Multisome lipid vesicles for delivery of cosmetic agents

Abstract

The present invention relates to compositions including formulations for delivering cosmetic agents comprising cosmetically active hyaluronic acid and anionic polymers such as peptides, and methods for using these formulations. These are useful in cosmetics and pharmaceuticals to prevent or improve the appearance of undesirable skin features, including wrinkles, or to enhance lip fullness.

Description

[Technical field]

[0001] cross reference This application claims the benefit of U.S. Provisional Patent Application No. 63 / 285,860, filed December 3, 2021, U.S. Provisional Patent Application No. 63 / 271,645, filed October 25, 2021, and U.S. Provisional Patent Application No. 63 / 165,603, filed March 24, 2021, which are incorporated by reference in their entireties. [Background technology]

[0002] Certain cosmetic agents are desirably delivered below the surface of the skin. There is a need for improved methods and compositions for the delivery of such cosmetic agents for a variety of cosmetic and medical purposes, including the prevention, reduction, or elimination of wrinkles. Summary of the Invention

[0003] The present invention relates to a composition for delivering cosmetic agents. In some embodiments, the composition is a lipid vesicle formulation of cosmetic agents that allows the agent to be delivered below the surface of the skin when topically applied. In some embodiments, the cosmetic agent is an anionic polymeric material such as hyaluronic acid, which is beneficial to the appearance of the skin, such as the skin of the lips of a subject. In some embodiments, the cosmetic agent is a peptide antagonist of muscle-type nicotinic acetylcholine receptors. In some embodiments, the cosmetic agent is delivered to a preferred or preselected layer of the skin or surrounding tissue, such as the epidermis, dermis, subcutaneous tissue, or muscle tissue.

[0004] In one aspect, provided herein is a pharmaceutical composition comprising: (a) lipid vesicles each comprising a lipid bilayer comprising a vesicle-forming lipid; (b) an oil-in-water emulsion encapsulated in the lipid vesicle and stabilized by one or more surfactants; and (c) a peptide antagonist of the muscle-type nicotinic acetylcholine receptor encapsulated in the lipid bilayer and / or the oil-in-water emulsion. In some embodiments, the peptide antagonist comprises an amino acid sequence having at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, the peptide antagonist comprises an amino acid sequence identical to the amino acid sequence of any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, the peptide antagonist has an amino acid sequence consisting of an amino acid sequence identical to the amino acid sequence of any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:60, 61, 73, 78, 82, 85, 91, or 95. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:60. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:61. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:73. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:78. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:82. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:85. In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO:91.In some embodiments, the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO: 95. In some embodiments, the peptide antagonist is present at a concentration of about 0.1 mg / mL to about 10 mg / mL. In some embodiments, the composition further comprises one or more penetration enhancers. In some embodiments, the one or more penetration enhancers comprise a non-ionic surfactant or a combination of non-ionic surfactants. In some embodiments, the non-ionic surfactant or combination of non-ionic surfactants is selected from polyethylene glycol ethers of fatty alcohols, sorbitan esters, polysorbates, sorbitan esters and polyethylene glycol fatty acid esters, and combinations thereof. In some embodiments, the polyethylene glycol ethers of fatty alcohols are C8-C. 22The polyethylene glycol ether of the aliphatic alcohol comprises an aliphatic alcohol and a polyethylene glycol group having about 2 to about 8 ethylene glycol subunits. In some embodiments, the polyethylene glycol ether of the aliphatic alcohol comprises diethylene glycol hexadecyl ether, 2-(2-octadecaoxyethoxy)ethanol, diethylene glycol monooleyl ether, polyoxyethylene(3) oleyl ether, or polyoxyethylene(5) oleyl ether, or any combination thereof. In some embodiments, the sorbitan ester comprises sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, or sorbitan isostearate, or any combination thereof. In some embodiments, the polyethylene glycol fatty acid ester comprises PEG-8 dilaurate, PEG-4 dilaurate, PEG-4 laurate, PEG-8 dioleate, PEG-8 distearate, PEG-8 distearate, PEG-7 glyceryl cocoate, and PEG-20 almond glycerides, or any combination thereof. In some embodiments, the polysorbate comprises polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, or any combination thereof. In some embodiments, each of the nonionic surfactants has a hydrophobic-lipophilic balance (HLB) of about 10 or less. In some embodiments, the nonionic surfactant or combination of nonionic surfactants is present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, at least one nonionic surfactant is present in an oil-in-water emulsion. In some embodiments, at least one nonionic surfactant is present in a lipid bilayer. In some embodiments, the one or more penetration enhancers include a combination of sorbitan esters, polysorbates, and polyethylene glycol fatty acid esters.In some embodiments, the one or more penetration enhancers include a combination of polyethylene glycol ethers of aliphatic alcohols, sorbitan esters, and polysorbates. In some embodiments, the one or more penetration enhancers include monolauroyllysine or dipalmitoyllysine, or a combination thereof. In some embodiments, the vesicle-forming lipids include phospholipids, glycolipids, lecithin, ceramide, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardiolipin, phosphatidic acid, cerebroside, or any combination thereof. In some embodiments, the vesicle-forming lipids include phospholipids. In some embodiments, the composition includes the vesicle-forming lipids in an amount of about 0.5% to about 25% (w / w) of the composition. In some embodiments, the composition includes a cationic surfactant. In some embodiments, the cationic surfactant is a monocationic surfactant. In some embodiments, the cationic surfactant comprises a propylene glycol diammonium phosphate ester derivatized with a fatty acid amide. In some embodiments, the cationic surfactant is present in an amount of about 1% to about 10%. In some embodiments, the oil-in-water emulsion comprises a triglyceride in the oil component. In some embodiments, the triglyceride comprises a medium chain triglyceride. In some embodiments, the triglyceride is present in an amount of about 1% to about 35% (w / w) of the composition. In some embodiments, the composition comprises a sterol. In some embodiments, the sterol is present in an amount of about 1% to about 5% (w / w) of the composition. In some embodiments, the composition comprises propylene glycol. In some embodiments, the propylene glycol is present in an amount of about 1% to about 25% (w / w) of the composition. In some embodiments, the composition comprises one or more viscosity enhancing agents. In some embodiments, the one or more viscosity enhancing agents are present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, the composition further comprises one or more additional agents.In some embodiments, the additional agent comprises one or more of a thickening agent, a preservative, a moisturizer, an emollient, a moisturizer, an antibacterial agent, or any combination thereof. In some embodiments, the composition is formulated for topical application to the skin of a subject. In some embodiments, the composition is formulated to deliver the peptide antagonist to a specific layer of the skin of a subject. In some embodiments, the composition is formulated as a cream, a lotion, a suspension, or an emulsion.

[0005] In one aspect, provided herein is a method of preparing a lipid vesicle composition provided herein, the method comprising the steps of: a) preparing an oil-in-water emulsion comprising a peptide antagonist of a muscle-type nicotinic acetylcholine receptor by mixing an oil component of the oil-in-water emulsion with an aqueous component of the oil-in-water emulsion, wherein the oil component and / or the aqueous component of the oil-in-water emulsion comprises one or more surfactants; b) solubilizing the vesicle-forming lipids in an acceptable solvent other than water; c) adding the oil-in-water emulsion to the solubilized vesicle-forming lipids; and d) mixing the oil-in-water emulsion and the solubilized vesicle-forming lipids under mixing conditions effective to form lipid vesicles comprising a lipid bilayer comprising the vesicle-forming lipids, and the water-in-oil emulsion encapsulated in the lipid vesicles.

[0006] In one aspect, provided herein is a method for providing one or more cosmetic effects by delivering a cosmetic agent below the skin surface of a subject, comprising administering a lipid vesicle composition provided herein to the skin surface. In some embodiments, the cosmetic agent is a polyanionic filler material. In some embodiments, the cosmetic agent is delivered to the dermis of the subject. In some embodiments, the one or more cosmetic effects include enhancing lip fullness, lip volume, lip smoothness, lip color, or a combination thereof. In some embodiments, the cosmetic agent is a peptide antagonist of the muscle-type nicotinic acetylcholine receptor. In some embodiments, the cosmetic agent is delivered to the muscle or subcutaneous tissue of the subject. In some embodiments, the one or more cosmetic effects include preventing or temporarily improving the appearance of one or more skin wrinkles. In some embodiments, the one or more skin wrinkles include moderate to severe glabellar lines associated with corrugator supercilii and / or procerus muscle activity, moderate to severe lateral gaze lines (crow's feet lines) associated with orbicularis oculi muscle activity, or moderate to severe forehead lines associated with frontalis muscle activity.

[0007] In one aspect, provided herein is a method of enhancing an individual's lip characteristics, comprising applying a composition to the individual's lips, the composition comprising a lipid vesicle comprising an oil-in-water emulsion and an anionic polymeric material. In some embodiments, the composition comprises a lipid vesicle comprising an oil-in-water emulsion and an anionic polymeric material. In some embodiments, the lipid vesicle is formulated according to the methods described herein. In some embodiments, the lipid vesicle comprises a phospholipid, a surfactant, a polymer, an emulsifier, a penetration enhancer, a triglyceride, a sterol, or any other material described herein. In some embodiments, the anionic polymeric material comprises an anionic polysaccharide. In some embodiments, the anionic polymeric material is hyaluronic acid. In some embodiments, the lip characteristic comprises lip fullness, lip volume, lip smoothness, lip color, or a combination thereof. In some embodiments, the composition is formulated for topical use. In some embodiments, the composition is delivered below the skin surface of the individual. In some embodiments, the composition is delivered below the skin surface of the individual's lips.

[0008] INCORPORATION BY REFERENCE All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. [Brief description of the drawings]

[0009] The novel features of the invention are set forth with particularity in the appended claims. To better understand the features and advantages of the present invention, reference should be made to the following detailed description that sets forth illustrative embodiments in which the principles of the invention are utilized and the accompanying drawings. [Figure 1]Panels A-C of FIG. 1 show the physicochemical characterization of multisome formulations (F1), (F2) and (F3). Panel A shows confocal microscopy images of multisome formulations (F1), (F2) and (F3) containing rhodamine red-labeled HA250K and green FITC-HA10K, and the FI traces show the co-localization of the two labels in the vesicles. Panel B of FIG. 1 shows optical microscopy images of multisome formulations (F1), (F2) and (F3). Panel C of FIG. 1 shows the particle size distribution of multisome formulations (F1), (F2) and (F3). [Diagram 2] Confocal microscopy images of human skin treated with cationic multisome formulations. The cationic multisome formulations were prepared with rhodamine red labeled HA250K and green FITC-HA150K or FITC-HA50K. The FI traces show the levels of rhodamine red labeled HA250K and green FITC-HA10K or FITC-HA50K in skin layers from the surface of the skin to the upper dermis. The plane of trace orientation is indicated on each micrograph. [Diagram 3] Confocal microscopy images of human skin treated with multisome formulations. Multisome formulations were prepared with rhodamine red labeled HA250K and green FITC-HA150K or FITC-HA50K. FI traces show the levels of rhodamine red labeled HA250K and green FITC-HA10K or FITC-HA50K in skin layers from the surface of the skin to the upper dermis. The plane of trace orientation is indicated on each micrograph. [Figure 4] Confocal microscopy images of human skin treated with multisome formulations. Multisome formulations were prepared with rhodamine red labeled HA250K and green FITC-HA150K or FITC-HA50K. FI traces show the levels of rhodamine red labeled HA250K and green FITC-HA10K or FITC-HA50K in skin layers from the surface of the skin to the upper dermis. The plane of trace orientation is indicated on each micrograph. [Diagram 5]Optical microscopy images of multiphase vesicle systems prepared with C7 peptide and their respective blank (no peptide) formulations are shown. Bar: 10 μm. [Figure 6A] Particle size distribution (left three graphs) and zeta potential (right three graphs) of the C7 peptide multiphasic vesicle delivery system. [Figure 6B] Particle size distribution (left three graphs) and zeta potential (right three graphs) of a blank multiphase vesicle delivery system are shown. [Figure 7] 1 shows an exemplary pictorial workflow for preparing lipid vesicles provided herein. [Figure 8] FIG. 1 shows an exemplary workflow for preparing lipid vesicles containing hyaluronic acid (HA) as provided herein. [Figure 9] 1 shows the results of topical application of lipid vesicles containing HA to the lips of subjects. [Figure 10A] 1 shows sensory properties and microscopic images of lipid vesicle formulations. [Figure 10B] 1 shows sensory properties and microscopic images of lipid vesicle formulations. [Figure 10C] 1 shows sensory properties and microscopic images of lipid vesicle formulations. [Figure 10D] 1 shows sensory properties and microscopic images of lipid vesicle formulations. [Figure 10E] 1 shows sensory properties and microscopic images of lipid vesicle formulations. [Figure 10F] 1 shows sensory properties and microscopic images of lipid vesicle formulations. [Figure 11A] 1 shows confocal microscopy images of lipid vesicle formulations. [Figure 11B] 1 shows confocal microscopy images of lipid vesicle formulations. [Figure 12] 1 shows confocal microscopy images of lead lipid vesicle formulations. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] definition As used herein, the term "comprise" or its variants "comprises" or "comprising" is intended to indicate the inclusion of any recited features, but is not meant to exclude any other features. Thus, as used herein, the term "comprising" is inclusive and does not exclude additional unrecited features. In some embodiments of any of the compositions and methods provided herein, "comprising" may be replaced with "consisting essentially of" or "consisting of." The phrase "consisting essentially of" is used to require specified characteristics or characteristics that do not substantially affect the characteristics or function of the invention. As used herein, the term "consisting of" is used to indicate the presence of only recited features.

[0011] As used in this specification and the appended claims, unless specified to the contrary, the following terms have the meanings specified below.

[0012] "Pharmaceutically acceptable salt" includes both acid and base addition salts. The pharmaceutically acceptable salt of any one of the compounds described herein is intended to include all pharmaceutically suitable salt forms. The preferred pharmaceutically acceptable salts of the compounds described herein are pharmaceutically acceptable acid and pharmaceutically acceptable base addition salts. In certain pharmaceutical embodiments of the present disclosure, "pharmaceutically acceptable salts" can be utilized.

[0013] As used herein, "treatment" or "treating" or "alleviating" or "ameliorating" are used interchangeably. Such terms refer to an approach to obtain beneficial or desired results, including, but not limited to, therapeutic benefit and / or prophylactic benefit. "Therapeutic benefit" refers to eradication or amelioration of the underlying disease being treated. Similarly, therapeutic benefit is achieved by eradication or amelioration of one or more of the physiological symptoms associated with the underlying disease, such that an improvement is observed in the patient, even though the patient still suffers from the underlying disease. For a prophylactic effect, the composition is administered, in some embodiments, to a patient at risk of contracting a particular disease, or to a patient who reports one or more of the physiological symptoms of the disease, even if a diagnosis of the disease has not been made. In certain pharmaceutical embodiments of the present disclosure, the terms "treatment of", or "treat", "apply", "alleviating", or "ameliorating" may be utilized.

[0014] As used herein, "conservative substitution" refers to replacing one amino acid with another amino acid that has similar properties, such as size, charge, and polarity. Substitutions can be for natural or modified (e.g., non-natural) amino acids. Non-limiting examples that can be replaced in conservative substitutions include the following groups: large hydrophobic (valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, methionine), small non-polar (alanine, glycine), polar (serine, threonine, glutamine, asparagine, cysteine, histidine), positively charged (lysine, arginine), and negatively charged (glutamic acid, aspartic acid).

[0015] Where % is used herein to refer to the amount of an ingredient, unless otherwise specified, % is intended to be % w / w.

[0016] The terms "penetration enhancer" and "penetration enhancers" are used interchangeably herein. As used herein, this refers to one or more ingredients that facilitate or enhance the penetration of one or more active ingredients (e.g., anionic polymeric materials such as hyaluronic acid or peptide antagonists) through one or more layers of the skin of a subject. In some embodiments, the penetration enhancer is a surfactant, for example, a nonionic surfactant having a hydrophilic-lipophilic balance (HLB) of about 10 or less, a cationic group, or another agent such as a terpene, an alkaloid, a salicylate derivative, a nicotinate derivative, or any combination thereof.

[0017] The term "multisome" as used herein refers to a lipid vesicle (such as a biphasic lipid vesicle) that contains one or more penetration enhancers, and in preferred embodiments, contains multiple penetration enhancers that act synergistically. In some embodiments, a multisome comprises a vesicle whose central core compartment is occupied by an oil-in-water emulsion composed of an aqueous continuous phase and a dispersed hydrophobic, hydrophilic or oil phase. In some embodiments, the space between adjacent bilayers of the lipid vesicle may be further occupied by an emulsion.

[0018] The term "lipid vesicle composition" as used herein refers to a composition comprising one or more lipid vesicles (e.g., polychromatic lipid vesicles, lipid bilayer vesicles, etc.). When a lipid vesicle composition is described as "comprising" one or more additional components (e.g., an anionic polymeric material or a peptide antagonist provided herein), it is intended that the composition includes the additional components in any manner within the composition (e.g., encapsulated within the lipid vesicle, etc.). For example, a lipid vesicle composition comprising an anionic polymeric material can include an anionic polymeric material encapsulated within the lipid bilayer of the lipid vesicle composition.

[0019] As used herein, the term "emulsion" refers to a mixture of two immiscible substances.

[0020] The term "bilayer" as used herein refers to a structure composed of amphiphilic lipid molecules arranged in two molecular layers, with the hydrophobic tails on the inner surface and the polar head groups on the outer surface.

[0021] The term "topical administration" or "topical delivery" as used herein refers to intradermal, transdermal and / or transmucosal delivery of a compound by administration of a compound or a composition containing multiple compounds to the skin and / or mucosa.

[0022] The term "gemini surfactant" as used herein refers to a surfactant molecule that contains multiple hydrophobic tails, each of which has a hydrophilic head, and the hydrophobic tails or hydrophilic heads are linked to each other by a spacer moiety. The hydrophobic tails can be the same or different. Similarly, the hydrophilic heads can be the same or different. The hydrophilic heads can be anionic, cationic, or neutral.

[0023] The term "HLB" or "Hydrophilic-Lipophilic Balance" value refers to the standard HLB according to Griffin, J. Soc. Cosm. Chem., vol. 5, 249 (1954), which indicates the degree of hydrophilicity and lipophilicity of a surfactant.

[0024] Lipid vesicle compositions of anionic polymeric materials such as hyaluronic acid for intradermal delivery - Patent Application 20070229933 In one aspect, a lipid vesicle composition comprising an anionic polymeric material is provided herein. In some embodiments, the lipid vesicle composition comprises lipid vesicles comprising lipid bilayers, each comprising a vesicle-forming lipid. In some embodiments, the lipid vesicle composition comprises an oil-in-water emulsion encapsulated in the lipid vesicle. In some embodiments, the oil-in-water emulsion is stabilized by one or more surfactants. In some embodiments, the anionic polymeric material is encapsulated in the lipid bilayer and / or the oil-in-water emulsion. In some embodiments, the anionic polymeric material is encapsulated in the lipid bilayer. In some embodiments, the anionic polymeric material is encapsulated in the oil-in-water emulsion.

[0025] Anionic Polymer Materials

[0026] In some aspects, the lipid vesicle composition provided herein comprises an anionic polymeric material. The anionic polymeric material is preferably adapted for delivery below the surface of the skin of a subject. In some embodiments, the anionic polymeric material acts as a volatilizing agent or filler after being delivered below the surface of the skin. In some embodiments, the anionic polymeric material acts as a support for another layer of the skin (e.g., the epidermis) to correct skin depressions or restore facial volume.

[0027] In some embodiments, the anionic polymeric material comprises an anionic polysaccharide. In some embodiments, the anionic polysaccharide is a non-sulfated glycosaminoglycan. In some embodiments, the anionic polymeric material is a naturally occurring substance. In some embodiments, the anionic polymeric material is naturally occurring in humans. In some embodiments, the anionic polymeric material is naturally occurring in human connective or epithelial tissue. In some embodiments, the anionic polymeric material is hyaluronic acid or a pharma- ceutically acceptable salt thereof. In some embodiments, the anionic polymeric material may not be crosslinked in the lipid vesicle compositions described herein.

[0028] In some embodiments, the hyaluronic acid is a pharma- ceutically acceptable salt of hyaluronic acid. In some embodiments, the salt is a sodium salt, a potassium salt, a magnesium salt, or any combination thereof. In some embodiments, the salt is a sodium salt.

[0029] In some embodiments, the anionic polymeric material has a molecular weight of about 5 kDa to about 500 kDa. In some embodiments, the molecular weight is a weight average molecular weight. In some embodiments, the anionic polymeric material has a molecular weight of about 5 kDa to about 500 kDa. In some embodiments, the anionic polymeric material has a molecular weight of about 5 kDa to about 10 kDa, about 5 kDa to about 20 kDa, about 5 kDa to about 50 kDa, about 5 kDa to about 100 kDa, about 5 kDa to about 200 kDa, about 5 kDa to about 250 kDa, about 5 kDa to about 300 kDa, about 5 kDa to about 400 kDa, about 5 kDa to about 500 kDa, about 10 kDa to about 20 kDa, about 10 kDa to about 50 kDa, about 10 kDa to about 100 kDa, about 10 kDa to about 200 kDa, about 10 kDa to about 250 kDa, about 10 kDa to about 300 kDa, about 10 kDa to about 400 kDa, about 10 kDa to about 500 kDa, about 20 kDa to about 50 kDa, about 20 kDa to about 100 kDa, about 20 kDa to about 200 kDa, about 20 kDa to about 250 kDa, about 20 kDa to about 300 kDa, about 20 kDa to about 400 kDa, About 20 kDa to about 500 kDa, about 50 kDa to about 100 kDa, about 50 kDa to about 200 kDa, about 50 kDa to about 250 kDa, about 50 kDa to about 300 kDa, about 50 kDa to about 400 kDa, about 50 kDa to about 500 kDa, about 100 kDa to about 200 kDa, about 100 kDa to about 250 kDa, about 100 kDa to about 300 kDa, about 100 kDa to about 400 kDa, about 100 kDa to about The polypeptide has a molecular weight of 500 kDa, about 200 kDa to about 250 kDa, about 200 kDa to about 300 kDa, about 200 kDa to about 400 kDa, about 200 kDa to about 500 kDa, about 250 kDa to about 300 kDa, about 250 kDa to about 400 kDa, about 250 kDa to about 500 kDa, about 300 kDa to about 400 kDa, about 300 kDa to about 500 kDa, or about 400 kDa to about 500 kDa. In some embodiments, the anionic polymeric material has a molecular weight of about 5 kDa, about 10 kDa, about 20 kDa, about 50 kDa, about 100 kDa, about 200 kDa, about 250 kDa, about 300 kDa, about 400 kDa, or about 500 kDa.In some embodiments, the anionic polymeric material has a molecular weight of at least about 5 kDa, about 10 kDa, about 20 kDa, about 50 kDa, about 100 kDa, about 200 kDa, about 250 kDa, about 300 kDa, or about 400 kDa. In some embodiments, the anionic polymeric material has a molecular weight of at most about 10 kDa, about 20 kDa, about 50 kDa, about 100 kDa, about 200 kDa, about 250 kDa, about 300 kDa, about 400 kDa, or about 500 kDa.

[0030] In some embodiments, the lipid vesicle composition comprises a first and a second anionic polymeric material. In some embodiments, the lipid vesicle composition further comprises a third anionic polymeric material.

[0031] In some embodiments, the first and second anionic polymeric materials are the same type. In some embodiments, each of the first and second anionic polymeric materials is an anionic polysaccharide. In some embodiments, each of the first and second anionic polymeric materials is hyaluronic acid.

[0032] When the first and second anionic polymeric materials are of the same type, each of the anionic polymeric materials has a different molecular weight. In some embodiments, the first anionic polymeric material has a molecular weight of up to about 75 kDa, and the second anionic polymeric material has a molecular weight greater than about 75 kDa. In some embodiments, the first anionic polymeric material has a molecular weight of up to about 75 kDa, and the second anionic polymeric material has a molecular weight greater than about 75 kDa.

[0033] When lipid vesicle comprises first and second anionic polymeric materials, each component can be comprised in different amounts.In some embodiments, the first and second anionic polymeric materials are present in approximately the same amount.In some embodiments, the ratio of the first and second anionic materials is about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 3:2, 2:1, or about 1:1.

[0034] In some embodiments, when the composition comprises a first, second, and third anionic polymeric material, each of the anionic polymeric materials can be of the same type (e.g., three different molecular weights of hyaluronic acid). In some embodiments, the composition comprises a first, second, and third anionic polymeric material, where the first anionic polymeric material has a molecular weight of about 5 kDa to about 20 kDa, the second anionic polymeric material has a molecular weight of about 20 kDa to about 75 kDa, and the third anionic polymeric material has a molecular weight greater than about 75 kDa. In some embodiments, each of the three anionic polymeric materials is present in about the same amount.

