty acids are referred to and described using conventional nomenclature as used by those skilled in the art. Saturated fatty acids do not contain carbon-carbon double bonds. Unsaturated fatty acids contain at least one carbon-carbon double bond. Monounsaturated fatty acids contain only one carbon-carbon double bond. Polyunsaturated fatty acids contain two or more carbon-carbon double bonds. Double bonds in fatty acids are generally cis, but trans double bonds are also possible. The position of a double bond may be indicated by Δn, where n indicates the lower-numbered carbon in each pair of double-bond carbon atoms. Total carbon number: number of double bonds, Δ二重結合位置 A simplified notation can be used to specify the value. For example, 20:4Δ 5,8,11,14 This refers to a fatty acid having 20 carbon atoms and 4 double bonds, where the double bonds are located between carbon atoms 5 and 6, 8 and 9, 11 and 12, and 14 and 15, with carbon atom 1 being the carbon of the carboxylic acid group. Stearate (octadecanoate) is a saturated fatty acid. Oleate (cis-Δ9-octadecanoate) is a monounsaturated fatty acid, and linoleate (total cis-Δ9,12,15-octadecanoate) is a polyunsaturated fatty acid. Fatty acids suitable for use may contain 5 to 30 carbon atoms, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 carbon atoms. Fatty acids can be completely saturated or contain as many double bonds as feasible relative to their chain length. Suitable fatty acids for functionalizing hectorite or other clays include palmitic acid and stearic acid. Examples of dialkyl quaternary cationic modifiers include dipalmoyldimonium chloride and distearyldimonium chloride. Examples of amidoamine quaternary cationic modifiers include palmitamidopropyltrimonium chloride cetearyl alcohol and palmitamidopropyltrimonium chloride.

[0069] In some embodiments, the peptide may be mixed with a suitable carrier, diluent, or excipient, and depending on the route of administration and the desired preparation, may contain auxiliary substances such as wetting or emulsifying agents, pH buffers, gelling or viscosity-enhancing additives, preservatives, flavoring agents, and coloring agents. See, for example, “Remington: The Science and Practice of Pharmacy”, Lippincott Williams & Wilkins; 20th edition (June 1, 2003) and “Remington's Pharmaceutical Sciences”, Mack Pub. Co.; 18th and 19th editions (December 1985 and June 1990, respectively). Such preparations may include complexes, metal ions, polyacetic acid, polyglycolic acid, hydrogels, polymer compounds such as dextran, liposomes, microemulsions, micelles, monolayer or multilayer vesicles, erythrocyte ghosts, or spheroblasts. Suitable lipids for the composition include, but are not limited to, monoglycerides, diglycerides, sulfatides, lysolecithins, phospholipids, saponins, and bile acids. In some embodiments, the compositions described herein include phosphatidylserine, phospholipids, tocopherol, ascorbyl palmitate, or a combination thereof. In some embodiments, phosphatidylserine, phospholipids, tocopherol, ascorbyl palmitate, or a combination thereof is provided in amounts of 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, phosphatidylserine, phospholipids, tocopherol, ascorbyl palmitate, or combinations thereof are provided in amounts ranging from about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight.In some embodiments, phosphatidylserine, phospholipids, tocopherol, ascorbyl palmitate, or combinations thereof are provided in amounts ranging from 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 5% by weight. In some embodiments, the additive is betaine. Betaine is provided in some embodiments in amounts ranging from 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 5% by weight. In some embodiments, the compositions described herein contain caprylyl glycol. In some embodiments, caprylyl glycol is provided in amounts ranging from 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 5% by weight. In some embodiments, the compositions described herein include caprylhydroxamic acid. In some embodiments, caprylhydroxamic acid is provided in amounts ranging from 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 5% by weight. The presence of such additional components may affect the physical state, solubility, stability, release rate, clearance rate, and penetration of the active ingredient.

[0070] A topical composition comprises a peptide composition as described herein and a dermatologically acceptable vehicle. The vehicle may be aqueous or non-aqueous. The dermatologically acceptable vehicle used in the topical composition may be in the form of a lotion, gel, ointment, liquid, cream, or emulsion. If the vehicle is an emulsion, the emulsion may have a continuous aqueous phase and a discontinuous non-aqueous or oil phase (oil-in-water emulsion), or a continuous non-aqueous or oil phase and a discontinuous aqueous phase (water-in-oil emulsion). When administered topically in liquid or gel form, a liquid carrier such as water, petroleum, animal or plant-derived oils, e.g., peanut oil, mineral oil, soybean oil, or sesame oil, or synthetic oil may be added to the active ingredient. Saline solution, dextrose, or other sugar solutions, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol are also suitable liquid carriers. The pharmaceutical composition may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil; a mineral oil, such as liquid paraffin; or a mixture thereof. Suitable emulsifiers include naturally occurring gums, such as acacia gum and tragacanth gum; naturally occurring phospholipids, such as soy lecithin; esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate; and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain colorants and flavorings.

[0071] In certain embodiments, silicone elastomers (e.g., dimethicone crosspolymers) are used to increase the delivery and penetration of peptides into the skin. An alternative to increasing the molecular weight (like silicone gum) or adding fillers (like silicone compounds) is to partially crosslink a siloxane polymer and disperse this material in a suitable silicone-supported fluid. The resulting dimethicone crosspolymer (also known as silicone elastomer in the personal care industry) differs from basic polydimethylsiloxane (PDMS) due to the crosslinking between linear polymers. These materials can be used in peptide compositions and also offer advantages in scar treatment, peri-wound protection, and enzyme delivery. In skincare applications, the aesthetics of silicone elastomers (including those with functional groups) and their ability to absorb various oils (e.g., using dimethicone / vinyl dimethicone crosspolymers such as Dow Corning® 9506 Elastomer Powder) are two of the desirable properties of elastomers. Silicone elastomers have a texture distinct from any of the silicone fluids described as "smooth," "soft," and "dry." This can be altered by controlling the amount of liquid phase in the formulation, and therefore the degree of swelling. Due to their film-forming properties, dimethicone crosspolymers can be used as a delivery system for active ingredients such as peptides described herein, or other compositional components such as oil-soluble vitamins and sunscreens. Sunscreens such as octyl methoxycinnamate can be delivered more efficiently from compositions containing silicone elastomers, resulting in a higher sun protection index (SPF). Silicone elastomer blends can be used to enhance the SPF in oil-in-water compositions containing organic sunscreens. For example, in tests conducted with respect to SPF, the addition of a 4% silicone elastomer blend to a sun care composition containing an organic sunscreen increased the SPF from 5.7 to 18. This property of silicone elastomers makes it possible to maximize the effectiveness of sunscreens in a composition while reducing the amount required to achieve the desired SPF.As a result, the cost of the composition can be reduced along with the potential irritation caused by the sunscreen activator. Therefore, a higher SPF can be achieved with the same amount of UV absorber, and as a result, performance can be improved without adding to the cost of the composition. Silicone elastomers can be produced from linear silicone polymers by various crosslinking reactions, for example, by hydrosilylation reactions in which vinyl groups react with silicon hydride. A common process involves a linear silicone polymer having reaction sites along the polymer chain that react with the crosslinking agent. Dimethicone crosspolymers can be produced either as a gel made from a suspension of elastomer particles swollen in a carrier fluid (e.g., a mixture of high molecular weight silicone elastomers in cyclopentasiloxane such as Dow Corning® 9040 Silicone Elastomer Blend) or as a spray-dried powder (dimethicone / vinyl dimethicone crosspolymer such as Dow Corning® 9506 Elastomer Powder). The gel form with the desired attributes is cyclomethicone, but low viscosity dimethicone and organic fluids can also be used. Examples of dimethicone crosspolymers in suspension or gel form include high molecular weight silicone elastomers (12%) in decamethylcyclopentasiloxane (e.g., Dow Corning® ST-Elastomer 10) and mixtures of high molecular weight silicone elastomers in cyclopentasiloxane (e.g., Dow Corning® 9040 Silicone Elastomer Blend), which typically have an elastomer content ranging from 10 to 20% by weight.

