CN116568290A - Compositions and methods related to stimulating hyaluronic acid - Google Patents

Abstract

Provided herein are compositions and methods for stimulating the production of intrinsic hyaluronic acid. Compositions and methods for stimulating the production of intrinsic hyaluronic acid can improve skin moisture and elasticity.

Description

Compositions and methods related to stimulating hyaluronic acid

Cross reference

The present application claims the benefit of U.S. provisional patent application No. 63/089,424 filed on 8 th 10/2020 and U.S. provisional patent application No. 63/218,068 filed on 2 7/2021, each of which is incorporated by reference in its entirety.

Background

Hyaluronic acid is a non-sulfated glycosaminoglycan that is widely distributed in connective tissue, epithelial tissue, and neural tissue such as skin. Hyaluronic acid has been used in topical compositions for skin because of its ability to promote skin hydration. However, low molecular weight hyaluronic acid may promote the production of pro-inflammatory mediators. Accordingly, there is a need to provide topical compositions that promote intrinsic hyaluronic acid production without negatively affecting, for example, pro-inflammatory mediator production.

Disclosure of Invention

The skin contains 50% of human Hyaluronic Acid (HA). HA is a major component of the extracellular matrix (ECM) of the skin, which occurs in the epidermis, dermis and basal lamina located therebetween. HA is also observed intracellularly and plays an important role in metabolism, cell renewal, differentiation, cell motility, tissue repair, hydration, nutrient exchange and prevention of free radical damage. The rapid renewal of HA suggests that HA may also serve as an important channel for the removal of toxic substances. Natural HA as well as modified crosslinked HA have been used to help skin retain and even restore elasticity, plumpness and moisture. Topical compositions are needed to promote intrinsic HA stimulation in the skin.

Aspects described herein are topical compositions for stimulating hyaluronic acid comprising: synthesizing tripeptide; an octapeptide; and hexapeptide, wherein the topical composition stimulates hyaluronic acid. In one feature, the synthetic tripeptide comprises tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate (tetradecyl-diaminobutyroylvalyldiaminobutyric urea trifluoroacetate). In one feature, the octapeptide is encapsulated in a liposome. 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 comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2). In one feature, the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3). In one feature, the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4). In one feature, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). In one feature, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6). In one feature, the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7). In one feature, the octapeptide comprises 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 a liposome. In one feature, the topical composition further comprises lactoferrin. In one feature, the lactoferrin is encapsulated in a liposome. In one feature, the topical composition further comprises phosphatidylserine. In one feature, the topical composition further comprises tremella (Tremella fuciformis) extract. In one feature, the topical composition further comprises a sodium hyaluronate crosslinked polymer. In one feature, the topical composition further comprises hydroxymethoxyphenyl decanone. 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. In one feature, the octapeptide comprises 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 extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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 extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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 extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. In one feature, the octapeptide comprises 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof.

Aspects described herein are methods for stimulating hyaluronic acid production comprising administering a topical composition comprising: synthesizing tripeptide; an octapeptide; and hexapeptides. In one feature, the synthetic tripeptide comprises tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate. In one feature, the octapeptide is encapsulated in a liposome. 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 comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2). In one feature, the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3). In one feature, the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4). In one feature, the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). In one feature, the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6). In one feature, the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7). In one feature, the octapeptide comprises 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 a liposome. In one feature, the topical composition further comprises lactoferrin. In one feature, the lactoferrin is encapsulated in a liposome. In one feature, the topical composition further comprises phosphatidylserine. In one feature, the topical composition further comprises tremella (Tremella fuciformis) extract. In one feature, the topical composition further comprises a sodium hyaluronate crosslinked polymer. In one feature, the topical composition further comprises hydroxymethoxyphenyl decanone. 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. In one feature, the octapeptide comprises 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 extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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 extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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 extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. In one feature, the octapeptide comprises 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, 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 per day. In one feature, the individual is a human.

Drawings

FIG. 1 depicts a graph of hyaluronate synthase 2 (HAS 2) expression in fibroblasts.

FIG. 2 depicts a graph of hyaluronan synthase 2 (HAS 2) expression in keratinocytes.

FIG. 3 depicts a graph of hyaluronidase 2 (HYAL 2) expression in keratinocytes.

FIG. 4 depicts SDS-PAGE gels showing the effect of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment in the first experiment.

FIG. 5 depicts SDS-PAGE gels showing the effect of octapeptide compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment in the first experiment.

FIG. 6 depicts SDS-PAGE gels showing the effect of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment in the second experiment.

FIG. 7 depicts SDS-PAGE gels showing the effect of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment in the second experiment.

FIGS. 8A-8B depict SDS-PAGE gels (FIG. 8A) and graphs of quantification results (FIG. 8B) of octapeptide-induced HA production in fibroblasts.

FIG. 9 is a graph showing the expression of the EGR3 gene after treatment with various compounds.

Citation reference

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

Detailed Description

Definition of the definition

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

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of any embodiments. As used herein, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and/or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Unless specifically stated or apparent from the context, as used herein, when referring to a number or range of numbers, the term "about" is understood to mean +/-10% of the stated number and its number, or 10% below the lower limit and 10% above the upper limit of the listed range of values.

Composition and method for producing the same

Hyaluronic Acid (HA) is a major component of the skin extracellular matrix (ECM). HA plays an important role in metabolism, cell renewal, differentiation, cell motility, tissue repair, hydration, nutrient exchange and prevention of free radical damage. Topical compositions are needed to promote intrinsic HA stimulation in the skin.

Described herein are compositions and methods for stimulating Hyaluronic Acid (HA). The compositions and methods described herein may comprise HA, such as high molecular weight HA. The compositions and methods described herein may further comprise one or more agents that stimulate cells in the skin to produce more HA. The compositions and methods described herein may promote intrinsic Hyaluronic Acid (HA) stimulation. The compositions and methods described herein can facilitate 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 gene expression of genes involved in HA synthesis (e.g., HAs2, HYAL2, EGR 3).

Hyaluronic acid

Compositions comprising hyaluronic acid are described herein. 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 comprises improved water binding capacity. In some embodiments, the hyaluronic acid is cross-linked.

In general, the term "hyaluronic acid" may also encompass all variants and combinations of variants of hyaluronic acid, hyaluronate or hyaluronan of various chain lengths and charge states, as well as various chemical modifications, including crosslinking. In some cases, the hyaluronic acid comprises hyaluronate of hyaluronic acid having various counter ions, such as sodium hyaluronate. The term also encompasses various modifications of hyaluronic acid, such as oxidation, e.g., -CH 2OH groups to-CHO and/or-COOH; periodate oxidation of the vicinal hydroxyl group, optionally followed by reduction, e.g., -CHO to-CH 2OH or coupling with an amine to form an imine, followed by reduction to a secondary amine; sulfating; deamidation, optionally followed by deamination with a new acid or amide formation; esterification; crosslinking; substitution with various compounds, for example, coupling using a crosslinker or carbodiimide assistance; including coupling different molecules such as proteins, peptides and active pharmaceutical ingredients to hyaluronic acid; and deacetylation. Other examples of modifications are isourea, hydrazide, cyanogen bromide (bromoxyan), monoepoxide and monosulfone couplings.

In some embodiments, the compositions described herein comprise sodium hyaluronate crosslinked polymer. The sodium hyaluronate cross-linked polymer is a high molecular weight synthetic hyaluronic acid derived from a non-animal source, having high water binding capacity and moisture retention. Sodium hyaluronate cross-linked polymers are also scavengers of destructive free radicals and have a unique gel structure with gel domains that hold tightly bound water.

In some embodiments, 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 greater than 4.0 wt% (wt%) of the sodium hyaluronate crosslinked polymer is provided. In some embodiments, about 0.5% by weight of the sodium hyaluronate crosslinked polymer is provided. In some embodiments, the sodium hyaluronate crosslinked polymer is provided in a 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%.

Peptides

In some embodiments, the peptides described herein promote intrinsic Hyaluronic Acid (HA) stimulation. In some embodiments, the peptides described herein promote HA synthesis. In some embodiments, a peptide described herein (e.g., an octapeptide) facilitates the production of high molecular weight HA (HMW-HA). In some embodiments, the peptides described herein promote gene expression of genes involved in HA synthesis (e.g., HAs2, HYAL2, EGR 3).

The compositions described herein comprise different concentrations of peptides. In some cases, the peptide is present at about 50ppm or less to 1000, 5000, 10000, 50000, 100000, 500000ppm or more, e.g., 100ppm 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 a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 ppm. In some cases, 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 a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 micrograms per milliliter. In some cases, the peptide is present in about 0.01 wt.% to about 10 wt.%, 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.%).

In some embodiments, the compositions described herein comprise a plurality of peptides. In some cases, the peptides of the plurality of peptides are present in about 50ppm or less to 1000, 5000, 10000, 50000, 100000, 500000ppm or more, such as 100ppm of the peptide, or any other suitable amount. In some cases, the peptide of the plurality of peptides is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or more than 1000 ppm. In some cases, the peptides of the plurality of peptides are present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 ppm. In some cases, the peptide of the plurality of peptides is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or more than 1000 micrograms per milliliter (ug/mL). In some cases, the peptides of the plurality of peptides are present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 micrograms per milliliter. In some cases, the peptides of the plurality of peptides are present at about 0.01 wt.% to about 10 wt.%, 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.%). In some embodiments, the peptides of the plurality of peptides are provided at least or about 0.00001 wt%, 0.0003 wt%, 0.0005 wt%, 0.001 wt%, 0.005 wt%, 0.0055 wt%, 0.01 wt%, 0.02 wt%, 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, the peptides of the plurality of peptides are provided in a range of 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, each peptide of the plurality of peptides is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%.

In some embodiments, the compositions described herein comprise hexapeptide-11, octapeptide, synthetic peptide, or a combination thereof. In some embodiments, the synthetic peptide is tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate.

In some embodiments, hexapeptide-11 is provided at least or about 0.00001 wt%, 0.0003 wt%, 0.0005 wt%, 0.001 wt%, 0.005 wt%, 0.0055 wt%, 0.01 wt%, 0.02 wt%, 0.05 wt%, 0.10 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%, 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, hexapeptide-11 is provided in a range of 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, hexapeptide-11 is provided in a 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 a range of about 0.005 wt% to about 0.02 wt%. In some embodiments, hexapeptide-11 is provided at least or about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or more than 250 ppm. In some embodiments, hexapeptide-11 is provided in a 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 a range of about 10 to about 100 ppm. In some embodiments, hexapeptide-11 is provided at least or about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or more than 250 micrograms per milliliter (ug/mL). In some embodiments, hexapeptide-11 is provided in a range of about 25 to about 250, about 50 to about 200, or about 75 to about 150 micrograms per milliliter.

In some embodiments, the octapeptide is provided at least or about 0.00001 wt%, 0.0003 wt%, 0.0005 wt%, 0.001 wt%, 0.005 wt%, 0.0055 wt%, 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, the octapeptide is provided in a range of about 0.25 wt% to about 10 wt%, about 0.5 wt% to about 8 wt%, about 0.75 wt% to about 6 wt%, or about 1 wt% to about 4 wt%. In some embodiments, the octapeptide is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm. In some embodiments, 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 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 per milliliter (ug/mL). In some embodiments, the octapeptide is provided 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 per milliliter (ug/mL). In some embodiments, the octapeptide is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter. In some embodiments, the octapeptide is provided in a range of about 25 to about 200, about 25 to about 150, about 50 to about 125, about 10 to about 60, or about 20 to about 40 micrograms per milliliter.