[0035] In some embodiments, the anionic polymeric material is present in an amount of about 0.01 mg / mL to about 10 mg / mL. In some embodiments, the anionic polymeric material is present in an amount of about 0.01 mg / mL to about 0.05 mg / mL, 0.01 mg / mL to about 0.1 mg / mL, about 0.01 mg / mL to about 0.5 mg / mL, about 0.01 mg / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.01 mg / mL to about 1.5 mg / mL, about 0.01 mg / mL to about 1.75 mg / mL, about 0.01 mg / mL to about 2 mg / mL, about 0.01 mg / mL to about 5 mg / mL, about 0.01 mg / mL to about 10 mg / mL, about 0. 05mg / mL~about 0.1mg / mL, about 0.05mg / mL~about 0.5mg / mL, about 0.05mg / mL~about 1mg / mL, about 0.05mg / mL~about 1.25mg / mL, about 0.05mg / mL~about 1.5mg / mL, about 0.05mg / mL~1.75 mg / mL, approximately 0.05 mg / mL to approximately 2 mg / mL, approximately 0.05 mg / mL to approximately 5 mg / mL, approximately 0.05 mg / mL to approximately 10 mg / mL, approximately 0.1 mg / mL to approximately 0.5 mg / mL, approximately 0.1 mg / mL to approximately 1 mg / mL, approximately 0.1 mg / mL to approximately 1.25 mg / mL, approximately 0.1 mg / mL to approximately 1.5 mg / mL, approximately 0.1 mg / mL to approximately 1.75 mg / mL, approximately 0.1 mg / mL to approximately 2 mg / mL, approximately 0.1 mg / mL to approximately 5 mg / mL, approximately 0.1 mg / mL to approximately 10 mg / mL, approximately 0.5 mg / mL to approximately 1 mg / mL, about 0.5mg / mL to about 1.25mg / mL, about 0.5mg / mL to about 1.5mg / mL, about 0.5mg / mL to about 1.75mg / mL, about 0.5mg / mL to about 2mg / mL, about 0.5mg / mL to about 5mg / mL, about 0.5mg / mL to about 10mg / mL, about 1mg / mL to 1.25mg / mL, about 1mg / mL to about 1.5mg / mL, about 1mg / mL to about 1.75mg / mL, about 1mg / ml to about 2mg / ml, about 1mg / ml to about 5mg / ml, about 1mg / ml to about 10mg / ml, about 1.25mg / mL ml~about 1.5mg / ml, about 1.25mg / ml~about 1.75mg / ml, about 1.25mg / ml~about 2mg / ml, about 1.25mg / ml~about 5mg / ml, about 1.25mg / ml~about 10mg / ml, about 1.5mg / ml~about 1.75mg / ml, about 1.The anionic polymeric material is present in an amount of about 5 mg / mL to about 2 mg / mL, about 1.5 mg / mL to about 5 mg / mL, about 1.5 mg / mL to about 10 mg / mL, about 1.75 mg / mL to about 2 mg / mL, about 1.75 mg / mL to about 5 mg / mL, about 1.75 mg / mL to about 10 mg / mL, about 2 mg / mL to 5 mg / mL, about 2 mg / mL to about 10 mg / mL, or about 5 mg / mL to about 10 mg / mL. In some embodiments, the anionic polymeric material is present in an amount of about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL. In some embodiments, the anionic polymeric material is present in an amount of at least about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, or about 5 mg / mL. In some embodiments, the anionic polymeric material is present in an amount of at most about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL.

[0036] vesicle-forming lipids In some embodiments, the vesicle composition comprises one or more vesicle-forming lipids. The vesicle-forming lipids act to encapsulate portions of the oil-in-water emulsion. In some embodiments, this allows the oil-in-water emulsion to remain stable for a period of time.

[0037] Vesicle-forming lipid can be any suitable lipid for such purpose.In some embodiments, vesicle-forming lipid comprises phospholipid, glycolipid, lecithin, ceramide, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardiolipin, phosphatidic acid, cerebroside, or any combination thereof.In some embodiments, vesicle-forming lipid comprises a combination of lipids.

[0038] In some embodiments, the vesicle-forming lipid comprises phospholipid. In some embodiments, the phospholipid is naturally occurring, semi-synthetic, or synthetically prepared, or a mixture thereof. In one embodiment, the phospholipid is one or more esters of glycerol with one or two (equal or different) residues of fatty acid and phosphoric acid, and the phosphoric acid residue is then linked to a hydrophilic group, such as choline (phosphatidylcholine--PC), serine (phosphatidylserine--PS), glycerol (phosphatidylglycerol--PG), ethanolamine (phosphatidylethanolamine--PE), or inositol (phosphatidylinositol). The esters of phospholipids with only one residue of fatty acid are generally referred to in the art as the "lyso" form of phospholipid or "lysophospholipid". The fatty acid residues present in phospholipids are generally long-chain fatty acids, typically containing 12-24 carbon atoms, or 14-22 carbon atoms, and the aliphatic chain may contain one or more unsaturations, or may be fully saturated. Examples of suitable fatty acids contained in phospholipids are, for example, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, oleic acid, linoleic acid, and linolenic acid. Saturated fatty acids such as myristic acid, palmitic acid, stearic acid, and arachidic acid may be used.

[0039] In some embodiments, the phospholipid comprises one or more natural phospholipids. In some embodiments, the phospholipid comprises one or more semi-synthetic phospholipids. In some embodiments, the semi-synthetic phospholipid is a partially or fully hydrogenated derivative of naturally occurring lecithin. In some embodiments, the phospholipid comprises a fatty acid diester of phosphatidylcholine, ethylphosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, or sphingomyelin. In some embodiments, the phospholipid comprises hydrogenated phosphatidylcholine (e.g., Sunlipon 90H). In some embodiments, the phospholipid may be, for example, dilauroyl-phosphatidylcholine (DLPC), dimyristoyl-phosphatidylcholine (DMPC), dipalmitoyl-phosphatidylcholine (DPPC), diarachidoyl-phosphatidylcholine (DAPC), distearoyl-phosphatidylcholine (DSPC), dioleoyl-phosphatidylcholine (DOPC), 1,2 distearoyl-sn-glycero-3-ethylphosphocholine (ethyl-DSPC), dipentadecanoyl-phosphatidylcholine (DPDPC), 1-myristoyl-2-palmitoyl-phosphatidylcholine (MPPC), 1-palmitoyl-2-myristoyl-phosphatidylcholine (PMPC), 1-palmitoyl-2-stearoyl-phosphatidylcholine ( PSPC), 1-stearoyl-2-palmitoyl-phosphatidylcholine (SPPC), 1-palmitoyl-2-oleyl-phosphatidylcholine (POPC), 1-oleyl-2-palmitoyl-phosphatidylcholine (OPPC), dilauroylphosphatidylglycerol (DLPG) and its alkali metal salts, diarachidoylphosphatidylglycerol (DAPG) and its alkali metal salts, dimyristoylphosphatidylglycerol (DMPG) and its alkali metal salts, dipalmitoylphosphatidylglycerol (DPPG) and its alkali metal salts, distearoylphosphatidylglycerol (DSPG) and its alkali metal salts, dioleoyl-phosphatidylglycerol (DOPG) and its alkali metal salts,Dimyristoyl phosphatidic acid (DMPA) and its alkali metal salts, dipalmitoyl phosphatidic acid (DPPA) and its alkali metal salts, distearoyl phosphatidic acid (DSPA), diarachidoyl phosphatidic acid (DAPA) and its alkali metal salts, dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), distearoyl phosphatidyl-ethanolamine (DSPE), dioleyl phosphatidylethanolamine (DOPE), diarachidoyl phosphatidylethanolamine (DAPE), dilinoleyl phosphatidylethanolamine (DLPE), dimyristoyl phosphatidylserine ( DMPS), diarachidoyl phosphatidylserine (DAPS), dipalmitoyl phosphatidylserine (DPPS), distearoyl phosphatidylserine (DSPS), dioleoyl phosphatidylserine (DOPS), dipalmitoyl sphingomyelin (DPSP), and distearoyl sphingomyelin (DSSP), dilauroyl-phosphatidylinositol (DLPI), diarachidoyl phosphatidylinositol (DAPI), dimyristoyl phosphatidylinositol (DMPI), dipalmitoyl phosphatidylinositol (DPPI), distearoyl phosphatidylinositol (DSPI), and dioleoyl-phosphatidylinositol (DOPI).

[0040] In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5% to about 25% (w / w) of the composition. In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5% to about 2%, about 0.5% to about 5%, about 0.5% to about 8%, about 0.5% to about 10%, about 0.5% to about 12%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 2% to about 5%, about 2% to about 8%, about 2% to about 10%, about 2% to about 12%, about 2% to about 15%, about 2% to about 20%, about 2% to about 25%, about 5% to about 8%, about 5% to about 10%, about 5% to about 10%, about 5% to about 20%, about 2% to about 25%, about 5% to about 8 ...0%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 2 In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, about 25%, about 8% to about 10%, about 8% to about 12%, about 8% to about 15%, about 8% to about 20%, about 8% to about 25%, about 10% to about 12%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 12% to about 15%, about 12% to about 20%, about 12% to about 25%, about 15% to about 20%, about 15% to about 25%, or about 20% to about 25% (w / w). In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25%. In some embodiments, the vesicle-forming lipids are present in an amount of at least about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, or about 20% (w / w) of the composition. In some embodiments, the vesicle-forming lipids are present in an amount of up to about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25% (w / w) of the composition.

[0041] In some embodiments, the vesicle-forming lipid is present in an amount of about 5% to about 15% (w / w) of the composition. In some embodiments, the vesicle-forming lipid is present in an amount of about 5% to about 8%, about 5% to about 9%, about 5% to about 10%, about 5% to about 11%, about 5% to about 12%, about 5% to about 13%, about 5% to about 14%, about 5% to about 15%, about 8% to about 9%, about 8% to about 10%, about 8% to about 11%, about 8% to about 12%, about 8% to about 13%, about 8% to about 14%, about 8% to about 15%, about 9% to about 10%, about 9% to about 11%, about 9% to about 12%, about 9% to about 12%, about 9% to about 14%, about 9% to about 15%, about 9% to about 10%, about 9% to about 11%, about 9% to about 12%, about 9% to about 14%, about 9% to about 15%, about 9% to about 16%, about 9% to about 18%, about 9% to about 19%, about 10% to about 20%, about 10% to about 21%, about 10% to about 22%, about 10% to about 23%, about 10% to about 24%, about 10% to about 25%, about 10% to about 26%, about 10% to about 27%, about 10% to about 28%, about 10% to about 29%, about 10% to about 30%, about 10% to about 31%, about 10% to about 32%, about 10% to about 33%, about 10% to about 34%, about 10 In some embodiments, the vesicle-forming lipid is present in an amount of about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% (w / w) of the composition. In some embodiments, the vesicle-forming lipids are present in an amount of at least about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14% (w / w) of the composition. In some embodiments, the vesicle-forming lipids are present in an amount of up to about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% (w / w) of the composition.

[0042] In some embodiments, the composition comprises a short-chain polyol. In some embodiments, the short-chain polyol acts to enhance the stability of the resulting lipid vesicle. In some embodiments, the short-chain polyol is a C2-C4 polyol containing two or three alcohol groups. In some embodiments, the short-chain polyol is propylene glycol. In some embodiments, the composition comprises propylene glycol.

[0043] In some embodiments, propylene glycol is present in an amount of about 0.5% to about 25% (w / w) of the composition. In some embodiments, propylene glycol is present in an amount of about 0.5% to about 2%, about 0.5% to about 5%, about 0.5% to about 8%, about 0.5% to about 10%, about 0.5% to about 12%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 2% to about 5%, about 2% to about 8%, about 2% to about 10%, about 2% to about 12%, about 2% to about 15%, about 2% to about 20%, about 2% to about 25%, about 5% to about 8%, about 5% to about 10%, about 5% to about 5%. In some embodiments, propylene glycol is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, about 25%, about 8% to about 10%, about 8% to about 12%, about 8% to about 15%, about 8% to about 20%, about 8% to about 25%, about 10% to about 12%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 12% to about 15%, about 12% to about 20%, about 12% to about 25%, about 15% to about 20%, about 15% to about 25%, or about 20% to about 25%. In some embodiments, propylene glycol is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25%. In some embodiments, propylene glycol is present in an amount of at least about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, or about 20%. In some embodiments, propylene glycol is present in an amount of up to about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25%. In some embodiments, propylene glycol is present in an amount of about 1% to about 10%. In some embodiments, propylene glycol is present in an amount of about 1% to about 2%, about 1% to about 4%, about 1% to about 6%, about 1% to about 8%, about 1% to about 10%, about 2% to about 4%, about 2% to about 6%, about 2% to about 8%, about 2% to about 10%, about 4% to about 6%, about 4% to about 8%, about 4% to about 10%, about 6% to about 8%, about 6% to about 10%, or about 8% to about 10%. In some embodiments, propylene glycol is present in an amount of about 1%, about 2%, about 4%, about 6%, about 8%, or about 10%. In some embodiments, propylene glycol is present in an amount of at least about 1%, about 2%, about 4%, about 6%, or about 8%.In some embodiments, propylene glycol is present in an amount of up to about 2%, about 4%, about 6%, about 8%, or about 10%. In some embodiments, propylene glycol is present in about the same amount as the vesicle-forming lipid. In some embodiments, the ratio of propylene glycol to the vesicle-forming lipid in the composition is about 2:1 to about 1:2 (w / w). In some embodiments, the ratio of propylene glycol to the vesicle-forming lipid in the composition is about 1:1 (w / w).

[0044] Oil Phase The lipid vesicle compositions provided herein include oil-in-water emulsions. The oil component is selected so that the material is liquid at operating temperatures (e.g., room temperature) and immiscible with water.

[0045] Any suitable oil can be used as oil phase.In some embodiments, the oil comprises naturally occurring oil.In some embodiments, the naturally occurring oil is derived from one or more plants or plant parts (e.g., seeds or nuts).In some embodiments, the oil is naturally occurring oil, such as olive oil, vegetable oil, sunflower oil, or other similar plant-derived oil.

[0046] In some embodiments, the oil phase is selected from the group consisting of vegetable oils, mono-, di-, and triglycerides, silicone fluids, mineral oils, and combinations thereof. In some embodiments, the oil phase comprises an emollient. In some embodiments, the emollient comprises caprylic and / or capric triglycerides (e.g., Labrafac CC). In some embodiments, the emollient comprises a natural oil-soluble emollient, such as an oil-soluble plant extract, aromatic oil, vegetable oil, vegetable butter, or any combination thereof. In some embodiments, the natural oil-soluble emollient comprises oil derived from sunflower. In some embodiments, the natural oil-soluble emollient comprises oil derived from avocado.

[0047] In some embodiments, the oil comprises a silicone oil, such as dimethicone, or a derivative. In some embodiments, the silicone oil comprises a siloxane polymer. In some embodiments, the siloxane polymer comprises C1-C3 substituents. In some embodiments, the siloxane is polydimethylsiloxane (PDMS). In some embodiments, the oil is a mixture that includes a silicone oil (e.g., dimethicone) as a smaller component. In some embodiments, the oil is a mixture that includes a natural emollient as an alternative to dimethicone (e.g., LexFeel™ N350MB). In some embodiments, the silicone oil is incorporated to enhance the feel of the resulting composition or as a moisturizer. In some embodiments, the oil comprises a silicone oil in an amount of up to about 5%, up to about 4%, up to about 3%, up to about 2%, or up to about 1% (w / w) of the composition. In some embodiments, the silicone oil is present in an amount of about 0.1% to about 2%.

[0048] In some embodiments, the oil is present in an amount of about 1% to about 35% (w / w) of the composition. In some embodiments, the oil is present in an amount of about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 1% to about 35%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 10% to about 15%, about 10% The oil is present in an amount of about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 10%, about 30%, about 10% to about 35%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 25% to about 30%, about 25% to about 35%, or about 30% to about 35%. In some embodiments, the oil is present in an amount of about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the oil is present in an amount of at least about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, or about 30%. In some embodiments, the oil is present in an amount of up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the oil is present in an amount of about 5% to about 15%. In some embodiments, the oil is present in an amount of about 5% to about 8%, about 5% to about 9%, about 5% to about 10%, about 5% to about 11%, about 5% to about 12%, about 5% to about 13%, about 5% to about 14%, about 5% to about 15%, about 8% to about 9%, about 8% to about 10%, about 8% to about 11%, about 8% to about 12%, about 8% to about 13%, about 8% to about 14%, about 8% to about 15%, about 9% to about 10%, about 9% to about 11%, about 9% to about 12%, about 9% to about 1 In some embodiments, the oil is present in an amount of about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15%.In some embodiments, the oil is present in an amount of at least about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14%. In some embodiments, the oil is present in an amount of up to about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15%.

[0049] In some embodiments, the oil comprises one or more triglycerides. In some embodiments, the triglycerides are medium chain triglycerides. In some embodiments, the medium chain triglycerides are C6-C 12 The fatty acid esters include those having a chain length of

[0050] In some embodiments, the triglyceride is present in an amount of about 1% to about 35% (w / w) of the composition. In some embodiments, the triglyceride is present in an amount of about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 1% to about 35%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 10% to about 15%, about The triglyceride is present in an amount of about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 25% to about 30%, about 25% to about 35%, or about 30% to about 35%. In some embodiments, the triglyceride is present in an amount of about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the triglyceride is present in an amount of at least about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, or about 30%. In some embodiments, triglycerides are present in an amount up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%.

[0051] In some embodiments, the lipid vesicles and / or the oil phase of the lipid vesicle portion of the composition contain a sterol. In some embodiments, the sterol is cholesterol. In some embodiments, the cholesterol can be plant-derived cholesterol. In some embodiments, the plant-derived cholesterol can be PhytoCho1®, SyntheCho1®, or any other plant-derived cholesterol (e.g., Avanti#700100), or any combination thereof. In some embodiments, the sterol can be a phytosterol, or a derivative thereof. In some embodiments, the phytosterol, or a derivative thereof can be phytosterol MM, Advasterol™ 90 IP or 95 IP F, NET Sterol-ISO, canola sterol, sitosterol 700095, lanosterol-95, brassicasterol, or any combination thereof.

[0052] In some embodiments, the sterol is present in an amount of about 1% to about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1% to about 3%, about 1% to about 4%, about 1% to about 5%, about 1.5% to about 2%, about 1.5% to about 2.5%, about 1.5% to about 3%, about 1.5% to about 4%, about 1.5% to about 5%, about 2% to about 2.5%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2.5% to about 3%, about 2.5% to about 4%, about 2.5% to about 5%, about 3% to about 4%, about 3% to about 5%, or about 4% to 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 4%, or about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of at least about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 4%, or about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of up to about 1.5%, about 2%, about 2.5%, about 3%, about 4%, or about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of about 2.6% (w / w) of the composition. In some embodiments, the ratio of the sterol to the vesicle-forming lipid is about 1:2 to about 1:5 (w / w). In some embodiments, the ratio of the sterol to the vesicle-forming lipid is about 1:2, 1:3, 1:4, or 1:5 (w / w).

[0053] Penetration enhancers In some embodiments, the lipid vesicle composition comprises one or more penetration enhancers that, when applied to the skin of an individual, act to increase the amount of penetration of the anionic polymeric material through one or more layers of the skin.

[0054] In some embodiments, the penetration enhancer is included in an oil-in-water emulsion of the composition, hi some embodiments, the penetration enhancer is included in the lipid bilayer of the composition.

[0055] There are many types of penetration enhancers that can be used, in some embodiments, the penetration enhancer comprises an ionic surfactant, a non-ionic surfactant, or a combination thereof.

[0056] In some embodiments, the penetration enhancer comprises a non-ionic surfactant or a combination of non-ionic surfactants. In some embodiments, the penetration enhancer is a single non-ionic surfactant. In some embodiments, the penetration enhancer is a combination of at least two, three, four, or more non-ionic surfactants. In some embodiments, the penetration enhancer is a combination of two non-ionic surfactants. In some embodiments, the penetration enhancer is a combination of three non-ionic surfactants.

[0057] In some embodiments, the nonionic surfactant or combination of nonionic surfactants is selected from polyethylene glycol ethers of fatty alcohols, sorbitan esters, polysorbates, sorbitan esters, and polyethylene glycol fatty acid esters, and combinations thereof.

[0058] In some embodiments, the nonionic surfactant comprises a polyethylene glycol (PEG) ether of an aliphatic alcohol. In some embodiments, the PEG ether of an aliphatic alcohol has from about 2 to about 8 PEG groups and a C 12 -C 22In some embodiments, the polyethylene glycol ether of the aliphatic alcohol comprises diethylene glycol hexadecyl ether, 2-(2-octadecaoxyethoxy)ethanol, diethylene glycol monooleyl ether, polyoxyethylene(2) oleyl ether, polyoxyethylene(3) oleyl ether, or polyoxyethylene(5) oleyl ether, or any combination thereof. In some embodiments, the polyethylene glycol ether of the aliphatic alcohol comprises 2-(2-octadecaoxyethoxy)ethanol. In some embodiments, the PEG ether of the aliphatic alcohol is super refined Brij® O2, or a derivative thereof.

[0059] In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5% to about 10%, about 0.5% to about 5%, about 0.5% to about 4%, or about 0.05% to about 3% (w / w) of the composition. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5% to about 2.5%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0060] In some embodiments, the nonionic surfactant comprises a sorbitan ester. In some embodiments, the sorbitan ester is a fatty acid ester. In some embodiments, the sorbitan ester is a C 12 -C 22A fatty acid ester. In some embodiments, the sorbitan ester comprises sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, or sorbitan isostearate, or any combination thereof. In some embodiments, the sorbitan ester comprises sorbitan monolaurate. In some embodiments, the sorbitan ester comprises sorbitan monopalmitate. In some embodiments, the sorbitan ester comprises sorbitan monostearate. In some embodiments, the sorbitan ester comprises sorbitan monooleate. In some embodiments, the sorbitan ester comprises sorbitan trioleate. In some embodiments, the sorbitan ester comprises sorbitan sesquioleate. In some embodiments, the sorbitan ester comprises sorbitan isostearate.

[0061] In some embodiments, the sorbitan ester is present in an amount of up to about 5% (w / w) of the composition. In some embodiments, the sorbitan ester is present in an amount of about 0.1% to about 0.5%. In some embodiments, the sorbitan ester is from about 0.1% to about 0.15%, from about 0.1% to about 0.2%, from about 0.1% to about 0.25%, from about 0.1% to about 0.3%, from about 0.1% to about 0.35%, from about 0.1% to about 0.4%, from about 0.1% to about 0.45%, from about 0.1% to about 0.5%, from about 0.15% to about 0.2%, from about 0.15% to about 0.25%, from about 0.15% to about 0.3%, from about 0.15% to about 0.35%, from about 0.15% to about 0.4%, from about 0.15% to about 0.45%, from about 0.15% to about 0.5%, from about 0.2% to about 0.25%, from about 0.2% to about 0.3%, from about 0.2% to about 0. The compound is present in an amount of about 0.35%, about 0.2% to about 0.4%, about 0.2% to about 0.45%, about 0.2% to about 0.5%, about 0.25% to about 0.3%, about 0.25% to about 0.35%, about 0.25% to about 0.4%, about 0.25% to about 0.45%, about 0.25% to about 0.5%, about 0.3% to about 0.35%, about 0.3% to about 0.4%, about 0.3% to about 0.45%, about 0.3% to about 0.5%, about 0.35% to about 0.4%, about 0.35% to about 0.45%, about 0.35% to about 0.5%, about 0.4% to about 0.45%, about 0.4% to about 0.5%, or about 0.45% to about 0.5%. In some embodiments, the sorbitan ester is present in an amount of from about 0.1%, about 0.15%, about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, or about 0.5%. In some embodiments, the sorbitan ester is present in an amount of at least about 0.1%, about 0.15%, about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, or about 0.45%. In some embodiments, the sorbitan ester is present in an amount of up to about 0.15%, about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, or about 0.5%.

[0062] In some embodiments, the sorbitan ester is present in an amount of about 0.1% to about 5%, 0.2% to about 5%, 0.3% to about 5%, 0.4% to about 5%, 0.5% to about 5%, about 0.5% to about 4%, or about 0.5% to about 3%. In some embodiments, the sorbitan ester is present in an amount of about 0.5% to about 2.5%. In some embodiments, the sorbitan ester is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the sorbitan esters are present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the sorbitan esters are present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the sorbitan esters are present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0063] In some embodiments, the non-ionic surfactant comprises a polysorbate. In some embodiments, the polysorbate comprises polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, or any combination thereof. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the polysorbate is polysorbate 20.

[0064] In some embodiments, the polysorbate is present in an amount up to about 5%. In some embodiments, the polysorbate is present in an amount of about 0.5% to about 5%, about 0.5% to about 4%, or about 0.5% to about 3% (w / w) of the composition. In some embodiments, the polysorbate is present in an amount of about 0.5% to about 2.5%. In some embodiments, the polysorbate is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the polysorbate is present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the polysorbate is present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the polysorbate is present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0065] In some embodiments, the non-ionic surfactant comprises a polyethylene glycol (PEG) fatty acid ester. In some embodiments, the PEG fatty acid ester comprises one C8-C hydroxyl group attached to each terminal hydroxyl to form a fatty acid ester. 22 The PEG chain is about 2-8 subunits containing fatty acids. In some embodiments, the PEG fatty acid ester includes PEG-8 dilaurate, PEG-4 dilaurate, PEG-4 laurate, PEG-8 dioleate, PEG-8 distearate, PEG-8 distearate, PEG-7 glyceryl cocoate, and PEG-20 almond glyceride, or any combination thereof. In some embodiments, the PEG fatty acid ester is PEG-4 dilaurate.