[0072] Pharmaceutical excipients used in topical preparations of peptide compositions may be selected from the group consisting of solvents, softeners and / or emulsifiers, oil bases, preservatives, antioxidants, tension modifiers, penetration enhancers and solubilizers, chelating agents, buffers, surfactants, one or more polymers, and combinations thereof.

[0073] Suitable solvents for aqueous or hydrophilic liposome compositions include water; ethyl alcohol; isopropyl alcohol; mixtures of water and ethyl and / or isopropyl alcohol; glycerin; ethylene, propylene, or butylene glycol; DMSO; pentylene glycol; and mixtures thereof. In some embodiments, glycerin is provided in amounts of at least or about 1% by weight, 2% by weight, 3% by weight, 4% by weight, 5% by weight, 6% by weight, 7% by weight, 8% by weight, 9% by weight, 10% by weight, 11% by weight, 12% by weight, or more than 12% by weight (wt.%). In some embodiments, glycerin is provided in at least or about 7%. In some embodiments, glycerin is provided in amounts ranging from about 1% by weight to about 12% by weight, about 2% by weight to about 11% by weight, or about 3% by weight to about 10% by weight. In some embodiments, butylene glycol is provided in amounts of at least or about 0.0025% by weight, 0.005% by weight, 0.075% by weight, 0.01% by weight, 0.025% by weight, 0.05% by weight, 0.75% by weight, 1% by weight, 2% by weight, 3% by weight, 4% by weight, 5% by weight, 6% by weight, 7% by weight, 8% by weight, 9% by weight, 10% by weight, 11% by weight, 12% by weight, or more than 12% by weight. In some embodiments, butylene glycol is provided in amounts ranging from about 0.01% by weight to about 10% by weight, from about 0.025% by weight to about 5% by weight, or from about 0.05% by weight to about 1.25% by weight. In some embodiments, pentylene glycol is provided in amounts of at least or about 0.0025% by weight, 0.005% by weight, 0.075% by weight, 0.01% by weight, 0.025% by weight, 0.05% by weight, 0.75% by weight, 1% by weight, 2% by weight, 3% by weight, 4% by weight, 5% by weight, 6% by weight, 7% by weight, 8% by weight, 9% by weight, 10% by weight, 11% by weight, 12% by weight, or more than 12% by weight. In some embodiments, pentylene glycol is provided in amounts ranging from about 0.01% by weight to about 10% by weight, from about 0.025% by weight to about 5% by weight, or from about 0.05% by weight to about 1.25% by weight. Suitable solvents for hydrophobic compositions include mineral oils, vegetable oils, and silicone oils.If necessary, the peptide compositions described herein may be dissolved or dispersed in a hydrophobic oil phase, and the oil phase may then be emulsified in an aqueous phase containing water, either alone or in combination with lower alcohols, glycerin, and / or glycols. In some embodiments, anhydrous compositions are used because the presence of water may cause irritation when administered to skin tissue undergoing laser treatment, chemical exfoliation, skin excision, etc. Anhydrous compositions may also act to prevent the development of aqueous irritant contact dermatitis in damaged or sensitive skin, which can cause rashes and skin irritation and delay wound healing and improvement of skin quality. (Tsai, TF, Maibach, HI: How irritant is water? An overview. Contact Dermatitis 41(6)(1999):311-314 (Describes contact dermatitis caused by water as an irritant)). However, in certain embodiments, it may be acceptable to provide an aqueous composition or allow the presence of a limited amount of water. For example, water may be present, but in an amount below the threshold that may cause irritation when applied to damaged skin. Osmotic shock or osmotic stress is a sudden change in solute concentration around a cell, causing a rapid change in the movement of water across the cell membrane. Under conditions of high concentrations of salt, substrate, or any solute in the supernatant, water is drawn out of the cell by osmosis. This also inhibits the transport of substrates and cofactors into the cell, thus "shocking" the cell. Alternatively, at low concentrations of solute, a large amount of water enters the cell, causing the cell to swell, burst, or undergo apoptosis. Certain compositions described herein can be advantageously used when it is desirable to minimize osmotic shock.

[0074] The compositions described herein may contain varying amounts of solvent. In some embodiments, the solvent is water. In some embodiments, the solvent is at least or about 10% by weight, 15% by weight, 20% by weight, 25% by weight, 30% by weight, 35% by weight, 40% by weight, 45% by weight, 50% by weight, 55% by weight, 60% by weight, 65% by weight, 70% by weight, 75% by weight, 80% by weight, 85% by weight, 90% by weight, 95% by weight, or more than 95% by weight (wt.%). In some embodiments, the solvent is in the range of about 10% by weight to about 95% by weight, about 20% by weight to about 90% by weight, about 30% by weight to about 85% by weight, about 40% by weight to about 80% by weight, or about 50% by weight to about 75% by weight.

[0075] The viscosity of the composition can be maintained at a selected level using a pharmaceutically acceptable thickener. Suitable viscosity improvers or thickeners that can be used to prepare viscous gels or creams using aqueous bases include sodium polyacrylate, xanthan gum, polyvinylpyrrolidone, acrylic polymers, carrageenan, hydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, ethylcellulose, propylcellulose, hydroxypropylmethylcellulose, polyethoxylated polyacrylamide, polyethoxylated acrylates, and polyethoxylated alkanethiols. Methylcellulose is preferred because it is readily and economically available and easy to handle. Other suitable thickeners include, for example, xanthan gum, carboxymethylcellulose, hydroxypropylcellulose, and carbomer. The preferred concentration of the thickener depends on the selected thickener. The amount used to achieve the desired viscosity is preferably used. Viscous compositions are usually prepared from solution by adding such thickeners or by using a base having an acceptable level of viscosity.

[0076] The viscosity of the compositions described herein is in the range of about 8,000 centipoise (cps) to about 30,000 cps in some embodiments. In some embodiments, the viscosity is at least or about 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000, 11,000, 12,000, 13,000, 14,000, 15,000, 16,000, 17,000, 18,000, 19,000, 20,000, 21,000, 22,000, 23,000, 24,000, 25,000, 26,000, 27,000, 28,000, 29,000, 30,000, 31,000, 32,000, 33,000, 34,000, 35,000, 36,000, 37,000, 38,000, 39,000, 40,000, or greater than 40,000 cps. In some embodiments, the composition has a viscosity in the range of about 4,000 to about 40,000, about 6,000 to about 38,000, about 8,000 to about 36,000, about 10,000 to about 34,000 cps, about 12,000 to about 32,000 cps, or about 14,000 to about 30,000 cps.

[0077] Suitable softeners include hydrocarbon oils and waxes, such as mineral oil, petrolatum, paraffin, ceresin, ozokerite, microcrystalline wax, polyethylene, squalene, perhydrosqualene, silicone oil, triglyceride esters, acetoglyceride esters, such as acetylated monoglycerides, ethoxylated glycerides, such as ethoxylated glyceryl monostearate, and alkyl esters of fatty acids or dicarboxylic acids. In some embodiments, the softener is tri(caprylic / capric acid)glyceryl.

[0078] In some embodiments, the softener is provided in an amount of at least or about 0.0025% by weight, 0.005% by weight, 0.075% by weight, 0.01% by weight, 0.025% by weight, 0.05% by weight, 0.75% by weight, 1% by weight, 2% by weight, 3% by weight, 4% by weight, 5% by weight, 6% by weight, 7% by weight, 8% by weight, 9% by weight, 10% by weight, 11% by weight, 12% by weight, or more than 12% by weight. In some embodiments, the softener is provided in an amount ranging from about 0.01% by weight to about 10% by weight, from about 0.01% by weight to about 2.5% by weight, from about 0.025% by weight to about 5% by weight, or from about 0.05% by weight to about 1.25% by weight. In some embodiments, caprylic / capric acid triglyceride is provided in amounts of at least or about 0.0025% by weight, 0.005% by weight, 0.075% by weight, 0.01% by weight, 0.025% by weight, 0.05% by weight, 0.75% by weight, 1% by weight, 2% by weight, 3% by weight, 4% by weight, 5% by weight, 6% by weight, 7% by weight, 8% by weight, 9% by weight, 10% by weight, 11% by weight, 12% by weight, or more than 12% by weight. In some embodiments, caprylic / capric acid triglyceride is provided in amounts ranging from about 0.01% by weight to about 10% by weight, from about 0.01% by weight to about 2.5% by weight, from about 0.025% by weight to about 5% by weight, or from about 0.05% by weight to about 1.25% by weight.