In some embodiments, the synthetic peptide is provided at least or about 0.00001 wt%, 0.0003 wt%, 0.0005 wt%, 0.001 wt%, 0.005 wt%, 0.0055 wt%, 0.01 wt%, 0.02 wt%, 0.05 wt%, 0.10 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%, 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 peptide is provided in a range of 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 a 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 a range of about 0.005 wt% to about 0.02 wt%. In some embodiments, the synthetic peptide is provided at least or 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 more than 250 ppm. In some embodiments, the synthetic peptide is provided in a 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 at least or about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, or more than 250 micrograms per milliliter (ug/mL). In some embodiments, the synthetic peptide is provided 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 per milliliter (ug/mL). In some embodiments, the synthetic peptide is provided in a 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 per milliliter (ug/mL). In some embodiments, the synthetic peptide is provided in a range of about 25 to about 250, about 50 to about 200, or about 75 to about 150 micrograms per milliliter (ug/mL). In some embodiments, the synthetic peptide is a synthetic tripeptide. In some embodiments, the synthetic peptide is tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate.

In some embodiments, tripeptide-1 is provided in at least or about 0.00001 wt.%, 0.0003 wt.%, 0.0005 wt.%, 0.001 wt.%, 0.005 wt.%, 0.0055 wt.%, 0.05 wt.%, 0.10 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.%. In some embodiments, tripeptide-1 is provided in a range of about 0.25 wt% to about 10 wt%, about 0.5 wt% to about 8 wt%, about 0.75 wt% to about 6 wt%, or about 1 wt% to about 4 wt%. In some embodiments, tripeptide-1 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm. In some embodiments, tripeptide-1 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, tripeptide-1 is provided in a range of about 1 to about 10 ppm. In some embodiments, tripeptide-1 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 micrograms per milliliter (ug/mL). In some embodiments, tripeptide-1 is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter.

In some embodiments, hexapeptide-12 is provided at least or about 0.00001 wt%, 0.0003 wt%, 0.0005 wt%, 0.001 wt%, 0.005 wt%, 0.0055 wt%, 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, hexapeptide-12 is provided in a range of 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, hexapeptide-12 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm. In some embodiments, hexapeptide-12 is provided in a range of about 1 to about 10 ppm. In some embodiments, hexapeptide-12 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, hexapeptide-12 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 micrograms per milliliter (ug/mL). In some embodiments, hexapeptide-12 is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 micrograms per milliliter.

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 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 the 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 (also referred to herein as "Ile" or "I"), leucine (also referred to herein as "Leu" or "L"), lysine (also referred to herein as "Lys" or "K"), methionine (also referred to herein as "Met" or "M"), phenylalanine (also referred to herein as "Phe" or "F"), proline (also referred to herein as "P"), serine (also referred to herein as "Ser" or "S"), threonine (also referred to herein as "Thr" or "T" or "Y"), tyrosine (also referred to herein as "tyrosine" Y "or" tyrosine ") or" tyrosine (also referred to herein as "W").

In some embodiments, the first peptide is a dipeptide. Suitable 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. Suitable 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. Suitable tetrapeptides include, but are not limited to, those having the following amino acid sequences: GQPR, KTFK, AQTR or RSRK. In some embodiments, the tetrapeptides have 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 gktks. 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, heptapeptides having the amino acid sequence RGYYLLE or heptapeptide-6 (pro-longevity protein peptide). The composition may include two or more peptides, for example two dipeptides and one pentapeptide; a tripeptide and a hexapeptide; a dipeptide, a tripeptide, a heptapeptide, etc., provided that the composition comprises at least one dipeptide, tripeptide or tetrapeptide and at least one pentapeptide, hexapeptide or heptapeptide. In some embodiments, the compositions described herein comprise hexapeptide, octapeptide, synthetic peptide, or a combination thereof. In some embodiments, the one or more hexapeptides is hexapeptide-11. In some embodiments, the one or more peptides is tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate.

The peptide may be functionalized. For example, the peptide may be functionalized with fatty acids such as myristoleic acid, palmitoleic acid, oleic acid, elaidic acid, isooleic acid, linoleic acid, trans-linoleic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, linolic acid, caproic acid, lauric acid, palmitic acid, stearic acid, arachic acid, behenic acid, lignoceric acid, cerotic acid, and the like. Examples include palmitoyl hexapeptide-12 (Pal-VGVAPG), palmitoyl tripeptide-1 (Pal-GHK), myristoyl hexapeptide-12 (Myr-VGVAPG), and myristoyl tripeptide-1 (Myr-GHK). In certain embodiments, palmitoyl or myristoyl functionalization may be desirable because it exhibits enhanced permeability when compared to other fatty acids. In some embodiments, the peptide is functionalized with a chemical group. For example, peptides are functionalized with acetyl groups. Examples include acetyl hexapeptide-38 and acetyl tetrapeptide-2. In some cases, the peptide is functionalized with a functional group comprising no more than 14 carbons. In some cases, the peptide is functionalized with a functional group comprising no more than 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more than 20 carbons. In some cases, the peptide is non-palmitoylated. Without wishing to be bound by a particular theory, in some embodiments, incorporating the peptide into the liposome increases the lipophilicity of the functionalized or unfunctionalized peptide.

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 commonly found rarely in proteins, but is often found in extracellular matrix proteins. The small size of GHK allows it to be more accessible to membrane receptors than larger peptides. In addition, its unique copper binding structure enhances copper transport into and out of cells and promotes wound healing through several different but related pathways. Because of its strong copper binding structure, GHK can be provided in the form of GHK-Cu (copper-bound GHK form).

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

The peptide may advantageously be provided in a matrix suitable for combination with other components of the liposome composition. The matrix may include one or more components such as a thickener/binder (e.g., pentaerythritol tetraisostearate), an emollient/dispersant (e.g., caprylic/capric triglyceride), a solvent (e.g., propylene carbonate), and/or a rheology modifier/anti-settling agent (e.g., distearyldimethylammonium hectorite).

Liposome

Liposome compositions for improving distribution, efficacy, bioavailability, and/or activity are described herein. 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, improved delivery and skin penetration results from the active ingredient being incorporated (e.g., encapsulated) in the liposome. In some cases, the active ingredient is a peptide encapsulated in a liposome.

The liposome compositions described herein can comprise a peptide encapsulated in a liposome. In some embodiments, the peptide is hexapeptide-11. In some embodiments, the peptide is functionalized with a palmitoyl group. In some embodiments, the peptide is functionalized with an acetyl group.

The liposome compositions described herein can comprise various ingredients encapsulated in liposomes. In some embodiments, the ingredient is lactoferrin. In some embodiments, the component is phosphatidylserine. In some embodiments, the ingredient is a juniper extract. In some embodiments, the ingredient is arnica extract. In some embodiments, the ingredient is sodium hyaluronate. In some embodiments, the component is greater than 50kDa.

Lecithin and other phospholipids can be used to prepare liposomes containing the peptide compositions described herein. In some embodiments, the liposomes are used to prepare one or more peptides. In some embodiments, the peptide is functionalized with an acetyl group. Lipid vesicles are formed when phospholipids, such as lecithin, are placed in water, and upon sufficient energy, a bilayer or series of bilayers are formed, each separated by water molecules. Liposomes can be produced by sonicating phospholipids in water. Low shear rates produce multilamellar liposomes. Continuous high shear sonication tends to form smaller unilamellar liposomes. Hydrophobic chemicals may be dissolved in the phospholipid bilayer membrane. Lipid bilayer delivery of liposomes peptide compositions as described herein.

The phospholipids used to prepare the liposome compositions described herein can comprise a transition phase temperature of about 10 ℃ to about 25 ℃. In some cases, the phospholipid comprises a transition phase temperature of about 10 ℃, 12 ℃, 14 ℃, 16 ℃, 18 ℃, 20 ℃, 22 ℃, 24 ℃, 26 ℃, 28 ℃, 30 ℃, 32 ℃, 34 ℃, 36 ℃, 38 ℃, 40 ℃, or more than 40 ℃. In some cases, the phospholipid comprises a transition phase temperature in a range of about 10 ℃ to about 40 ℃, about 12 ℃ to about 36 ℃, about 14 ℃ to about 32 ℃, about 16 ℃ to about 20 ℃, or about 21 ℃ to about 25 ℃.

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

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

In some embodiments, described herein are methods for preparing a composition comprising a peptide encapsulated in a liposome, comprising: combining the peptide and a solvent to form a mixture; and contacting the mixture with an aqueous solution comprising liposomes. In some cases, the contacting occurs at a temperature between about 10 ℃ and about 25 ℃. In some cases, the contacting occurs at a temperature of about 10 ℃, 12 ℃, 14 ℃, 16 ℃, 18 ℃, 20 ℃, 22 ℃, 24 ℃, 26 ℃, 28 ℃, 30 ℃, 32 ℃, 34 ℃, 36 ℃, 38 ℃, 40 ℃, or more than 40 ℃. In some cases, the contacting occurs at a temperature in a range of about 10 ℃ to about 40 ℃, about 12 ℃ to about 36 ℃, about 14 ℃ to about 32 ℃, about 16 ℃ to about 20 ℃, or about 21 ℃ to about 25 ℃.

The method for preparing the composition comprising the peptide encapsulated in the liposome may comprise 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, methylene chloride, chloroform, diethyl ether, diisopropyl ether, ethyl acetate, butanol, n-propanol, ethanol, methanol, polyethylene glycol, propylene glycol, and pyridine. In some cases, the solvent is a glycol. In some cases, the solvent is butanediol. In some cases, the solvent is octanoyl glycol. In some cases, the solvent is propylene glycol (propylene glycol).

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

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

Methods for generating a liposome composition as described herein can result in an entrapment rate of no more than 100%. In some cases, the entrapment rate is no more than 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5%.

Liposome compositions are described herein, wherein the peptides comprise a percentage of the composition. In some embodiments, the peptide is provided 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 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 a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 5 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, the peptide is provided at about 0.03% of the composition.

Liposome compositions are described herein, wherein the liposomes comprise a percentage of the composition. In some embodiments, the liposome is provided 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 liposomes are provided in a 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, the liposomes are provided at about 30%. In some embodiments, the liposomes are provided at 27%.

In some embodiments, the liposome compositions described herein comprise an average particle size of up to 220 nanometers (nm). In some cases, the average particle size is up to 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 about 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 about 50nm to about 500nm, about 100nm to about 400nm, about 150nm to about 220nm, about 180nm to about 220nm, or about 190nm to about 210 nm.

In some cases, the liposome composition comprises an active agent having a molecular weight of no more than about 600 daltons (Da). In some cases, the active agent has a molecular weight of at least or about 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, or more than 1000 daltons (Da). In some cases, the active agent has a molecular weight of at least or about 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 4000, 5000, 6000, or more than 6000 daltons (Da). In some cases, the active agent has a molecular weight in the range of about 50 to about 1000, about 100 to about 900, about 200 to about 800, about 300 to about 700, or about 400 to about 600 daltons (Da). In some cases, the active agent is a peptide. In some cases, the active agent is a peptide encapsulated in a liposome.

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

In some cases, the intercept of a liposome composition as described herein is in the range of about 0.85 to about 0.95. In some cases, the intercept is the amplitude. In some cases, the intercept is at least or about 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, or 0.95.

In some embodiments, the liposome comprises propylene glycol, lecithin, or a combination thereof. In some embodiments, at least or about 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.% propylene glycol is provided. In some embodiments, propylene glycol is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, at least or about 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.% lecithin is provided. In some embodiments, lecithin is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, the liposome comprises propylene glycol and lecithin. In some embodiments, propylene glycol and lecithin are provided at least or about 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, propylene glycol and lecithin are provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, propylene glycol and lecithin are provided at about 0.90 wt.%.

Liposome compositions comprising improved distribution, efficacy, bioavailability, and/or activity are described herein. The liposome compositions can comprise improved distribution, efficacy, bioavailability, and/or activity as compared to compositions that do not comprise liposomes. In some cases, the distribution is improved by at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 4.0X, 4.5X, 5X, or more than 5X as compared to a composition that does not comprise liposomes. In some cases, the efficacy is improved by at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 4.0X, 4.5X, 5X, or more than 5X as compared to a composition that does not comprise liposomes. In some cases, the bioavailability is improved by at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 4.0X, 4.5X, 5X, or more than 5X as compared to a composition that does not comprise liposomes. In some cases, the activity is improved by at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 4.0X, 4.5X, 5X, or more than 5X as compared to a composition that does not comprise liposomes. The distribution, efficacy, bioavailability, and/or activity may be improved by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90% as compared to a composition that does not comprise liposomes.