[0066] In some embodiments, the PEG fatty acid ester is present in an amount of up to about 5% (w / w) of the composition. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5% to about 5%, about 0.5% to about 4%, or about 0.5% to about 3%. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5% to about 2.5%. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the PEG fatty acid ester is present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the PEG fatty acid ester is present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the nonionic surfactant has a hydrophobic-lipophilic balance (HLB) of about 10 or less. In some embodiments, the nonionic surfactant comprises glyceryl monostearate (e.g., Cithrol GMS 40). In some embodiments, the nonionic surfactant comprises oleyl alcohol (e.g., Lipocol O-95). In some embodiments, the nonionic surfactant comprises a polyoxyethylene oleyl ether (e.g., Oleth-2). In some embodiments, the nonionic surfactant comprises a propylene glycol monocaprylate (e.g., Capryol® 90). In some embodiments, the composition comprises a plurality of nonionic surfactants, each having an HLB of about 10 or less.In some embodiments, the nonionic surfactant having an HLB of 10 or less is selected from Table 1 below, or any combination thereof.

[0067] [Table 1]

[0068] In some embodiments, the nonionic surfactant has a hydrophobic-lipophilic balance (HLB) of about 10 or greater. In some embodiments, the composition comprises multiple nonionic surfactants, each having an HLB of about 10 or greater.

[0069] In some embodiments, the nonionic surfactant or combination of nonionic surfactants is present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, the nonionic surfactant or combination of nonionic surfactants is from about 0.5% to about 1%, from about 0.5% to about 1.5%, from about 0.5% to about 2%, from about 0.5% to about 3%, from about 0.5% to about 4%, from about 0.5% to about 5%, from about 0.5% to about 6%, from about 0.5% to about 7%, from about 0.5% to about 8%, from about 0.5% to about 10%, from about 1% to about 1.5%, from about 1% to about 2%, from about 1% to about 3%, from about 1% to about 4%, from about 1% to about 5%, from about 1% to about 6%, from about 1% to about 7%, from about 1% to about 8%, from about 1% to about 10%, from about 1.5% to about 2%, from about 1.5% to about 3%, from about 1.5% to about 4%, from about 1.5% to about 5%, from about 1.5% to about 6%, About 1.5% to about 7%, about 1.5% to about 8%, about 1.5% to about 10%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2% to about 6%, about 2% to about 7%, about 2% to about 8%, about 2% to about 10%, about 3% to about 4%, about 3% to about 5%, about 3% to about 6%, about 3% to about 7%, about 3% to about 8%, about 3% to about 10% , about 4% to about 5%, about 4% to about 6%, about 4% to about 7%, about 4% to about 8%, about 4% to about 10%, about 5% to about 6%, about 5% to about 7%, about 5% to about 8%, about 5% to about 10%, about 6% to about 7%, about 6% to about 8%, about 6% to about 10%, about 7% to about 8%, about 7% to about 10%, or about 8% to about 10%. In some embodiments, the nonionic surfactant or combination of nonionic surfactants is present in an amount of about 0.5%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, or about 10%. In some embodiments, the nonionic surfactant is present in an amount of 1.4%. In some embodiments, the non-ionic surfactant or combination of non-ionic surfactants is present in an amount of at least about 0.5%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, or about 8%. In some embodiments, the non-ionic surfactant or combination of non-ionic surfactants is present in an amount of up to about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, or about 10%.

[0070] In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion, the lipid bilayer, or both. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion. In some examples, the composition comprises a non-ionic surfactant in the lipid bilayer. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion and the lipid bilayer, and the composition comprises two or more different non-ionic surfactants.

[0071] In some embodiments, the penetration enhancer comprises a salicylate ester or a nicotinate ester. In some embodiments, the ester is a C1-C6 alkyl ester or a benzyl ester. In some embodiments, the penetration enhancer comprises methyl salicylate or benzyl nicotinate. In some embodiments, the penetration enhancer is a nicotinate ester present in an amount of up to about 0.1%, 0.5%, 1%, 2%, or 3% (w / w) of the composition. In some embodiments, the nicotinate ester is present in an amount of about 0.1% to about 3%, about 0.1% to about 2%, or about 0.1% to about 1%.

[0072] Cationic Surfactants In some embodiments, the composition comprises an ionic surfactant. In some embodiments, the ionic surfactant is a cationic surfactant. In some embodiments, the cationic surfactant is a monocationic surfactant, a dicationic surfactant, or a polycationic surfactant.

[0073] In some embodiments, the monocationic surfactant is used in the composition to form a submicron emulsion prior to the formation of the final lipid vesicle composition provided herein (e.g., prior to the addition of lipid forming vesicles). In some embodiments, the monocationic surfactant is net monocationic (e.g., a phosphate salt containing two side chains, each with a single cationic functionality, partially neutralized by a phosphate anion).

[0074] In some embodiments, the monocationic surfactant is a fatty amide-derived propylene glycol-diammonium phosphate ester. A fatty amide-derived propylene glycol-diammonium phosphate ester is a phospholipid that contains at least one propylene glycol phosphoester bound to a quaternary ammonium group, which is then bound to a fatty acid amide. One non-limiting example of a fatty amide-derived propylene glycol-diammonium phosphate ester is linoleamidopropyl PG-dimonium chloride phosphate. Similar compounds with different fatty acid amide groups bound are also known. In some embodiments, a fatty amide-derived propylene glycol-diammonium phosphate ester has the following structure:

[0075] [ka] wherein n is an integer from 1 to 3, m is an integer from 0 to 2, the sum of m and n is 3, X is a cation selected from a proton, sodium, potassium, magnesium, and calcium, and R is a C8-C 30 (the acyl group of a fatty acid).

[0076] In some embodiments, the fatty acid is 12 -C 24 In some embodiments, the monocationic penetration enhancer is a fatty acid. In some embodiments, the fatty acid is an unsaturated fatty acid. In some embodiments, the fatty acid is linoleic acid. In some embodiments, the monocationic penetration enhancer is linoleamidopropyl PG-dimonium chloride phosphate (e.g., Arlasilk™ PTM, Arlasilk™ EFA).

[0077] In some embodiments, the fatty amide-derived propylene glycol-dimonium chloride phosphate ester is present in an amount of about 1% to about 10% (w / w) of the composition. In some embodiments, the fatty amide-derived propylene glycol-dimonium chloride phosphate ester is present in an amount of about 1% to about 2%, about 1% to about 3%, about 1% to about 4%, about 1% to about 5%, about 1% to about 6%, about 1% to about 7%, about 1% to about 8%, about 1% to about 9%, about 1% to about 10%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2% to about 6%, about 2% to about 7%, about 2% to about 8%, about 2% to about 9%, about 2% to about 10%, about 3% to about 4%, about 3% to about 5%, about 3% to about 6%, about 3% about 7%, about 3% to about 8%, about 3% to about 9%, about 3% to about 10%, about 4% to about 5%, about 4% to about 6%, about 4% to about 7%, about 4% to about 8%, about 4% to about 9%, about 4% to about 10%, about 5% to about 6%, about 5% to about 7%, about 5% to about 8%, about 5% to about 9%, about 5% to about 10%, about 6% to about 7%, about 6% to about 8%, about 6% to about 9%, about 6% to about 10%, about 7% to about 8%, about 7% to about 9%, about 7% to about 10%, about 8% to about 9%, about 8% to about 10%, or about 9% to about 10%. In some embodiments, the propylene glycol-dimonium chloride phosphate ester derived from a fatty amide is present in an amount of about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%. In some embodiments, the propylene glycol-dimonium chloride phosphate ester derived from a fatty amide is present in an amount of 7%. In some embodiments, the propylene glycol-dimonium chloride phosphate ester derived from a fatty amide is present in an amount of at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, or about 9%. In some embodiments, the propylene glycol-dimonium chloride phosphate ester derived from a fatty amide is present in an amount of up to about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%. In some embodiments, the propylene glycol-dimonium chloride phosphate ester derived from a fatty amide is present in an amount of at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%.

[0078] In some embodiments, the ratio of the non-ionic surface (e.g., propylene glycol monocaprylate) to the cationic penetration enhancer (e.g., linoleamidopropyl PG-dimonium chloride phosphate) is about 1:1 to about 1:10 (w / w). In some embodiments, the ratio of nonionic surface to cationic penetration enhancer is about 1:1 to 1:2, 1:1 to 1:3, 1:1 to 1:4, 1:1 to 1:5, 1:1 to 1:6, 1:1 to 1:7, 1:1 to 1:8, 1:1 to 1:9, 1:1 to 1:10, 1:2 to 1:3, 1:2 to 1:4, 1:2 to 1:5, 1:2 to 1:6, 1:2 to 1:7, 1:2 to 1:8, 1:2 to 1:9, 1:2 to 1:10, 1:3 to 1:4, 1:3 to 1:5, 1:3 to 1:6, 1:3 to 1:7, 1:3 to 1:8, 1:2 to 1:9, 1:2 to 1:10, 1:3-1:8, 1:3-1:9, 1:3-1:10, 1:4-1:5, 1:4-1:6, 1:4-1:7, 1:4-1:8, 1:4-1:9, 1:4-1:10, 1:5-1:6, 1:5-1:7, 1:5-1:8, 1:5-1:9, 1:5-1:10, 1:6-1:7, 1:6-1:8, 1:6-1:9, 1:6-1:10, 1:7-1:8, 1:7-1:9, 1:7-1:10, 1:8-1:9, 1:8-1:10, or 1:9-1:10 (w / w). In some embodiments, the ratio of non-ionic surface to cationic penetration enhancer is about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10 (w / w). In some embodiments, the ratio of non-ionic surface to cationic penetration enhancer is at least about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10 (w / w). In some embodiments, the ratio of non-ionic surface to cationic penetration enhancer is at most about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10 (w / w).

[0079] In some embodiments, the cationic surfactant is a dicationic penetration enhancer. In some embodiments, the dicationic surfactant is a gemini surfactant. In some embodiments, the gemini surfactant is a surfactant comprising two quaternary amines represented by the formula AN(R)2-BN(R)2-C, where each of A and C is independently an optionally substituted C6-C 24 alkyl group, each R is independently an optionally substituted C1-C6 alkyl, and B is an optionally substituted C2-C6 alkyl group. 10 In some embodiments, each of A and C is a C-C alkylene chain. 24 In some embodiments, each of A and C is a C6-C 24 In some embodiments, B is a saturated C-C hydrocarbon. In some embodiments, each R is methyl. In some embodiments, B is a saturated C-C 10 is an alkylene chain. In some cases, gemini surfactants follow the nomenclature XYZ, where X, Y, and Z are each integers representing the number of carbon atoms in each substituent, and Y is a spacer between two quaternary amines. Thus, for example, a 12-3-12 gemini surfactant has the formula CH3(CH2) 11 -[N + (CH3)2]-(CH2)3-[N + (CH3)2]-(CH2) 11 In some embodiments, the gemini surfactant is a 10-2-10, 12-2-12, 14-2-14, 10-3-10, 12-3-12, 14-3-14, 10-4-10, 12-4-12, or 14-4-14 gemini surfactant. In some embodiments, the gemini surfactant is a 12-3-12 gemini surfactant.

[0080] In some embodiments, the gemini surfactant is present in an amount of about 0.1% to about 1.5% (w / w) of the composition. In some embodiments, the gemini surfactant is present in an amount of about 0.1% to about 0.2%, about 0.1% to about 0.3%, about 0.1% to about 0.5%, about 0.1% to about 0.7%, about 0.1% to about 0.9%, about 0.1% to about 1%, about 0.1% to about 1.2%, about 0.1% to about 1.5%, about 0.2% to about 0.3%, about 0.2% to about 0.5%, about 0.2% to about 0.7%, about 0.2% to about 0.9%, about 0.2% to about 1%, about 0.2% to about 1.2%, about 0.2% to about 1.5%, about 0.3% to about 0.5%, about 0.3% to about 0.7%, about The compound is present in an amount of from about 0.3% to about 0.9%, from about 0.3% to about 1%, from about 0.3% to about 1.2%, from about 0.3% to about 1.5%, from about 0.5% to about 0.7%, from about 0.5% to about 0.9%, from about 0.5% to about 1%, from about 0.5% to about 1.2%, from about 0.5% to about 1.5%, from about 0.7% to about 0.9%, from about 0.7% to about 1%, from about 0.7% to about 1.2%, from about 0.7% to about 1.5%, from about 0.9% to about 1%, from about 0.9% to about 1.2%, from about 0.9% to about 1.5%, from about 1% to about 1.2%, from about 1% to about 1.5%, or from about 1.2% to about 1.5%. In some embodiments, the gemini surfactant is present in an amount of about 0.1%, about 0.2%, about 0.3%, about 0.5%, about 0.7%, about 0.9%, about 1%, about 1.2%, or about 1.5%. In some embodiments, the gemini surfactant is present in an amount of at least about 0.1%, about 0.2%, about 0.3%, about 0.5%, about 0.7%, about 0.9%, about 1%, or about 1.2%. In some embodiments, the gemini surfactant is present in an amount of up to about 0.2%, about 0.3%, about 0.5%, about 0.7%, about 0.9%, about 1%, about 1.2%, or about 1.5%.

[0081] In some embodiments, the cationic surfactant comprises a polycationic group. In some embodiments, the polycationic group is a polymer in which each monomer of the polymer comprises a charged group (e.g., an amino group). In some embodiments, the polycationic group is polylysine. In some embodiments, the polycationic group is polyarginine.

[0082] In some embodiments, the polylysine has a molecular weight of about 1 kDa to about 10 kDa, about 1 kDa to about 5 kDa, or about 3 kDa to about 5 kDa. In some embodiments, the polylysine is present in an amount of about 0.01% to about 1%, about 0.01% to about 0.5%, about 0.01% to about 0.2%, about 0.05% to about 1%, about 0.05% to about 0.5%, or about 0.05% to about 0.2% (w / w) of the composition.

[0083] Additional Ingredients In some embodiments, the vesicle composition includes additional components that, in some embodiments, improve one or more properties of the vesicle without dramatically altering the delivery of the anionic polymeric material.

[0084] In some embodiments, the vesicle composition further comprises one or more viscosity enhancing agents. In some embodiments, the viscosity enhancing agent thickens the composition to enhance the stability and / or feel of the vesicle composition to the user. In some embodiments, the viscosity enhancing agent further acts as a surfactant. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof. In some embodiments, the fatty alcohol is a C8-C 20 In some embodiments, the fatty alcohol is cetyl alcohol. In some embodiments, the cetyl alcohol is Crodacol C95. In some embodiments, the wax is a natural or synthetic wax. In some embodiments, the wax is beeswax. In some embodiments, the wax is synthetic beeswax. In some embodiments, the synthetic beeswax is Syncrowax™ BB4. In some embodiments, the synthetic beeswax is a non-animal derived beeswax. In some embodiments, the non-animal derived beeswax is Syncrowax™ SB1. In some embodiments, the fatty acid ester of glycerol is a monoester. In some embodiments, the monoester is a C8-C 24An ester of a fatty acid. In some embodiments, the fatty acid ester of glycerol is glycerol monostearate.

[0085] In some embodiments, the viscosity enhancing agent is present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, the viscosity enhancing agent is present in an amount of about 0.5% to about 5%, about 0.5% to about 5%, about 0.5% to about 4%, about 0.5% to about 3%, or about 0.5% to about 2%. In some embodiments, the viscosity enhancing agent comprises a fatty alcohol in an amount of up to about 2%, a wax in an amount of up to about 2%, and a fatty acid ester of glycerol in an amount of up to about 5%. In some embodiments, the fatty alcohol is present in an amount of about 0.1 to about 1.5%. In some embodiments, the wax is present in an amount of about 0.1% to about 1%. In some embodiments, the fatty acid ester of glycerol is present in an amount of about 0.5% to about 2%.

[0086] In some embodiments, the vesicle composition further comprises one or more of a thickening agent, a preservative, a moisturizer, an emollient, a moisturizing agent, or any combination thereof. In some embodiments, the vesicle composition further comprises a thickening agent. In some embodiments, the vesicle composition further comprises a preservative. In some embodiments, the vesicle composition further comprises a moisturizing agent. In certain embodiments, the vesicle composition further comprises an emollient. In some embodiments, the vesicle composition further comprises a moisturizing agent.

[0087] In some embodiments, the vesicle composition further comprises an antimicrobial agent. In some embodiments, the antimicrobial agent is a paraben ester. In some embodiments, the antimicrobial agent is methylparaben or propylparaben, or a combination thereof. In some embodiments, the antimicrobial agent is present in an amount of up to about 1%, up to about 0.9%, up to about 0.8%, up to about 0.7%, up to about 0.6%, up to about 0.5%, up to about 0.4%, up to about 0.3%, or about 0.2% (w / w) of the composition.

[0088] In some embodiments, the vesicle composition further comprises a thickening agent. In some embodiments, the thickening agent is an inert polymeric material. In some embodiments, the thickening agent is a siloxane polymer. In some embodiments, the thickening agent is polydimethylsiloxane (PDMS). In some embodiments, the PDMS is present in an amount of up to about 5%, up to about 4%, up to about 3%, up to about 2%, or up to about 1%. In some embodiments, the PDMS is present in an amount of about 0.1% to about 2% (w / w) of the composition.

[0089] In some embodiments, the composition further comprises a humectant. In some embodiments, the composition comprises glycerol. In some embodiments, the glycerol is present in an amount of about 0.5% to about 25%, about 0.5% to about 20%, about 0.5% to about 15%, or about 0.5% to about 10%. In some embodiments, the glycerol is present in an amount of about 1% to about 10%. In some embodiments, the glycerol is present in an amount of about 1% to about 2%, about 1% to about 4%, about 1% to about 6%, about 1% to about 8%, about 1% to about 10%, about 2% to about 4%, about 2% to about 6%, about 2% to about 8%, about 2% to about 10%, about 4% to about 6%, about 4% to about 8%, about 4% to about 10%, about 6% to about 8%, about 6% to about 10%, or about 8% to about 10%. In some embodiments, glycerol is present in an amount of about 1%, about 2%, about 4%, about 6%, 8%, or about 10%. In some embodiments, glycerol is present in an amount of at least about 1%, about 2%, about 4%, about 6%, or about 8%. In some embodiments, glycerol is present in an amount of up to about 2%, about 4%, about 6%, about 8%, or about 10% (w / w) of the composition.

[0090] In some embodiments, the vesicle composition includes a preservative. In some embodiments, the preservative is a cosmetic preservative such as Euxyl® PE 9010, or Spectrastat®. Euxyl® PE 9010 is a phenoxyethanol / ethylhexylglycerin mixture. Spectrastat® is a blend of caprylhydroxamic acid, caprylyl glycol, and glycerin. In some embodiments, the preservative is present in an amount of up to about 2%, up to about 1.5%, or up to about 1% (w / w) of the composition. In some embodiments, the preservative is present in an amount of about 0.1% to about 2%, about 0.1% to about 1.5%, or about 0.1% to about 1%. In some embodiments, the preservative is present in an amount of about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, or 1.5%.

[0091] In some embodiments, the additional component comprises purified water. In some embodiments, the purified water is present in an amount of about 50% to about 90% (w / w). In some embodiments, the purified water is present in an amount of about 50% to about 55%, about 50% to about 60%, about 50% to about 65%, about 50% to about 70%, about 50% to about 75%, about 50% to about 80%, about 50% to about 85%, about 50% to about 90%, about 55% to about 60%, about 55% to about 65%, about 55% to about 70%, about 55% to about 75%, about 55% to about 80%, about 55% to about 85%, about 55% to about 90%, about 60% to about 65%, about 60% to about 70%, about 60% to about In some embodiments, purified water is present in an amount of about 75%, about 60% to about 80%, about 60% to about 85%, about 60% to about 90%, about 65% to about 70%, about 65% to about 75%, about 65% to about 80%, about 65% to about 85%, about 65% to about 90%, about 70% to about 75%, about 70% to about 80%, about 70% to about 85%, about 70% to about 90%, about 75% to about 80%, about 75% to about 85%, about 75% to about 90%, about 80% to about 85%, about 80% to about 90%, or about 85% to about 90%. In some embodiments, purified water is present in an amount of about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or about 90%. In some embodiments, purified water is present in an amount of at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, or about 85%. In some embodiments, purified water is present in an amount of up to about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or about 90%.

[0092] Exemplary Compositions for Delivering Anionic Polymeric Materials Exemplary compositions for delivering anionic polymeric materials are provided below. The embodiments described below may further include any of the other ingredients or components provided herein.

[0093] Hyaluronic Acid Composition 1: In one aspect, provided herein is a lipid vesicle composition, the composition comprising: (a) lipid vesicles, each of which comprises a lipid bilayer comprising a vesicle-forming lipid, the vesicle-forming lipid being present in an amount of about 5% to about 20%; (b) oil-in-water emulsions encapsulated in lipid vesicles and stabilized by one or more surfactants, the one or more surfactants comprising cationic surfactants; oil-in-water emulsions; (c) comprising hyaluronic acid in an amount of about 0.1 mg / mL to about 10 mg / mL, encapsulated in a lipid bilayer and / or an oil-in-water emulsion.

[0094] In some embodiments, the oil component is present in an amount of about 2.5% to about 20%.

[0095] In some embodiments, the lipid vesicle composition comprises hyaluronic acid at about 0.01 mg / mL to about 0.05 mg / mL, at about 0.01 mg / mL to about 0.1 mg / mL, at about 0.01 mg / mL to about 0.5 mg / mL, at about 0.01 mg / mL to about 1 mg / mL, at about 0.01 mg / mL to about 1.25 mg / mL, at about 0.01 mg / mL to about 1.5 mg / mL, at about 0.01 mg / mL to about 1.75 mg / mL, at about 0.01 mg / mL to about 2 mg / mL, at about 0.01 mg / mL to about 5 mg / mL, at about 0.01 mg / mL to about 10 mg / mL, at about 0.1 mg / mL to about 0. Contains an amount of about 5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 1.25 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.1 mg / mL to about 1.75 mg / mL, about 0.1 mg / mL to 2 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 1 mg / mL, about 1 mg / mL to about 1.5 mg / mL, about 1 mg / mL to 1.75 mg / mL, about 1 mg / mL to about 2 mg / mL, about 1 mg / mL to about 5 mg / mL, about 1 mg / mL to about 10 mg / mL. In some embodiments, the lipid vesicle composition comprises hyaluronic acid in an amount of about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL.

[0096] In some embodiments, the lipid vesicle composition further comprises a viscosity enhancing agent in an amount of about 0.5% to about 5%. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof.

[0097] In some embodiments, the lipid vesicle composition further comprises a non-ionic surfactant in an amount of about 0.1% to about 3%. In some embodiments, the non-ionic surfactant is a PEG ether of an aliphatic alcohol.

[0098] In some embodiments, the cationic surfactant is a propylene glycol-diammonium phosphate ester derived from a fatty amide, hi some embodiments, the cationic surfactant is present in an amount of about 1% to about 10%.

[0099] In some embodiments, the lipid vesicle composition further comprises a peptide antagonist of the muscle-type nicotinic acetylcholine receptor encapsulated in the lipid bilayer and / or oil-in-water emulsion in an amount of about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 3 mg / mL, about 0.1 mg / mL to about 4 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, or about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, or about 5 mg / mL.

[0100] Hyaluronic Acid Composition 2: In one aspect, provided herein is a lipid vesicle composition, the composition comprising: (a) lipid vesicles, each of which comprises a lipid bilayer comprising a vesicle-forming lipid, the vesicle-forming lipid being present in an amount of about 2% to about 20%; (b) oil-in-water emulsions encapsulated in lipid vesicles and stabilized by one or more surfactants; (c) hyaluronic acid in an amount of about 0.1 mg / mL to about 10 mg / mL encapsulated in a lipid bilayer and / or an oil-in-water emulsion, a gemini surfactant in an amount of about 0.01% to about 0.5%, and Further included is polysorbate in an amount of about 0.1% to about 2%.

[0101] In some embodiments, the oil component is present in an amount of about 2.5% to about 20%.

[0102] In some embodiments, the lipid vesicle composition comprises hyaluronic acid at about 0.01 mg / mL to about 0.05 mg / mL, at about 0.01 mg / mL to about 0.1 mg / mL, at about 0.01 mg / mL to about 0.5 mg / mL, at about 0.01 mg / mL to about 1 mg / mL, at about 0.01 mg / mL to about 1.25 mg / mL, at about 0.01 mg / mL to about 1.5 mg / mL, at about 0.01 mg / mL to about 1.75 mg / mL, at about 0.01 mg / mL to about 2 mg / mL, at about 0.01 mg / mL to about 5 mg / mL, at about 0.01 mg / mL to about 10 mg / mL, at about 0.1 mg / mL to about 0. Contains an amount of about 5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 1.25 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.1 mg / mL to about 1.75 mg / mL, about 0.1 mg / mL to 2 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 1 mg / mL, about 1 mg / mL to about 1.5 mg / mL, about 1 mg / mL to 1.75 mg / mL, about 1 mg / mL to about 2 mg / mL, about 1 mg / mL to about 5 mg / mL, about 1 mg / mL to about 10 mg / mL. In some embodiments, the lipid vesicle composition comprises hyaluronic acid in an amount of about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL.

[0103] In some embodiments, the lipid vesicle composition further comprises a viscosity enhancing agent in an amount of about 0.5% to about 5%. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof.

[0104] In some embodiments, the polysorbate is polysorbate 80.

[0105] In some embodiments, the lipid vesicle composition further comprises a peptide antagonist of the muscle-type nicotinic acetylcholine receptor encapsulated in the lipid bilayer and / or oil-in-water emulsion in an amount of about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 3 mg / mL, about 0.1 mg / mL to about 4 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, or about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, or about 5 mg / mL.