[0079] Suitable silicone oils for use as softeners include dimethylpolysiloxane, methyl(phenyl)polysiloxane, and water-soluble and alcohol-soluble silicone glycol copolymers. Suitable triglyceride esters for use as softeners include vegetable and animal fats and oils, including castor oil, safflower oil, cottonseed oil, corn oil, olive oil, liver oil, almond oil, avocado oil, coconut oil, sesame oil, and soybean oil.

[0080] Suitable carboxylic acid or diacid esters for use as softening agents include methyl, isopropyl, and butyl esters of fatty acids. Specific examples of alkyl esters include hexyl laurate, isohexyl laurate, isohexyl palmitate, isopropyl palmitate, decyl oleate, isodecyl oleate, hexadecyl stearate, decyl stearate, isopropyl isostearate, dilauryl lactate, myristyl lactate, and cetyl lactate, as well as alkenyl esters of fatty acids, such as oleyl myristate, oleyl stearate, and oleyl oleate. Specific examples of alkyl esters of diacids include diisopropyl adipate, diisohexyl adipate, bis(hexyldecyl) adipate, and diisopropyl sebacate.

[0081] Other suitable classes of softeners or emulsifiers that can be used in the composition include fatty acids, fatty alcohols, fatty alcohol ethers, ethoxylated fatty alcohols, fatty acid esters of ethoxylated fatty alcohols, and waxes.

[0082] Specific examples of fatty acids for use as emollients include pelargonic acid, lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, hydroxystearic acid, oleic acid, linoleic acid, ricinoleic acid, arachidic acid, behenic acid, and erucic acid. Specific examples of fatty alcohols for use as emollients include lauryl, myristyl, cetyl, hexadecyl, stearyl, isostearyl, hydroxystearyl, oleyl, ricinoleyl, behenyl, and erucyl alcohol, as well as 2-octyldodecanol.

[0083] Specific examples of waxes suitable for use as softening agents include lanolin oil, lanolin wax, lanolin alcohol, lanolin fatty acids, isopropyl lanolate, ethoxylated lanolin, ethoxylated lanolin alcohol, ethoxylated cholesterol, propoxylated lanolin alcohol, acetylated lanolin, acetylated lanolin alcohol, lanolin alcohol linoleate, lanolin alcohol resinolate, lanolin alcohol resinolate acetate, lanolin alcohol resinolate acetate, ethoxylated alcohol ester acetate, hydrogenolysates of lanolin, hydrogenated lanolin, ethoxylated hydrogenated lanolin, ethoxylated sorbitol lanolin, and lanolin and its derivatives, including liquid and semi-solid lanolin. Furthermore, hydrocarbon waxes, ester waxes, and amide waxes can also be used as waxes. Useful waxes include wax esters such as beeswax, whale wax, myristyl myristate, and stearyl stearate, as well as beeswax derivatives such as polyoxyethylene sorbitol beeswax, and vegetable waxes including carnauba and candela wax.

[0084] Polyhydric alcohols and polyether derivatives can be used as solvents and / or surfactants in compositions. Suitable polyhydric alcohols and polyethers include propylene glycol, dipropylene glycol, polypropylene glycol 2000 and 4000, poly(oxyethylene-co-oxypropylene) glycol, glycerol, sorbitol, ethoxylated sorbitol, hydroxypropyl sorbitol, polyethylene glycol 200-6000, methoxypolyethylene glycol 350, 550, 750, 2000 and 5000, poly[ethylene oxide] homopolymer (100,000-5,000,000), polyalkylene glycols and derivatives, hexylene glycol, 2-methyl-2,4-pentanediol, 1,3-butylene glycol, 1,2,6-hexanetriol, 2-ethyl-1,3-hexanediol, adjacent glycols having 15-18 carbon atoms, and polyoxypropylene derivatives of trimethylpropane.

[0085] Polyhydric alcohol esters may be used as emulsifiers or softeners. Suitable polyhydric alcohol esters include ethylene glycol mono and di fatty acid esters, diethylene glycol mono and di fatty acid esters, polyethylene glycol (200-6000) mono and di fatty acid esters, propylene glycol mono and di fatty acid esters, polypropylene glycol 2000 monooleate, polypropylene glycol 2000 monostearate, ethoxylated propylene glycol monostearate, glyceryl mono and di fatty acid esters, polyglycerol poly fatty acid esters, ethoxylated glyceryl monostearate, 1,3-butylene glycol monostearate, 1,3-butylene glycol distearate, polyoxyethylene polyol fatty acid esters, sorbitan fatty acid esters, and polyoxyethylene sorbitan fatty acid esters.

[0086] Suitable emulsifiers for use in compositions include anionic, cationic, nonionic, and zwitterionic surfactants. Preferred ionic emulsifiers include phospholipids such as lecithin and its derivatives.

[0087] For example, asterols containing cholesterol and cholesterol fatty acid esters, fatty acid amides, ethoxylated fatty acid amides, and fatty acid alkanolamides can also be used as blunting agents and / or penetration enhancers.

[0088] The shelf life of a composition can be increased by using pharmaceutically acceptable preservatives. Other suitable preservatives and / or antioxidants for use in a composition include, but can be used, benzalkonium chloride, benzyl alcohol, phenol, urea, parabens, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), tocopherol, thimerosal, chlorobutanol, and mixtures thereof. When preservatives such as antioxidants are used, the concentration is typically about 0.02% to about 2% based on the total weight of the composition, but a larger or smaller amount may be desirable depending on the selected agent. The reducing agents described herein can be advantageously used to maintain a good shelf life of the composition. In general, since the anhydrous compositions of the embodiments are observed to exhibit sufficient stability, preservatives can be omitted from the composition.

[0089] Suitable chelating agents for use in compositions include ethylenediaminetetraacetic acid, its alkali metal salts, its alkaline earth metal salts, its ammonium salts, and its tetraalkylammonium salts. In some embodiments, the chelating agent is disodium ethylenediaminetetraacetic acid (EDTA). In some embodiments, disodium EDTA is provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, or more than 4% by weight (wt.%). In some embodiments, disodium EDTA is provided in the range of about 0.25% to about 10% by weight, about 0.1% to about 2.5% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, disodium EDTA 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.

[0090] The carrier preferably has a pH of about 4.0 to 10.0, more preferably about 4.8 to 7.8, and more preferably about 5.0 to 6.5. The pH can be controlled using a buffer or other pH adjuster. Suitable pH adjusters include phosphoric acid and / or phosphates, citrate and / or citrates, hydroxide salts (i.e., calcium hydroxide, sodium hydroxide, potassium hydroxide), and amines such as triethanolamine. Suitable buffers include a buffer containing a solution of monopotassium phosphate and dipotassium phosphate maintaining a pH of 5.8 to 8, and a buffer containing a solution of monosodium phosphate and disodium phosphate maintaining a pH of 6 to 7.5. Other buffers include citrate / sodium citrate and dibasic sodium phosphate / citric acid. The peptide composition of the embodiment is preferably isotonic with the recipient's blood or other body fluids. The isotonicity of the composition can be achieved using sodium tartrate, propylene glycol, or other inorganic or organic solutes. Sodium chloride is particularly preferred. Buffering agents, such as acetic acid and its salts, citric acid and its salts, boric acid and its salts, and phosphoric acid and its salts, can be used. It may be desirable to include reducing agents in the composition, such as vitamin C, vitamin E, or other reducing agents known in the pharmaceutical field.

[0091] Surfactants, such as anionic detergents, such as sodium lauryl sulfate, sodium dioctyl sulfosuccinate, and sodium dioctyl sulfonate; cationic detergents, such as benzalkonium chloride or benzethonium chloride; or nonionic detergents, such as polyoxyethylene hydrogenated castor oil, glycerol monostearate, polysorbate, sucrose fatty acid ester, methylcellulose, or carboxymethylcellulose, can be used as excipients.