In some embodiments, the liposome compositions and methods described herein are topical compositions. In some cases, the liposome composition is oil-free. In some cases, the liposome composition is free of preservatives. In some embodiments, the liposome formulation is an aqueous formulation. In some embodiments, the liposome formulation is an anhydrous formulation. In some cases, the liposome composition comprises a pH in the range of about 5 to about 8. In some cases, the liposome composition comprises a pH of at least or about 2, 3, 4, 5, 6, 7, 8, 9, or 10.

Methods and compositions as described herein may result in improved follicular penetration. In some cases, follicular penetration is improved by at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 4.0X, 4.5X, 5X, or more than 5X. 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 composition results in a follicular penetration depth of at least or about 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 7, 8, 9, 10, or more than 10 millimeters.

Phosphatidylserine

In some embodiments, the compositions as described herein comprise phosphatidylserine. Exposure of phosphatidylserine from the inner cell membrane of erythrocytes can induce phagocytosis of erythrocytes. See Chang CF, good BA, askenase MH et al, "recovery of erythrocyte-mediated macrophages after cerebral hemorrhage (Erythrocyte efferocytosis modulates macrophages towards recovery after intracerebral hemorrhage.)" journal of clinical research (The Journal of clinical research.)) "2018; 128 (2): 607-624.

In some embodiments, phosphatidylserine is provided at least or about 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 wt% (wt%). In some embodiments, phosphatidylserine is provided in a range of 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, phosphatidylserine is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.005 wt% to about 0.1 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, phosphatidylserine is provided in a range of about 0.005 wt% to about 0.02 wt%. In some embodiments, phosphatidylserine is provided at about 0.05 wt%. In some embodiments, phosphatidylserine is provided at about 0.25 wt%. In some embodiments, phosphatidylserine is provided at about 1 wt%. In some embodiments, phosphatidylserine is provided at least or 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 more than 1000 micrograms per milliliter (ug/mL). In some embodiments, phosphatidylserine is provided in a 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 per milliliter (ug/mL).

Lactoferrin protein

In some embodiments, the compositions described herein comprise transferrin. In some embodiments, the transferrin is lactoferrin. In some embodiments, the lactoferrin is encapsulated in liposomes. Lactoferrin HAs wound healing properties, promoting fibroblast proliferation and increasing HA secretion. See Saito S, takayama Y, mizumachi K, suzuki c, lactoferrin promotes synthesis of hyaluronic acid in human skin fibroblasts (Lactoferrin promotes hyaluronan synthesis in human dermal fibriplasts.) "Biotechnology rapid report (Biotechnology letters.)" 2011;33 (1): 33-39; takayama y (influence of lactoferrin on skin wound healing (Effects of Lactoferrin on Skin Wound health.)): lactoferrin and its role in wound healing (Lactoferrin and its Role in Wound health.)) (2012: 87-100.

In some embodiments, lactoferrin is provided in at least or about 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 wt.% (wt.%). In some embodiments, the lactoferrin is provided in a range of about 0.005 wt% to about 0.1 wt%, about 0.25 wt% to about 10 wt%, about 0.5 wt% to about 8 wt%, about 0.75 wt% to about 6 wt%, or about 1 wt% to about 4 wt%. In some embodiments, lactoferrin is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 2.5 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, lactoferrin is provided at about 0.025%. In some embodiments, lactoferrin is provided at about 0.05%. In some embodiments, lactoferrin is provided at about 0.10%. In some embodiments, lactoferrin is provided at least or 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 more than 1000 micrograms per milliliter (ug/mL). In some embodiments, lactoferrin is provided in a 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 per milliliter (ug/mL).

Hydroxy methoxy benzene decanone

In some embodiments, the compositions described herein comprise hydroxymethoxyphenyl decanone. In some embodiments, the hydroxymethoxyphenyl decanone is a potent intrinsic hyaluronic acid accelerator, an antioxidant, an anti-irritant, or a combination thereof.

In some embodiments, the hydroxymethoxyphenyl decanone is provided at least or about 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 wt.% (wt.%). In some embodiments, the hydroxymethoxyphenyl decanone is provided in a range of from about 0.25 wt.% to about 10 wt.%, from about 0.1 wt.% to about 2.5 wt.%, from about 0.5 wt.% to about 8 wt.%, from about 0.75 wt.% to about 6 wt.%, or from about 1 wt.% to about 4 wt.%. In some embodiments, the hydroxymethoxyphenyl decanone is provided in a range of from about 0.001 wt% to about 6 wt%, from about 0.002 wt% to about 4 wt%, from about 0.01 wt% to about 3 wt%, or from about 0.02 wt% to about 2 wt%.

Tremella fuciformis berk

In some embodiments, the compositions described herein comprise tremella extract. In some embodiments, the tremella extract is derived from an edible fungus. In some embodiments, the tremella extract provides moisture and antioxidant properties. In some embodiments, the tremella extract provides moisture from natural hyaluronic acid-containing sources.

In some embodiments, the tremella extract is provided at least or about 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 wt% (wt%). In some embodiments, the tremella extract is provided in a range of about 0.25 wt% to about 10 wt%, about 0.1 wt% to about 2.5 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 tremella extract is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, the tremella extract is provided at least or 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 more than 1000 micrograms per milliliter (ug/mL). In some embodiments, the tremella extract is provided in a 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 per milliliter (ug/mL).

Chemically cross-linked hyaluronic acid

In some embodiments, the compositions as described herein comprise chemically cross-linked hyaluronic acid (e.g., hyalastome ^TM ). In some embodiments, the chemically cross-linked hyaluronic acid is provided at least or about 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 wt% (wt%). In some embodiments, the chemically cross-linked hyaluronic acid is provided in a range of from about 0.005 wt% to about 0.1 wt%, from about 0.25 wt% to about 10 wt%, from about 0.5 wt% to about 8 wt%, from about 0.75 wt% to about 6 wt%, or from about 1 wt% to about 4 wt%. In some embodiments, the chemically cross-linked hyaluronic acid is provided in a range of from about 0.001% to about 6%, from about 0.002% to about 4%, from about 0.01% to about 2.5%, or from about 0.02% to about 2% by weight. In some embodiments, the chemically cross-linked hyaluronic acid is provided at about 0.025%. In some embodiments, the chemically cross-linked hyaluronic acid is provided at about 0.05%. In some embodiments, the chemically cross-linked hyaluronic acid is provided at about 0.10%. In some embodiments, the chemically cross-linked hyaluronic acid is present in an amount of at least or about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300 Provided are, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, or more than 1000 micrograms per milliliter (ug/mL). In some embodiments, the chemically cross-linked hyaluronic acid is provided in a 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 per milliliter (ug/mL).

Other components

In some embodiments, the compositions as described herein comprise symdecenox ^TM . In some embodiments, symDecanox ^TM Provided in at least or about 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 wt.% (wt.%). In some embodiments, symDecanox ^TM Provided in a range of about 0.005 wt% to about 0.1 wt%, about 0.25 wt% to about 10 wt%, about 0.5 wt% to about 8 wt%, about 0.75 wt% to about 6 wt%, or about 1 wt% to about 4 wt%. In some embodiments, symDecanox ^TM Provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 2.5 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, symDecanox ^TM Provided at about 0.025%. In some embodiments, symDecanox ^TM Provided at about 0.05%. In some embodiments, symDecanox ^TM Provided at about 0.10%. In some embodiments, symDecanox ^TM Provided at least or 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 more than 1000 micrograms per milliliter (ug/mL). In some embodiments, symDecanox ^TM Is provided in a 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 per milliliter (ug/mL).

In some embodiments, a combination as described hereinThe material comprises Aquaxyl ^TM . In some embodiments, aquaxyl ^TM Provided in at least or about 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 wt.% (wt.%). In some embodiments, aquaxyl ^TM Provided in a range of about 0.005 wt% to about 0.1 wt%, about 0.25 wt% to about 10 wt%, about 0.5 wt% to about 8 wt%, about 0.75 wt% to about 6 wt%, or about 1 wt% to about 4 wt%. In some embodiments, aquaxyl ^TM Provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 2.5 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, aquaxyl ^TM Provided at about 0.025%. In some embodiments, aquaxyl ^TM Provided at about 0.05%. In some embodiments, aquaxyl ^TM Provided at about 0.10%. In some embodiments, aquaxyl ^TM Provided at least or 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 more than 1000 micrograms per milliliter (ug/mL). In some embodiments, aquaxyl ^TM Is provided in a 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 per milliliter (ug/mL).

Other components may include anti-inflammatory agents, antioxidants, and solubilizing agents. Exemplary anti-irritants include, but are not limited to, panthenol triacetate and naringenin. Panthenol triacetate and naringenin are natural plant extracts that reduce skin redness and loss of water. Typical amounts of anti-irritants, when used in a composition, are 1 to 4 weight percent (wt.%).

Exemplary antioxidants include, but are not limited to, dunaliella salina (Dunaliella salina) extract and squalane. The dunaliella salina extract comprises components such as beta-carotene. It can exhibit an antioxidant effect. Typical amounts of anti-inflammatory agents, when used in a composition, are 0.1 wt.% to 2.5 wt.% (wt.%). In some embodiments, the dunaliella salina extract is provided at least or about 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 wt%. In some embodiments, the dunaliella salina extract is provided in a 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 at least or about 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 wt%. In some embodiments, squalane is provided in a 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, the dunaliella salina extract and squalane are provided at least or about 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 wt%. In some embodiments, the dunaliella salina and squalane extract is provided in a 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, the composition comprises a silicone polymer. In some embodiments, the silicone polymer is octanoyl polymethylsiloxane. In some embodiments, the octanoyl polymethylsiloxane is provided at least or about 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.% (wt.%). In some embodiments, the octanoyl polymethylsiloxane is provided at about 0.5 wt.%. In some embodiments, the octanoyl polymethylsiloxane is provided in a range of 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%. In some embodiments, the octanoyl polymethylsiloxane is provided at about 0.25 wt.%. In some embodiments, the octanoyl polymethylsiloxane is provided at about 1 wt%.

Bentonite may be employed in combination with peptides to impart osmotic and adsorptive properties to the composition and may help stabilize the emulsion. Other clays such as hectorite and magnesium aluminum silicate may also be used. Bentonite or other clays can be modified to produce organically modified clay compounds. Salts (e.g., quaternary ammonium salts) of fatty acids (e.g., hydrogenated fatty acids) may be reacted with hectorite or other clays. As provided herein, fatty acids are mentioned and described using conventional nomenclature employed by those skilled in the art. Saturated fatty acids do not include carbon-carbon double bonds. Unsaturated fatty acids include at least one carbon-carbon double bond. The single unsaturated fatty acid comprises only one carbon-carbon double bond. Polyunsaturated fatty acids comprise two or more carbon-carbon double bonds. The double bond in fatty acids is typically cis; however, trans double bonds are also possible. The position of the double bond may be indicated by an, where n indicates the lower numbered carbon of each pair of double bond carbon atoms. The total carbon number of the double bond and delta can be specified _{Double bond position} Is a shorthand notation for (c). For example, 20:4. Delta _5,8,11,14 Refers to fatty acids having 20 carbon atoms and four double bonds, wherein the double bonds are located between 5 and 6 carbon atoms, 8 and 9 carbon atoms, 11 and 12 carbon atoms, and 14 and 15 carbon atoms, wherein carbon atom 1 is the carbon of the carboxylic acid group. Stearates (stearates) are saturated fatty acids. Oleic acid ester (cis- Δ9-octadecenoic acid ester) is a single unsaturated fatty acid, and linolenic acid ester (all-cis- Δ9,12, 15-octadecatrienoic acid ester) is a polyunsaturated fatty acid. Suitable fatty acids may comprise from 5 to 30 carbonsAn atom, 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. The fatty acids may be fully saturated or may include as many double bonds as are feasible for chain length. Fatty acids suitable for functionalizing hectorite or other clays include palmitic acid and stearic acid. Dialkyl quaternary cation modifiers include dipalmitoyl diammonium chloride and distearyl dimethyl ammonium chloride. The amidoamine quaternary cationic modifier comprises palmitamidopropyl trimethyl ammonium chloride cetyl stearyl alcohol and palmitamidopropyl trimethyl ammonium chloride.