[0106] Lipid vesicle compositions of muscle-type nicotinic acetylcholine receptor antagonist peptides for intradermal delivery - Patent Application 20070233334 In one aspect, provided herein is a lipid vesicle composition comprising a peptide antagonist of muscle-type nicotinic acetylcholine receptor. In some embodiments, the lipid vesicle composition comprises lipid vesicles comprising lipid bilayers, each comprising vesicle-forming lipids. In some embodiments, the lipid vesicle composition comprises an oil-in-water emulsion encapsulated in the lipid vesicle. In some embodiments, the oil-in-water emulsion is stabilized by one or more surfactants. In some embodiments, the peptide antagonist is encapsulated in the lipid bilayer and / or the oil-in-water emulsion. In some embodiments, the peptide antagonist is encapsulated in the lipid bilayer. In some embodiments, the peptide antagonist is encapsulated in the oil-in-water emulsion.

[0107] Muscle-type nicotinic acetylcholine receptors In one aspect, the present disclosure relates to lipid vesicle compositions comprising peptide antagonists of the muscle-type nicotinic acetylcholine receptor, also referred to as muscle nAChR.

[0108] Muscle nAChRs are ligand-activated ion channel receptors with a structure commonly described as a heteropentamer of four related but genetically and immunologically distinct subunits. The subunits are organized around a central pore in the membrane with a stoichiometry of two α subunits and one each of β, δ and γ. Muscle nAChRs are activated by the endogenous neurotransmitter acetylcholine (ACh, the natural receptor agonist), which is released by nerves at the neuromuscular junction. ACh binds to the receptor, resulting in the transduction of a signal for channel activation or gating.

[0109] The peptide antagonists of the present disclosure bind at the active site of muscle nAChR and inhibit ACh binding to the receptor. As a result, the neuromuscular postsynaptic membrane is depolarized and blocked so that signals from nerves (ACh release) are no longer effective in stimulating muscle contraction. See, for example, Albuquerque, et al., 2009, "Mammalian Nicotinic Acetylcholine Receptors: From Structure to Function," Physiol.Rev.89(1):73-120, and Kalamida, et al., 2007, "Muscle and neuronal nicotinic acetylcholine receptors," The FEBS Journal 274:3799-3845, each of which is incorporated herein by reference in its entirety.

[0110] Intentional muscle denervation has been achieved using the following anticholinergic botulinum toxin products: onabotulinumtoxinA (BTX-A, sold as BOTOX®), abotulinumtoxinA (Dysport®), incobotulinumtoxinA (Xeomin®), rimabotulinumtoxinB (Myobloc®) and prabotulinumtoxinA-xvfs (Jeuveau®). BTX-A prevents the release of ACh, present in neuronal vesicles, from synaptic neurons. The result is the absence of ACh at the synapse and the inability to innervate muscle cells. Thus, the mechanism of action of these toxins is presynaptic. Botulinum toxins are indicated for use, for example, in preventing or improving the appearance of skin wrinkles, skin laxity, moderate to severe glabellar lines associated with corrugator supercilii and / or procerus muscle activity, moderate to severe lateral canthal lines (crow's feet lines) associated with orbital corneal activity, moderate to severe forehead lines associated with frontalis muscle activity, treatment of overactive bladder (OAB), treatment of urinary incontinence, prevention of headaches in adult patients suffering from chronic migraine, prevention or treatment of episodic migraine, treatment of upper and lower limb spasticity, treatment of cervical dystonia, treatment of hypersalivation (also called hypersalivation or sialorrhea), treatment of blepharospasm associated with dystonia, and treatment and treatment of strabismus. (See, e.g., Botox Cosmetic BLA 103000, Botox Cosmetic Product Insert Revision 5 / 2018, Botox Product Insert Revision 4 / 2017, Dysport BLA 125274, Dysport Product Insert Revision 6 / 2017, Xeomin BLA 125360, Xeomin Product Insert Revision 7 / 2018, Myobloc Product Insert Revision 8 / 2019, and Jeuveau BLA 761085 Product Insert Revision 7 / 2019, each of which is incorporated herein by reference.)

[0111] In contrast, the peptide antagonists of the present disclosure occupy the ACh active site on the muscle cell AChR (postsynaptic). Upon binding, the peptide antagonists of the present disclosure block the binding of ACh secreted from the neuron.

[0112] Peptide antagonists of muscle-type nicotinic acetylcholine receptors The present disclosure provides lipid vesicle compositions comprising peptide antagonists of mammalian muscle nAChRs, including human muscle nAChRs. In some embodiments, the peptide antagonists provided herein have desirable or improved properties compared to muscle nAChR antagonists known in the art. Such properties include, for example, pharmacokinetic properties (including, but not limited to, absorption, bioavailability, distribution, metabolism, and excretion), pharmacodynamic properties (including, but not limited to, receptor binding properties, e.g., binding half-life, post-receptor effects, and chemical interactions), enhanced activity (e.g., IC 50 The properties of the peptide antagonists may include their activity (e.g., as represented by half-life), stability (e.g., as represented by half-life), solubility (e.g., in a formulation), or permeability (e.g., permeability of the skin by a formulation containing the peptide antagonist). In some embodiments, a formulation containing a peptide antagonist of the present disclosure has desirable or improved properties compared to a formulation containing a muscle nAChR antagonist known in the art. In some embodiments, the desirable or improved properties of the formulation of the present disclosure are properties related to the use of the formulation for an indication as described elsewhere herein, e.g., for reducing or improving the appearance of wrinkles in the skin.

[0113] Peptide antagonists In some embodiments, the peptide antagonist of the lipid vesicle composition comprises a conotoxin peptide. In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist comprises an amino acid sequence identical to the amino acid sequence of any one of SEQ ID NOs: 1-52 or 60-99. In some embodiments, the peptide antagonist has an amino acid sequence identical to any one of SEQ ID NOs: 1-52 or 60-99.

[0114] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:1. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:1. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:1. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:1.

[0115] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:3. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:3. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:3. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:3.

[0116] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:60. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:60. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:60. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:60.

[0117] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:61. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:61. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:61. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:61.

[0118] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO: 73. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO: 73. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO: 73. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO: 73.

[0119] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:78. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:78. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:78. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:78.

[0120] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:82. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:82. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:82. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:82.

[0121] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:85. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:85. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:85. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:85.

[0122] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:91. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:91. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:91. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:91.

[0123] In some embodiments, the peptide antagonist comprises an amino acid sequence with at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% sequence homology to the amino acid sequence of SEQ ID NO:95. In some embodiments, the peptide antagonist comprises one or more amino acid substitutions relative to SEQ ID NO:95. In some embodiments, the one or more amino acid substitutions are selected from the amino acids defined as Xaa1-Xaa14. In some embodiments, the peptide antagonist comprises 1, 2, 3, 4, or 5 amino acid substitutions relative to SEQ ID NO:95. In some embodiments, at least 1, 2, or 3 of the 1, 2, 3, 4, or 5 amino acid substitutions are conservative substitutions. In some embodiments, the peptide antagonist has an amino acid sequence consisting of the identical sequence of SEQ ID NO:95.

[0124] In some embodiments, the peptidic antagonist comprises up to about 20 amino acids, up to about 18 amino acids, up to about 16 amino acids, up to about 14 amino acids, In some embodiments, the peptidic antagonist has a molecular weight of up to about 2500 Da, up to about 2200 Da, up to about 2000 Da, up to about 1800 Da, up to about 1700 Da, up to about 1600 Da, up to about 1500 Da.

[0125] In some embodiments, the muscle-type nicotinic acetylcholine receptor peptide antagonist of the lipid vesicle composition has 12-14 residues and is

[0126] comprising the amino acid sequence Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14,

[0127] During the ceremony,

[0128] Xaa1 is absent or selected from Ala, Gly, Val, Leu, Ile, and derivatives of Ala, Gly, Val, Leu, or Ile;

[0129] Xaa2 is selected from absent or Asn, Asp, Gln, Glu, Arg, His, Lys, Phe, Trp, Tyr, Ala, Gly, Val, Leu, He, and derivatives of Asn, Asp, Gln, Glu, Arg, His, Lys, Phe, Trp, Tyr, Ala, Gly, Val, Leu, or He;

[0130] Xaa3 and Xaa8 form the bond Xaa3-Xaa8;

[0131] Xaa4 and Xaa14 form the bond Xaa4-Xaa14;

[0132] Xaa5 is selected from Asn, Asp, Gln, Glu, Arg, His, Lys, and derivatives of Asn, Asp, Gln, Glu, Arg, His, or Lys;

[0133] Xaa6 is selected from Pro and derivatives thereof;

[0134] Xaa7 is selected from Ala, Gly, Val, Leu, Ile, and derivatives of Ala, Gly, Val, Leu, or Ile;

[0135] Xaa9 is selected from Ala, Gly, Val, Leu, Ile, and derivatives of Ala, Gly, Val, Leu, or Ile;

[0136] Xaa10 is selected from Arg, His, Lys, and derivatives of Arg, His, or Lys;

[0137] Xaa11 is selected from Asn, Asp, Gln, Glu, Arg, His, Lys, and derivatives of Asn, Asp, Gln, Glu, Arg, His, or Lys;

[0138] Xaa12 is selected from Phe, Trp, Tyr, and derivatives of Phe, Trp, or Tyr;

[0139] Xaa13 is selected from Cys, Met, Sec, Ser, Thr, Arg, His, Lys, and derivatives of Cys, Met, Sec, Ser, Thr, Arg, His, or Lys;

[0140] the N-terminus is optionally modified; and

[0141] The C-terminus is optionally modified.

[0142] In some embodiments, the muscle-type nicotinic acetylcholine receptor peptide antagonist of the lipid vesicle composition has 12-14 residues and is as follows:

[0143] comprising the amino acid sequence Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14,

[0144] During the ceremony,

[0145] Xaa1 is absent;

[0146] Xaa2 is absent;

[0147] Xaa3 and Xaa8 form the bond Xaa3-Xaa8;

[0148] Xaa4 and Xaa14 form the bond Xaa4-Xaa14;

[0149] Xaa5 is selected from Asp, Gln, Glu, Arg, His, and Lys;

[0150] Xaa6 is selected from Pro and hydroxyproline;

[0151] Xaa7 is selected from Ala, Gly, Val, Leu, and Ile;

[0152] Xaa9 is selected from Ala, Gly, Val, Leu, and Ile;

[0153] Xaa10 is selected from Arg and His;

[0154] Xaa11 is selected from Asn, Asp, Gln, Glu, Arg, His, and Lys;

[0155] Xaa12 is selected for Trp and Tyr;

[0156] Xaa13 is selected from Cys, Met, Sec, Ser, Thr, Arg, His, and Lys;

[0157] the N-terminus is optionally modified; and

[0158] The C-terminus is optionally modified.

[0159] In some embodiments, the peptide antagonist of the present disclosure has the following amino acid sequence:

[0160] does not consist of Glu-Cys-Cys-Asn-Pro-Ala-Cys-Gly-Arg-His-Tyr-Ser-Cys (SEQ ID NO: 1),

[0161] Here, the first and third cysteine ​​residues (Xaa3-Xaa8) are linked and the second and fourth cysteine ​​residues (Xaa4-Xaal4) are linked (α-conotoxin GI, or CGI).

[0162] In some embodiments, the peptide antagonist of the present disclosure has the following amino acid sequence:

[0163] does not consist of Glu-Cys-Cys-Asn-Pro-Ala-Cys-Gly-Lys-His-Phe-Ser-Cys (SEQ ID NO: 2),

[0164] Here, the first and third cysteine ​​residues (Xaa3-Xaa8) are linked and the second and fourth cysteine ​​residues (Xaa4-Xaal4) are linked.

[0165] In some embodiments, the peptide antagonist of the present disclosure has the following amino acid sequence:

[0166] Glu-Cys-Cys-His-Pro-Ala-Cys-Gly-Lys-His-Phe-Ser-Cys (SEQ ID NO: 56),

[0167] Here, the first and third cysteine ​​residues (Xaa3-Xaa8) are linked and the second and fourth cysteine ​​residues (Xaa4-Xaal4) are linked (α-conotoxin GII sequence).

[0168] In some embodiments, the peptide antagonist of the present disclosure has the following amino acid sequence:

[0169] does not consist of Gly-Arg-Cys-Cys-His-Pro-Ala-Cys-Gly-Lys-Asn-Tyr-Ser-Cys (SEQ ID NO: 3),

[0170] Here, the first and third cysteine ​​residues (Xaa3-Xaa8) are linked and the second and fourth cysteine ​​residues (Xaa4-Xaal4) are linked (α-conotoxin MI, or CMI).

[0171] In some embodiments, the number of amino acid residues in the peptide antagonists of the disclosure is no more than 12, no more than 13, or no more than 14. In embodiments, the peptide antagonists of the disclosure consist of 12, 13, or 14 amino acid residues.

[0172] Non-limiting examples of peptide antagonists of the present disclosure are shown in Table 2.

[0173] [Table 2-1]

[0174] [Table 2-2]

[0175] 1 It contains a cystathionine (Cyt-Cyt) bond at (Xaa3-Xaa8). 2 (Xaa4-Xaal4) containing a cystathionine (Cyt-Cyt) bond. 3 (Xaa3-Xaa8) contains a disulfide (Cys-Cys) bond. 4 (Xaa4-Xaal4) contains a disulfide (Cys-Cys) bond. 5 (Xaa3-Xaa8) contains a Sec-Sec bond. 6 (Xaa4-Xaal4) including the Sec-Sec bond. All refer to the Xaal-Xaal4 numbering provided herein.

[0176] Unless otherwise indicated in the table, the peptides listed in Table 1 may contain all L-amino acids or all D-amino acids.

[0177] restraint structure

[0178] In some embodiments, the peptide antagonists of the present disclosure include a constrained structure, including, but not limited to, a bond, a cross-link, or a ligation of any means between residues at two positions. In some embodiments, the peptide is constrained by its termini or by a position within the peptide, or both. In some embodiments, the constrained structure enhances peptide antagonist properties, such as pharmacokinetic properties (including, but not limited to, absorption, bioavailability, distribution, metabolism, and excretion), pharmacodynamic properties (including, but not limited to, receptor binding properties, such as binding half-life, post-receptor effects, and chemical interactions), enhanced activity (e.g., IC 50The constrained structure may affect the stability (e.g., as represented by the half-life), solubility (e.g., in a formulation), or permeability (e.g., permeability of the skin by a formulation containing the peptide antagonist). In certain embodiments, the constrained structure enhances the stability of the peptide antagonist. In certain embodiments, the constrained structure enhances the permeability of the peptide antagonist through the skin. In certain embodiments, the constrained structure enhances the solubility of the peptide antagonist in a formulation, e.g., a topical formulation.

[0179] In embodiments, the peptide antagonist constrained as described herein is referred to as a macrocyclic peptide or structure. A macrocyclic peptide refers to a linear peptide closed ring structure formed in a molecule by a bond between two positions in the peptide, and is referred to as a bonded amino acid, bonded amino acid derivative, bonded molecule, bonded moiety, bonded residue, bonded entity, etc., as appropriate. Two bonded amino acids, bonded amino acid derivatives, bonded molecules, bonded moieties, bonded residues, or bonded entities are separated from each other by two or more amino acid residues, are directly bonded to each other, or are bonded via a linker, etc.

[0180] In embodiments, the bonds of the peptide antagonists of the present disclosure are formed by two linked amino acids, linked amino acid derivatives, linked molecules, linked moieties, linked residues, or linked entities linked to each other by, for example, a disulfide bond, a peptide bond, an alkyl bond, an alkenyl bond, an ester bond, a thioester bond, an ether bond, a thioether bond, a phosphonate ether bond, an azo bond, a C--S--C bond, a C=N--C bond, a C=N--C bond, an amide bond, a lactam bridge, a carbamoyl bond, a urea bond, a thiourea bond, an amine bond, a thioamide bond, etc. Macrocyclization may be formed by a bond between the N-terminal and C-terminal amino acids of the peptide, by a bond between a terminal amino acid and a non-terminal amino acid, or by a bond between non-terminal amino acids.

[0181] For convenience, reference to a particular amino acid involved in a bond can use the nomenclature of the non-bonded amino acid (e.g., the structure it may have prior to the formation of the bond). It is also understood that a particular bond, e.g., a synthetic bond, may not be formed by connecting two amino acids or derivatives that are commonly referred to in the art. Thus, reference to a bonded amino acid herein can use the most closely approximating word to describe each participating chemical entity at a given residue position in the peptide antagonist. Correspondingly, the bonded entities in a peptide sequence, e.g., Xaa3, Xaa4, Xaa8, and Xaal4, can be referred to as bonded amino acids, but they are not the amino acids that are commonly referred to in the art. In some embodiments, Xaa3 and Xaa8, and Xaa4 and Xaal4, in the case of linked entities (e.g., forming Xaa3-Xaa8 and Xaa4-Xaal4 bonds), may be referred to as linked amino acids, linked amino acid derivatives, linked molecules, linked moieties, linked residues, or alternatively linked entities. These terms may be used to refer to amino acids, molecules, moieties, residues, or entities present at either Xaa3, Xaa4, Xaa8, or Xaal4, and are alternatively either linked or unlinked. For example, when not linked in the peptide antagonists of the present disclosure, but intended to be linked, the two linked amino acids may also be referred to as linked (or bond-forming) amino acids, linked (or bond-forming) amino acid derivatives, linked (or bond-forming) molecules, linked (or bond-forming) moieties, linked (or bond-forming) residues, or alternatively linked (or bond-forming) entities. When linked, the two linked amino acids may alternatively be referred to as linked (or bond-forming) amino acids, linked (or bond-forming) amino acid derivatives, linked (or bond-forming) molecules, linked (or bond-forming) moieties, linked (or bond-forming) residues, or linked (or bond-forming) entities.When not bound and not intended to be bound, the two amino acids may be referred to as non-bound (or non-bond-forming) amino acids, non-bound (or non-bond-forming) amino acid derivatives, non-bound molecules, non-bound moieties, non-bound residues, or non-bound entities. In some embodiments, the disclosed peptide antagonists are not bound. Each residue at a non-bound amino acid position in the disclosed peptide antagonists may be referred to as an amino acid, amino acid derivative, molecule, moiety, residue, or entity, or may be referred to as a non-bound (or non-bond-forming) amino acid, non-bound (or non-bond-forming) amino acid derivative, non-bound (or non-bond-forming) molecule, non-bound (or non-bond-forming) moiety, non-bound (or non-bond-forming) residue, or non-bound (or non-bond-forming) entity.

[0182] Any constraint structure known to those skilled in the art is contemplated for linking residues. Examples of constraint structures and their respective linking residues include, but are not limited to, bonds or bridges selected from the following: disulfide bridges (e.g., Cys-Cys bond, where each linking amino acid is Cys), Sec-Sec bond (selenocysteine ​​bond, where each linking amino acid is selenocysteine), cystathionine bond or bridge (e.g., Ser-homocysteine ​​bond), lactam bridge (e.g., Asp-Lys or Glu-Lys bond), also referred to herein as Cyt-Cyt (e.g., CH2-CH2-S-CH2), thioether bond (e.g., Cys-lanthionine bond, including but not limited to dehydroalanine or methyl variants), and dicarba bond (e.g., bond of olefin-containing amino acids, e.g., allylglycine or prenylglycine). In some embodiments, the linkage is selected from a disulfide bridge with linkage residues Cys-Cys, a selenocysteine ​​linkage with linkage residues Sec-Sec, a cystathionine linkage with linkage residues Ser-homocysteine, a lactam bridge with residues Asp-Lys or Glu-Lys, a lanthionine linkage with linkage residues Cys-dehydroalanine or methyl variants, and a dicarba linkage with linkage residues allylglycine or prenylglycine. In embodiments, the linkage amino acid, linkage amino acid derivative, linkage molecule, linkage moiety, linkage residue, or linkage entity is selected from Cys, Sec, Ser, homocysteine, Asp, Lys, Glu, dehydroalanine, or an olefin-containing amino acid (e.g., allylglycine or prenylglycine).

[0183] In some embodiments, each of the Xaa3-Xaa8 and Xaa4-Xaal4 bonds of the peptide antagonists of the present disclosure is a bond independently selected from a disulfide bridge formed by two Cys linking residues, a Sec-Sec bond formed by two selenocysteine ​​linking residues, a cystathionine bond formed by Ser and a homocysteine ​​linking residue, a lactam bridge formed by Asp and Lys linking residues or Glu and Lys linking residues, a thioether bond that is a lanthionine bond formed by Cys and a dehydroalanine or methyl variant residue, a dicarba bond formed by an olefin-containing linking residue, such as an allylglycine or prenylglycine linking residue, or any of these bonds formed by linking residues as known and described in the art. In some embodiments, the Xaa3-Xaa8 or Xaa4-Xaal4 bonds are the same as or different from each other.

[0184] (For example, Knerr et al., 2011, “Synthesis and activity of thioether-containing analogues of the complement inhibitor compstatin,” ACS Chem Biol. 6(7):753-760; DiMarco et al., 2006, “Discovery of novel, highly potent and selective b-hairpin mimetic CXCR4 inhibitors with excellent anti-HIV activity and pharmacokinetic profiles”, Bioorganic & Medicinal Chemistry 14:8396-8404;Dekan et al.,2011,“α-Conotoxin ImI incorporating stable cystathionine bridges maintains full potency and identical three-dimensional structure,”J.Am.Chem.Soc.2011,133:15866-15869;Nguyen and Wong,2017,“Making circles:recent advance in chemical and enzymatic approaches in peptide macrocyclization,”Journal of Biochemistry and Chemical Sciences 1(1):1-13; Tam and Wong, 2012, "Chemical Synthesis of Circular Proteins," The Journal of Biological Chemistry 287(32):27020-27025, each of which is incorporated by reference herein in its entirety.) In some embodiments, any suitable constrained structure suitable for use with a linking residue known in the art is contemplated for use in the peptide antagonists of the present disclosure.

[0185] In some embodiments, a particular constraint structure is selected based on its resistance to degradation, for example degradation caused by reduction of disulfide bond constraint structure. In some embodiments, the peptide antagonist comprises a constraint structure that resists degradation by reduction. For example, in a reducing environment, disulfide bond may be susceptible to degradation and the resulting loss of activity or other desired peptide antagonist properties. In some embodiments, cystathionine bond or at least two C1-C6 heterocycloalkyl rings confer increased stability compared to disulfide bond.

[0186] In some embodiments, two amino acids in the chain are joined by a bond to create a macrocyclic ring structure. In some embodiments, the bond mimics a hairpin turn in a peptide. In some embodiments, the bond comprises a covalent bond between standard or non-standard amino acids, such as a cystathionine bond, a lactam bridge, or a thioether bridge (e.g., a lanthionine bond). In some embodiments, the bond comprises a dipeptide. In some embodiments, the bond comprises a covalent bond between standard or non-standard amino acids, such as a lanthionine or methyllanthionine bond. In some embodiments, the bond comprises at least one aromatic or non-aromatic ring. In some embodiments, the bond comprises at least one cycloalkyl ring. In some embodiments, the bond comprises at least one heterocyclic ring. In some embodiments, the bond comprises at least two heterocyclic rings. In some embodiments, the bond comprises at least one nitrogen-containing heterocycloalkyl ring.

[0187] In some embodiments, the bond is

[0188] [ka] wherein A and B are heterocycles. In some embodiments, the bond is

[0189] [ka] wherein A and B are heterocycles.

[0190] In some embodiments, the bond comprises pyrrolidine, piperidine, dehydropyrrolidine, dehydropiperidine, aziridine, azetidine, oxazolidine, or thiazolidine. In some embodiments, the bond comprises two C1-C6 heterocycloalkyl rings. In some embodiments, the bond comprises at least one 5-membered heterocycloalkyl ring. In some embodiments, the bond comprises at least one 6-membered heterocycloalkyl ring. In some embodiments, the bond comprises two 5-membered heterocycloalkyl rings. In some embodiments, the bond comprises two 5-membered heterocycloalkyl rings, each ring comprising at least one nitrogen atom. In some embodiments, the bond comprises two 5-membered heterocycloalkyl rings, at least one ring comprising at least one nitrogen atom. In some embodiments, the bond comprises two 6-membered heterocycloalkyl rings. In some embodiments, the bond comprises two C1-C6 heterocycloalkyl rings connected by an amide bond. In some embodiments, the bond comprises two C1-C6 heterocycloalkyl rings connected by -C(=O)NH-. In some embodiments, the bond comprises two pyrrolidine rings. In some embodiments, the bond comprises at least one non-standard amino (unnatural) acid residue. In some embodiments, the bond comprises two (standard or non-standard) amino acids, the first amino acid having an (S) configuration at the alpha position and the second amino acid having an (R) configuration at the alpha position. In some embodiments, the bond comprises two (standard or non-standard) amino acids connected by a peptide bond. In some embodiments, the bond comprises two proline residues (diproline bond). In some embodiments, the bond comprises two proline residues connected by a peptide bond. In some embodiments, the bond comprises D-proline and L-proline (D-proline-L-proline, or L-proline-D-proline).

[0191] In some embodiments, the linkage comprises D-proline and L-proline, or a derivative thereof. In some embodiments, such derivatives comprise substitution of proline into the pyrrolidine ring. In some embodiments, the linkage comprises a non-standard amino acid residue selected from 3-fluoroproline, 4-fluoroproline, 3-hydroxyproline, 4-hydroxyproline, 3-aminoproline, 4-aminoproline, 3,4-dehydroproline, aziridine-2-carboxylic acid, azetidine-2-carboxylic acid, pipecolic acid, 4-oxa-proline, 3-thiaproline, or 4-thiaproline. In some embodiments, the linkage comprises two amino acids selected from proline, 3-fluoroproline, 4-fluoroproline, 3-hydroxyproline, 4-hydroxyproline, 3-aminoproline, 4-aminoproline, 3,4-dehydroproline, aziridine-2-carboxylic acid, azetidine-2-carboxylic acid, pipecolic acid, 4-oxa-proline, 3-thiaproline, or 4-thiaproline.