[0092] In certain embodiments, it may be advantageous to include additional agents having pharmacological activity. Examples of antiinfective agents include anthelmintics (mebendazole), aminoglycoside antibiotics (gentamicin, neomycin, tobramycin), antifungal antibiotics (amphotericin b, fluconazole, griseofulvin, itraconazole, ketoconazole, nistatin, mikatin, tolnaftate), cephalosporins (cefaclor, cefazolin, cefotaxime, ceftazidime, ceftriaxone, cefuroxime, cephalexin), beta-lactam antibiotics (cefotetan, meropenem), chloramphenicol, macrolides (azithromycin, clarithromycin, erythromycin), penicillins (penicillin G sodium salt, amoxiline, ampicillin, dicloxacillin, nafcillin, piperacillin, etc.). Examples of antimicrobial agents include, but are not limited to, calcilin, tetracyclines (doxycycline, minocycline, tetracycline), bacitracin, clindamycin, colistimethate sodium, polymyxin B sulfate, vancomycin, antimicrobial agents (including acyclovir, amantadine, didanosine, efavirenz, foscarnet, ganciclovir, indinavir, lamivudine, nelfinavir, ritonavir, saquinavir, stabudine, valacyclovir, valganciclovir, and zidovudine), quinolones (ciprofloxacin, levofloxacin), sulfonamides (sulfadiazine, sulfisoxazole), sulfones (dapsone), furazolidone, metronidazole, pentamidine, sulfanilamidum crystallinum, gatifloxacin, and sulfamethoxazole / trimethoprim.Examples of anesthetics include, but are not limited to, ethanol, bupivacaine, chloroprocaine, levobupivacaine, lidocaine, mepivacaine, procaine, ropivacaine, tetracaine, desflurane, isoflurane, ketamine, propofol, sevoflurane, codeine, fentanyl, hydromorphone, marcaine, meperidine, methadone, morphine, oxycodone, remifentanil, sufentanil, butorphanol, nalbufine, tramadol, benzocaine, dibucaine, ethyl chloride, xylocaine, and phenazopyridine. Anti-inflammatory agents include non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin, celecoxib, choline magnesium trisalicylate, diclofenac potassium, diclofenac sodium, diflunisal, etodolac, fenoprofen, fluviprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, melenamic acid, nabumetone, naproxen, naproxen sodium, oxaprozin, piroxicam, rofecoxib, sarsalate, sulindac, and tolmetin, as well as corticosteroids. Examples include, but are not limited to, cortisone, hydrocortisone, methylprednisolone, prednisone, prednisolone, betamethasone, beclomethasone dipropionate, budesonide, dexamethasone sodium phosphate, flunisolide, fluticasone propionate, triamcinolone acetonide, betamethasone, fluocinonide, betamethasone dipropionate, betamethasone valerate, desonide, desoxymethasone, fluocinolone, triamcinolone, clobetasol propionate, and dexamethasone.

[0093] In certain embodiments, the addition of softeners, emulsion stabilizers, humectants, excipients, and other compounds may be modified to enhance the sensory properties of the topical composition, including but not limited to skin feel (silky softness, lightness, creaminess, etc.), absorbency (the time required for the product to lose its moistness and become undetectable on the skin), consistency, hardness, diffusivity (e.g., viscosity, flow onset, shear rate), tackiness, shape integrity, gloss, hydrophilicity or hydrophobicity, and others. Preferably, the composition has high diffusivity and low viscosity properties. Compositions possessing such properties have been demonstrated to have an enhanced "silky" or "light" skin feel evaluation (see, for example, Bekker, M., Webber, G., Louw, N., Relating rheological measurements to primary and secondary skin feeling when mineral-based and Fischer-Tropsch wax-based cosmetic emulsions and jellies are applied to the skin, International Journal of Cosmetic Science 2013, 35(4), pp.354-61).

[0094] In some embodiments, the composition comprises phenoxyethanol, ethylhexylglycerin, or a combination thereof. In some embodiments, phenoxyethanol is provided in at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, phenoxyethanol is provided in the range of about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, ethylhexylglycerin is provided in amounts of at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, ethylhexylglycerin is provided in amounts ranging from about 0.25% by weight to about 10% by weight, from about 0.5% by weight to about 8% by weight, from about 0.75% by weight to about 6% by weight, or from about 1% by weight to about 4% by weight. In some embodiments, phenoxyethanol and ethylhexylglycerin are provided in amounts of at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, phenoxyethanol and ethylhexylglycerin are provided in amounts ranging from about 0.25% by weight to about 10% by weight, about 0.1% by weight to about 4% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight.

[0095] In some embodiments, the composition comprises polyacrylate-13, polyisobutene, polysorbate 20, or a combination thereof. In some embodiments, polyacrylate-13 is provided in at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, polyacrylate-13 is provided in amounts ranging from about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, polyisobutene is provided in amounts of at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, polyisobutene is provided in amounts ranging from about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, polyacrylate-13 is provided in amounts ranging from about 0.25% to about 10% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight. In some embodiments, the polysorbate 20 is provided in an amount of at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%).In some embodiments, polysorbate 20 is provided in amounts ranging from about 0.25% by weight to about 10% by weight, about 0.5% by weight to about 8% by weight, about 0.75% by weight to about 6% by weight, or about 1% by weight to about 4% by weight. In some embodiments, polyacrylate-13, polyisobutene, and polysorbate 20 are provided in amounts of at least or about 0.05% by weight, 0.10% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, polyacrylate-13, polyisobutene, and polysorbate 20 are provided in amounts ranging from about 0.25% to about 10% by weight, about 0.1% to about 4% by weight, about 0.5% to about 8% by weight, about 0.75% to about 6% by weight, or about 1% to about 4% by weight (wt.%).

[0096] In some embodiments, the compositions described herein contain potassium sorbate. In some embodiments, potassium sorbate is provided in amounts of at least or about 0.001% by weight, 0.00175% by weight, 0.0025% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, potassium sorbate is provided in amounts ranging from 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.

[0097] In some embodiments, the liposomes comprise propanediol, lecithin, or a combination thereof. In some embodiments, the propanediol is provided in at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, the 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 in an amount of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, lecithin is provided in amounts ranging from 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, the liposomes contain propanediol and lecithin.In some embodiments, propanediol and lecithin are provided in amounts of at least or about 0.001% by weight, 0.005% by weight, 0.01% by weight, 0.02% by weight, 0.05% by weight, 0.10% by weight, 0.20% by weight, 0.25% by weight, 0.50% by weight, 0.75% by weight, 1.0% by weight, 1.5% by weight, 2.0% by weight, 2.5% by weight, 3.0% by weight, 3.5% by weight, 4.0% by weight, 4.5% by weight, 5.0% by weight, 5.5% by weight, 6.0% by weight, 6.5% by weight, 7.0% by weight, 8% by weight, 9% by weight, 10% by weight, or more than 10% by weight (wt.%). In some embodiments, propanediol and lecithin are provided in amounts ranging from 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, propanediol and lecithin are provided in about 0.90% by weight.

[0098] Topical compositions may contain micelles dispersed in an aqueous solution, or aggregates of surfactant molecules. Micelles can be prepared by dispersing an oil solvent in an aqueous solution containing a surfactant, where the concentration of the surfactant exceeds the critical micelle concentration. The resulting composition contains micelles, i.e., spherical oil droplets.

[0099] Penetration enhancer Fatty acids and alcohols are used to enhance the penetration of peptides, and the composition includes, for example, methane acid, ethane acid, propanoic acid, butanoic acid, isobutyric acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, myristoleic acid, isovaleric acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoleidic acid, α-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, medium-chain fatty acids, for example, C 6~12 It can provide fatty acids and other substances. The typical amount used in a composition is 1% to 4% by weight.