In some embodiments, the peptide may be mixed with suitable carriers, diluents or excipients, and may contain auxiliary substances such as wetting or emulsifying agents, pH buffering agents, gelling or viscosity enhancing additives, preservatives, flavoring agents, coloring agents and the like, depending on the route of administration and the desired formulation. See, for example, ramington: pharmaceutical science and practice (Remington: the Science and Practice of Pharmacy), lippincott Williams and Wilkins corporation (Lippincott Williams & Wilkins); version 20 (month 1 of 2003) and Remington's pharmaceutical science (Remington's Pharmaceutical Sciences), mike publishing company (Mack pub. Co.); 18 th edition and 19 th edition (month 12 in 1985 and month 6 in 1990, respectively). Such formulations may include complexing agents; a metal ion; polymeric compounds such as polyacetic acid, polyglycolic acid, hydrogels, dextran, and the like; a liposome; a microemulsion; a micelle; unilamellar or multilamellar vesicles; erythrocyte ghosts or bulbar buds. Suitable lipids for use in the composition include, but are not limited to, monoglycerides, diglycerides, sulfatides, lysolecithins, phospholipids, saponins, bile acids, and the like. In some embodiments, the compositions described herein comprise phosphatidylserine, phospholipids, tocopherols, ascorbyl palmitate, or a combination thereof. In some embodiments, phosphatidylserine, phospholipids, tocopherols, ascorbyl palmitate, or combinations thereof, is provided at 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 wt.% (wt.%). In some embodiments, phosphatidylserine, phospholipids, tocopherols, ascorbyl palmitate, or combinations thereof, are provided in a range of 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, phosphatidylserine, phospholipids, tocopherols, ascorbyl palmitate, or combinations thereof, are provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 5 wt%. In some embodiments, the additive is betaine. In some embodiments, the betaine is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 5 wt%. In some embodiments, the compositions as described herein comprise octanoyl glycol. In some embodiments, the octanoyl glycol is provided in a range of about 0.001 wt.% to about 6 wt.%, about 0.002 wt.% to about 4 wt.%, about 0.01 wt.% to about 3 wt.%, or about 0.02 wt.% to about 5 wt.%. In some embodiments, the compositions described herein comprise octanoyl hydroxamic acid. In some embodiments, octanoyl hydroxamic acid is provided in a range of about 0.001 wt.% to about 6 wt.%, about 0.002 wt.% to about 4 wt.%, about 0.01 wt.% to about 3 wt.%, or about 0.02 wt.% to about 5 wt.%. The presence of such additional components can affect the physical state, solubility, stability, release rate, clearance rate, and penetration of the active ingredient.

Compositions for topical administration comprise a peptide composition as described herein and a dermatologically acceptable vehicle. The vehicle may be aqueous or non-aqueous. The dermatologically acceptable vehicle for 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 applied topically in liquid or gel form, a liquid carrier such as water, petroleum, an oil of animal or vegetable origin such as peanut oil, mineral oil, soybean oil or sesame oil, or a synthetic oil may be added to the active ingredient. Saline solution, dextrose or other saccharide solution, 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 oily phase may be a vegetable oil such as olive oil or arachis oil, a mineral oil such as liquid paraffin, or a mixture thereof. Suitable emulsifying agents include naturally-occurring gums such as gum acacia and gum tragacanth, naturally-occurring phosphatides such as soy bean, 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 fragrances.

In certain embodiments, a silicone elastomer (e.g., a dimethicone cross-linked polymer) is employed to increase peptide delivery and penetration into the skin. An alternative to increasing the molecular weight (as with silicone gums) or adding fillers (as with silicone compounds) is to partially crosslink the silicone polymer and disperse the material in a suitable silicone carrier liquid. The resulting dimethicone crosslinked polymer (also known in the personal care industry as a silicone elastomer) differs from the base Polydimethylsiloxane (PDMS) due to the crosslinking between the linear polymers. These materials can be used in peptide compositions and also provide benefits in scar treatment, wound perimeter protection, and enzyme delivery. In skin care applications, the aesthetics of silicone elastomers (including those with functional groups) and their absorption of various oils (e.g., with dimethicone/vinyl dimethicone cross polymers, such as dakaning 9506 elastomer powder) is a desire for elastomersTwo of the characteristics. Silicone elastomers have a skin feel that is different from any silicone fluid, described as "smooth", "soft" and "powdery". It can be varied by controlling the amount of liquid phase in the formulation and thus the degree of swelling. Due to its film-forming properties, the dimethicone crosslinked polymer may be used as a delivery system for active ingredients such as peptides described herein or other composition components such as oil soluble vitamins and sunscreens. Sunscreens such as octyl methoxycinnamate can be more effectively delivered from compositions containing silicone elastomers, yielding higher Sun Protection Factor (SPF). The silicone elastomer blends may be used to enhance SPF in oil-in-water compositions containing organic sunscreens. For example, in tests conducted with respect to SPF, adding 4% of the silicone elastomer blend to a sunscreen composition containing an organic sunscreen agent increased the SPF from 5.7 to 18. This property of the silicone elastomer allows the effectiveness of the sunscreen in the composition to be maximized while reducing the amount required to achieve the desired SPF. Thus, the cost of the composition can be reduced while reducing the potential irritation caused by the activity of the sunscreens. Thus, higher SPF can be achieved with the same amount of UV absorber, resulting in enhanced performance without increasing the cost of the composition. The silicone elastomer may be produced from a linear silicone polymer by various crosslinking reactions, for example by hydrosilylation reactions in which vinyl groups react with silicon tetrahydrolate. The general process involves linear siloxane polymers having reactive sites along the polymer chain that react with a crosslinking agent. The dimethicone crosslinked polymer may be as a gel made from a suspension of elastomer particles swollen in a carrier liquid (e.g., a mixture of a high molecular weight silicone elastomer in cyclopentasiloxane, such as 9040 silicone elastomer blends), or as spray dry powders (dimethicone/vinyl dimethicone cross polymers, such as +.>9506 elastomerPowder) is produced. The gel form with the desired properties is cyclomethicone, but low viscosity dimethicones and organic fluids may also be used. Examples of dimethicone crosslinked polymers in suspension or gel form are high molecular weight silicone elastomers (12%) in decamethyl cyclopentasiloxane (e.g.,ST-elastomer 10) and a mixture of a high molecular weight silicone elastomer in cyclopentasiloxane (e.g.,9040 silicone elastomer blend) having an elastomer content in the range of typically 10 to 20% by weight.

Pharmaceutical excipients for topical formulation of the peptide composition may be selected from the group consisting of: solvents, emollients and/or emulsifiers, oil bases, preservatives, antioxidants, tonicity modifiers, permeation promoters and solubilizers, chelating agents, buffers, surfactants, one or more polymers and combinations thereof.

Suitable solvents for aqueous or hydrophilic liposome compositions include water; ethanol; isopropyl alcohol; a mixture of water and ethanol and/or isopropanol; glycerol; ethylene glycol, propylene glycol or butylene glycol; DMSO; pentanediol; and mixtures thereof. In some embodiments, the glycerol is provided at least or about 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, or more than 12 wt% (wt%). In some embodiments, the glycerol is provided at least or about 7%. In some embodiments, the glycerol is provided in a range of about 1 wt% to about 12 wt%, about 2 wt% to about 11 wt%, or about 3 wt% to about 10 wt%. In some embodiments, the butanediol is provided at least or about 0.0025 wt%, 0.005 wt%, 0.075 wt%, 0.01 wt%, 0.025 wt%, 0.05 wt%, 0.75 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, or more than 12 wt%. In some embodiments, the butanediol is provided in a range of about 0.01 wt% to about 10 wt%, about 0.025 wt% to about 5 wt%, or about 0.05 wt% to about 1.25 wt%. In some embodiments, the pentanediol is provided at least or about 0.0025 wt%, 0.005 wt%, 0.075 wt%, 0.01 wt%, 0.025 wt%, 0.05 wt%, 0.75 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, or more than 12 wt%. In some embodiments, the pentanediol is provided in a range of from about 0.01 wt% to about 10 wt%, from about 0.025 wt% to about 5 wt%, or from about 0.05 wt% to about 1.25 wt%. Suitable solvents for the hydrophobic composition include mineral oil, vegetable oil and silicone oil. If desired, the peptide compositions described herein can be dissolved or dispersed in a hydrophobic oil phase, and then the oil phase can be emulsified in an aqueous phase comprising water alone or in combination with lower alcohols, glycerol, and/or glycols. In some embodiments, anhydrous compositions are applied because the presence of water can result in stinging when applied to skin tissue subjected to laser treatment, chemical peeling, and the like. Anhydrous compositions can also be used to prevent the development of water-based irritant contact dermatitis in damaged or sensitive skin, which can produce rashes and skin irritation, which can slow wound healing and improve skin quality. Tsai, T.F, maibach, h.i. how stimulating water is? Summary (How irritant is waterAn overview.) contact dermatitis 41 (6) (1999): 311-314 (describing contact dermatitis caused by water as a stimulus). However, in certain embodiments, it may be acceptable to provide a water-based composition, or to allow for the presence of a limited amount of water. For example, water may be present, but the amount of water is below a threshold that may cause a tingling sensation when applied to damaged skin. Osmotic shock or osmotic stress is a sudden change in the concentration of the extracellular solute, causing rapid changes in the movement of water across its cell membrane. Water is removed from the cells by osmosis under conditions of high concentration of salts, substrates or any solutes in the supernatant. This also inhibits the transport of substrates and cofactors into the cell, thereby "shocking" the cell. Alternatively, at low concentrations of solutes, water enters the cell in large amounts, causing it to swell and rupture or undergo apoptosis. Certain compositions described herein may be advantageously employed when minimizing osmotic shock is desired.

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

Pharmaceutically acceptable thickeners may be used to maintain the viscosity of the composition at a selected level. Suitable viscosity enhancers or thickeners that may be used to prepare the viscous gel or cream with an aqueous base include sodium polyacrylate, xanthan gum, polyvinylpyrrolidone, acrylic polymers, carrageenan, hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, propyl cellulose, hydroxypropyl methyl cellulose, polyethoxylated polyacrylamide, polyethoxylated acrylates, and polyethoxylated alkanethiols. Methylcellulose is preferred because it is readily and economically available and easy to handle. Other suitable thickening agents include, for example, xanthan gum, carboxymethyl cellulose, hydroxypropyl cellulose, carbomers, and the like. The preferred concentration of thickener depends on the thickener selected. Preferably, an amount is used to achieve the selected viscosity. Viscous compositions are typically prepared from solutions by adding such thickeners, or by employing a matrix with acceptable viscosity levels.

In some embodiments, 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, 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 in excess of 40,000cps. In some embodiments, the viscosity of the composition is 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 34,000cps, about 12,000 to about 32,000cps, or about 14,000 to about 30,000 cps.

Suitable emollients include hydrocarbon oils and waxes, such as mineral oil, petrolatum, paraffin wax, ceresin, microcrystalline wax, polyethylene, squalene, perhydro squalene, silicone oil, triglycerides, acetylglycerides, such as acetylated monoglycerides; ethoxylated glycerides, such as ethoxylated glyceryl monostearate; alkyl esters of fatty acids or dicarboxylic acids. In some embodiments, the emollient is caprylic/capric triglyceride.