[0192] In some embodiments, the bond comprises a covalent bond between a standard or non-standard amino acid lactam bridge.

[0193] [ka] Includes the structure of.

[0194] In some embodiments, the bond comprises a covalent bond between a standard or non-standard amino acid thioether bridge.

[0195] [ka] Includes the structure of.

[0196] These and similar constraint structures can be used to link residues at terminal and / or non-terminal positions in the peptide. In some embodiments, Xaa3 and Xaa8 of the peptide antagonists of the present disclosure are linked. In some embodiments, Xaa4 and Xaal4 of the peptide antagonists of the present disclosure are linked. In some embodiments, Xaa3 and Xaa8, and Xaa4 and Xaal4 of the peptide antagonists of the present disclosure are linked.

[0197] Bond Spacing In some embodiments, the constrained structures as described herein are selected based on the spatial separation that occurs between the constrained residues. In some embodiments, the spatial separation affects the peptide antagonist properties described above. The peptide antagonists of the present disclosure can include a constrained structure that imparts a spatial separation of about 3.5 to about 10 angstroms between the alpha carbons of two linked amino acid residues or between the geometric centers of two linked residues (e.g., amino acid derivatives). In some embodiments, the spatial separation between the alpha carbons of two linked amino acid residues or between the geometric centers of two linked residues is about 3.5 angstroms to about 10 angstroms. In some embodiments, the spatial separation between the alpha carbons of two linked amino acid residues or between the geometric centers of two linked residues is at least about 3.5 angstroms. In some embodiments, the spatial separation between the alpha carbons of two linked amino acid residues or between the geometric centers of two linked residues is at most about 10 angstroms. In some embodiments, the spatial separation between the alpha carbons of two binding amino acid residues, or the spatial separation between the geometric centers of two binding residues, is between about 3.5 angstroms and about 4.5 angstroms, between about 3.5 angstroms and about 5 angstroms, between about 3.5 angstroms and about 5.5 angstroms, between about 3.5 angstroms and about 6 angstroms, between about 3.5 angstroms and about 6.5 angstroms, between about 3.5 angstroms and about 7 angstroms, between about 3.5 angstroms and about 7.5 ... angstroms to about 8 angstroms, about 3.5 angstroms to about 8.5 angstroms, about 3.5 angstroms to about 9 angstroms, about 3.5 angstroms to about 10 angstroms, about 4.5 angstroms to about 5 angstroms, about 4.5 angstroms to about 5.5 angstroms, about 4.5 angstroms to about 6 angstroms, about 4.5 angstroms to about 6.5 angstroms, about 4.5 angstroms to about 7 angstroms, about 4.5 angstroms to about 7.5 angstroms, about 4.5 angstroms to about 8 angstroms, about 4.5 angstroms to about 8.5 angstroms, about 4.5 angstroms to about 9 angstroms, about 4.5 angstroms to about 10 angstroms, about 5 angstroms to about 5.5 angstroms, about 5 angstroms to about 6 angstroms, about 5 angstroms to about 6.5 angstroms, about 5 angstroms to about 7 angstroms, about 5 angstroms to about 7.5 angstroms, about 5 angstroms to about 8 angstroms, about 5 angstroms to about about 8.5 angstroms, about 5 angstroms to about 9 angstroms, about 5 angstroms to about 10 angstroms, about 5.5 angstroms to about 6 angstroms, about 5.5 angstroms to about 6.5 angstroms, about 5.5 angstroms to about 7 angstroms, about 5.5 angstroms to about 7.5 angstroms, about 5.5 angstroms to about 8 angstroms, about 5.5 angstroms to about 8.5 angstroms, about 5.5 angstroms to about 9 angstroms, about 5.5 angstroms to about about 10 angstroms, about 6 angstroms to about 6.5 angstroms, about 6 angstroms to about 7 angstroms, about 6 angstroms to about 7.5 angstroms, about 6 angstroms to about 8 angstroms, about 6 angstroms to about 8.5 angstroms, about 6 angstroms to about 9 angstroms, about 6 angstroms to about 10 angstroms, about 6.5 angstroms to about 7 angstroms, about 6.5 angstroms to about 7.5 angstroms, about 6.5 angstroms to about 8 angstroms , about 6.5 angstroms to about 8.5 angstroms, about 6.5 angstroms to about 9 angstroms, about 6.5 angstroms to about 10 angstroms, about 7 angstroms to about 7.5 angstroms, about 7 angstroms to about 8 angstroms, about 7 angstroms to about 8.5 angstroms, about 7 angstroms to about 9 angstroms, about 7 angstroms to about 10 angstroms, about 7.5 angstroms to about 8 angstroms, about 7.5 angstroms to about 8 angstroms, about 7.5 angstroms to about 8.5 angstroms, about 7.5 angstroms to about 9 angstroms, about 7.5 angstroms to about 10 angstroms, about 8 angstroms to about 8.5 angstroms, about 8 angstroms to about 9 angstroms, about 8 angstroms to about 10 angstroms, about 8.5 angstroms to about 9 angstroms, about 8.5 angstroms to about 10 angstroms, or about 9 angstroms to about 10 angstroms. In some embodiments, the spatial separation between the alpha carbons of two linked amino acid residues, or between the geometric centers of two linked residues, is about 3.5 angstroms, about 4.5 angstroms, about 5 angstroms, about 5.5 angstroms, about 6 angstroms, about 6.5 angstroms, about 7 angstroms, about 7.5 angstroms, about 8 angstroms, about 8.5 angstroms, about 9 angstroms, or about 10 angstroms. In embodiments, the specific spatial separation is achieved using linker or spacer molecules known in the art.

[0198] Amino Acid Derivatives The present disclosure contemplates the use of any amino acid derivative or analogue in any of the peptide antagonists of the present disclosure.In some embodiments, amino acid modification can be chemically performed using any known method.Selective protein modification is described in the literature, for example, Spicer and Davis, 2014, "Selective chemical protein modification", Nature Communications 5:4740, which is incorporated herein by reference.

[0199] In some embodiments, the amino acid derivative is a non-standard amino acid. In some embodiments, the non-standard amino acid has an (S) configuration at the alpha position. In some embodiments, the non-standard amino acid has an (R) configuration at the alpha position. In some embodiments, the non-standard amino acid is an alpha amino acid. In some embodiments, the non-standard amino acid is a beta or gamma amino acid. In some embodiments, the non-standard amino acid is selected from the group consisting of aromatic side chain amino acids, non-aromatic side chain amino acids, aliphatic side chain amino acids, side chain amide amino acids, side chain ester amino acids, heteroaromatic side chain amino acids, side chain thiol amino acids, beta amino acids, and backbone modified amino acids. In some embodiments, the non-standard amino acid is a derivative of tyrosine, histidine, tryptophan, or phenylalanine. In some embodiments, the derivative of an amino acid includes an ester, amide, disulfide, carbamate, urea, phosphate, or ether of an amino acid. In some embodiments, the non-aromatic side chain amino acid is a derivative of serine, threonine, cysteine, methionine, arginine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, proline, glycine, alanine, valine, isoleucine, or leucine. In some embodiments, the non-standard amino acid is selected from the group consisting of 2-aminoadipic acid, 3-aminoadipic acid, β-alanine, β-aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, piperidinic acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, 2,4-diaminobutyric acid, desmosine, 2,2'-diaminopimelic acid, 2,3-diaminopropionic acid, N-ethylglycine, N-ethylasparagine, hydroxylysine, allo-hydroxylysine, 3-hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylglycine, sarcosine, n-methylisoleucine, 6-N-methyllysine, N-methylallyl, norvaline, norleucine, and ornithine. In some embodiments, the non-standard amino acid is a proline derivative.In some embodiments, the proline derivative is 3-fluoroproline, 4-fluoroproline, 3-hydroxyproline, 4-hydroxyproline, 3-aminoproline, 4-aminoproline, 3,4-dehydroproline, aziridine-2-carboxylic acid, azetidine-2-carboxylic acid, pipecolic acid, 4-oxa-proline, 3-thiaproline, or 4-thiaproline. In some embodiments, the non-standard amino acid comprises a lipid.

[0200] In some embodiments, the peptide antagonists of the disclosure include one or more amino acid derivatives or analogs known to those of skill in the art and described in the literature or herein, for example, In some embodiments, the peptide antagonists of the disclosure include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 1-11, 1-12, or 1-13 amino acid derivatives.

[0201] In some embodiments, each amino acid derivative present in a peptidic antagonist of the disclosure is a non-standard amino acid independently selected from the group consisting of aromatic side chain amino acids, non-aromatic side chain amino acids, aliphatic side chain amino acids, side chain amide amino acids, side chain ester amino acids, heteroaromatic side chain amino acids, side chain thiol amino acids, beta amino acids, and backbone modified amino acids, e.g., selected from non-standard amino acids described herein or known in the art and described in the published literature.

[0202] In some embodiments, the peptide antagonist comprises one or more amino acids having the D-amino acid configuration, with the remaining amino acids in the peptide having the L-amino acid configuration.

[0203] In some embodiments, the non-standard amino acid is a proline derivative. In some embodiments, the proline derivative comprises one or more substitutions on the pyrrolidine ring. In some embodiments, the proline derivative comprises one or more substitutions on the pyrrolidine ring, the substitutions include halogen, alkoxy, amino, hydroxyl, alkyl (methyl, ethyl), thiol, or alkylthio. In some embodiments, the proline derivative comprises one or more substitutions on the pyrrolidine ring, the substitutions include halogen, or alkyl (methyl, ethyl). In some embodiments, the proline derivative comprises one or more substitutions on the pyrrolidine ring, the substitutions include halogen. In some embodiments, the proline derivative comprises one or more substitutions on the pyrrolidine ring, the substitutions include alkoxy, hydroxyl, amino. In some embodiments, the proline derivative comprises one or more substitutions on the pyrrolidine ring, the substitutions include halogen, alkoxy, alkyl (methyl, ethyl), thiol, or alkylthio.

[0204] N-Terminal Modification of Peptide Antagonists In some embodiments, the N-terminal amino group of the peptide antagonists of the present disclosure is modified (N-terminal modification). In some embodiments, the N-terminus of the peptide antagonist is not modified with additional amino acids or amino acid derivatives. In some embodiments, the unmodified N-terminus comprises hydrogen. In some embodiments, the N-terminal modification is C1-C6 acyl, C1-C8 alkyl, C6-C 12 Aralkyl, C5-C 10 In some embodiments, the N-terminal modification is a C6-C 12In some embodiments, the N-terminal modification comprises a C1-C6 aralkyl. In some embodiments, the N-terminal modification comprises a C1-C6 acyl. In some embodiments, the N-terminal modification comprises an acetyl (Ac). In some embodiments, the N-terminal modification comprises a C1-C6 alkyl. In some embodiments, the N-terminal modification comprises a methyl, ethyl, propyl, or tert-butyl. In some embodiments, the N-terminal modification comprises a C1-C6 aralkyl. In some embodiments, the N-terminal modification comprises a benzyl. In some embodiments, the N-terminal modification comprises a formyl. In some embodiments, the peptides described herein, for example, any peptide having an amino acid sequence listed in Table 2 (regardless of the N-terminus shown in the table), have any of these N-terminal modifications or an unmodified N-terminus.

[0205] C-Terminal Modifications of Peptide Antagonists In some embodiments, the C-terminal acid group of the peptide antagonists of the present disclosure is modified (C-terminal modification). In some embodiments, the C-terminus is not modified with additional amino acids or amino acid derivatives. In some embodiments, the C-terminus is not modified with a glycine residue. In some embodiments, the unmodified C-terminus comprises -OH. In some embodiments, the C-terminal modification comprises an amino group, which is optionally substituted. In some embodiments, the C-terminal modification comprises an amino group, which is unsubstituted (-NH2). In some embodiments, the C-terminal modification comprises an amino group, which is substituted. In some embodiments, the C-terminal modification comprises an amino group, which is substituted. In some embodiments, the C-terminal modification comprises -NH2, -amino-acyl, -amino-C1-C8 alkyl, -amino-C6-C 12 Aralkyl, -amino-C5-C 10 aryl, or -amino-C4-C8 heteroaryl, -amino-C4-C8 heteroaryl, or -O-(C1-C8 alkyl). In some embodiments, the C-terminal modification is -amino-C6-C 12 In some embodiments, the C-terminal modification comprises -O-(C1-C8 alkyl). In some embodiments, the C-terminal modification comprises -amino-C6-C 12In some embodiments, the C-terminal modification comprises -NH-CH2phenyl. In some embodiments, the C-terminal modification comprises -OEt. In some embodiments, the C-terminal modification comprises -OMe. In some embodiments, the peptides described herein, for example, any peptide having an amino acid sequence listed in Table 2 (regardless of the C-terminus shown in the table), have any of these C-terminal modifications or an unmodified C-terminus.

[0206] In some embodiments, both the N-terminal amino group and the C-terminal acidic group of the disclosed peptide antagonist are modified.In some embodiments, the peptides described herein, for example, any peptide having an amino acid sequence listed in Table 2 (regardless of the C-terminus and N-terminus shown in the table), have an N-terminus and a C-terminus independently selected from those described herein.In some embodiments, the peptides described herein, for example, any peptide having an amino acid sequence listed in Table 2 (regardless of the C-terminus and N-terminus shown in the table), have an N-terminus and a C-terminus independently selected from Ac, NH2 and H.

[0207] Concentration of peptide antagonist in the composition In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 3 mg / mL, about 0.1 mg / mL to about 4 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, about 0.1 mg / mL to about 50 mg / mL, about 0.5 mg / mL to about 1 mg / mL. / mL, approximately 0.5 mg / mL to approximately 2 mg / mL, approximately 0.5 mg / mL to approximately 3 mg / mL, approximately 0.5 mg / mL to approximately 4 mg / mL, approximately 0.5 mg / mL to approximately 5 mg / mL, approximately 0.5 mg / mL to approximately 10 mg / mL, approximately 0.5 mg / m L ~ about 20mg / mL, about 0.5mg / mL - about 50mg / mL, about 1mg / ml - about 2mg / ml, about 1mg / ml - about 3mg / ml, about 1mg / ml - about 4mg / ml, about 1mg / ml - about 5mg / ml, about 1mg / ml - about 10mg / ml, about 1mg / ml to about 20mg / ml, about 1mg / ml to about 50mg / ml, about 2mg / mL to about 3mg / mL, about 2mg / mL to about 4mg / mL, about 2mg / mL to about 5mg / mL, about 2mg / mL to about 10mg / mL, about 2 mg / mL to about 20 mg / mL, about 2 mg / mL to about 50 mg / mL, about 3 mg / mL to about 4 mg / mL, about 3 mg / mL to about 5 mg / mL, about 3 mg / mL to about 10 mg / mL, about 3 mg / mL to 20 mg / mL, is present in the vesicle composition in an amount of about 3 mg / mL to about 50 mg / mL, about 4 mg / mL to about 5 mg / mL, about 4 mg / mL to about 10 mg / mL, about 4 mg / mL to about 20 mg / mL, about 4 mg / mL to about 50 mg / mL, about 5 mg / mL to about 10 mg / mL, about 5 mg / mL to about 20 mg / mL, about 5 mg / mL to about 50 mg / mL, about 10 mg / mL to about 20 mg / mL, about 10 mg / mL to about 50 mg / mL, or about 20 mg / mL to about 50 mg / mL.In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of at least about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, or about 20 mg / mL. In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of up to about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the peptide antagonist is present in the vesicle composition in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, or about 5 mg / mL.

[0208] vesicle-forming lipids In some embodiments, the vesicle composition comprises one or more vesicle-forming lipids. The vesicle-forming lipids act to encapsulate a portion of the oil-in-water emulsion. In some embodiments, this allows the oil-in-water emulsion to remain stable for a period of time.

[0209] The vesicle-forming lipid can be any lipid suitable for such purpose.In some embodiments, the vesicle-forming lipid comprises phospholipid, glycolipid, lecithin, ceramide, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, cardiolipin, phosphatidic acid, cerebroside, or any combination thereof.In some embodiments, the vesicle-forming lipid comprises a combination of lipids.

[0210] In some embodiments, the vesicle-forming lipid comprises a phospholipid. In some embodiments, the phospholipid is naturally occurring, semi-synthetic, or synthetically prepared, or a mixture thereof. In one embodiment, the phospholipid is one or more esters of fatty acid and glycerol with one or two (equal or different) residues of phosphoric acid, where the phosphoric acid residue is in turn bound to a hydrophilic group, such as, for example, choline (phosphatidylcholine--PC), serine (phosphatidylserine--PS), glycerol (phosphatidylglycerol--PG), ethanolamine (phosphatidylethanolamine--PE), or inositol (phosphatidylinositol). Phospholipid esters with only one fatty acid residue are commonly referred to in the art as the "lyso" form of phospholipid, or "lysophospholipid". Usually, the fatty acid residues present in phospholipids are long chain aliphatic acids, typically containing 12-24 carbon atoms, or 14-22 carbon atoms, and the aliphatic chain may contain one or more unsaturations, or may be fully saturated. Examples of suitable fatty acids contained in phospholipids are, for example, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, oleic acid, linoleic acid, and linolenic acid. Saturated fatty acids such as myristic acid, palmitic acid, stearic acid, and arachidic acid may be used.

[0211] In some embodiments, the phospholipid comprises one or more natural phospholipids. In some embodiments, the phospholipid comprises one or more semi-synthetic phospholipids. In some embodiments, the semi-synthetic phospholipid is a partially or fully hydrogenated derivative of naturally occurring lecithin. In some embodiments, the phospholipid comprises a fatty acid diester of phosphatidylcholine, ethylphosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, phosphatidylserine, or sphingomyelin. In some embodiments, the phospholipid comprises hydrogenated phosphatidylcholine (e.g., Sunlipon 90H). In some embodiments, the phospholipid may be, for example, dilauroyl-phosphatidylcholine (DLPC), dimyristoyl-phosphatidylcholine (DMPC), dipalmitoyl-phosphatidylcholine (DPPC), diarachidoyl-phosphatidylcholine (DAPC), distearoyl-phosphatidylcholine (DSPC), dioleoyl-phosphatidylcholine (DOPC), 1,2 distearoyl-sn-glycero-3-ethylphosphocholine (ethyl-DSPC), dipentadecanoyl-phosphatidylcholine (DPDPC), 1-myristoyl-2-palmitoyl-phosphatidylcholine (MPPC), 1-palmitoyl-2-myristoyl-phosphatidylcholine (PMPC), 1-palmitoyl-2-stearoyl-phosphatidylcholine (P SPC), 1-stearoyl-2-palmitoyl-phosphatidylcholine (SPPC), 1-palmitoyl-2-oleylphosphatidylcholine (POPC), 1-oleyl-2-palmitoyl-phosphatidylcholine (OPPC), dilauroylphosphatidylglycerol (DLPG) and its alkali metal salts, diarachidoylphosphatidylglycerol (DAPG) and its alkali metal salts, dimyristoylphosphatidylglycerol (DMPG) and its alkali metal salts, dipalmitoylphosphatidylglycerol (DPPG) and its alkali metal salts, distearoylphosphatidylglycerol (DSPG) and its alkali metal salts, dioleoyl-phosphatidylglycerol (DOPG) and its alkali metal salts,Dimyristoyl phosphatidic acid (DMPA) and its alkali metal salts, dipalmitoyl phosphatidic acid (DPPA) and its alkali metal salts, distearoyl phosphatidic acid (DSPA), diarachidoyl phosphatidic acid (DAPA) and its alkali metal salts, dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), distearoyl phosphatidylethanolamine (DSPE), dioleoyl phosphatidylethanolamine (DOPE), diarachidoyl phosphatidylethanolamine (DAPE), dilinoleyl phosphatidylethanolamine (DLPE), dimyristoyl phosphatidylserine (D MPS), diarachidoyl phosphatidylserine (DAPS), dipalmitoyl phosphatidylserine (DPPS), distearoyl phosphatidylserine (DSPS), dioleoyl phosphatidylserine (DOPS), dipalmitoyl sphingomyelin (DPSP), as well as distearoyl sphingomyelin (DSSP), dilauroyl-phosphatidylinositol (DLPI), diarachidoyl phosphatidylinositol (DAPI), dimyristoyl phosphatidylinositol (DMPI), dipalmitoyl phosphatidylinositol (DPPI), distearoyl phosphatidylinositol (DSPI), and dioleoyl-phosphatidylinositol (DOPI).

[0212] In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5% to about 25% (w / w) of the composition. In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5% to about 2%, about 0.5% to about 5%, about 0.5% to about 8%, about 0.5% to about 10%, about 0.5% to about 12%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 2% to about 5%, about 2% to about 8%, about 2% to about 10%, about 2% to about 12%, about 2% to about 15%, about 2% to about 20%, about 2% to about 25%, about 5% to about 8%, about 5% to about 10%, about 5% to about 10%, about 5% to about 20%, about 2% to about 25%, about 5% to about 8 ...0%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 20%, about 2% to about 2 In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, about 25%, about 8% to about 10%, about 8% to about 12%, about 8% to about 15%, about 8% to about 20%, about 8% to about 25%, about 10% to about 12%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 12% to about 15%, about 12% to about 20%, about 12% to about 25%, about 15% to about 20%, about 15% to about 25%, or about 20% to about 25% (w / w). In some embodiments, the vesicle-forming lipid is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25%. In some embodiments, the vesicle-forming lipids are present in an amount of at least about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, or about 20% (w / w) of the composition. In some embodiments, the vesicle-forming lipids are present in an amount of up to about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25% (w / w) of the composition.

[0213] In some embodiments, the vesicle-forming lipid is present in an amount of about 5% to about 15% (w / w) of the composition. In some embodiments, the vesicle-forming lipid is present in an amount of about 5% to about 8%, about 5% to about 9%, about 5% to about 10%, about 5% to about 11%, about 5% to about 12%, about 5% to about 13%, about 5% to about 14%, about 5% to about 15%, about 8% to about 9%, about 8% to about 10%, about 8% to about 11%, about 8% to about 12%, about 8% to about 13%, about 8% to about 14%, about 8% to about 15%, about 9% to about 10%, about 9% to about 11%, about 9% to about 12%, about 9% to about 13%, about 9% to about 14%, about 9% to about 15%, about 9% to about 10%, about 9% to about 11%, about 9% to about 12%, about 9% to about 15%, about 9% to about 16%, about 9% to about 17%, about 9% to about 18%, about 9% to about 19%, about 10% to about 20%, about 10% to about 21%, about 10% to about 22%, about 10% to about 23%, about 10% to about 24%, about 10% to about 25%, about 10% to about 26%, about 10% to about 27%, about 10% to about 28%, about 10% to about 29%, about 10% to about 30%, about 10% to about 31%, about 10% to about 32%, about 10% to about 33%, about 10% to about 34%, about 10 In some embodiments, the vesicle-forming lipid is present in an amount of about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% (w / w) of the composition. In some embodiments, the vesicle-forming lipids are present in an amount of at least about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14% (w / w) of the composition. In some embodiments, the vesicle-forming lipids are present in an amount of up to about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15% (w / w) of the composition.

[0214] In some embodiments, the composition comprises a short-chain polyol. In some embodiments, the short-chain polyol acts to enhance the stability of the resulting lipid vesicle. In some embodiments, the short-chain polyol is a C2-C4 polyol that comprises two or three alcohol groups. In some embodiments, the short-chain polyol is propylene glycol. In some embodiments, the composition comprises propylene glycol.

[0215] In some embodiments, propylene glycol is present in an amount of about 0.5% to about 25% (w / w) of the composition. In some embodiments, propylene glycol is present in an amount of about 0.5% to about 2%, about 0.5% to about 5%, about 0.5% to about 8%, about 0.5% to about 10%, about 0.5% to about 12%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 2% to about 5%, about 2% to about 8%, about 2% to about 10%, about 2% to about 12%, about 2% to about 15%, about 2% to about 20%, about 2% to about 25%, about 5% to about 8%, about 5% to about 10%, about In some embodiments, propylene glycol is present in an amount of about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, about 25%, about 8% to about 10%, about 8% to about 12%, about 8% to about 15%, about 8% to about 20%, about 8% to about 25%, about 10% to about 12%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 12% to about 15%, about 12% to about 20%, about 12% to about 25%, about 15% to about 20%, about 15% to about 25%, or about 20% to about 25%. In some embodiments, propylene glycol is present in an amount of at least about 0.5%, about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, or about 20%. In some embodiments, propylene glycol is present in an amount of up to about 2%, about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25%. In some embodiments, propylene glycol is present in an amount of about 1% to about 10%. In some embodiments, propylene glycol is present in an amount of about 1% to about 2%, about 1% to about 4%, about 1% to about 6%, about 1% to about 8%, about 1% to about 10%, about 2% to about 4%, about 2% to about 6%, about 2% to about 8%, about 2% to about 10%, about 4% to about 6%, about 4% to about 8%, about 4% to about 10%, about 6% to about 8%, about 6% to about 10%, or about 8% to about 10%. In some embodiments, propylene glycol is present in an amount of about 1%, about 2%, about 4%, about 6%, about 8%, or about 10%. In some embodiments, propylene glycol is present in an amount of at least about 1%, about 2%, about 4%, about 6%, or about 8%.In some embodiments, propylene glycol is present in an amount of up to about 2%, about 4%, about 6%, about 8%, or about 10%. In some embodiments, propylene glycol is present in about the same amount as the vesicle-forming lipid. In some embodiments, the ratio of propylene glycol to vesicle-forming lipid in the composition is about 2:1 to 1:2 (w / w).