[0100] Antibacterial effectiveness In some embodiments, the compositions described herein are chemically and physically stable at physiological pH. In some embodiments, the compositions are sterile and safe for human administration. In some embodiments, the compositions comply with or pass necessary antimicrobial efficacy tests, such as the Antimicrobial Effectiveness Test. In some embodiments, the compositions result in the complete or substantially complete eradication of bacteria, yeasts, fungi, or combinations thereof.

[0101] How to use This specification describes compositions and methods for stimulating hyaluronic acid (HA). In some embodiments, the compositions and methods stimulate HA, thereby improving skin moisture and maintaining and improving skin elasticity and firmness. In some embodiments, the compositions and methods described herein promote or improve skin hydration.

[0102] The compositions and methods described herein, in some embodiments, improve aged skin or promote the recovery of aged skin. In some embodiments, the compositions and methods described herein improve the elasticity of aged skin. In some cases, the compositions and methods described herein improve the appearance of aged skin. In some cases, the compositions and methods improve the appearance of aged skin by promoting or improving skin hydration. In some embodiments, the compositions and methods improve the appearance of age spots. In some cases, the compositions and methods improve the appearance of white pseudoscars. In some cases, the compositions and methods improve the appearance of uneven skin tone. In some cases, the compositions and methods improve the appearance of wrinkles.

[0103] In some embodiments, the compositions and methods described herein improve the appearance of the skin before, during, and after the injection of the filler. In some embodiments, the compositions and methods described herein improve skin hydration before, during, and after the injection of the filler. In some embodiments, the filler is a soft tissue filler product. For example, the soft tissue filler is an injectable skin or subcutaneous filler. In some embodiments, the filler is a breast augmentation or breast reconstruction filler, a lip filler, or a filler suitable for the restoration or enhancement of other soft tissues. In some embodiments, the filler is a dermal filler. In some cases, the dermal filler is administered by injection into or under the skin of the target.

[0104] In some embodiments, the compositions and methods described herein improve the appearance of the skin by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more than 95%. In some embodiments, the compositions and methods described herein improve the appearance of the skin by at least or about 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10, or more than 10 times.

[0105] The compositions and methods described herein simulate endogenous hyaluronic acid (HA) production in some embodiments. The compositions and methods described herein can improve the penetration of high molecular weight HA into the dermis of the skin. In some embodiments, compositions comprising hydroxymethoxyphenyldecanone, Trellera fusiformis extract, lactoferrin, sodium hyaluronate crosspolymer, phosphatidylserine, or a combination thereof stimulate endogenous hyaluronic acid production.

[0106] In some embodiments, the compositions described herein stimulate endogenous hyaluronic acid production by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more than 95%. In some embodiments, the compositions described herein stimulate endogenous hyaluronic acid production by at least or about 0.5 times, 1.0 times, 1.5 times, 2.0 times, 2.5 times, 3.0 times, 3.5 times, 4.0 times, 5.0 times, 6.0 times, 7.0 times, 8.0 times, 9.0 times, 10 times, or more than 10 times.

[0107] The compositions described herein may be used in conjunction with various skin regimens. In some cases, the topical compositions described herein are administered once daily, twice daily, or three or more times daily. In some cases, the topical compositions described herein are administered twice daily. In some embodiments, the topical compositions described herein are administered daily, every day, every other day, five days a week, once a week, every other week, two weeks a month, three weeks a month, once a month, twice a month, or three or more times a month. In some embodiments, the topical compositions described herein are administered twice daily, for example, in the morning and evening. In some embodiments, the topical compositions described herein are administered for at least one day, two days, three days, four days, five days, six days, seven months, eight months, nine months, ten months, eleven months, twelve months, eighteen months, two years, three years, four years, five years, or more than ten years. In some embodiments, the topical compositions described herein are administered twice daily for at least one week, two weeks, three weeks, one month, two months, three months, four months, five months, or six months or longer. In some embodiments, the topical compositions described herein are administered once daily, twice daily, three times daily, four times daily, or more than four times daily for at least one week, two weeks, three weeks, one month, two months, three months, four months, five months, or six months or longer.

[0108] Stability testing Stability testing of the composition can be carried out as follows:

[0109] 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 a product is stored at 45°C for 3 months (and exhibits acceptable stability), it should be stable at room temperature for 2 years. A good control temperature is 4°C (39°F), and most products exhibit excellent stability at this temperature. In some cases, products are also subjected to -10°C (14°F) for 3 months.

[0110] In some cases, product stability is evaluated by undergoing a three-cycle temperature test from -10°C (14°F) to 25°C (77°F). In such cases, the product is placed at -10°C for 24 hours, and then at room temperature (25°C) for 24 hours. This completes one cycle. A more rigorous test is a five-cycle test from -10°C to 45°C. This places significant stress on the emulsion.

[0111] The dispersed phase (of an oil-in-water emulsion) tends to 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 that emulsion instability is imminent. A test method to predict creaming is centrifugation. The emulsion is heated to 50°C (122°F) and centrifuged at 3000 rpm for 30 minutes. The resulting product is then inspected for signs of creaming.

[0112] Both the formulation and the packaging may be sensitive to ultraviolet light. Place the product in a glass and the actual packaging in a lightbox with broad-spectrum output. Another glass bottle completely covered with aluminum foil will serve as a control. Discoloration of the product may be observed.

[0113] For all of the above tests, color, odor / fragrance, viscosity, pH value, and, if available, particle size uniformity and / or particle aggregation can be observed under a microscope.

[0114] Kit for non-invasive use and use with invasive procedures Some embodiments of the methods and compositions provided herein include kits comprising hyaluronic acid-stimulating peptides and agents. In some embodiments, the kit may be provided to an administering physician, other healthcare professional, patient, or caregiver. In some embodiments, the kit includes a container containing the composition in a suitable topical composition and instructions for administering the composition to the target. The kit may optionally include one or more additional therapeutic or other agents. For example, a kit containing a peptide composition in topical form may be provided together with other skincare agents such as cleansers, occlusive moisturizers, penetrating moisturizers, sunscreens, and sunscreens. The kit may contain the peptide composition in bulk form or the peptide composition in divided doses for continuous or sequential administration. The kit may optionally include one or more diagnostic tools, administration tools, and / or instructions for use. The kit may include a suitable delivery device such as a syringe, pump dispenser, or single-dose packet, along with instructions for administering the peptide composition and any other therapeutic or beneficial agent. The kit may optionally include instructions for the storage, reconstitution (if applicable), and administration of any or all of the therapeutic or beneficial drugs contained. The kit may include multiple containers that reflect the number of doses to be administered to the subject or the different products to be administered to the subject.

[0115] In some embodiments, the composition is configured to support the skin before, during, and after cosmetic procedures such as filler injections, and to work in conjunction with the skin's own natural regeneration process to help improve skin appearance and reduce skin tightness. Topical compositions can be applied immediately after the procedure for faster recovery or for skin that generally looks healthier. The composition can increase hyaluronic acid and promote skin hydration. Topical compositions are suitable for all skin types and post-operative skin. Topical compositions may be provided to the patient in bulk form to allow the patient to self-administer a suitable amount of peptide. For example, the patient may apply a sufficient amount of the composition to evenly cover the affected area, or as separately instructed by the physician. In certain embodiments, it may be desirable to incorporate additional therapeutic or activating agents into the topical composition. Alternatively, adjunctive therapies or agents may be administered separately. For example, cleansers, sunscreens, sunscreens, penetrating moisturizers, and / or occlusive moisturizers may be provided for administration before or after the topical composition of the embodiment. The kit may include a topical peptide composition, an occlusive moisturizer, a mild cleanser, a penetrating moisturizer, and a broad-spectrum SPF30+ sunscreen.

[0116] 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 activators that act synergistically on the skin to promote wound healing or wound closure, or to treat chronic skin wounds.