In some embodiments, the emollient is provided at least or about 0.0025 wt%, 0.005 wt%, 0.075 wt%, 0.01 wt%, 0.025 wt%, 0.05 wt%, 0.75 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, or more than 12 wt%. In some embodiments, the emollient is provided in a range of about 0.01 wt.% to about 10 wt.%, about 0.01 wt.% to about 2.5 wt.%, about 0.025 wt.% to about 5 wt.%, or about 0.05 wt.% to about 1.25 wt.%. In some embodiments, the caprylic/capric triglyceride is provided at least or about 0.0025 wt%, 0.005 wt%, 0.075 wt%, 0.01 wt%, 0.025 wt%, 0.05 wt%, 0.75 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, or more than 12 wt%. In some embodiments, the caprylic/capric triglyceride is provided in a range of about 0.01 wt% to about 10 wt%, about 0.01 wt% to about 2.5 wt%, about 0.025 wt% to about 5 wt%, or about 0.05 wt% to about 1.25 wt%.

Suitable silicone oils for use as emollients include dimethylpolysiloxane, methyl (phenyl) polysiloxane, and water-soluble and alcohol-soluble silicon glycol copolymers. Suitable triglycerides for use as emollients include vegetable and animal fats and oils including castor oil, safflower oil, cottonseed oil, corn oil, olive oil, cod liver oil, almond oil, avocado oil, palm oil, sesame oil and soybean oil.

Suitable esters of carboxylic or dicarboxylic acids for use as emollients include the methyl, isopropyl and butyl esters of fatty acids. Specific examples of the alkyl esters include hexyl laurate, isohexyl palmitate, isopropyl palmitate, decyl oleate, isodecyl oleate, cetyl stearate, decyl stearate, isopropyl isostearate, dilauryl lactate, myristyl lactate and cetyl lactate; and 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.

Other suitable classes of emollients or emulsifiers that may be used in the compositions include fatty acids, fatty alcohols, fatty alcohol ethers, ethoxylated fatty alcohols, fatty acid esters of ethoxylated fatty alcohols, and waxes.

Specific examples of fatty acids useful as emollients include pelargonic, lauric, myristic, palmitic, stearic, isostearic, hydroxystearic, oleic, linoleic, ricinoleic, arachidic, behenic and erucic acids. Specific examples of fatty alcohols useful as emollients include lauryl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol, isostearyl alcohol, hydroxystearyl alcohol, oleyl alcohol, ricinoleic alcohol, behenyl alcohol, erucyl alcohol, and 2-octyldodecanol.

Specific examples of waxes suitable for use as emollients include lanolin and its derivatives, including lanolin oil, lanolin wax, lanolin alcohol, lanolin fatty acids, isopropyl lanolate, ethoxylated lanolin alcohol, ethoxylated cholesterol, propoxylated lanolin alcohol, acetylated lanolin alcohol, lanolin alcohol linoleate, lanolin alcohol ricinoleate, acetate of ethoxylated alcohol ester, lanolin hydrogenolytes, hydrogenated lanolin, ethoxylated sorbitol lanolin, and liquid and semisolid lanolin. Also useful as waxes are hydrocarbon waxes, ester waxes and amide waxes. Useful waxes include wax esters such as beeswax, spermaceti, myristyl myristate and stearyl stearate; beeswax derivatives such as polyoxyethylene sorbitol beeswax; and vegetable waxes, including carnauba and candelilla waxes.

Polyols and polyether derivatives may be used as solvents and/or surfactants in the composition. Suitable polyols and polyethers include propylene glycol, dipropylene glycol, polypropylene glycol 2000 and 4000, poly (oxyethylene-co-oxypropylene) glycol, glycerin, 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, o-glycol having 15 to 18 carbon atoms, and polyoxypropylene derivatives of trimethylolpropane.

Polyol esters may be used as emulsifiers or emollients. Suitable polyol 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, glycerol mono-and di-fatty acid esters, polyglycerol poly-fatty acid esters, ethoxylated glycerol monostearate, 1, 3-butanediol distearate, polyoxyethylene polyol fatty acid esters, sorbitol fatty acid esters, and polyoxyethylene sorbitol fatty acid esters.

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

Sterols include, for example, cholesterol and cholesterol fatty acid esters; amides such as fatty acid amides, ethoxylated fatty acid amides, and fatty acid alkanolamides may also be used as emollients and/or penetration enhancers.

Pharmaceutically acceptable preservatives can be used to increase the shelf life of the composition. Other suitable preservatives and/or antioxidants for use in the composition may be employed including benzalkonium chloride, benzyl alcohol, phenol, urea, parabens, butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), tocopherols, thimerosal, chlorobutanol, and the like, and mixtures thereof. If a preservative, such as an antioxidant, is employed, the concentration is typically from about 0.02% to about 2% by total weight of the composition, although greater or lesser amounts may be desirable depending on the agent selected. As described herein, reducing agents may be advantageously used to maintain good shelf life of the composition. It is generally observed that the anhydrous compositions of the examples exhibit satisfactory stability such that preservatives can be omitted from the composition.

Suitable chelating agents for use in the composition include ethylenediamine tetraacetic 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 ethylenediamine tetraacetate (EDTA). In some embodiments, the disodium EDTA is provided at least or about 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 wt.% (wt.%). In some embodiments, the disodium EDTA is provided in a range of about 0.25 wt% to about 10 wt%, about 0.1 wt% to about 2.5 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 disodium EDTA is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%.

The carrier preferably has a pH of between about 4.0 and about 10.0, more preferably between about 4.8 and about 7.8, most preferably between about 5.0 and about 6.5. The pH may be controlled using a buffer solution or other pH adjusting agent. Suitable pH adjusting agents include phosphoric acid and/or phosphates, citric acid and/or citrates, hydroxide salts (i.e., calcium hydroxide, sodium hydroxide, potassium hydroxide), and amines such as triethanolamine. Suitable buffer solutions include buffers comprising solutions of monopotassium phosphate and dipotassium phosphate, maintaining a pH between 5.8 and 8; and a buffer comprising a monosodium phosphate and disodium phosphate solution, maintaining a pH between 6 and 7.5. Other buffers include citric acid/sodium citrate and disodium phosphate/citric acid. The peptide composition of the embodiments is preferably isotonic with the blood or other body fluid of the recipient. Isotonicity of the composition may be achieved using sodium tartrate, propylene glycol or other inorganic or organic solutes. Sodium chloride is particularly preferred. Buffers such as acetic acid and salts, citric acid and salts, boric acid and salts, and phosphoric acid and salts may be employed. It may be desirable to include a reducing agent in the composition, such as vitamin C, vitamin E, or other reducing agents known in the pharmaceutical arts.

Surfactants may also be used as excipients, for example anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate; cationic detergents such as benzalkonium chloride or benzethonium chloride; or nonionic detergents such as polyoxyethylene hydrogenated castor oil, glyceryl monostearate, polysorbate, sucrose fatty acid esters, methylcellulose or carboxymethylcellulose.

In certain embodiments, it may be advantageous to include additional agents that have pharmacological activity. The anti-infective agents include, but are not limited to, insect repellents (mebendazole), antibiotics (gentamicin), neomycin (neomycin), tobramycin (tobramycin), antifungal antibiotics (amphotericin b (amphotericinb), fluconazole (fluconazole), griseofulvin, itraconazole (itraconazole), ketoconazole (ketoconazole), nystatin (statin), miconazole (tolnaftate), tolnaftate (tolnaftate), cephalosporins (cefamandole), cefaclor (cefazolin), ceftazidime (ceftazidime), ceftazidime (cinnolamine), cyclosporin (cyclosporin), and penicillin (cyclosporin), and the like, bacitracin (bacitracin), clindamycin mesylate (colistimethate sodium), colistin b (polymyxin b sulfate), vancomycin (vancomycin), antiviral drugs including acyclovir (acyclovir), amantadine (amantadine), didanosine (didanosine), efavirenz (efavirenz), foscarnet acid (foscarnet), ganciclovir (ganciclovir), indinavir (indinavir), lamivudine (lamivudine), nelfinavir (nelfinavir), ritonavir (ritonavir), saquinavir (saquinavir), stavudine (stavudine), valacyclovir (valacyclovir), quinolones (sulfadiazine), sulfadiazine (sulfadiazine), and sulfadiazine (sulfadiazine). The anesthetic may include, but is not limited to, ethanol, bupivacaine (bupivacaine), chloroprocaine (chloroprocaine), levobupivacaine (levobupivacaine), lidocaine (lidocaine), mepivacaine (mepivacaine), procaine (procaine), ropivacaine (ropivacaine), tetracaine (tetracaine), desflurane (isoflurane), isoflurane, ketamine (ketamine), propofol (propofol), sevoflurane (sevoflurane), codeine (codeine), fentanyl (fenyl), hydromorphone (hydromorphone), tetracaine (marcaine), meperidine (meperidine), sand one (methyl), morphine (mopaine), oxycodone (oxycodone), fentanyl (tetracaine), sulfenamide (suprofen), buprofaine (buprofaine), buprofaine (buprofen), buprofen (buprofaine), buprofen (buprofen). Anti-inflammatory agents include, but are not limited to, non-steroidal anti-inflammatory drugs (NSAIDs), such as aspirin (aspirin), celecoxib (celecoxib), choline magnesium trisalicylate (choline magnesium trisalicylate), potassium diclofenac (diclofenac potassium), sodium diclofenac (diclofenac sodium), diflunisal (diflunisal), etodolac (etodolac), fenoprofen (fenoprofen), flurbiprofen (flubiprofen), ibuprofen (ibuprofen), indomethacin (indomethacin), ketoprofen (ketoprofen), ketorolac (keytorol), melenamic acid, nabumetone (nabumetone), naproxen (naproxen), naproxen sodium (naproxen soundium), oxazine (oxaprozin), piroxicam (piroxicam), rofecoxib (rofecoxib), bisquate (salixalate), sulindac (sulindac) and tolamine (etoposide); and corticosteroids such as cortisone (coristolone), hydrocortisone (hydrocortisone), methylprednisolone (methylprednisolone), prednisone (prednisone), prednisolone (prednisolone), betamethasone, beclomethasone dipropionate (beclomethasone dipropionate), budesonide (budesonide), dexamethasone sodium phosphate (dexamethasone sodium phosphate), flunisolide (flunisolide), fluticasone propionate (fluticasone propionate), triamcinolone acetonide (triamcinolone acetonide), betamethasone (betamethasone), fluocinolone acetate (fluocinonide), betamethasone dipropionate (betamethasone dipropionate), betamethasone valerate (betamethasone valerate), desonide (desoximetasone), desoximetasone (desoximetasone), fluocinolone, triamcinolone), clobetasol propionate (clobetasol propionate) and dexamethasone.

In certain embodiments, emollients, emulsion stabilizers, humectants, excipients, and other compounds may be added to modify to enhance the sensory properties of the topical composition, including but not limited to: skin feel (silky, light, creamy, etc.), absorbency (desired time when the product loses moistness and no longer feels on the skin), consistency, hardness, spreadability (e.g., viscosity, flow onset, shear rate, etc.), tackiness, shape integrity, gloss, hydrophilicity or hydrophobicity, etc. Preferably, the composition will have high coverage and low viscosity characteristics. Compositions having such characteristics have been shown to have enhanced "silky" or "light" skin feel ratings (see, e.g., bekker, m.webber, G, low, n. "when mineral and fischer-Tropsch wax based cosmetic emulsions and pectins are applied to the skin, rheological measurements are correlated with primary and secondary skin feel (Relating rheological measurements to primary and secondary skin feeling when mineral-based and Fischer-tropschwax-based cosmetic emulsions and jellies are applied to the skin)," journal of cosmetic science (International Journal of Cosmetic Science), "2013, 35 (4), pages 354-61).