[0216] Oil Phase The lipid vesicle compositions provided herein include oil-in-water emulsions. The oil component is selected so that the material is liquid at the operating temperature (e.g., room temperature) and is immiscible with water.

[0217] Any suitable oil can be used as oil phase.In some embodiments, oil comprises naturally occurring oil.In some embodiments, naturally occurring oil is derived from one or more plants or plant parts (e.g., seeds or nuts).In some embodiments, oil is naturally occurring oil such as olive oil, vegetable oil, sunflower oil, or other similar plant-derived oil.

[0218] In some embodiments, the oil phase is selected from the group consisting of vegetable oils, mono-, di-, and triglycerides, silicone fluids, mineral oils, and combinations thereof.

[0219] In some embodiments, the oil comprises a silicone oil or derivative, such as dimethicone. In some embodiments, the silicone oil comprises a siloxane polymer. In some embodiments, the siloxane polymer comprises C1-C3 substituents. In some embodiments, the siloxane is polydimethylsiloxane (PDMS). In some embodiments, the oil is a mixture that includes a silicone oil (e.g., dimethicone) as a smaller component. In some embodiments, the silicone oil is incorporated to improve the feel of the resulting composition or as a moisturizer. In some embodiments, the oil comprises an amount of silicone oil up to about 5%, up to about 4%, up to about 3%, up to about 2%, or up to about 1%. In some embodiments, the silicone oil is present in an amount of about 0.1% to about 2% (w / w) of the composition.

[0220] In some implementations, the oil formulation is present in an amount of about 1% to about 35% (w / w) of the composition. In some embodiments, the oil is present in an amount of about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 1% to about 35%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 10% to about 15%, about 10% to about 5%. The oil is present in an amount of about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 20% to about, 25%, about 20% to about 30%, about 20% to about 35%, about 25% to about 30%, about 25% to about 35%, or about 30% to about 35%. In some embodiments, the oil is present in an amount of about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the oil is present in an amount of at least about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, or about 30%. In some embodiments, the oil is present in an amount up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the oil is present in an amount of about 5% to about 15%. In some implementations, the oil is present in an amount of about 5% to about 8%, about 5% to about 9%, about 5% to about 10%, about 5% to about 11%, about 5% to about 12%, about 5% to about 13%, about 5% to about 14%, about 5% to about 15%, about 8% to about 9%, about 8% to about 10%, about 8% to about 11%, about 8% to about 12%, about 8% to about 13%, about 8% to about 14%, about 8% to about 15%, about 9% to about 10%, about 9% to about 11%, about 9% to about 12%, about 9% to about 13%. %, about 9% to about 14%, about 9% to about 15%, about 10% to about 11%, about 10% to about 12%, about 10% to about 13%, about 10% to about 14%, about 10% to about 15%, about 11% to about 12%, about 11% to about 13%, about 11% to about 14%, about 11% to about 15%, about 12% to about 13%, about 12% to about 14%, about 12% to about 15%, about 13% to about 14%, about 13% to about 15%, or about 14% to about 15%. In some embodiments, the oil is present in an amount of about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15%.In some embodiments, the oil is present in an amount of at least about 5%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14%. In some embodiments, the oil is present in an amount of up to about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, or about 15%.

[0221] In some embodiments, the oil comprises one or more triglycerides. In some embodiments, the triglycerides are medium chain triglycerides. In some embodiments, the medium chain triglycerides are C6-C 12 The fatty acid esters include those having a chain length of

[0222] In some embodiments, the triglyceride is present in an amount of about 1% to about 35% (w / w) of the composition. In some embodiments, the triglyceride is present in an amount of about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 30%, about 1% to about 35%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 10% to about 15%, about 1 The triglyceride is present in an amount of about 0% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 20% to about, 25%, about 20% to about 30%, about 20% to about 35%, about 25% to about 30%, about 25% to about 35%, or about 30% to about 35%. In some embodiments, the triglyceride is present in an amount of about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the triglyceride is present in an amount of at least about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, or about 30%. In some embodiments, the triglyceride is present in an amount of up to about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, or about 35%. In some embodiments, the lipid vesicle and / or the oil phase of the lipid vesicle portion of the composition comprises a sterol. In some embodiments, the sterol is cholesterol. In some embodiments, the cholesterol may be plant-derived cholesterol. In some embodiments, the plant-derived cholesterol may be PhytoChol®, SyntheChol®, or any other plant-derived cholesterol (e.g., Avanti#700100), or any combination thereof. In some embodiments, the sterol may be a phytosterol or a derivative thereof. In some embodiments, the phytosterol or a derivative thereof may be phytosterol MM, Advasterol™ 90 IP or 95 IP F, NET sterol-ISO, canola sterol, sitosterol 700095, lanosterol-95, brassicasterol, or any combination thereof.

[0223] In some embodiments, the sterol is present in an amount of about 1% to about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1% to about 3%, about 1% to about 4%, about 1% to about 5%, about 1.5% to about 2%, about 1.5% to about 2.5%, about 1.5% to about 3%, about 1.5% to about 4%, about 1.5% to about 5%, about 2% to about 2.5%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2.5% to about 3%, about 2.5% to about 4%, about 2.5% to about 5%, about 3% to about 4%, about 3% to about 5%, or about 4% to about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 4%, or about 5% (w / w) of the composition. In some embodiments, the sterol is present in an amount of at least about 1%, about 1.5%, about 2%, about 2.5%, about 3%, or about 4% (w / w) of the composition. In some embodiments, the sterol is present in an amount of up to about 1.5%, about 2%, about 2.5%, about 3%, about 4%, or about 5% (w / w) of the composition.

[0224] Penetration enhancers In some embodiments, the lipid vesicle composition comprises one or more penetration enhancers that, when applied to the skin of an individual, act to increase the amount of penetration of the anionic polymeric material through one or more layers of the skin.

[0225] In some embodiments, the penetration enhancer is included in an oil-in-water emulsion of the composition, hi some embodiments, the penetration enhancer is included in the lipid bilayer of the composition.

[0226] There are various types of penetration enhancers that can be used. In some embodiments, the penetration enhancer comprises an ionic surfactant, a non-ionic surfactant, or a combination thereof.

[0227] In some embodiments, the penetration enhancer comprises a non-ionic surfactant or a combination of non-ionic surfactants. In some embodiments, the penetration enhancer is a single non-ionic surfactant. In some embodiments, the penetration enhancer is a combination of at least two, three, or four or more non-ionic surfactants. In some embodiments, the penetration enhancer is a combination of two non-ionic surfactants. In some embodiments, the penetration enhancer is a combination of three non-ionic surfactants.

[0228] In some embodiments, the nonionic surfactant or combination of nonionic surfactants is selected from polyethylene glycol ethers of fatty alcohols, sorbitan esters, polysorbates, and polyethylene glycol fatty acid esters, and combinations thereof.

[0229] In some embodiments, the combination of nonionic surfactants is a combination of polyethylene glycol ethers and sorbitan esters of aliphatic alcohols. In some embodiments, the combination of nonionic surfactants is a combination of polyethylene glycol ethers and polysorbates of aliphatic alcohols. In some embodiments, the combination of nonionic surfactants is a combination of polyethylene glycol ethers and sorbitan esters of aliphatic alcohols. In some embodiments, the combination of nonionic surfactants is a combination of polyethylene glycol ethers and polyethylene glycol fatty acid esters of aliphatic alcohols. In some embodiments, the combination of nonionic surfactants is a combination of polyethylene glycol ethers, sorbitan esters, and polysorbates of aliphatic alcohols. In some embodiments, the combination of nonionic surfactants is a combination of polyethylene glycol ethers, sorbitan esters, and polyethylene glycol fatty acid esters ... polysorbates, and polyethylene glycol fatty acid esters of aliphatic alcohols.

[0230] In some embodiments, the combination of non-ionic surfactants comprises a polyethylene glycol fatty acid ester and a sorbitan ester. In some embodiments, the combination of non-ionic surfactants comprises a polyethylene glycol fatty acid ester and a polysorbate. In some embodiments, the combination of non-ionic surfactants is a combination of a polyethylene glycol fatty acid ester, a polysorbate, and a sorbitan ester.

[0231] In some embodiments, the nonionic surfactant comprises a polyethylene glycol (PEG) ether of an aliphatic alcohol. In some embodiments, the PEG ether of an aliphatic alcohol has from about 2 to about 8 PEG groups and a C 12 -C 22In some embodiments, the polyethylene glycol ether of the aliphatic alcohol comprises diethylene glycol hexadecyl ether, 2-(2-octadecaoxyethoxy)ethanol, diethylene glycol monooleyl ether, polyoxyethylene(2) oleyl ether, polyoxyethylene(3) oleyl ether, or polyoxyethylene(5) oleyl ether, or any combination thereof. In some embodiments, the polyethylene glycol ether of the aliphatic alcohol comprises 2-(2-octadecaoxyethoxy)ethanol. In some embodiments, the PEG ether of the aliphatic alcohol is super refined Brij® O2 or a derivative thereof.

[0232] In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5% to about 2.5%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.5% to about 2.5%, about 1.2% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the PEG ether of an aliphatic alcohol is present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0233] In some embodiments, the non-ionic surfactant comprises a sorbitan ester. In some embodiments, the sorbitan ester is a fatty acid ester. In some embodiments, the sorbitan ester is a C 12 -C 22 A fatty acid ester. In some embodiments, the sorbitan ester comprises sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, or sorbitan isostearate, or any combination thereof. In some embodiments, the sorbitan ester comprises sorbitan monolaurate. In some embodiments, the sorbitan ester comprises sorbitan monopalmitate. In some embodiments, the sorbitan ester comprises sorbitan monostearate. In some embodiments, the sorbitan ester comprises sorbitan monooleate. In some embodiments, the sorbitan ester comprises sorbitan trioleate. In some embodiments, the sorbitan ester comprises sorbitan sesquioleate. In some embodiments, the sorbitan ester comprises sorbitan isostearate.

[0234] In some embodiments, the sorbitan ester is present in an amount of about 0.5% to about 2.5% (w / w) of the composition. In some embodiments, the sorbitan ester is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the sorbitan esters are present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the sorbitan esters are present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the sorbitan esters are present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0235] In some embodiments, the non-ionic surfactant comprises a polysorbate. In some embodiments, the polysorbate comprises polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, or any combination thereof. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the polysorbate is polysorbate 20.

[0236] In some embodiments, the polysorbate is present in an amount of about 0.5% to about 2.5% (w / w) of the composition. In some embodiments, the polysorbate is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the polysorbate is present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the polysorbate is present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the polysorbate is present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0237] In some embodiments, the non-ionic surfactant comprises a polyethylene glycol (PEG) fatty acid ester. In some embodiments, the PEG fatty acid ester comprises a C8-C hydroxyl group attached to each terminal hydroxyl group to form a fatty acid ester. 22 The PEG chain is about 2-8 subunits containing fatty acids. In some embodiments, the PEG fatty acid ester comprises PEG-8 dilaurate, PEG-4 dilaurate, PEG-4 laurate, PEG-8 dioleate, PEG-8 distearate, PEG-8 distearate, PEG-7 glyceryl cocoate, and PEG-20 almond glyceride, or any combination thereof. In some embodiments, the PEG fatty acid ester is PEG-4 dilaurate.

[0238] In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5% to about 2.5% (w / w) of the composition. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5% to about 0.8%, about 0.5% to about 1%, about 0.5% to about 1.2%, about 0.5% to about 1.5%, about 0.5% to about 2%, about 0.5% to about 2.5%, about 0.8% to about 1%, about 0.8% to about 1.2%, about 0.8% to about 1.5%, about 0.8% to about 2%, about 0.8% to about 2.5%, about 1% to about 1.2%, about 1% to about 1.5%, about 1% to about 2%, about 1% to about 2.5%, about 1.2% to about 1.5%, about 1.2% to about 2%, about 1.2% to about 2.5%, about 1.5% to about 2%, about 1.5% to about 2.5%, or about 2% to about 2.5%. In some embodiments, the PEG fatty acid ester is present in an amount of about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%. In some embodiments, the PEG fatty acid ester is present in an amount of at least about 0.5%, about 0.8%, about 1%, about 1.2%, about 1.5%, or about 2%. In some embodiments, the PEG fatty acid ester is present in an amount of up to about 0.8%, about 1%, about 1.2%, about 1.5%, about 2%, or about 2.5%.

[0239] In some embodiments, the non-ionic surfactant has a hydrophobic-lipophilic balance (HLB) of about 10 or less. In some embodiments, the non-ionic surfactant may be Cithrol GMS40. In some embodiments, the composition comprises a plurality of non-ionic surfactants, each having an HLB of about 10 or less. In some embodiments, the non-ionic surfactants having an HLB of 10 or less are selected from Table 1, or any combination thereof.

[0240] In some embodiments, the nonionic surfactant or combination of nonionic surfactants is present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, the nonionic surfactant or combination of nonionic surfactants is from about 0.5% to about 1%, from about 0.5% to about 1.5%, from about 0.5% to about 2%, from about 0.5% to about 3%, from about 0.5% to about 4%, from about 0.5% to about 5%, from about 0.5% to about 6%, from about 0.5% to about 7%, from about 0.5% to about 8%, from about 0.5% to about 10%, from about 1% to about 1.5%, from about 1% to about 2%, from about 1% to about 3%, from about 1% to about 4%, from about 1% to about 5%, from about 1% to about 6%, from about 1% to about 7%, from about 1% to about 8%, from about 1% to about 10%, from about 1.5% to about 2%, from about 1.5% to about 3%, from about 1.5% to about 4%, from about 1.5% to about 5%, from about 1.5% to about 6%, About 1.5% to about 7%, about 1.5% to about 8%, about 1.5% to about 10%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2% to about 6%, about 2% to about 7%, about 2% to about 8%, about 2% to about 10%, about 3% to about 4%, about 3% to about 5%, about 3% to about 6%, about 3% to about 7%, about 3% to about 8%, about 3% to about 10% , about 4% to about 5%, about 4% to about 6%, about 4% to about 7%, about 4% to about 8%, about 4% to about 10%, about 5% to about 6%, about 5% to about 7%, about 5% to about 8%, about 5% to about 10%, about 6% to about 7%, about 6% to about 8%, about 6% to about 10%, about 7% to about 8%, about 7% to about 10%, or about 8% to about 10%. In some embodiments, the nonionic surfactant or combination of nonionic surfactants is present in an amount of about 0.5%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, or about 10%. In some embodiments, the non-ionic surfactant or combination of non-ionic surfactants is present in an amount of at least about 0.5%, about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, or about 8%. In some embodiments, the non-ionic surfactant or combination of non-ionic surfactants is present in an amount of up to about 1%, about 1.5%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, or about 10%.

[0241] In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion, the lipid bilayer, or both. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion. In some embodiments, the composition comprises a non-ionic surfactant in the lipid bilayer. In some embodiments, the composition comprises a non-ionic surfactant in the oil-in-water emulsion and the lipid bilayer, where the composition comprises two or more different non-ionic surfactants.

[0242] In some embodiments, the penetration enhancer comprises a salicylate or a nicotinate. In some embodiments, the ester is a C1-C6 alkyl ester or a benzyl ester. In some embodiments, the penetration enhancer comprises methyl salicylate or benzyl nicotinate. In some embodiments, the penetration enhancer is a nicotinate present in an amount up to about 0.1%, 0.5%, 1%, 2%, or 3% (w / w) of the composition. In some embodiments, the nicotinate is present in an amount of about 0.1% to about 3%, about 0.1% to about 2%, about 0.1% to about 1%.

[0243] In some embodiments, the penetration enhancer comprises a fatty acylated amino acid. In some embodiments, the fatty acylated amino acid is lysine. In some embodiments, the lysine is monoacylated with a fatty acid. In some embodiments, the penetration enhancer is monolauryl lysine. In some embodiments, the lysine is diacylated. In some embodiments, the penetration enhancer is dipalmitoyl lysine. In some embodiments, the fatty acylated amino acid is present in an amount of up to about 1%, up to about 2%, up to about 3%, up to about 4%, or up to about 5% (w / w) of the composition. In some embodiments, the fatty acylated amino acid is present in an amount of about 0.1% to about 5%, about 0.1% to about 4%, about 0.1% to about 3%, about 0.1% to about 2%, about 0.5% to about 5%, about 0.5% to about 4%, about 0.5% to about 3%, about 0.5% to about 2%, about 1% to about 5%, about 1% to about 4%, about 1% to about 3%, about 1% to about 2%, or about 1.5% to about 2.5%.

[0244] Cationic Surfactants In some embodiments, the composition further comprises a cationic surfactant. In some embodiments, the cationic surfactant is used to stabilize the water-in-oil emulsion (e.g., at the submicron emulsion stage before lipid vesicle formation). In some embodiments, the cationic surfactant is a monocationic surfactant. In some embodiments, the monocationic surfactant is a net monocationic (e.g., a phosphate salt that contains two side chains, each of which has a single cationic functionality, and is partially neutralized by phosphate anion).

[0245] In some embodiments, the monocationic surfactant is propylene glycol diammonium phosphate ester derived from a fatty amide. In some embodiments, the monocationic surfactant is linoleamidopropyl PG-dimonium chloride phosphate.

[0246] In some embodiments, the propylene glycol diammonium phosphate ester derived from a fatty amide is present in an amount of about 1% to about 10% (w / w) of the composition. In some embodiments, the propylene glycol diammonium phosphate ester derived from a fatty amide is present in an amount of about 1% to about 2%, about 1% to about 3%, about 1% to about 4%, about 1% to about 5%, about 1% to about 6%, about 1% to about 7%, about 1% to about 8%, about 1% to about 9%, about 1% to about 10%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2% to about 6%, about 2% to about 7%, about 2% to about 8%, about 2% to about 9%, about 2% to about 10%, about 3% to about 4%, about 3% to about 5%, about 3% to about 6%, about 3% about 7%, about 3% to about 8%, about 3% to about 9%, about 3% to about 10%, about 4% to about 5%, about 4% to about 6%, about 4% to about 7%, about 4% to about 8%, about 4% to about 9%, about 4% to about 10%, about 5% to about 6%, about 5% to about 7%, about 5% to about 8%, about 5% to about 9%, about 5% to about 10%, about 6% to about 7%, about 6% to about 8%, about 6% to about 9%, about 6% to about 10%, about 7% to about 8%, about 7% to about 9%, about 7% to about 10%, about 8% to about 9%, about 8% to about 10%, or about 9% to about 10%. In some implementations, the propylene glycol diammonium phosphate ester derived from a fatty amide is present in an amount of about 1%, 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% (w / w) of the composition. In some embodiments, the propylene glycol diammonium phosphate ester derived from a fatty amide is present in an amount of at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, or about 9%. In some embodiments, the propylene glycol diammonium phosphate ester derived from a fatty amide is present in an amount of up to about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%.

[0247] Additional Ingredients In some embodiments, the vesicle composition comprises additional components that, in some embodiments, improve one or more properties of the vesicle without dramatically altering the delivery of the anionic polymeric material.

[0248] In some embodiments, the vesicle composition comprises one or more viscosity enhancing agents. In some embodiments, the viscosity enhancing agent increases the viscosity of the composition for improved stability of the vesicle composition and / or user feel. In some embodiments, the viscosity enhancing agent further acts as a surfactant. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof. In some embodiments, the fatty alcohol is a C8-C 20 In some embodiments, the fatty alcohol is cetyl alcohol. In some embodiments, the cetyl alcohol is Crodacol C95. In some embodiments, the wax is a natural or synthetic wax. In some embodiments, the wax is beeswax. In some embodiments, the wax is synthetic beeswax. In some embodiments, the synthetic beeswax is Syncrowax™ BB4. In some embodiments, the synthetic beeswax is a non-animal derived beeswax. In some embodiments, the non-animal derived beeswax is Syncrowax™ SB1. In some embodiments, the fatty acid ester of glycerol is a monoester. In some embodiments, the monoester is a C8-C 24 An ester of a fatty acid. In some embodiments, the fatty acid ester of glycerol is glycerol monostearate.

[0249] In some embodiments, the viscosity enhancing agent is present in an amount of about 0.5% to about 10% (w / w) of the composition. In some embodiments, the viscosity enhancing agent is present in an amount of about 0.5% to about 5%, about 0.5% to about 5%, about 0.5% to about 4%, about 0.5% to about 3%, or about 0.5% to about 2% (w / w) of the composition. In some embodiments, the viscosity enhancing agent comprises a fatty alcohol in an amount of up to about 2%, a wax in an amount of up to about 2%, and a fatty acid ester of glycerol in an amount of up to about 5%. In some embodiments, the fatty alcohol is present in an amount of about 0.1 to about 1.5%. In some embodiments, the wax is present in an amount of about 0.1% to about 1%. In some embodiments, the fatty acid ester of glycerol is present in an amount of about 0.5% to about 2%.

[0250] In some embodiments, the vesicle composition further comprises one or more of a thickening agent, a preservative, a moisturizer, an emollient, a humectant, or any combination thereof. In some embodiments, the vesicle composition further comprises a thickening agent. In some embodiments, the vesicle composition further comprises a preservative. In some embodiments, the vesicle composition further comprises a humectant (e.g., isopropyl myristate). In some embodiments, the vesicle composition further comprises an emollient. In some embodiments, the vesicle composition further comprises a humectant.

[0251] In some embodiments, the vesicle composition further comprises a humectant. In some embodiments, the composition comprises glycerol. In some embodiments, the glycerol is present in an amount of about 0.5% to about 25%, about 0.5% to about 20%, about 0.5% to about 15%, or about 0.5% to about 10% (w / w) of the composition. In some embodiments, the glycerol is present in an amount of about 1% to about 10%. In some embodiments, the glycerol is present in an amount of about 1% to about 2%, about 1% to about 4%, about 1% to about 6%, about 1% to about 8%, about 1% to about 10%, about 2% to about 4%, about 2% to about 6%, about 2% to about 8%, about 2% to about 10%, about 4% to about 6%, about 4% to about 8%, about 4% to about 10%, about 6% to about 8%, about 6% to about 10%, or about 8% to about 10%. In some embodiments, glycerol is present in an amount of about 1%, about 2%, about 4%, about 6%, about 8%, or about 10%. In some embodiments, glycerol is present in an amount of at least about 1%, about 2%, about 4%, about 6%, or about 8%. In some embodiments, glycerol is present in an amount of up to about 2%, about 4%, about 6%, about 8%, or about 10%.

[0252] In some embodiments, the vesicle composition further comprises a preservative. In some embodiments, the preservative is a paraben ester. In some embodiments, the preservative is methylparaben or propylparaben or a combination thereof. In some embodiments, the preservative is present in an amount of up to about 1%, up to about 0.9%, up to about 0.8%, up to about 0.7%, up to about 0.6%, up to about 0.5%, up to about 0.4%, up to about 0.3%, up to about 0.2% (w / w) of the composition.

[0253] In some embodiments, the additional component comprises purified water. In some embodiments, the purified water is present in an amount of about 50% to about 90% (w / w). In some embodiments, the purified water is present in an amount of about 50% to about 55%, about 50% to about 60%, about 50% to about 65%, about 50% to about 70%, about 50% to about 75%, about 50% to about 80%, about 50% to about 85%, about 50% to about 90%, about 55% to about 60%, about 55% to about 65%, about 55% to about 70%, about 55% to about 75%, about 55% to about 80%, about 55% to about 85%, about 55% to about 90%, about 60% to about 65%, about 60% to about 70%, about 60% to about In some embodiments, purified water is present in an amount of about 75%, about 60% to about 80%, about 60% to about 85%, about 60% to about 90%, about 65% to about 70%, about 65% to about 75%, about 65% to about 80%, about 65% to about 85%, about 65% to about 90%, about 70% to about 75%, about 70% to about 80%, about 70% to about 85%, about 70% to about 90%, about 75% to about 80%, about 75% to about 85%, about 75% to about 90%, about 80% to about 85%, about 80% to about 90%, or about 85% to about 90%. In some embodiments, purified water is present in an amount of about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or about 90%. In some embodiments, purified water is present in an amount of at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, or about 85%. In some embodiments, purified water is present in an amount of up to about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or about 90%.

[0254] Exemplary Compositions for Delivery of Peptide Antagonists Exemplary compositions for delivery of peptide antagonists are provided below. The embodiments described below may additionally further comprise other ingredients or components as provided herein.

[0255] Peptide Composition 1: In one embodiment, (a) lipid vesicles, each of which comprises a lipid bilayer comprising a vesicle-forming lipid, the vesicle-forming lipid being present in an amount of about 5% to about 20%; (b) an oil-in-water emulsion encapsulated in lipid vesicles and stabilized by one or more surfactants; (c) a peptide antagonist of the muscle-type nicotinic acetylcholine receptor in an amount of about 0.1 mg / mL to about 50 mg / mL encapsulated in a lipid bilayer and / or an oil-in-water emulsion; Provided herein is a lipid vesicle composition comprising: The composition comprises: propylene glycol diammonium phosphate ester derived from a fatty acid amide in an amount of about 1% to about 10%; a nonionic surfactant in an amount of about 0.1% to about 3%; Further includes:

[0256] In some embodiments, the oil component is present in an amount of about 2.5% to about 20%.

[0257] In some embodiments, the lipid vesicle composition comprises an amount of peptide antagonist of about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 3 mg / mL, about 0.1 mg / mL to about 4 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises an amount of peptide antagonist of about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, or about 5 mg / mL.