[0117] Numbered Embodiments Numbered Embodiment 1 comprises a topical composition for stimulating hyaluronic acid, comprising a synthetic tripeptide, an octapeptide, and a hexapeptide, wherein the topical composition stimulates hyaluronic acid. Numbered Embodiment 2 comprises the topical composition according to Numbered Embodiment 1, wherein the synthetic tripeptide comprises tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate. Numbered Embodiment 3 comprises the topical composition according to Numbered Embodiments 1-2, wherein the octapeptide is encapsulated in liposomes. Numbered Embodiment 4 comprises the topical composition according to Numbered Embodiments 1-3, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). Numbered Embodiment 5 includes the topical composition described in Numbered Embodiments 1 to 4, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1). Numbered Embodiment 6 includes the topical composition described in Numbered Embodiments 1 to 5, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2). Numbered Embodiment 7 includes the topical composition described in Numbered Embodiments 1 to 6, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3). Numbered Embodiment 8 includes the topical composition described in Numbered Embodiments 1 to 7, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4). Numbered Embodiment 9 includes the topical composition described in Numbered Embodiments 1 to 8, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). Numbered Embodiment 10 includes the topical composition described in Numbered Embodiments 1 to 9, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6). Numbered Embodiment 11 comprises a topical composition according to Numbered Embodiments 1 to 10, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7).Numbered Embodiment 12 comprises the topical composition described in Numbered Embodiments 1 to 11, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8). Numbered Embodiment 13 comprises the topical composition described in Numbered Embodiments 1 to 12, wherein the hexapeptide is hexapeptide-11. Numbered Embodiment 14 comprises the topical composition described in Numbered Embodiments 1 to 13, wherein the hexapeptide-11 is encapsulated in liposomes. Numbered Embodiment 15 comprises the topical composition described in Numbered Embodiments 1 to 14, further comprising lactoferrin. Numbered Embodiment 16 comprises the topical composition described in Numbered Embodiments 1 to 15, wherein the lactoferrin is encapsulated in liposomes. Numbered Embodiment 17 comprises the topical composition described in Numbered Embodiments 1 to 16, further comprising phosphatidylserine. Numbered Embodiment 18 comprises the topical composition described in Numbered Embodiments 1 to 17, further comprising Trellis fusiformis extract. Numbered Embodiment 19 comprises the topical composition described in Numbered Embodiments 1 to 18, further comprising sodium hyaluronate crosspolymer. Numbered Embodiment 20 comprises the topical composition described in Numbered Embodiments 1 to 19, further comprising hydroxymethoxyphenyldecanone. Numbered Embodiment 21 comprises the topical composition described in Numbered Embodiments 1 to 20, which is aqueous. Numbered Embodiment 22 comprises the topical composition described in Numbered Embodiments 1 to 21, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2) and the hexapeptide is hexapeptide-11. Numbered Embodiment 23 comprises the topical composition described in Numbered Embodiments 1 to 22, further comprising lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyldecanone, or a combination thereof. Numbered Embodiment 24 comprises the topical composition described in Numbered Embodiments 1 to 23, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3) and the hexapeptide is hexapeptide-11.Numbered Embodiment 25 comprises the topical composition described in Numbered Embodiments 1 to 24, further comprising lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 26 comprises the topical composition described in Numbered Embodiments 1 to 25, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4) and the hexapeptide is hexapeptide-11. Numbered Embodiment 27 comprises the topical composition described in Numbered Embodiments 1 to 26, further comprising lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 28 comprises the topical composition described in Numbered Embodiments 1 to 27, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5) and the hexapeptide is hexapeptide-11. Numbered Embodiment 29 comprises the topical composition described in Numbered Embodiments 1 to 28, further comprising lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 30 comprises the topical composition described in Numbered Embodiments 1 to 29, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6) and the hexapeptide is hexapeptide-11. Numbered Embodiment 31 comprises the topical composition described in Numbered Embodiments 1 to 30, further comprising lactoferrin, phosphatidylserine, Trellera fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 32 comprises the topical composition described in Numbered Embodiments 1 to 31, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7) and the hexapeptide is hexapeptide-11.Numbered Embodiment 33 comprises a topical composition according to Numbered Embodiments 1 to 32, further comprising lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 34 comprises a topical composition according to Numbered Embodiments 1 to 33, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8) and the hexapeptide is hexapeptide-11. Numbered Embodiment 35 comprises a topical composition according to Numbered Embodiments 1 to 34, further comprising lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 36 comprises a method for stimulating hyaluronic acid production, comprising the step of administering a topical composition comprising a synthetic tripeptide, an octapeptide, and a hexapeptide. Numbered Embodiment 37 comprises the method according to any one of Numbered Embodiments 1 to 36, wherein the synthetic tripeptide comprises tetradecyl-diaminobutyroylvalyldiaminobutyric acid urea trifluoroacetate. Numbered Embodiment 38 comprises the method according to any one of Numbered Embodiments 1 to 37, wherein the octapeptide is encapsulated in liposomes. Numbered Embodiment 39 comprises the method according to any one of Numbered Embodiments 1 to 38, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8). Numbered Embodiment 40 comprises the method according to any one of Numbered Embodiments 1 to 39, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1). Numbered Embodiment 41 comprises the method according to any one of Numbered Embodiments 1 to 40, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2).Numbered Embodiment 42 includes the method according to any one of Numbered Embodiments 1 to 41, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3). Numbered Embodiment 43 includes the method according to any one of Numbered Embodiments 1 to 42, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4). Numbered Embodiment 44 includes the method according to any one of Numbered Embodiments 1 to 43, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). Numbered Embodiment 45 includes the method according to any one of Numbered Embodiments 1 to 44, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6). Numbered Embodiment 46 includes the method according to any one of Numbered Embodiments 1 to 45, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7). Numbered Embodiment 47 includes the method according to any one of Numbered Embodiments 1 to 46, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8). Numbered Embodiment 48 includes the method according to any one of Numbered Embodiments 1 to 47, wherein the hexapeptide is hexapeptide-11. Numbered Embodiment 49 includes the method according to any one of Numbered Embodiments 1 to 48, wherein the hexapeptide-11 is encapsulated in liposomes. Numbered Embodiment 50 includes the method according to any one of Numbered Embodiments 1 to 49, wherein the topical composition further comprises lactoferrin. Numbered Embodiment 51 includes the method according to any one of Numbered Embodiments 1 to 50, wherein the lactoferrin is encapsulated in liposomes. Numbered Embodiment 52 includes the method according to any one of Numbered Embodiments 1 to 51, wherein the topical composition further comprises phosphatidylserine. Numbered Embodiment 53 includes the method according to any one of Numbered Embodiments 1 to 52, wherein the topical composition further comprises Trellera fusiformis extract. Numbered Embodiment 54 includes the method according to any one of Numbered Embodiments 1 to 53, wherein the topical composition further comprises sodium hyaluronate crosspolymer. Numbered Embodiment 55 comprises the method according to any one of Numbered Embodiments 1 to 54, wherein the topical composition further comprises hydroxymethoxyphenyldecanone. Numbered Embodiment 56 comprises the method according to any one of Numbered Embodiments 1 to 55, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2) and the hexapeptide is hexapeptide-11. Numbered Embodiment 57 comprises the method according to any one of Numbered Embodiments 1 to 56, wherein the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyldecanone, or a combination thereof. Numbered Embodiment 58 comprises the method according to any one of Numbered Embodiments 1 to 57, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3) and the hexapeptide is hexapeptide-11. Numbered Embodiment 59 comprises the method according to any one of Numbered Embodiments 1 to 58, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof.Numbered Embodiment 60 includes the method according to any one of Numbered Embodiments 1 to 59, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4) and the hexapeptide is hexapeptide-11. Numbered Embodiment 61 includes the method according to any one of Numbered Embodiments 1 to 60, wherein the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 62 includes the method according to any one of Numbered Embodiments 1 to 61, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5) and the hexapeptide is hexapeptide-11. Numbered Embodiment 63 includes the method according to any one of Numbered Embodiments 1 to 62, wherein the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 64 includes the method according to any one of Numbered Embodiments 1 to 63, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6) and the hexapeptide is hexapeptide-11. Numbered Embodiment 65 includes the method according to any one of Numbered Embodiments 1 to 64, wherein the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 66 includes the method according to any one of Numbered Embodiments 1 to 65, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7) and the hexapeptide is hexapeptide-11. Numbered Embodiment 67 includes the method according to any one of Numbered Embodiments 1 to 66, wherein the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof.Numbered Embodiment 68 includes the method according to any one of Numbered Embodiments 1 to 67, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8) and the hexapeptide is hexapeptide-11. Numbered Embodiment 69 includes the method according to any one of Numbered Embodiments 1 to 68, wherein the topical composition further comprises lactoferrin, phosphatidylserine, Tremella fusiformis extract, sodium hyaluronate crosspolymer, hydroxymethoxyphenyl decane, or a combination thereof. Numbered Embodiment 70 includes the method according to any one of Numbered Embodiments 1 to 69, wherein the topical composition is aqueous. Numbered Embodiment 71 includes the method according to any one of Numbered Embodiments 1 to 70, wherein the topical composition improves skin moisture. Numbered Embodiment 72 includes the method according to any one of Numbered Embodiments 1 to 71, wherein the topical composition reduces the appearance of bruises, age spots, or wrinkles. Numbered Embodiment 73 includes the method according to any one of Numbered Embodiments 1 to 72, wherein the topical composition is administered 1, 2, 3, 4, 5, 6, 7, or 8 times per day. Numbered Embodiment 74 includes the method according to any one of Numbered Embodiments 1 to 73, wherein the individual is a human. [Examples]