In some embodiments, the composition comprises phenoxyethanol, ethylhexyl glycerol, or a combination thereof. In some embodiments, the phenoxyethanol is provided at least or about 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, the phenoxyethanol is provided in a range of 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 ethylhexyl glycerol is provided at least or about 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, the ethylhexyl glycerin is provided in a range from about 0.25 wt% to about 10 wt%, from about 0.5 wt% to about 8 wt%, from about 0.75 wt% to about 6 wt%, or from about 1 wt% to about 4 wt%. In some embodiments, phenoxyethanol and ethylhexyl glycerol are provided at least or about 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, phenoxyethanol and ethylhexyl glycerin are provided in a range from about 0.25 wt% to about 10 wt%, from about 0.1 wt% to about 4 wt%, from about 0.5 wt% to about 8 wt%, from about 0.75 wt% to about 6 wt%, or from about 1 wt% to about 4 wt%.

In some embodiments, the composition comprises polyacrylate-13, polyisobutylene, polysorbate 20, or a combination thereof. In some embodiments, polyacrylate-13 is provided at least or about 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, polyacrylate-13 is provided in a range of 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 polyisobutylene is provided at least or about 0.05 wt.%, 0.10 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.% (wt.%). In some embodiments, the polyisobutylene is provided in a range of 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, polyacrylate-13 is provided in a range of 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, polysorbate 20 is provided at least or about 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, polysorbate 20 is provided in a range of 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, polyacrylate-13, polyisobutylene, and polysorbate 20 are provided at least or about 0.05 wt%, 0.10 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, polyacrylate-13, polyisobutylene, and polysorbate 20 are provided in a range of about 0.25 wt.% to about 10 wt.%, about 0.1 wt.% to about 4 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.% (wt.%).

In some embodiments, the compositions as described herein comprise potassium sorbate. In some embodiments, potassium sorbate is provided in at least or about 0.001 wt.%, 0.00175 wt.%, 0.0025 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.%. In some embodiments, potassium sorbate is provided in the range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%.

In some embodiments, the liposome comprises propylene glycol, lecithin, or a combination thereof. In some embodiments, at least or about 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.% propylene glycol is provided. In some embodiments, propylene glycol is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, at least or about 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.%, 4.5 wt.%, 5.0 wt.%, 5.5 wt.%, 6.0 wt.%, 6.5 wt.%, 7.0 wt.%, 8 wt.%, 9 wt.%, 10 wt.%, or more than 10 wt.% lecithin is provided. In some embodiments, lecithin is provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, the liposome comprises propylene glycol and lecithin. In some embodiments, propylene glycol and lecithin are provided at least or about 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%, 4.5 wt%, 5.0 wt%, 5.5 wt%, 6.0 wt%, 6.5 wt%, 7.0 wt%, 8 wt%, 9 wt%, 10 wt%, or more than 10 wt% (wt%). In some embodiments, propylene glycol and lecithin are provided in a range of about 0.001 wt% to about 6 wt%, about 0.002 wt% to about 4 wt%, about 0.01 wt% to about 3 wt%, or about 0.02 wt% to about 2 wt%. In some embodiments, propylene glycol and lecithin are provided at about 0.90 wt.%.

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

Penetration enhancer

Fatty acids and alcohols may be used to enhance peptide penetration and provide silky feel to the composition, e.g., formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, myristoleic acid, isovaleric acid, palmitoleic acid, hexadecenoic acid, oleic acid, elaidic acid, isooleic acid, linoleic acid, trans-linoleic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, linolic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachic acid, behenic acid, cerotic acid, medium chain fatty acidsFor example C _6-12 Fatty acids, and the like. When used in a composition, typical amounts are from 1 to 4 wt.%.

Antibacterial efficacy

In some embodiments, described herein are compositions that are chemically and physically stable at physiological pH. In some embodiments, the composition is sterile and safe for human administration. In some embodiments, the composition meets or passes a desired antimicrobial efficacy test, such as an antimicrobial efficacy test. In some embodiments, the composition results in complete or substantially complete eradication of bacteria, yeast, mold, or a combination thereof.

Application method

Described herein are compositions and methods for stimulating Hyaluronic Acid (HA). In some embodiments, the compositions and methods stimulate HA and thereby improve skin moisture and maintain and improve skin elasticity and plumpness. In some embodiments, the compositions and methods described herein promote or improve skin hydration.

In some embodiments, the compositions and methods described herein improve aged skin or promote 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 pseudo-scarring. 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.

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

In some embodiments, the compositions and methods described herein improve the appearance of 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 skin by at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 3.5X, 4.0X, 5.0X, 6.0X, 7.0X, 8.0X, 9.0X, 10X, or more than 10X.

In some embodiments, the compositions and methods as described herein mimic intrinsic Hyaluronic Acid (HA) production. The compositions and methods described herein can improve skin-to-dermis high molecular weight HA penetration. In some embodiments, a composition comprising hydroxymethoxyphenyl decanone, tremella extract, lactoferrin, sodium hyaluronate cross-linked polymer, phosphatidylserine, or a combination thereof stimulates the production of intrinsic hyaluronic acid.

In some embodiments, a composition as described herein stimulates production of 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% of intrinsic hyaluronic acid. In some embodiments, a composition as described herein stimulates the production of at least or about 0.5X, 1.0X, 1.5X, 2.0X, 2.5X, 3.0X, 3.5X, 4.0X, 5.0X, 6.0X, 7.0X, 8.0X, 9.0X, 10X, or more than 10X of intrinsic hyaluronic acid.

The compositions described herein may be used with a variety of skin regimens. In some cases, the topical compositions described herein are administered once a day, twice a day, three times a day, or more. In some cases, the topical compositions described herein are administered twice daily. In some embodiments, the topical compositions described herein are administered daily, every other day, five days per week, once per week, every other week, three weeks per month, once per month, twice per month, three times per month, or more. In some embodiments, the topical compositions described herein are administered twice daily, e.g., in the morning and evening. In some embodiments, the topical compositions described herein are administered for at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 18 months, 2 years, 3 years, 4 years, 5 years, 10 years, or more. In some embodiments, the topical compositions described herein are administered twice daily for at least or about 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or more. 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 or about 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or more.

Stability test

The stability test of the composition can be performed as follows.

High temperature testing is now commonly used as a predictor of long term stability. High temperature testing can be performed at 37 ℃ (98°f) and 45 ℃ (113°f). If the product is stored at 45 ℃ for 3 months (and exhibits acceptable stability), it should be stable for two years at room temperature. The good control temperature is 4 ℃ (39°f), with most products exhibiting excellent stability. Sometimes, the product is also subjected to-10 ℃ (14°f) for 3 months.

In some cases, the stability of the product was evaluated by three temperature test cycles from-10 ℃ (14°f) to 25 ℃ (77°f). In such cases, the product is left at-10℃for 24 hours, and then at room temperature (25 ℃) for 24 hours. This completes one cycle. Even more stringent tests are five cycle tests at-10 to 45 ℃. This subjects the emulsion to extreme pressure.

The dispersed phase (of the oil-in-water emulsion) has a tendency to separate and rise to the top of the emulsion forming a layer of oil droplets. This phenomenon is known as emulsion delamination. Emulsion stratification is one of the first signs of impending emulsion instability. The test method for predictive emulsion stratification is centrifugation. The emulsion was heated to 50 ℃ (122°f) and centrifuged at 3000rpm for 30 minutes. The resulting product was then checked for signs of creaming.

Both the formulation and the package may be sensitive to UV radiation. The product was placed in glass and the actual package was placed in a light box with a broad spectrum output. Another glass jar fully covered with aluminum foil was used as a control. A discoloration of the product can be observed.

For all of the above tests, the color, odor/flavor, viscosity, pH, and, if available, particle size uniformity and/or particle agglomeration can be observed under a microscope.

Kit for non-invasive use and use with invasive surgery

Some embodiments of the methods and compositions provided herein include kits comprising a peptide and an agent that stimulates hyaluronic acid. In some embodiments, the kit may be provided to a managing physician, other health care professional, patient, or caregiver. In some embodiments, the kit comprises a container containing the composition in a suitable topical composition, and instructions for administering the composition to a subject. The kit may optionally also contain one or more additional therapeutic agents or other agents. For example, kits containing the topical form of the peptide composition may be provided with other skin care agents, such as cleansers, occlusive moisturizers, penetrating moisturizers, sunscreens, sunblocks, and the like. The kit may contain the peptide composition in bulk form, or may contain separate doses of the peptide composition for continuous or sequential administration. The kit may optionally contain one or more diagnostic tools, administration tools, and/or instructions for use. The kit contains a suitable delivery device, such as a syringe, pump dispenser, single dose pack, etc., and instructions for administering the peptide composition and any other therapeutic or beneficial agent. The kit may optionally contain instructions for storing, reconstituting (if applicable), and administering any or all of the included therapeutic or beneficial agents. The kit may comprise a plurality of containers reflecting the number of administrations to the subject, or different products to be administered to the subject.

In some embodiments, the composition is configured to support the skin before, during, and after cosmetic surgery, such as injection of fillers, and also to act with natural regenerative processes of the skin itself, and to help improve skin appearance and skin firmness. The topical composition may be applied immediately after surgery for faster recovery, or generally for skin that appears healthier. The composition can increase hyaluronic acid and promote skin hydration. The topical compositions are suitable for all skin types and post-operative skin. The topical composition may be provided to the patient in bulk form to allow the patient to self-administer the appropriate amount of peptide. For example, the patient may apply an amount of the composition sufficient to provide a uniform coating over the affected area or as otherwise indicated by the physician. In certain embodiments, it may be desirable to incorporate additional therapeutic or active agents into the topical composition. Alternatively, the adjuvant therapy or agent may be administered alone. For example, cleansers, sunscreens, sunblocks, permeable moisturizers, and/or occlusive moisturizers may be provided for application before or after the topical compositions of the embodiments. The kit may include topical peptide compositions, occlusive moisturizers, mild cleansers, permeable moisturizers, and broad spectrum spf30+ sunscreens.