[0258] In some embodiments, the composition further comprises a fatty acylated amino acid in an amount of about 0.5% to about 3%. In some embodiments, the fatty acylated amino acid is monolauryl lysine.

[0259] In some embodiments, the lipid vesicle composition further comprises a viscosity enhancing agent in an amount of about 0.5% to about 5%. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof.

[0260] In some embodiments, the non-ionic surfactant comprises a PEG ether of an aliphatic alcohol.

[0261] In some embodiments, the lipid vesicle composition further comprises an anionic polymeric material in an amount of about 10 mg / mL encapsulated in the lipid bilayer, oil-in-water emulsion, or a combination thereof. In some embodiments, the lipid vesicle composition comprises an anionic polymeric material in an amount of about 0.01 mg / mL to about 0.05 mg / mL, about 0.01 mg / mL to about 0.1 mg / mL, about 0.01 mg / mL to about 0.5 mg / mL, about 0.01 mg / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.01 mg / mL to about 1.5 mg / mL, about 0.01 mg / mL to about 1.75 mg / mL, about 0.01 mg / mL to about 2 mg / mL, about 0.01 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 ... g / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.1 mg / mL to about 1.75 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 1 mg / mL, about 1 mg / mL to about 1.5 mg / mL, about 1 mg / mL to about 1.75 mg / mL, about 1 mg / mL to about 2 mg / mL, about 1 mg / mL to about 5 mg / mL, or about 1 mg / mL to about 10 mg / mL of the anionic polymer material. In some embodiments, the lipid vesicle composition comprises anionic polymeric material in an amount of about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL.

[0262] Peptide Composition 2: In one embodiment, (a) lipid vesicles, each of which comprises a lipid bilayer comprising a vesicle-forming lipid, the vesicle-forming lipid being present in an amount of about 2% to about 20%; (b) an oil-in-water emulsion encapsulated in lipid vesicles and stabilized by one or more surfactants; (c) a peptide antagonist of the muscle-type nicotinic acetylcholine receptor in an amount of about 0.1 mg / mL to about 50 mg / mL encapsulated in a lipid bilayer and / or an oil-in-water emulsion; Provided herein is a lipid vesicle composition comprising: The composition comprises: PEG fatty acid ester in an amount of about 0.1% to about 2%; Polysorbate in an amount of about 0.5% to about 3%; A sorbic acid ester in an amount of about 0.1% to about 2%; Further includes:

[0263] In some embodiments, the oil component is present in an amount of about 2.5% to about 20%.

[0264] In some embodiments, the lipid vesicle composition comprises an amount of peptide antagonist of about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 3 mg / mL, about 0.1 mg / mL to about 4 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises an amount of peptide antagonist of about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, or about 5 mg / mL.

[0265] In some embodiments, the lipid vesicle composition further comprises a viscosity enhancing agent in an amount of about 0.5% to about 5%. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof.

[0266] In some embodiments, the PEG fatty acid ester comprises PEG4-dilaurate. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the sorbic acid ester is sorbitan palmitate.

[0267] In some embodiments, the lipid vesicle composition further comprises an anionic polymeric material in an amount of about 10 mg / mL encapsulated in the lipid bilayer, oil-in-water emulsion, or a combination thereof. In some embodiments, the lipid vesicle composition comprises an anionic polymeric material in an amount of about 0.01 mg / mL to about 0.05 mg / mL, about 0.01 mg / mL to about 0.1 mg / mL, about 0.01 mg / mL to about 0.5 mg / mL, about 0.01 mg / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.01 mg / mL to about 1.5 mg / mL, about 0.01 mg / mL to about 1.75 mg / mL, about 0.01 mg / mL to about 2 mg / mL, about 0.01 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 ... g / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.1 mg / mL to about 1.75 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 1 mg / mL, about 1 mg / mL to about 1.5 mg / mL, about 1 mg / mL to about 1.75 mg / mL, about 1 mg / mL to about 2 mg / mL, about 1 mg / mL to about 5 mg / mL, or about 1 mg / mL to about 10 mg / mL of the anionic polymer material. In some embodiments, the lipid vesicle composition comprises anionic polymeric material in an amount of about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL.

[0268] Peptide Composition 3: In one embodiment, (a) lipid vesicles, each of which comprises a lipid bilayer comprising a vesicle-forming lipid, the vesicle-forming lipid being present in an amount of about 5% to about 20%; (b) an oil-in-water emulsion encapsulated in lipid vesicles and stabilized by one or more surfactants; (c) a peptide antagonist of the muscle-type nicotinic acetylcholine receptor in an amount of about 0.1 mg / mL to about 50 mg / mL encapsulated in a lipid bilayer and / or an oil-in-water emulsion; Provided herein is a lipid vesicle composition comprising: The composition comprises: propylene glycol diammonium phosphate ester derived from a fatty amide in an amount of about 1% to about 10%; a PEG ether of an aliphatic alcohol in an amount of about 0.1% to about 3%; Polysorbate in an amount of about 0.5% to about 3%; A sorbic acid ester in an amount of about 0.1% to about 2%; Further includes:

[0269] In some embodiments, the oil component is present in an amount of about 2.5% to about 20%.

[0270] In some embodiments, the lipid vesicle composition comprises an amount of peptide antagonist of about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 3 mg / mL, about 0.1 mg / mL to about 4 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 20 mg / mL, about 0.1 mg / mL to about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises an amount of peptide antagonist of about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 10 mg / mL, about 20 mg / mL, or about 50 mg / mL. In some embodiments, the lipid vesicle composition comprises the peptide antagonist in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, or about 5 mg / mL.

[0271] In some embodiments, the lipid vesicle composition further comprises a viscosity enhancing agent in an amount of about 0.5% to about 5%. In some embodiments, the viscosity enhancing agent comprises one or more of a fatty alcohol, a wax, a fatty acid ester of glycerol, or any combination thereof.

[0272] In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the sorbic acid ester is sorbitan palmitate. In some embodiments, the PEG ether of an aliphatic alcohol is diethylene glycol monooleyl ether.

[0273] In some embodiments, the lipid vesicle composition further comprises an anionic polymeric material in an amount of about 10 mg / mL encapsulated in the lipid bilayer, oil-in-water emulsion, or a combination thereof. In some embodiments, the lipid vesicle composition comprises an anionic polymeric material in an amount of about 0.01 mg / mL to about 0.05 mg / mL, about 0.01 mg / mL to about 0.1 mg / mL, about 0.01 mg / mL to about 0.5 mg / mL, about 0.01 mg / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.01 mg / mL to about 1.5 mg / mL, about 0.01 mg / mL to about 1.75 mg / mL, about 0.01 mg / mL to about 2 mg / mL, about 0.01 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.1 mg / mL to about 0.5 mg / mL, about 0.1 mg / mL to about 1 ... g / mL to about 1 mg / mL, about 0.01 mg / mL to about 1.25 mg / mL, about 0.1 mg / mL to about 1.5 mg / mL, about 0.1 mg / mL to about 1.75 mg / mL, about 0.1 mg / mL to about 2 mg / mL, about 0.1 mg / mL to about 5 mg / mL, about 0.1 mg / mL to about 10 mg / mL, about 0.5 mg / mL to about 1 mg / mL, about 1 mg / mL to about 1.5 mg / mL, about 1 mg / mL to about 1.75 mg / mL, about 1 mg / mL to about 2 mg / mL, about 1 mg / mL to about 5 mg / mL, or about 1 mg / mL to about 10 mg / mL of the anionic polymer material. In some embodiments, the lipid vesicle composition comprises anionic polymeric material in an amount of about 0.01 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.25 mg / mL, about 1.5 mg / mL, about 1.75 mg / mL, about 2 mg / mL, about 5 mg / mL, or about 10 mg / mL.

[0274] Methods of Using the Lipid Vesicle Compositions Provided herein The lipid vesicle compositions provided herein are intended for cosmetic use in a subject for indications including, but not limited to, prevention or temporary improvement in the appearance of one or more of skin wrinkles, skin laxity, moderate to severe glabellar wrinkles associated with activity of the corrugator supercilii and / or procerus muscles, moderate to severe lateral canthal wrinkles (crow's feet lines) associated with activity of the orbicularis oculi muscle, and moderate to severe frontal wrinkles associated with activity of the frontalis muscle.

[0275] In certain embodiments, including pharmaceutical embodiments, the lipid vesicle compositions provided herein are contemplated for pharmaceutical use in a subject for indications including, but not limited to, prevention or temporary improvement in the appearance of one or more skin wrinkles, such as facial wrinkles, skin laxity, moderate to severe glabellar wrinkles associated with activity of the corrugator supercilii and / or procerus muscles, moderate to severe lateral canthal wrinkles (crow's feet lines) associated with activity of the orbicularis oculi muscle, and moderate to severe frontal wrinkles associated with activity of the frontalis muscle.

[0276] In some embodiments, the subject is a mammal. In specific embodiments, the mammal is a human. In some embodiments, the human subject is a pediatric or adult subject of any age.

[0277] Method of using cosmetic or pharmaceutical composition In certain embodiments, including pharmaceutical embodiments, the present disclosure also relates to a method of using a cosmetic or pharmaceutical composition comprising a peptide antagonist or an anionic polymeric material such as hyaluronic acid. In some embodiments, the present disclosure relates to a method of using a cosmetic or pharmaceutical composition to prevent or temporarily improve the appearance of one or more skin wrinkles in a subject, including facial wrinkles, skin laxity, moderate to severe glabellar wrinkles associated with activity of the corrugator supercilii and / or procerus muscles, moderate to severe lateral canthal wrinkles (crow's feet) associated with activity of the orbicularis oculi muscle, and moderate to severe frontal wrinkles associated with activity of the frontalis muscle, comprising applying an effective amount of the cosmetic or pharmaceutical composition to the skin of the subject. In some embodiments, the present disclosure relates to a method of using the cosmetic or pharmaceutical composition to improve the appearance of a subject's lips, for example, by making the lips appear fuller. In some embodiments, the cosmetic or pharmaceutical composition is used to enhance lip fullness, lip volume, lip smoothness, lip color, or a combination thereof. In some embodiments, the cosmetic or pharmaceutical composition provides fuller and / or natural looking lips to a subject. In some embodiments, the cosmetic or pharmaceutical composition is used to restore any one of volume, definition, softness, or fullness to the lips of a subject. In some embodiments, the cosmetic or pharmaceutical composition is used to visibly remove or reduce lines or wrinkles on the lips of a subject. In some embodiments, the cosmetic or pharmaceutical composition enhances lip color (e.g., rosy redness). In some embodiments, the cosmetic or pharmaceutical composition provides one or more of volume, softness, and definition to the lips of a subject.

[0278] In some embodiments, the lipid vesicle composition is applied topically to a subject. Topical application as referred to herein can refer to application onto one or more surfaces, for example, keratinous tissue. In some embodiments, the topical composition is administered to the skin of a subject. In some embodiments, the skin is the facial skin of a subject. In some embodiments, the skin includes the lips of a subject. Topical application can refer to direct application to a desired area. In certain embodiments, including pharmaceutical embodiments, the topical cosmetic or pharmaceutical composition or preparation can be applied, for example, by pouring, dripping, or spraying, if present as a liquid or aerosol composition, by spreading, rubbing, painting, etc., if present as an ointment, lotion, cream, gel, or similar composition, by sprinkling, if present as a powder, or by any other suitable means.

[0279] In some embodiments, the lipid vesicle composition is formulated in a form suitable for topical application. In some embodiments, the lipid vesicle composition is formulated as a cream, a lotion, a suspension, or an emulsion. In some embodiments, the lipid vesicle composition is formulated as a cream. In some embodiments, the lipid vesicle composition is formulated as a lotion. In some embodiments, the lipid vesicle composition is formulated as a suspension.

[0280] In some embodiments, the subject uses or is treated with a topical application comprising an effective amount of the lipid vesicle composition one or more times during the course of use or treatment, for example, 1-3 times per day, 1-21 times per week, once per day, twice per day, or three times per day. In some embodiments, the subject uses or is treated with an effective amount of the lipid vesicle composition about once per week to about 12 times per week. In some embodiments, the subject uses or is treated with an effective amount of the lipid vesicle composition at least about once per week. In some embodiments, the subject uses or is treated with an effective amount of the lipid vesicle composition up to about 12 times per week. In some embodiments, the subject administers an effective amount of the lipid vesicle composition about once per week to about twice per week, about once per week to about three times per week, about once per week to about four times per week, about once per week to about five times per week, about once per week to about six times per week, about once per week to about seven times per week, about once per week to about eight times per week, about once per week to about nine times per week, about once per week to about ten times per week, about once per week to about eleven times per week, about once per week to about twelve times per week, about twice per week to about three times per week, about twice per week to about four times per week, about twice per week to about five times per week, about twice per week to about six ... 2 times to about 7 times per week, about 2 times to about 8 times per week, about 2 times to about 9 times per week, about 2 times to about 10 times per week, about 2 times to about 11 times per week, about 2 times to about 12 times per week, about 3 times to about 4 times per week, about 3 times to about 5 times per week, about 3 times to about 6 times per week, about 3 times to about 7 times per week, about 3 times to about 8 times per week, about 3 times to about 9 times per week, about 3 times to about 10 times per week, about 3 times to about 11 times per week, about 3 times to about 12 times per week, about 4 times to about 5 times per week, about 4 times to about 6 times per week,About 4 times per week to about 7 times per week, about 4 times per week to about 8 times per week, about 4 times per week to about 9 times per week, about 4 times per week to about 10 times per week, about 4 times per week to about 11 times per week, about 4 times per week to about 12 times per week, about 5 times per week to about 6 times per week, about 5 times per week to about 7 times per week, about 5 times per week to about 8 times per week, about 5 times per week to about 9 times per week, about 5 times per week to about 10 times per week, about 5 times per week to about 11 times per week, about 5 times per week to about 12 times per week, about 6 times per week to about 7 times per week, about 6 times per week to about 8 times per week, about 6 times per week to about 9 times per week, about 6 times per week to about 10 times per week, about 6 times per week to about 10 times per week 11 times per week, 6 times to 12 times per week, 7 times to 8 times per week, 7 times to 9 times per week, 7 times to 10 times per week, 7 times to 11 times per week, 7 times to 12 times per week, 8 times to 9 times per week, 8 times to 10 times per week, 8 times to 11 times per week, 7 times to 12 times per week, 8 times to 9 times per week, 8 times to 10 times per week, 8 times to 10 times per week about 8 times per week to about 12 times per week, about 9 times per week to about 10 times per week, about 9 times per week to about 11 times per week, about 9 times per week to about 12 times per week, about 10 times per week to about 11 times per week, about 10 times per week to about 12 times per week, about 11 times per week to about 12 times per week. In some embodiments, a subject uses or is treated with an effective amount of a lipid vesicle composition about once per week, about twice per week, about three times per week, about four times per week, about five times per week, about six times per week, about seven times per week, about eight times per week, about nine times per week, about ten times per week, about eleven times per week, about 12 times per week, about 13 times per week, or about 14 times per week.

[0281] In some embodiments, one or more layers of the lipid vesicle composition of the present disclosure are applied to the skin of a subject at a given time. In some embodiments, a subsequent layer may be applied after a previous layer of lipid vesicle composition has been sufficiently absorbed into the skin of the subject. In some embodiments, the lipid vesicle composition may take several seconds (e.g., 1 second, 2 seconds, 3 seconds, 5 seconds, 10 seconds, 15 seconds, 30 seconds, etc.) to be sufficiently absorbed into the skin of the subject. In some embodiments, 1, 2, 3, 4, 5, 6, or 7 layers of lipid vesicle composition are applied to the skin of the subject at a given time. In some embodiments, the lipid vesicle composition is applied to the skin of the subject one or more times per day (e.g., 1-3 times per day, 1 time per day, 2 times per day, 3 times per day, etc.). In some embodiments, the lipid vesicle composition is applied to the skin of the subject one or more times per week (e.g., 1-21 times per week, 1-14 times per week, 1-7 times per week, etc.). In some embodiments, the lipid vesicle composition is applied to the skin of the subject daily. In some embodiments, one or more layers of the lipid vesicle composition are applied to the subject's skin once a day for one or more days. In some embodiments, two or more layers of the lipid vesicle composition are applied to the subject's skin once a day for one or more days. In some embodiments, three or more layers of the lipid vesicle composition are applied to the subject's skin once a day for one or more days. In some embodiments, one or more layers of the lipid vesicle composition are applied to the subject's skin twice a day for one or more days. In some embodiments, two or more layers of the lipid vesicle composition are applied to the subject's skin twice a day for one or more days. In some embodiments, three or more layers of the lipid vesicle composition are applied to the subject's skin twice a day for one or more days. In some embodiments, the lipid vesicle composition is applied to the subject's skin for at least one day, two days, three days, four days, five days, six days, one week, two weeks, three weeks, one month, two months, three months, six months, one year. In some embodiments, the lipid vesicle composition is applied to the subject's skin for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, or a year or more. In some embodiments, one or more layers of the lipid vesicle composition are applied to the subject's skin twice daily for several days, and then three times daily.In some embodiments, five layers of the lipid vesicle composition are applied to the subject's skin twice daily (e.g., morning and night) for five days, after which one to three layers of the lipid vesicle composition are applied to the subject's skin three times daily (e.g., morning, midday, and night).

[0282] In some embodiments, the lipid vesicle compositions of the present disclosure are administered to a subject for indications including, but not limited to, prevention or temporary improvement in the appearance of one or more skin wrinkles, such as facial wrinkles, skin laxity, moderate to severe glabellar wrinkles associated with activity of the corrugator supercilii and / or procerus muscles, moderate to severe lateral canthal wrinkles (crow's feet lines) associated with activity of the orbicularis oculi muscle, and moderate to severe frontal wrinkles associated with activity of the frontalis muscle.

[0283] In some embodiments, the lipid vesicle compositions of the present disclosure are administered to a subject for indications including, but not limited to, temporary improvement in the appearance of fullness of the lips. In some embodiments, the lipid vesicle compositions of the present disclosure are used with other products including, but not limited to, petrolatum, lip balms, lipsticks, lip tints, lip glosses, medicated lip creams, lip conditioners, sunscreens, etc.

[0284] In some embodiments, the topical cosmetic compositions of the present disclosure are self-applied or administered by a subject. In certain embodiments, including pharmaceutical embodiments, the disclosed cosmetic or pharmaceutical compositions are applied or administered by a medical professional, for example, in a clinic setting.

[0285] Methods of Making the Lipid Vesicle Compositions Provided herein Further, a method of making lipid vesicle composition is provided herein.In some embodiments, the composition of the present disclosure as described above is prepared by mixing the oil component of an oil-in-water emulsion with the aqueous component of an oil-in-water emulsion, and either the component of the oil-in-water emulsion or the emulsion component comprises one or more surfactants for emulsification with the component of the oil component of the oil-in-water emulsion.In one embodiment, surfactants are mixed with the aqueous component and added to the oil to form an emulsion.The oil-in-water emulsion is then mixed with the solubilized vesicle-forming lipids and, if added, other lipid components under effective mixing conditions to form lipid vesicles (e.g., multisomes).

[0286] In some embodiments, one or more penetration enhancers and one or more compounds (e.g., anionic polymeric materials, one or more peptides, etc.) are added to the oil component of the oil-in-water emulsion, the aqueous component of the oil-in-water emulsion, or both. Alternatively, or in addition, one or more penetration enhancers and / or one or more compounds can be added to the lipid component.

[0287] In one aspect, provided herein is a method for preparing a lipid vesicle composition provided herein, comprising: a) preparing an oil-in-water emulsion comprising an active ingredient by mixing an oil component of an emulsion-in-water emulsion with an aqueous component of an oil-in-water emulsion; b) solubilizing vesicle-forming lipids in an acceptable solvent other than water; c) adding the oil-in-water emulsion to the solubilized vesicle-forming lipids; and d) mixing the oil-in-water emulsion with the solubilized vesicle-forming lipids under mixing conditions effective to form lipid vesicles comprising a lipid bilayer comprising vesicle-forming lipids and the emulsion-in-water emulsion encapsulated in the lipid vesicles. In some embodiments, the active ingredient is a peptide provided herein. In some embodiments, the active ingredient is an anionic polymeric material provided herein.

[0288] In some embodiments, the method further comprises the step of adding one or more additional ingredients provided herein (eg, penetration enhancers, viscosity enhancers, etc.).

[0289] In some embodiments, the step of mixing the oil component of the oil-in-water vesicle in step a) with the oil component of the oil-in-water emulsion and / or the mixing conditions in step e) include using stirring such as homogenization or emulsification, or microemulsion techniques without stirring. In one embodiment, mixing includes high pressure homogenization. High pressure homogenization provides relatively precise control of the composition of lipid vesicles. High pressure homogenization is suitable for small molecules, peptides, or proteins that are resistant to shear. In one embodiment, the composition formed is any one of the lipid vesicle compositions described herein.

[0290] In some embodiments, other lipid components are added at any one of the steps. EXAMPLES

[0291] Example 1. Preparation of Multisome Lipid Vesicle Compositions of Hyaluronic Acid Biphasic vesicles with multiple / synergistic penetration enhancers (multisomes) were formulated with three different molecular weights of hyaluronic acid: 250K, 50K, and 10K (Creative PEGWorks, Chapel Hill, NC) at either 1 mg / mL or 1.5 mg / mL concentrations. Unlabeled HA was used in formulation development. For diffusion cell experiments, vesicles were prepared with labeled HA (rhodamine-HA250K, FITC-HA50K, and FITC-HA-10K; Creative PEGWorks).

[0292] The general procedure for multisome preparation was as follows.

[0293] 1) Oil phase and water phase ingredients were weighed out into separate beakers.

[0294] 2) Both beakers were heated to approximately 70°C to completely dissolve and incorporate all ingredients.

[0295] 3) The aqueous phase was added to the oil phase with vigorous stirring using a spatula to form a crude o / w emulsion, effectively obtaining a homogeneous milky solution in a 70°C water bath (approximately 2-6 min). The temperature of the solution was approximately 55-65°C.

[0296] 4) The formulation was batch processed in triplicate at approximately 20,000 psi using an LV1 Microfluidizer equipped with a Z5 module or a Nano DeBee Homogenizer.

[0297] Procedure for vesicle formation (applicable to all formulations):

[0298] 1) The lipid phase ingredients were weighed into a 20 mL glass vial.

[0299] 2) The vial was heated in a water bath to approximately 70°C until all components were completely dissolved and incorporated.

[0300] 3) The aqueous phase (system A) was added to the liquid phase with vigorous stirring for approximately 10 to 20 minutes until the temperature of the solution was approximately 60°C.

[0301] In some cases, the mixture was subjected to 8-10 cycles of intermittent vortexing for 5 seconds and heating for 5 seconds until a uniform creamy lotion was produced.

[0302] A pictorial representation of this process is shown in Figure 7. A flow chart of this exemplary process is shown in Figure 8.

[0303] The various lipid phases used throughout the experiments are described in Table A below, and the aqueous phases are described in Table B below.

[0304] [Table 3]

[0305] [Table 4-1]

[0306] [Table 4-2]

[0307] Example 2. Methods of analysis and characterization The following methods were used to characterize the formulations described in the Examples below, as well as the performance of the formulations.

[0308] Physicochemical characterization - Sensory observation, optical microscopy, and confocal microscopy (Zeiss710 laser scanning confocal microscope (CLSM)) were performed to characterize the formulations. Confocal microscopy images of the formulations were obtained using a Zeiss LSM710 CLSM using the Argon laser 488 and HeNe laser 543 lines for FITC (495 / 525) and Rhodamine (570 / 590) and either a Plan-Apochromat 20x / 0.80 dry objective or a 63x / 1.40 oil immersion objective. Optical zoom selection was applied for selected cases. Laser intensity, pinhole, and gain settings were kept constant between sample sets to allow comparison of relative fluorescence intensity measurements between samples. Images were captured and processed using Zen2009 software.

[0309] Size (hydrodynamic diameter), polydispersity index, and zeta (ζ) potential measurements were performed on the formulations using a Nano ZS Zetasizer (Malvern Instruments, Worcestershire, UK), which uses dynamic light scattering (DLS) to measure the hydrodynamic diameter of particles. Aliquots of the formulations were diluted 20-fold with water and 100 μL and 1000 μL of each formulation were prepared for size and zeta potential measurements, respectively. Measurements were performed in triplicate.

[0310] In vitro diffusion cell study - Full thickness human breast skin was obtained from female donors undergoing elective mammoplasty surgery at the Royal University Hospital, University of Saskatchewan (Saskatoon, Saskatchewan, Canada). Approval for skin collection was obtained from the University of Saskatchewan Human Research Ethics Committee. Skin was harvested within 2 hours postoperatively, trimmed of subcutaneous fat, and stored at -20°C until use. A 9 mm orifice diameter (0.63 cm 2 An in-line Bronaugh flow-through diffusion cell with a 100% ethanol solution was mounted on a waterproof cell warmer (PermeGear, Inc., Hellertown, PA) and set at a constant temperature of 32°C. A pre-cut 1 cm2 skin section was placed in the diffusion cell with the stratum corneum side facing up. Perfusion buffer (100 mM phosphate buffer containing 0.05% Na azide) at 37°C was circulated through the lower half of the diffusion cell at a rate of 1 mL / h using a peristaltic pump. The surface of the skin was dosed with 0.1 mL of formulation. After 24 h of incubation, the skin samples were removed from the cell, washed, and processed for analysis.