[0118] The following embodiments are provided for illustrative purposes to illustrate various embodiments of the Disclosure and are not intended to limit the Disclosure in any way. These embodiments, together with the methods described herein, represent and are exemplary of currently preferred embodiments and are not intended to be limitations on the scope of the Disclosure. Modifications and other uses encompassed within the spirit of the Disclosure as defined by the claims will arise for those skilled in the art.

[0119] Example 1: Gene expression study This example demonstrates the effects of different activators on gene expression in fibroblasts and keratinocytes.

[0120] method Cell lines. Human adult dermal fibroblast and keratinocyte cell lines were treated with 11 different compound treatments (+DMSO control as treatment #12). Primary fibroblasts, keratinocytes supplied from ZenBio. Primary cells were prepared according to ZenBio's instructions, 1 cm². 2 Cells were seeded in cell-specific medium at 5K or 10K per well. Cells were seeded in three ways in 48-well plates (Greiner), one cell line per plate. The medium volume was 500 μl per well. The two outer tandem rows of wells were not used. All horizontal rows were used. Cells were cultured for 2 days in a 37°C 5% CO2 incubator. After 48 hours, all cell cultures were homogeneous and appeared healthy, with no significant number of suspended dead cells or evidence of cellular vacuolation that could indicate apoptosis or death of cells.

[0121] Compounds. Stocks of compounds 1-8 on the list below were prepared in PBS at 100-fold dilutions of the concentrations indicated in red. A 50 mg / ml stock of compound 9 (PS, phosphatidylserine) was prepared in DMSO, but it did not completely dissolve in the solution even with extensive vortexing and heating. This stock was 100-fold dilution of the final concentration. A 20 mg / ml stock of compound 10 (CBD) was prepared in DMSO. No undissolved particles were observed, but the stock solution was slightly cloudy and not completely clear. This stock was 200-fold dilution of the final concentration.

[0122] 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% 8. Octapeptide (Octa) 9. Phosphatidylserine (Phos)

[0123] Administration. After 48 hours of adherent culture, fibroblasts and keratinocytes were treated with the test compound. The compound was resuspended in appropriate cell medium at the final concentrations shown in the table below, the adherent culture medium was removed, and the compound-containing medium was added.

[0124] RNA lysate preparation. After 24 hours of compound exposure, the medium was removed and the cells were washed once with PBS. 100 μl of RNA lysis buffer (Takara Bio catalog number 635013, 1x diluted "10x RNA lysis buffer") was added to the wells, thoroughly mixed by grinding, combined in RNAse-free microcentrifuge tubes, and immediately frozen at -30°C. Samples were prepared from one cell line (one plate) at a time. The plate array was 12 × 4 with each treatment in a row of three wells. The three wells were lysed and combined in tubes in PCR tube strips. All samples were sent to MedGenome frozen on dry ice for RNA extraction, library construction, and sequencing against 25M paired-end 100bp reads per sample.

[0125] Sequencing. Library preparation and sequencing were completed in MedGenome.

[0126] result Figure 1 shows data on hyaluronic acid synthase 2 (HAS2), the primary stimulant of hyaluronic acid (HA) in fibroblasts treated with various compounds. Octapeptides showed superior stimulation of hyaluronic acid synthase 2 (HAS2) (Figure 1).

[0127] Figure 2 shows data for HAS2 in keratinocytes treated with various compounds. Figure 3 shows data for hyaluronidase 2 (HYAL2), an HA reductase, in keratinocytes treated with various compounds. Hexapeptide-11 showed excellent upregulation of HAS2 and potent downregulation of HYAL2 in keratinocytes.

[0128] This example demonstrates that the peptides described herein are involved in regulating the gene expression of genes involved in hyaluronic acid stimulation.

[0129] Example 2: Evaluation of HA production This example assays whether octapeptides, SymDecanox®, Tremella, lactoferrin, phosphatidylserine, Hylasome®, Aquaxyl®, and complete formulations stimulate the secretion of high molecular weight (HMW) hyaluronic acid (HA) from dermal fibroblasts (and keratinocytes). This example also assays HA synthesis.

[0130] In short, dermal fibroblasts and keratinocytes were cultured in growth medium (6-well dish). When they reached confluence, the growth medium was replaced with serum-free medium for 24 hours. The following test reagents were added as experimental treatments.

[0131] Test drug for HA research (Experiment 1): 1. Octapeptide - 10 ug / mL 2. Lactoferrin - 500 ug / mL 3. SynHycan - 500 ug / mL 4. Phosphatidylserine - 500 ug / mL 5.Hylasome(trademark)-500ug / mL 6. Tremella - 500 ug / mL 7.SymDecanox(TM)-250ug / mL 8.Aquaxyl(trademark)-500ug / mL 9. A mixture of all compounds (12.5% ​​of the above concentration for each compound) 10. A mixture of all compounds (40% of the above concentration for each compound)

[0132] Supernatants from treated cells were collected at 24, 48, and 72 hours and divided equally before storage to prevent freeze-thaw degradation. 60K, 100K, and 2MDa HA were used as reference proteins. SDS-PAGE was used to isolate MW size from proteins isolated from the supernatant, along with reference proteins as comparison factors. Gels were stained with ALL-STAINS to compare molecular weight (MW). Figure 4 shows data illustrating the effects of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment. A: Negative control (PBS), B: Loading dye, C: 60k HA marker, D: 100k HA marker, E: 2M HA marker. Numbers 1-10 are the names of the compounds listed above. Different MW-HA and HMW-HA in 2M Dalton are shown as reference points. Figure 4 shows fibroblasts producing HA in the 2M Dalton / HMW HA range and those not producing HMW-HA.

[0133] The effects of the octapeptide compounds were also tested for HA production in human fibroblasts by adding hyaluronidase 72 hours after treatment to confirm the disappearance of the band. The compounds tested are listed below. Briefly, 100 μL of medium was taken from human fibroblast cultures and concentrated in a SpeedVac concentrator to 10 μL. The samples were treated with octapeptides along with 5 μL (1 mg / μL) of hyaluronidase enzyme at 37°C for 90 minutes. The data can be seen in Figure 5.

[0134] Compounds tested: 1. Loading buffer (negative control) 2. Octapeptide - 1 ug / mL 3. Octapeptide - 10 ug / mL 4. Octapeptide - 100 ug / mL 5. Collected culture medium 6. Collected culture medium 7. Octapeptide-100ug / mL + Hyaluronidase enzyme 8. HA 2M + Hyaluronidase Enzyme Control 9. HA 2M (positive control) 10. HA 2M (positive control)

[0135] In the second experiment, HA production was also evaluated using the following test compounds. Briefly, primary adult human dermal fibroblasts were cultured in 6-well plates until nearly confluent. The cells were treated with the compounds and concentrations shown on the right. After 72 hours, 100 μL of medium was collected from each treatment condition and concentrated to a final volume of approximately 10 μL in a SpeedVac concentrator.