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

Numbered examples

Numbered example 1 comprises a topical composition for stimulating hyaluronic acid comprising: synthesizing tripeptide; an octapeptide; and hexapeptide, wherein the topical composition stimulates hyaluronic acid. Numbering example 2 comprises the topical composition of numbering example 1 wherein the synthetic tripeptide comprises tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate. Numbering example 3 comprises the topical composition of numbering examples 1 to 2 wherein the octapeptide is encapsulated in a liposome. Numbering example 4 contains the topical compositions of numbering examples 1 to 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), GPGKNGGDD (SEQ ID NO: 7) or GPMGPRGP (SEQ ID NO: 8). Numbering example 5 contains the topical compositions of numbering examples 1 to 4, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1). Numbering example 6 contains the topical compositions of numbering examples 1 to 5, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2). Numbering example 7 contains the topical compositions of numbering examples 1 to 6, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3). Numbering example 8 contains the topical compositions of numbering examples 1 to 7, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4). Numbering example 9 contains the topical compositions of numbering examples 1 to 8, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). Numbering example 10 contains the topical compositions of numbering examples 1 to 9, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6). Numbering example 11 comprises the topical compositions of numbering examples 1 to 10, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7). Numbering example 12 comprises the topical compositions of numbering examples 1 to 11, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8). Numbered example 13 comprises the topical compositions of numbered examples 1 through 12 wherein the hexapeptide is hexapeptide-11. Numbering example 14 comprises the topical compositions of numbering examples 1 to 13 wherein hexapeptide-11 is encapsulated in a liposome. Numbered example 15 comprises the topical compositions of numbered examples 1-14, further comprising lactoferrin. Numbered example 16 comprises the topical compositions of numbered examples 1-15 wherein lactoferrin is encapsulated in liposomes. Numbered example 17 comprises the topical compositions of numbered examples 1-16, further comprising phosphatidylserine. Numbered example 18 comprises the topical compositions of numbered examples 1-17, wherein the topical composition further comprises tremella extract. Numbered example 19 comprises the topical compositions of numbered examples 1-18, wherein the topical composition further comprises a sodium hyaluronate crosslinked polymer. Numbered example 20 comprises the topical compositions of numbered examples 1 through 19, wherein the topical composition further comprises hydroxymethoxyphenyl decanone. Numbered example 21 comprises the topical compositions of numbered examples 1 through 20, wherein the topical composition is aqueous. Numbering example 22 comprises the topical compositions of numbering examples 1 to 21, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2) and the hexapeptide is hexapeptide-11. Numbered example 23 comprises the topical composition of numbered examples 1-22 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 24 contains the topical compositions of numbering examples 1 to 23, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3) and the hexapeptide is hexapeptide-11. Numbered example 25 comprises the topical compositions of numbered examples 1-24 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 26 comprises the topical compositions of numbering examples 1 to 25, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4) and the hexapeptide is hexapeptide-11. Numbered example 27 comprises the topical compositions of numbered examples 1-26 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 28 comprises the topical compositions of numbering examples 1 to 27, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5) and the hexapeptide is hexapeptide-11. Numbered example 29 comprises the topical compositions of numbered examples 1-28 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 30 comprises the topical compositions of numbering examples 1 to 29, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6) and the hexapeptide is hexapeptide-11. Numbered example 31 comprises the topical compositions of numbered examples 1-30 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 32 contains the topical compositions of numbering examples 1 to 31, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7) and the hexapeptide is hexapeptide-11. Numbered example 33 comprises the topical compositions of numbered examples 1-32 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 34 contains the topical compositions of numbering examples 1 to 33, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8) and the hexapeptide is hexapeptide-11. Numbered example 35 comprises the topical composition of numbered examples 1-34 further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbered example 36 comprises a method of stimulating hyaluronic acid production comprising administering a topical composition comprising: synthesizing tripeptide; an octapeptide; and hexapeptides. Numbering example 37 includes the method of any one of numbering examples 1 to 36, wherein the synthetic tripeptide comprises tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate. Numbering example 38 comprises the method of any one of numbering examples 1 to 37, wherein the octapeptide is encapsulated in a liposome. Numbering example 39 includes the method of any one of numbering examples 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), GPGKNGGDD (SEQ ID NO: 7) or GPMGPRGP (SEQ ID NO: 8). Numbering example 40 includes the method of any one of numbering examples 1 to 39, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1). Numbering example 41 includes the method according to any one of numbering examples 1 to 40, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2). Numbering example 42 contains the method according to any one of numbering examples 1 to 41, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3). Numbering example 43 comprises the method of any one of numbering examples 1 to 42, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4). Numbering example 44 contains the method according to any one of numbering examples 1 to 43, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5). Numbering example 45 includes the method according to any one of numbering examples 1 to 44, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6). Numbering example 46 includes the method according to any one of numbering examples 1 to 45, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7). Numbering example 47 includes the method of any one of numbering examples 1 to 46, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8). Numbering example 48 includes the method of any one of numbering examples 1 to 47, wherein the hexapeptide is hexapeptide-11. Numbering example 49 comprises the method of any one of numbering examples 1 to 48, wherein the hexapeptide-11 is encapsulated in a liposome. Numbering example 50 comprises the method of any one of numbering examples 1 to 49, wherein the topical composition further comprises lactoferrin. Numbering example 51 comprises the method of any one of numbering examples 1 to 50, wherein lactoferrin is encapsulated in liposomes. Numbering example 52 includes the method of any one of numbering examples 1 to 51, wherein the topical composition further comprises phosphatidylserine. Numbering example 53 includes the method of any one of numbering examples 1 to 52, wherein the topical composition further comprises tremella extract. Numbering example 54 comprises the method according to any one of numbering examples 1 to 53, wherein the topical composition further comprises a sodium hyaluronate crosslinked polymer. Numbering example 55 includes the method of any one of numbering examples 1 to 54, wherein the topical composition further comprises hydroxymethoxyphenyl decanone. Numbering example 56 contains the method of any one of numbering examples 1 to 55, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2) and the hexapeptide is hexapeptide-11. Numbering example 57 comprises the method of any one of numbering examples 1 to 56, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxybenzene decanone, or a combination thereof. Numbering example 58 includes the method of any one of numbering examples 1 to 57, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3) and the hexapeptide is hexapeptide-11. Numbering example 59 includes the method of any one of numbering examples 1 to 58, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 60 includes the method of any one of numbering examples 1 to 59, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4) and the hexapeptide is hexapeptide-11. Numbering example 61 comprises the method of any one of numbering examples 1 to 60, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 62 includes the method of any one of numbering examples 1 to 61, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5) and the hexapeptide is hexapeptide-11. Numbering example 63 comprises the method of any one of numbering examples 1 to 62, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxybenzene decanone, or a combination thereof. Numbering example 64 includes the method of any one of numbering examples 1 to 63, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6) and the hexapeptide is hexapeptide-11. Numbering example 65 includes the method of any of numbering examples 1 through 64, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy phenyl decanone, or a combination thereof. Numbering example 66 includes the method according to any one of numbering examples 1 to 65, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7) and the hexapeptide is hexapeptide-11. Numbering example 67 comprises the method of any one of numbering examples 1 to 66, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxybenzene decanone, or a combination thereof. Numbering example 68 includes the method of any one of numbering examples 1 to 67, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8) and the hexapeptide is hexapeptide-11. Numbering example 69 includes the method of any one of numbering examples 1 through 68, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxybenzene decanone, or a combination thereof. Numbered embodiment 70 comprises the method of any one of numbered embodiments 1 through 69, wherein the topical composition is aqueous. Numbered example 71 comprises the method of any one of numbered examples 1-70, wherein the topical composition improves skin moisture. Numbering example 72 includes the method of any one of numbering examples 1 to 71, wherein the topical composition reduces the appearance of bruises, age spots, or wrinkles. Numbering example 73 comprises the method of any one of numbering examples 1 to 72, wherein the topical composition is administered 1, 2, 3, 4, 5, 6, 7, or 8 times per day. Numbering embodiment 74 comprises the method of any one of numbering embodiments 1 to 73, wherein the individual is a human.

Examples

The following examples are presented for the purpose of illustrating various embodiments of the disclosure and are not meant to limit the disclosure in any way. This example, together with the methods described herein, is presently representative of the preferred embodiments, is exemplary, and is not intended as limiting the scope of the present disclosure. Variations and other uses thereof will occur to those skilled in the art and are encompassed within the spirit of the disclosure as defined by the scope of the claims.

Example 1: gene expression studies

This example shows the effect of different agents on gene expression in fibroblasts and keratinocytes.

Method

A cell line. Human adult dermal fibroblasts and keratinocyte cell lines were treated with 11 different compound treatments (plus DMSO control as treatment No. 12). Primary fibroblasts, keratinocytes were derived from ZenBio. Primary cells were grown at 5K or 10K cells per cm according to ZenBio specifications ² Plated in cell-specific medium. Cells were plated in triplicate in 48-well plates (Greiner), one cell line per plate. The volume of medium was 500ul per well. The outer two rows of holes are not used. All rows are used. Cells at 37℃5% CO ₂ Culturing in an incubator for 2 days. After 48 hours, all cell cultures were grown Even and seemingly healthy, there was no evidence of a significant number of floating dead cells or cell vacuolation, which could indicate apoptosis or death.

A compound. Stock solutions of compounds 1 to 8 in the following list were prepared in PBS at a concentration of 100X, indicated in red. Stock solutions of compound 9 (PS, phosphatidylserine) were prepared at 50mg/ml in DMSO, but did not go completely into solution under vigorous vortexing and warming. The stock solution was at a final concentration of 100X. A stock solution of 20mg/ml of Compound 10 (CBD) was prepared in DMSO. Undissolved particles were not observed, but the stock solution was slightly cloudy and not completely clear. The stock solution was at a final concentration of 200X.

Cell treatment:

1. lactoferrin (Lacton)

2.TCVRRAF(LCV)

3. Tripeptide-1 (Tri)

4. Hexapeptide-12 (Hex 12)

5. Tripeptide-1 and hexapeptide-12 (TriHex)

6. Hexapeptide-11 (Hex 11)

7. Tranexamic acid 5% (tranexamic acid)

8. Octapeptide (Octa)

9. Phosphatidylserine (Phos)

And (5) administration. After 48 hours of attachment culture, fibroblasts and keratinocytes were treated with the test compound. The compounds were resuspended in the appropriate cell culture medium at the final concentrations in the table below, and the adherent medium was removed and the medium containing the compounds was added.

RNA lysate preparation. After 24 hours of compound exposure, the medium was removed and the cells were washed 1X with PBS. 100ul of RNA lysis buffer (Takara Bio, takara Shuzo Co., ltd. (Takara Bio) catalog No. 635013, "10X RNA lysis buffer", diluted to 1X) was added to the wells, and thoroughly mixed by milling, combined in an RNAse-free microcentrifuge tube and immediately frozen at-30C. Samples were prepared from one cell line (one plate) at a time. The plate array was 12x 4, processed in 3 wells per row. Triplicate wells were lysed and pooled into tubes in PCR tube bands. All samples were frozen on dry ice to MedGenome for RNA extraction, library construction and sequencing, each 25M paired end 100bp read.

Sequencing. Library preparation and sequencing was done at MedGenome.

Results

Fig. 1 shows data for hyaluronate synthase 2 (HAS 2), which is the major stimulus for HA in fibroblasts, in fibroblasts treated with various compounds. Octapeptide showed excellent stimulation of hyaluronate synthase 2 (HAS 2) (fig. 1).

Fig. 2 shows data for HAS2 in keratinocytes treated with various compounds. Fig. 3 shows data for hyaluronidase 2 (HYAL 2), which is HA reductase, in keratinocytes treated with various compounds. Hexapeptide-11 shows excellent up-regulation of HAS2 in keratinocytes and effective down-regulation of HYAL2 in keratinocytes.

This example shows that the peptides described herein are involved in regulating gene expression of genes involved in hyaluronic acid stimulation.

Example 2: HA production assessment

This example determines octapeptide, symDecanox ^TM Tremella, lactoferrin, phosphatidylserine, hylasome ^TM 、Aquaxyl ^TM And whether the whole formulation stimulates the secretion of High Molecular Weight (HMW) Hyaluronic Acid (HA) by dermal fibroblasts (and keratinocytes). HA synthesis was also determined in this example.

Briefly, 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 test reagents listed below were added as experimental treatments.

Test reagent for HA study (experiment 1):

1. octapeptide-10 ug/mL

2. Lactoferrin-500 ug/mL

3.SynHycan–500ug/mL

4. Phosphatidylserine-500 ug/mL

5.Hylasome ^TM –500ug/mL

6. Tremella 500ug/mL

7.SymDecanox ^TM –250ug/mL

8.Aquaxyl ^TM –500ug/mL

9. Mixtures of all compounds (12.5% of each compound at the concentrations indicated above)

10. Mixtures of all compounds (40% of each compound above)

Supernatants from treated cells were collected at 24, 48 and 72 hours and aliquoted prior to storage to prevent freeze-thaw degradation. 60K, 100K and 2MDa HA were used as reference proteins. The MW size was separated from the proteins separated from the supernatant along with the reference protein as a comparison using SDS-PAGE. The gels were stained with ALL-STAINS to compare Molecular Weight (MW). Data see fig. 4, showing the effect of several compounds on hyaluronic acid production in human fibroblasts 72 hours after treatment. A. Negative control (PBS), b.supported dye, HA tag of c.60k, HA tag of d.100k, HA tag of e.2 m. The numbers 1 to 10 are the names of the compounds listed above. Different MW-HA and HMW-HA are shown with 2M daltons as a reference point. FIG. 4 shows fibroblasts producing HA in the 2M daltons/HMW HA range and no HMW-HA is produced.

The effect of octapeptide compounds on HA production in human fibroblasts was also tested at 72 hours post-treatment and with hyaluronidase added to confirm the disappearance of the band. The compounds tested are listed below. Briefly, 100uL of medium was harvested from human fibroblast cultures and concentrated down to 10uL in a SpeedVac concentrator. Samples were treated with octapeptide with 5uL (1 mg/uL) hyaluronidase for 90 min at 37 ℃. See fig. 5 for data.

Compounds tested:

1. sample buffer (negative control)

2. Octapeptide-1 ug/mL

3. Octapeptide-10 ug/mL

4. Octapeptide-100 ug/mL

5. Harvested culture medium

6. Harvested culture medium

7. Octapeptide-100 ug/mL+hyaluronidase

Ha 2m+ hyaluronidase control

HA 2M (positive control)

HA 2M (positive control)

In a second experiment, HA production was also assessed with the following test compounds. Briefly, primary adult dermal fibroblasts were cultured in 6-well plates until near confluence. Cells were treated with compounds and concentrations indicated on the right. After 72 hours, 100uL of medium was collected from each treatment condition and concentrated in a SpeedVac concentrator to a final volume of about 10uL.

Test reagent for HA study (experiment 2):

1. Octapeptide-100 ug/mL

2. Lactoferrin-500 ug/mL

3.SynHycan–500ug/mL

4. Phosphatidylserine-500 ug/mL

5.Hylasome ^TM –500ug/mL

6. Tremella 500ug/mL

7.SymDecanox ^TM –250ug/mL

8.Aquaxyl ^TM –500ug/mL

9. Mixtures of all compounds (12.5% of each compound at the concentrations indicated above)

10. Mixtures of all compounds (40% of each compound above)

11. Negative control (PBS)

HA 2M (positive control)

All 10uL of supernatant was loaded onto an acrylamide gel (NuPAGE 4 to 12% Bis-Tris protein gel) and electrophoresed at 200V for 3 hours. The gel was stained to detect the presence of HA. See fig. 6 for data. Lane 12 is a positive control and shows the size of the band representing HA molecules of size 2 megadaltons (2M). Lane 11 is a negative control, representing untreated cells. The bands represent cellular constituent levels of HA. Treatment of cells with all indicated compounds (lanes 1 to 10) produced increased levels of high molecular weight HA. Treatment with octapeptide alone showed the strongest bands.

8 compounds from experiment 2, and then the supernatant was digested with hyaluronidase at 37 for 2 hours (lanes 1 to 8 of fig. 7). The fact that the band disappeared after digestion indicates that the band was indeed HA. Lane 9 is the undigested supernatant after octapeptide (100 ug/ml) treatment. Lane 10 is a positive control with 2M HA.

Experiments were then performed to quantify octapeptide-induced HA production in fibroblasts. Four replicate wells were treated with two concentrations of octapeptide. After 72 hours, the supernatant was collected, concentrated and electrophoresed on a gel. Repeated comparisons in lanes 6 to 9 and lanes 2 to 5 indicate the presence of dose response for octapeptide-induced HA production (fig. 8A). The signal strength is quantified as seen in fig. 8B.

This example shows that fibroblasts are treated with octapeptide, lactoferrin, synHycan, phosphatidylserine, hylasome ^TM Tremella, symDecanox ^TM 、Aquaxyl ^TM Or a combination of all of these, they secrete high molecular weight HA when stimulated. Confirming the presence of HA; after digestion of the fibroblast-derived supernatant with hyaluronidase, the bands disappeared. Octapeptide alone induced the strongest HA band, which showed a dose dependent increase. In summary, these compounds are potent stimulators of high molecular weight HA in fibroblasts, particularly octapeptides.

Example 3: research on EGR3 gene

This example evaluates EGR3 gene expression. EGR3, in particular the transcription factor present in the granular layer of the epidermis, is the gene responsible for the formation of the skin barrier. Selectively increasing EGR3 expression in keratinocytes enhances the skin barrier.

Briefly, human adult dermal fibroblasts and keratinocyte cell lines were treated with 11 different compounds. Triplicate wells were lysed and pooled into tubes in PCR tube bands. All samples were frozen on dry ice to MedGenome for RNA extraction, library construction and sequencing, each 25M paired end 100bp read.

See fig. 9 for data. Hexapeptide-11 shows an effective up-regulation of EGR3 in keratinocytes.

Example 4: hyaluronic Acid (HA) production assessment

The effect of tripeptide-1 and hexapeptide-12 (TriHex), octapeptide, hexapeptide-11, tetradecyl-diaminobutyryl valyl diaminobutyrate urea trifluoroacetate, syndecanox, tremella, lactoferrin, phosphatidylserine alone and in combination on stimulation of secretion of High Molecular Weight (HMW) HA by dermal fibroblasts and keratinocytes was determined.

Dermal fibroblasts and keratinocytes were cultured in growth medium (6-well plate). When they reached confluence, the growth medium was replaced with serum-free medium for 24 hours. The cells are then treated with various compounds. For fibroblasts, tfg+inf was added to stimulate HMW HA, and tfg+il-1 was added to stimulate LMW HA as controls. After the cells reached confluence (up to 2MDa in size), the keratinocytes secreted HA themselves and, therefore, the endogenous secretion was compared with experimental conditions. Other controls included no serum starvation and serum starvation+vehicle controls.

The supernatants of the treated cells were collected at 24, 48 and 72 hours. The supernatant was aliquoted before storage to prevent freeze-thaw degradation. ELISA standards were prepared using 6.4, 66 and 132kDa from Lifecore Biomedical and 2MDa powder form HA. The supernatants were tested for the presence of HMW HA and LMW HA using the HA AlphaLISA kit (Perkin Elmer) according to the manufacturer's instructions.

Example 5: size of secreted HA

This example describes determining the size of secreted HA. The supernatant was used for SDS-PAGE analysis following an experimental procedure similar to that described in example 2. 6.4, 66 and 132kDa and 2MDa HA (Lifecore) were used as Molecular Weight (MW) controls. SDS-PAGE was stained with Coomassie blue (Coomassie blue) or Western blot was performed to determine MW of secreted HA.

Example 6: HA mimicking of keratinocytes

This example describes HA mimics secreted by fibroblasts of keratinocytes.

Dermal fibroblasts are cultured in a growth medium. After confluence, fibroblasts were serum starved for 24 hours. The compound is then added. As a control, tgf+inf was added to fibroblasts. At the same time, keratinocytes are cultured. Other controls included no serum starvation and serum starvation+vehicle controls.

Conditioned Medium (CM) was collected and added to serum starved keratinocytes (CM may be frozen if keratinocytes were not available at the same time). CD44 expression was determined by immunofluorescence. Downstream CD44 signaling was also determined.

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

Claims (74)

1. A topical composition for stimulating hyaluronic acid, comprising:

synthesizing tripeptide;

an octapeptide; and

a hexapeptide which is used as a source of the hexapeptide,

wherein the topical composition stimulates hyaluronic acid.

2. A topical composition according to claim 1, wherein the synthetic tripeptide comprises tetradecyl-diaminobutyryl valyl diaminobutyrate urea trifluoroacetate.

3. The topical composition of claim 1, wherein the octapeptide is encapsulated in a liposome.

4. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence gdgdgdgasa (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).

5. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1).

6. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2).

7. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3).

8. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4).

9. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5).

10. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6).

11. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7).

12. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8).

13. The topical composition of claim 1, wherein the hexapeptide is hexapeptide-11.

14. The topical composition of claim 13, wherein the hexapeptide-11 is encapsulated in a liposome.

15. The topical composition of claim 1, further comprising lactoferrin.

16. The topical composition of claim 15, wherein the lactoferrin is encapsulated in liposomes.

17. The topical composition of claim 1, further comprising phosphatidylserine.

18. The topical composition of claim 1, wherein the topical composition further comprises tremella (Tremella fuciformis) extract.

19. The topical composition of any one of claims 1-18, wherein the topical composition further comprises a sodium hyaluronate crosslinked polymer.

20. The topical composition of any one of claims 1-19, wherein the topical composition further comprises hydroxymethoxyphenyl decanone.

21. The topical composition of any one of claims 1-20, wherein the topical composition is aqueous.

22. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2) and the hexapeptide is hexapeptide-11.

23. The topical composition of claim 22, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

24. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3) and the hexapeptide is hexapeptide-11.

25. The topical composition of claim 24, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

26. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4) and the hexapeptide is hexapeptide-11.

27. The topical composition of claim 26, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

28. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5) and the hexapeptide is hexapeptide-11.

29. The topical composition of claim 28, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

30. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6) and the hexapeptide is hexapeptide-11.

31. The topical composition of claim 30, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

32. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7) and the hexapeptide is hexapeptide-11.

33. The topical composition of claim 32, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

34. The topical composition of claim 1, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8) and the hexapeptide is hexapeptide-11.

35. The topical composition of claim 34, further comprising lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxymethoxyphenyl decanone, or a combination thereof.

36. A method for stimulating hyaluronic acid production comprising administering a topical composition comprising: synthesizing tripeptide; an octapeptide; and hexapeptides.

37. A method according to claim 36, wherein the synthetic tripeptide comprises tetradecyl-diaminobutyryl valyl diaminobutyric acid urea trifluoroacetate.

38. The method of claim 36, wherein the octapeptide is encapsulated in a liposome.

39. The method of claim 36, wherein the octapeptide comprises the amino acid sequence gdgdgdgasa (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).

40. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GDGDGASA (SEQ ID NO: 1).

41. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2).

42. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3).

43. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4).

44. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5).

45. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6).

46. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7).

47. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8).

48. The method of claim 36, wherein the hexapeptide is hexapeptide-11.

49. The method of claim 48, wherein the hexapeptide-11 is encapsulated in a liposome.

50. The method of claim 36, wherein the topical composition further comprises lactoferrin.

51. The method of claim 50, wherein the lactoferrin is encapsulated in a liposome.

52. The method of claim 36, wherein the topical composition further comprises phosphatidylserine.

53. The method of claim 36, wherein the topical composition further comprises tremella extract.

54. The method of any one of claims 36 to 53, wherein the topical composition further comprises a sodium hyaluronate crosslinked polymer.

55. The method of any one of claims 36 to 54, wherein the topical composition further comprises hydroxymethoxyphenyl decanone.

56. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPMGPSGP (SEQ ID NO: 2) and the hexapeptide is hexapeptide-11.

57. The method of claim 56, wherein said topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

58. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GLGPGARA (SEQ ID NO: 3) and the hexapeptide is hexapeptide-11.

59. The method of claim 58, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

60. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPQGFQGP (SEQ ID NO: 4) and the hexapeptide is hexapeptide-11.

61. The method of claim 60, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

62. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPHGVREA (SEQ ID NO: 5) and the hexapeptide is hexapeptide-11.

63. The method of claim 62, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

64. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 6) and the hexapeptide is hexapeptide-11.

65. The method of claim 64, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

66. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPGKNGDD (SEQ ID NO: 7) and the hexapeptide is hexapeptide-11.

67. The method of claim 66, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

68. The method of claim 36, wherein the octapeptide comprises the amino acid sequence GPMGPRGP (SEQ ID NO: 8) and the hexapeptide is hexapeptide-11.

69. The method of claim 68, wherein the topical composition further comprises lactoferrin, phosphatidylserine, tremella extract, sodium hyaluronate cross-linked polymer, hydroxy methoxy benzene decanone, or a combination thereof.

70. The method according to any one of claims 36 to 69, wherein the topical composition is aqueous.

71. The method of any one of claims 36 to 70, wherein the topical composition improves skin moisture.

72. The method of any one of claims 36 to 70, wherein the topical composition reduces the appearance of bruises, age spots, or wrinkles.

73. The method of any one of claims 36 to 72, wherein the topical composition is administered 1, 2, 3, 4, 5, 6, 7, or 8 times per day.

74. The method of any one of claims 36-73, wherein the individual is a human.