[0136] Test drug for HA research (Experiment 2): 1. Octapeptide - 100 ug / mL 2. Lactoferrin - 500 ug / mL 3. SynHycan - 500 ug / mL 4. Phosphatidylserine - 500 ug / mL 5.Hylasome(trademark)-500ug / mL 6. Tremella - 500 ug / mL 7.SymDecanox(TM)-250ug / mL 8.Aquaxyl(trademark)-500ug / mL 9. A mixture of all compounds (12.5% ​​of the above concentration for each compound) 10. A mixture of all compounds (40% of the above concentration for each compound) 11. Negative control (PBS) 12. HA 2M (positive control)

[0137] All 10 μL of supernatant was loaded onto an acrylamide gel (NuPAGE 4-12% Bis-Tris protein gel) and electrophoresed at 200 V for 3 hours. The gel was stained to detect the presence of HA. The data are shown in Figure 6. Lane 12 is the positive control, showing the size of the band representing HA molecules with a size of 2 megadaltons (2 M). Lane 11 is the negative control, representing untreated cells. This band represents the cellular composition level of HA. Treatment of cells with all the indicated compounds (lanes 1-10) produced increased levels of high molecular weight HA. Treatment with octapeptides alone showed the strongest band.

[0138] From the eight compounds in Experiment 2, the supernatant was then digested with hyaluronidase enzyme at 37°C for 2 hours (lanes 1-8 in Figure 7). The fact that the bands disappeared after digestion indicates that the bands were indeed HA. Lane 9 is the undigested supernatant after treatment with octapeptide (100 ug / ml). Lane 10 is a positive control using 2M HA.

[0139] Next, experiments were conducted to quantify octapeptide-induced HA production in fibroblasts. Four replication wells were treated with two concentrations of octapeptide. After 72 hours, the supernatant was collected, concentrated, and run on a gel. Comparison of replications in lanes 6-9 and lanes 2-5 shows a dose-response to octapeptide-induced HA production (Figure 8A). Signal intensity was quantified, as shown in Figure 8B.

[0140] This example demonstrates that fibroblasts secrete high molecular weight HA when stimulated with octapeptide, lactoferrin, SynHycan, phosphatidylserine, Hylasome®, Tremella, SymDecanox®, Aquaxyl®, or any combination thereof. The presence of HA was verified, and the band disappeared after digestion of the fibroblast supernatant with hyaluronidase. Octapeptide alone induced the strongest HA band, showing a dose-dependent increase. Overall, these compounds, particularly octapeptide, are potent stimulants of high molecular weight HA in fibroblasts.

[0141] Example 3: EGR3 gene research This example evaluates EGR3 gene expression. EGR3, a transcription factor specifically present in the granular layer of the epidermis, is a gene involved in the formation of the skin barrier. Selectively increasing EGR3 expression in keratinocytes strengthens the skin barrier.

[0142] In short, human adult dermal fibroblast and keratinocyte cell lines were treated with 11 different compound treatments. Three wells were lysed and combined in tubes within a PCR tube strip. All samples were sent to MedGenome frozen on dry ice for RNA extraction, library construction, and sequencing of 25M paired-end 100bp reads per sample.

[0143] The data can be seen in Figure 9. Hexapeptide-11 showed potent upregulation of EGR3 in keratinocytes.

[0144] Example 4: Evaluation of Hyaluronic Acid (HA) Production The effects of tripeptide-1 and hexapeptide-12 (TriHex), octapeptide, hexapeptide-11, tetradecyl-diaminobutyroyl valyldiaminobutyric acid urea trifluoroacetate, syndecanox, tremella fusiformis, lactoferrin, and phosphatidylserine on stimulation and secretion of high molecular weight (HMW) HA from dermal fibroblasts and keratinocytes are determined individually and in combination.

[0145] Skin fibroblasts and keratinocytes are cultured in growth medium (6-well dish). Once they reach confluence, the growth medium is replaced with serum-free medium for 24 hours. The cells are then treated with various compounds. For fibroblasts, TFG+INF and TFG+IL-1 are added to stimulate HMW HA, while LMW HA is added as a control. Keratinocytes spontaneously secrete HA after the cells reach confluence (maximum size of 2MDa), and thus endogenous secretion is compared to the experimental conditions. Other controls include no serum starvation and serum starvation + vehicle controls.

[0146] The supernatant from the treated cells is collected at 24, 48, and 72 hours. To prevent freeze-thaw degradation, the supernatant is divided into equal portions before storage. Standards for ELISA are prepared using 6.4, 66, and 132 kDa, as well as 2 MDa, HA in powder form from Lifecore Biomedical. The supernatant is tested for the presence of HMW HA and LMW HA using the HA AlphaLISA kit (Perkin Elmer) according to the manufacturer's instructions.

[0147] Example 5: Size of secreted HA This example describes the determination of the size of secreted HA. Following a similar experimental procedure as described in Example 2, the supernatant is used for SDS-PAGE analysis. HA (Lifecore) of 6.4, 66, and 132 kDa, as well as 2 MDa, are used as molecular weight (MW) controls. The MW of the secreted HA is determined by staining the SDS-PAGE with Coomassie blue or by performing a Western blot.

[0148] Example 6: HA simulation of keratinocytes This example illustrates a simulation of fibroblast-secreted HA in keratinocytes.

[0149] Skin fibroblasts are cultured in growth medium. Once confluent, the fibroblasts are subjected to serum starvation for 24 hours. Then, the compound is added. As a control, TGF + INF is added to the fibroblasts. Keratinocytes are cultured simultaneously. Other controls include no serum starvation and serum starvation + vehicle controls.

[0150] The conditioned medium (CM) is collected and added to serum-starved keratinocytes (the CM can be frozen if keratinocytes are not available at the same time). CD44 expression is determined by immunofluorescence. Downstream CD44 signaling is also determined.

[0151] While preferred embodiments of the Disclosure are shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided merely as examples. Numerous variations, modifications, and substitutions will arise therefrom without departing from the Disclosure. It should be understood that various alternatives to the embodiments of the Disclosure described herein may be used in the practice of the Disclosure. The appended claims define the scope of the Disclosure and are intended to encompass the methods and structures of these claims and their equivalents.

Claims

1. An octapeptide consisting of the sequence GPHGVREA (sequence number: 5).

2. An octapeptide consisting of the sequence GPHGVREA (SEQ ID NO: 5), and Pharmaceutically acceptable carrier A composition comprising:

3. The composition described in claim 2, comprising 0.25 ppm to 100 ppm of the octapeptide.

4. A composition described in claim 2 or 3, containing 1 ppm to 30 ppm of the octapeptide.

5. A composition described in any one of claims 2 to 4, further comprising a hexapeptide.

6. The composition of claim 5, wherein the hexapeptide is hexapeptide-11.

7. The composition according to any one of claims 2 to 6, further comprising a synthetic tripeptide, preferably tetradecyl-diaminobutyroylvalyldiaminobutyric acid urea trifluoroacetate.

8. The topical composition of any one of claims 2 to 7, comprising lactoferrin, phosphatidylserine, or a combination thereof.

9. A composition described in any one of claims 2 to 8, wherein one or more of the phosphatidylserine, lactoferrin, octapeptide, or hexapeptide are encapsulated in a liposome.

10. A topical composition for treating skin, comprising an octapeptide consisting of the sequence GPHGVREA (SEQ ID NO: 5).

11. The topical composition of claim 10, further comprising a hexapeptide.

12. The composition of claim 10 or 11, further comprising a synthetic tripeptide, preferably tetradecyl-diaminobutyroylvalyldiaminobutyric acid urea trifluoroacetate.

13. The topical composition of any one of claims 10 to 12, wherein the octapeptide or the hexapeptide is encapsulated in a liposome.

14. A topical composition described in any one of claims 10 to 13, wherein the hexapeptide is hexapeptide-11.

15. 15. The topical composition of any one of claims 10 to 14, which stimulates the production of hyaluronic acid having a molecular weight of about 2 MDa.

Images