[0311] Skin analysis - After removing the skin samples from the diffusion cell, the formulation remaining on the skin surface was first removed. Each skin sample was subjected to a washing protocol and a tape stripping protocol to remove the remaining bound cream and stratum corneum as follows: the skin sample was washed three times with 10 mL water, patted dry with a Kimwipe, and split in half. One skin was tape stripped twice (to remove the formulation bound to the surface), embedded in OCT compound on dry ice, and cryosectioned. The frozen sections were examined by confocal microscopy.

[0312] Skin samples were cryosectioned into 10 μm sections using a Leica CM1850 cryostat. Sections on slides were left unstained. Confocal microscopy images of skin sections were obtained using a Zeiss LSM710CLSM using the Argon Laser 488 and HeNe Laser 543 lines for FITC (495 / 525) and Rhodamine (570 / 590) with either a Plan-Apochromat 20x / 0.80 dry objective or a 63x / 1.40 oil immersion objective. Optical zoom selection was applied for selected cases. Laser intensity, pinhole, and gain settings were kept constant between sample sets to allow comparison of relative fluorescence intensity measurements between different treatments. Images were captured and processed using Zen2009 software.

[0313] An "untreated" skin sample was used to verify gain and pinhole settings and to eliminate noise and autofluorescence background prior to subsequent analysis on the treated samples.

[0314] Example 3. Evaluation of cationic penetration enhancers The first approach to formulate 1 mg / mL of the HA250K+HA10K combination is to incorporate them into multisomes, next generation biphasic vesicles (synergistic enhancer type), using monocationic, dicationic, or polycationic building blocks to enhance the encapsulation and delivery of negatively charged hyaluronic acid to the skin layers. The components of these formulations are shown in Table C below.

[0315] [Table 5]

[0316] HA encapsulation in vesicles was analyzed using confocal microscopy studies on multisomes showing the distribution of Rho-HA250K (red) and FITC-HA10K (green) fluorescence in the formulations (images in Figure 1, panel A). The final concentration of HA in these samples was 1 mg / mL. Profile tracing by confocal microscopy confirmed the association of red Rho-HA250K and green FITC-HA10K with vesicles in formulations F1, F2, and F3 (Figure 1, panel A traces). Fluorescence intensity (FI) curves tracing vesicles along selected planes show the colocalization of red and green fluorescence, confirming the coencapsulation of two different molecular weights of HA. Images of the formulations taken by light microscopy showed the formation of multisomes (next generation biphasic vesicles) in each type of formulation (Figure 1, panel B). Zetasizer studies were performed on system A (submicron emulsion components and biphasic vesicles) (Figure 1, panel C). These formulations were shown to be polydisperse, with vesicle sizes generally ranging between 0.3-10 μm. Zetasizer data show consistent results with microscopy (Panel B of Figure 1). This is typical of multisomes. The zeta potentials of F1, F2, and F3 were +33.6 ± 0.6, +13.0 ± 0.71, and -5.78 ± 0.31, respectively. Similar data was observed with other formulations described in subsequent examples (data not shown).

[0317] The physicochemical properties of the multisome formulations were evaluated for color, consistency, and homogeneity. All formulations were of a lotion or cream consistency suitable for topical application. The formulations were physically stable without signs of separation, precipitation, or other stability issues for more than 3 months of storage at 4° C. Microscopic observation confirmed that the multisomes remained intact and were uniformly distributed upon storage. Similar observations were made for other formulations described in subsequent examples (data not shown).

[0318] Frozen sections of human skin samples treated in vitro in diffusion cells with topical formulations containing fluorescently labeled HA were evaluated for the presence of fluorescent protein. Enhanced delivery of (negatively charged) HA compounds is shown using three basic vesicle formulations (Table C) utilizing three cationic vesicle building blocks.

[0319] These studies demonstrated that all three cationic formulations increased HA delivery (Table D, Figure 2). The order of enhancement was dicationic, followed by monocationic, followed by polycationic.

[0320] [Table 6]

[0321] Example 4. Evaluation of hyaluronic acid concentration and the presence of additional cosmetic ingredients Next, the effect of HA concentration in the basic vesicle formulation was evaluated. To evaluate this effect, 1 mg / mL and 1.5 mg / mL of 250 kDa and 10 kDa hyaluronic acid (total weight of HA combined, mass of each molecular weight is equivalent) and 1 mg / mL and 1.5 mg / mL of 250 kDa and 50 kDa hyaluronic acid (total weight of HA combined, mass of each molecular weight is equivalent) were prepared in the formulations provided in Table E. In addition, solution or gel formulations of HA according to formulations E11 and E12 of Table E, made from 1% hydroxypropylmethylcellulose (HPMC) gel, were prepared.

[0322] [Table 7]

[0323] After administration to human skin as provided in Example 2, the results shown in Table F below were obtained. As shown by comparing formulations E1 vs. E2 and E4 vs. E5 (Table F), increasing the concentration of HA250 / 10K or HA250 / 50K vesicle formulations at total concentrations of 1 mg / mL to 1.5 mg / mL increases delivery, especially evident from the increase in the HA250K component. Upon further optimization to obtain cosmetic vesicle formulations, it was found that these changes did not affect delivery, see formulations E2 vs. E3 and E5 vs. E6 (Table F).

[0324] A comparison of formulations E7, E8, E9, and E10 (Table F) shows that these compositions did not achieve enhanced delivery compared to the other formulations. In formulations E7 and E10 (Table 7), the strategy of using both polycationic and monocationic agents reduced delivery compared to the use of monocationic agents alone. Gel formulations in which HA250 / 10K or 250 / 50K was incorporated in a "free" (unencapsulated) form (formulations E11 and E12 in Table F) showed very low / negligible levels of delivery.

[0325] All multiphasic vesicle formulations showed enhanced delivery compared with solution or gel formulations of HA. Differences in the formulation of multiphasic vesicles demonstrated that HA delivery could be tuned.

[0326] [Table 8]

[0327] Additionally, multisome compositions with additional cosmetic properties were evaluated, including formulations containing Lipovol GBT (tribehenin) or benzyl nicotinate. Hyaluronic acid with combined molecular weights of 250 / 10 kDa and 250 / 50 kDa were evaluated for their effect on transdermal penetration. The formulations tested are shown in Table G below.

[0328] [Table 9]

[0329] Administration to human skin as provided in Example 2 produced the results shown in Table H below.

[0330] [Table 10]

[0331] Comparison of delivery efficiency of compositions with ingredients added for cosmeceutical effects showed that tribehenin and benzyl nicotinate (BN) affect delivery. For example, for the delivery of HA250 / 10K formulation, G1 vs. G2 (Table H) and formulations G3 and G4 (Table H and Figure 4) show that BN enhanced delivery with or without T.

[0332] Comparing formulations G5 and G7 (Table H) and formulations G6 and G8 (Table H and Figure 4) shows that the presence or absence of tribehenin alone (no BN) did not affect delivery, but in the absence of tribehenin, BN enhanced delivery, which may mean that tribehenin inhibits the effect of BN to deliver enhancement. This is also noticeable when comparing formulations G7 and G8 (Table H and Figure 3) which show that in the absence of tribehenin, BN increases delivery.

[0333] Comparing formulations G4 and G8 (Table H and Figure 4), it is noted that using BN but without tribehenin was the preferred formulation for delivering both the HA250 / 10K or 250 / 50K combinations, with an overall increase in efficiency of 200%.

[0334] Another formulation composition that was tested and found to be less effective in delivering HA250 / 50K was formulation G9 (Table H and FIG. 4), where the phospholipid component was replaced with another type of phospholipid. Additionally, increasing the concentration of lipid phase components and inclusion of hyaluronic acid of 250 kDa, 50 kDa, and 10 kDa MW each (formulation G10; Table H) were also found to be less effective. For example, compare G10 in Table H with E2 and E5 in Table F.

[0335] Furthermore, there was no substantial difference in the delivery of two different HA mwt combinations 250 / 10K or 250 / 50K from different sets of equivalent formulations with the same overall composition, see for example G1 vs. G5 and G3 vs. G7.

[0336] In Tables F and H, the ratio of Ch1 / Ch2 fluorescence values ​​is also shown for skin samples treated with the various formulations. Similar ratios indicate that the delivery of FITC-HA10 or FITC-HA50K and Rho-HA250K is similar to the original ratio of these two actives in the multisome formulation, i.e., they are delivered simultaneously to the same extent. Lower ratios indicate that the delivery of the HA250K component is further enhanced compared to the smaller polymers.

[0337] Example 5. Evaluation of Multisome Formulations to Enhance Penetration of Nicotinic Acetylcholine Receptor Peptide Antagonists (Conotoxin Peptide Analogs) The purpose of this study was to investigate the skin penetration properties of a conotoxin peptide analog (hereafter referred to as "C7 peptide"; Glo Pharma) from a next generation biphasic vesicle formulation (multisome). The C7 peptide has an amino acid sequence similar to that of SEQ ID NO: 3, a natural conotoxin peptide antagonist of the muscle-type nicotinic acetylcholine receptor. As with all of the peptides provided herein (e.g., SEQ ID NOs: 1-52 and 60-99), the C7 peptide has similar properties (e.g., similar size, conformation, charge, etc.) to the natural conotoxin of SEQ ID NO: 3 for purposes of formulating lipid vesicle delivery compositions. Thus, lipid vesicle compositions that work for the C7 peptide are expected to work similarly for the other peptides provided herein.

[0338] Three multisome vesicles (F6A-C7, F1B-C7, F1C-C7) were formulated with C7 peptide at a loading concentration of 2 mg / mL and compared with C7 peptide solution. The formulations were characterized for physicochemical properties.

[0339] In vitro diffusion cell studies were performed with multiphasic vesicle formulations F6A-C7, F1B-C7, and F1C-C7, and transdermal sections were collected for further analysis by mass spectrometry at Climax Laboratories.

[0340] Overall, the results indicate that a suitable C7 peptide-containing multiphasic vesicular cream formulation can be prepared for intradermal / transdermal delivery.

[0341] The specific objectives of these experiments were to evaluate the dermal delivery of C7 peptide from the various formulations developed as follows: 1) The uptake of C7 peptide through human skin was evaluated in experiments using Bronaugh-type in-line diffusion cells and human breast skin. Skin samples were treated with formulations containing C7 peptide, and the permeation through the skin was evaluated in transdermal sections by mass spectrometry. 2) Free C7 peptide solution was used for comparison as a reference, and blank vehicle was used as a control. 3) Transdermal sections were collected every hour for analysis. 4) The sections were pooled, concentrated by Pall filtration, and shipped for analysis at Climax Laboratories, Inc. (San Jose, CA).

[0342] material and method

[0343] Formulations: Biphasic vesicles (multisomes) with multiple / synergistic penetration enhancers - 5 different vesicles were formulated. From these, 3 formulations were selected for testing. For formulation development, C7 peptide (Anaspec, code: 74337, lot no. 1958617) was used. For diffusion cell experiments, multisomes without peptide and multisomes with 2 mg / mL of C7 peptide were used. The diffusion cell dose was 0.1 g of formulation with 0.2 mg of peptide. The pH of the formulations was 6.2-6.7.

[0344] Physicochemical characterization - Sensory observation, optical microscopy, and confocal microscopy (Zeiss710 confocal laser scanning microscope (CLSM; Carl Zeiss GmbH, Germany) were performed to characterize the formulations.

[0345] Size (hydrodynamic diameter) and polydispersity index and zeta (ζ) potential measurements were performed for formulations F6A-C7, F1B-C7, F1C-C7 prepared with unlabeled C7 peptide using a Zetasizer NanoZS (Malvern Instruments, Worcestershire, UK), which uses dynamic light scattering (DLS) to measure the hydrodynamic diameter of particles. An aliquot of 80 μL formulation was evaluated for particle size distribution and subsequently diluted 10-fold with water for zeta potential measurements. Measurements were performed in triplicate.

[0346] In vitro diffusion cell study - Full thickness human breast skin was obtained with permission from female donors undergoing elective mammoplasty surgery at the Royal University Hospital, University of Saskatchewan (Saskatoon, Saskatchewan, Canada). Approval for tissue collection was obtained from the University of Saskatchewan Human Research Ethics Committee. Skin was harvested within 2 hours post-surgery, trimmed of subcutaneous fat, and stored at -20°C until use.

[0347] In vitro absorption of C7 peptide from formulations into excised human skin was measured using a 0.636 cm 2 The cellular responses were evaluated using a 9 mm diameter Bronaugh-type Teflon flow-through diffusion cell (PermeGear, Inc., Hellertown, PA) with an exposed surface area of ​​100 μl. The cell holder was maintained at 32° C. by a circulating water bath heater. Degassed phosphate-buffered saline (PBS) buffer pH 7.2 containing 0.05% sodium azide maintained at 37° C. was used as the perfusion fluid at a flow rate of 1 mL / h. Skin samples were removed from the freezer, cut into square pieces of approximately 1 cm×1 cm, and mounted epidermis-side up in the diffusion cell. F6A-C7F1B-C7, F1C-C7 multisome formulations bearing C7 peptide or a control blank formulation (100 μl per cell) were applied to the skin at t=0 and the cell was covered with a Teflon cap to result in occlusion. Treatments were performed for 24 hours. Transdermal fragments were collected into 3 mL tissue culture tubes using a programmed fraction collector, and fragments were collected at 1 mL / cell every hour for up to a total of 24 h.

[0348] Transdermal Section Analysis - Transdermal sections were taken every hour for 24 hour analysis. For each cell, 24 1 mL sections were collected and labeled 1 / 1h, 1 / 2h, 1 / 3h, ... 1 / 24h, etc. Samples were sent to Glo / Climax Analytical Labs for analysis.

[0349] The skin samples for the diffusion cell test were washed to remove residual bound cream by the usual protocol, i.e., the skin samples were removed from the diffusion cell, washed three times with 10 mL of water, and then patted dry with a Kimwipe. The washed skin discs were stored at -20°C.

[0350] Results and Discussion

[0351] Optimization and characterization of multisome formulations

[0352] Tables I and J show the composition of the formulations developed with and without C7 peptide. Initially, formulations were prepared using Phospholipon 90H (soybean phosphatidylcholine). It was then replaced with Sunlipon 90H (sunflower phosphatidylcholine) (SunL) based on sensory and physicochemical observations. Optical microscopy images of the resulting lipid vesicle formulations are shown in Figure 5.

[0353] [Table 11]

[0354] [Table 12]

[0355] Table K summarizes the sensory properties, physical stability particle size range, and physical stability. All formulations were of a lotion or cream consistency suitable for topical application. The formulations were physically stable for over one month of storage at 4° C. without showing any signs of separation, settling, or other stability issues.

[0356] [Table 13]

[0357] These formulations were shown to be polydisperse with vesicle sizes generally ranging from 0.3 to 5 µm, as shown on optical microscopy images (Figure 1 and Table 4). Microscopic observations confirmed the formation of multisomes with typical biphasic vesicle morphology and uniform distribution of vesicles throughout the formulation for both the C7 peptide-containing formulations and the blank (no peptide) formulation.

[0358] Each of the C7 peptide-containing formulations was similar in terms of size distribution compared to their respective blank formulations, but overall, the blank formulations had a narrower size distribution compared to the peptide formulations (Figures 6A and 6B).

[0359] The Zetasizer data showed results consistent with the microscopy observations (Figure 6B), which are typical of multisomes. The zeta potential of F6A-C7 was positive, whereas the other two, F1B-C7 and F1C-C7, were negatively charged (Table L and Figures 6A and 6B).

[0360] [Table 14]

[0361] Lipid vesicle formulations were prepared at a C7 concentration of 2 mg / mL and administered to skin samples at 200 micrograms per skin sample. Blank versions of each formulation were prepared as controls, and mg / mL solutions of C7 peptide in water were prepared as additional controls. Each formulation was tested in triplicate, and blank formulations, untreated skin, and C7 peptide solutions as free, unencapsulated peptide were used as background fractions for the analysis.

[0362] Diffusion Cell Study-Transdermal Delivery of C7

[0363] In this study, skin sections were taken for further analysis by mass spectrometry by Climax Analytical Laboratories.

[0364] These studies demonstrated that all three multisome formulations delivered C7 peptide deep into and through human skin in vitro. Among these multisome formulations, formulations F6A-C7 and F1B-C7 delivered approximately two-fold more C7 peptide compared to F1C-C7.

[0365] Diffusion cell (0.636 cm 2 The total amount of C7 peptide delivered (Qt(24h)) through a 9mm diameter skin disk treated in 100μg of skin surface area (20μg / cm) was 599.62±265.62, 600.46±402.77, and 276.56±111.47ng / 24h for F6A-C7, F1B-C7, and F1C-C7, respectively, consistent with delivery rates of 0.3, 0.3, and 0.14% for each formulation. Qt per unit surface area of ​​skin, i.e., ng / cm 2 and percent C7 peptide delivery are shown in Table M.

[0366] [Table 15]

[0367] These experiments showed that the levels of enhancement by the synergistic components were as follows for C7 peptide: PEFA / Oleth-2=Tween80 / Span40 / PEG-4-dilaurate>Tween80 / Span40 / Oleth-2.

[0368] Example 6. Evaluation of the safety and efficacy of a muscle-type nAChR peptide antagonist compared to placebo for the treatment of facial wrinkles Lipid vesicle formulations of the disclosed muscle-type nAChR peptide antagonists are tested for safety and efficacy compared to placebo in the treatment of facial wrinkles (skin wrinkles) and glabellar lines in a randomized, double-blind, human clinical trial. Patients (50 per group) are treated with an amount of muscle-type nAChR peptide antagonist or placebo (blank lipid vesicle) applied to the bilateral frontal and glabellar lines areas on day 1.

[0369] Primary outcome: Proportion of participants achieving no or mild investigator-assessed score on the Photo-Simulated Facial Wrinkle Scale (FWS) for frontal wrinkles at maximum brow lift.

[0370] At day 30, patients will be assessed for severity of frontal wrinkles at maximum brow lift using a 4-point Facial Wrinkle Scale (FWS) with photo-quantification guide: 0=none, 1=mild, 2=moderate, or 3=severe. The percentage of participants with no or mild scores will be determined.

[0371] Primary outcome: Proportion of participants achieving no or mild scores on subjective assessment of the Facial Wrinkle Scale (FWS) using a photographic quantification guide (FWS) for frontal wrinkles at maximum brow lift.

[0372] Additionally, on day 30, patients will assess the severity of frontal wrinkles at maximum brow lift using a 4-point Facial Wrinkle Scale (FWS) with a photo-quantification guide: 0=none, 1=mild, 2=moderate, or 3=severe, to determine the percentage of participants with no or mild scores.

[0373] Secondary outcomes: Proportion of participants achieving satisfied or very satisfied by subjective assessment (participant assessment) of satisfaction with the appearance of frontal wrinkles.

[0374] At day 30, participants will rate their overall satisfaction with the appearance of their frontal wrinkle area using a 5-point scale: 1=very dissatisfied, 2=dissatisfied, 3=neutral, 4=satisfied, or 5=very satisfied. The percentage of participants who rated themselves as satisfied or very satisfied will be determined.

[0375] Secondary Outcome: Proportion of participants with =1 grade improvement from baseline by investigator-assessed FWS in frontal crease at rest.

[0376] At baseline and day 30, the investigator will assess the severity of patients' frontal wrinkles at rest using a 4-point FWS: 0=none, 1=mild, 2=moderate, or 3=severe. The percentage of participants with a 1-grade improvement from baseline will be determined.

[0377] Secondary outcome: Proportion of participants with =1 grade improvement from baseline by subject-rated FWS in frontal crease at rest.

[0378] At baseline and day 30, participants will rate the severity of their frontal wrinkles at rest using a 4-point FWS: 0=none, 1=mild, 2=moderate, or 3=severe. The percentage of participants with a 1-grade improvement from baseline will be determined.

[0379] Example 7. Evaluation of the safety and efficacy of topical application of a lipid vesicle composition containing hyaluronic acid compared to placebo for enhancing lip features The lipid vesicle formulation of hyaluronic acid of the present disclosure is tested for safety and efficacy in comparison with placebo when applied to lips in a randomized double-blind human clinical trial.On day 1, a certain amount of lipid vesicle formulation containing hyaluronic acid or placebo (blank lipid vesicle) is applied to the upper and lower lips of subjects (50 in each group).

[0380] Primary outcome: An increase in lip fullness of at least 1 grade on the Medicis Lip Fullness Scale (MLFS) after 14 and 30 days of use. The MLFS is adjudicated by an independent dermatologist: the MLFS is a validated 5-point scale for lip fullness (1=very thin; 2=thin; 3=medium; 4=full; 5=very plump).

[0381] On day 14, subjects' lip fullness is assessed using a 5-point MLFS scale: 1=very thin; 2=thin; 3=medium; 4=full; 5=very plump, and compared to baseline (before use of the lipid vesicle composition). The number of subjects with an improvement in at least one MLFS level is determined.

[0382] At day 30, subjects' lip fullness is assessed again using a 5-point MLFS to determine longevity of effect after use. MLFS scores are compared to baseline and day 14 results. The number of subjects with improvement of at least one MLFS level is determined.

[0383] Secondary outcomes: Proportion of participants achieving satisfied or very satisfied by subjective assessment of satisfaction with lip appearance (participant assessment).

[0384] At day 30, participants rate their overall satisfaction with the appearance of their lips using a 5-point scale: 1=very dissatisfied, 2=dissatisfied, 3=neutral, 4=satisfied, or 5=very satisfied. The percentage of participants who rated themselves as satisfied or very satisfied is determined.

[0385] Secondary outcomes: Dermatologist assessment of filler effectiveness and durability: Dermatologist opinion of the effectiveness and durability of the filler effect will be assessed using a 5-point Physician's Global Assessment (IGA) scale, where 1 = worse, 2 = slightly improved, 3 = improved, 4 = very improved, 5 = very well improved.

[0386] An independent dermatologist will compare images of the subjects' lips taken on days 0 (before use), 14, and 30. The number of subjects rating improved, very improved, or very much improved will be assessed and compared for days 14 and 30.

[0387] Example 8. Evaluation of the safety and efficacy of topical application of lipid vesicle compositions containing hyaluronic acid for enhancement of lip features Hyaluronic acid lipid vesicle formulations as described herein (e.g., hyaluronic acid lipid vesicle formulations as described in Example 1) were tested for safety and effectiveness of application to lips in human clinical trials.A certain amount of hyaluronic acid-containing lipid vesicle formulations was applied to the upper and lower lips of subjects (55 in total) for 5 days.Subjects were aged 35 years or older and included various ethnic backgrounds and skin types.

[0388] Subjects applied five layers of the formulation twice daily (morning and night). Each layer of the formulation was reapplied once the previous layer was completely absorbed into the subject's upper and lower lips. The increase in upper and lower lip height of each subject was measured before and after five days, as measured from a baseline between the upper and lower lips.

[0389] Figure 9 shows the results after 5 days of application, where subjects across ethnicities and skin types showed significantly fuller lips. As shown, subjects had anywhere from a 2.7% to 45% increase in upper lip height measured from baseline, and anywhere from a 3.6% to 85% increase in lower lip height measured from baseline. Additionally, subjects felt that their lips were smoother and more vibrant in color 5 days after applying the formulation.

[0390] Example 9: Development of lipid vesicle formulations The starting compositions for the development of lipid vesicle formulations were based on the earlier F6A-C7 formulation as well as the F-C10-5 and F-C10-6 formulations, whose main enhancer combinations are shown in Table N.

[0391] [Table 16]

[0392] Using these formulations as a starting point, variations of System A (aqueous phase) (Table O) were developed as well as variations of the phospholipid phase (Table P).

[0393] [Table 17]

[0394] [Table 18]

[0395] The various formulations were evaluated using viscosity (sensory attribute) as well as microscopy (Figure 10A-10F). The various formulations were assigned a Cosmetic Rating Scale (CRS): 1 (slightly unattractive), 2 (acceptable) and 3 (excellent). Of the formulations evaluated in Figures 10A-10F, five sets of multiphasic vesicle formulations containing C7 (2 mg / g) were selected for diffusion cell testing to evaluate the best enhancer combination. The F1 series represented the cholesterol-containing formulations in the phospholipid phase and the F5P series represented the vegan substitute phytosterol-containing formulations. The five sets selected include F5P-F15COSM and F1-F15COSM, F5P-F15-4COSM and F1-F15-4COSM, F5P-F16COSM and F1-F16COSM, F5P-F19COSM and F1-F19COSM, and F5P-F18COSM and F1-F18COSM.

[0396] The first diffusion cell study involved analysis of the average amount of peptide absorbed into the viable skin as well as the average percent absorbed into the viable skin. The results are shown in Table Q.

[0397] [Table 19]

[0398] Based on the results shown in Table Q, three formulations were selected for further evaluation: F5P-G15-4COSM, F5P-F16COSM, and F5P-F18COSM. These formulations were prioritized based on delivery efficiency and vegan composition.

[0399] A second diffusion test was performed using confocal microscopy to evaluate peptide delivery in skin sections. Figure 11A shows confocal microscopy images of skin samples with F5P2.6-F16COSM, F5P2.6-F15-4COSM, and F5P2.6-F18COSM. These formulations contained phytosterol MM. Figure 11B shows confocal microscopy images of skin samples with F5P2.6-F16MCOSM, F5P2.6-F15-4MCOSM, and F5P2.6-F18MCOSM. The M in the formulation code indicates a "modified" composition in which isopropyl myristate was added to system A to evaluate whether this addition produced an enhancing effect in the formulation.

[0400] Based on the results shown in Figures 11A-11B, the F5P-F16COSM formulation showed the highest level of peptide delivery. Furthermore, we observed penetration of the peptide deep into the skin layers. Also, the peptide was delivered across the full thickness skin. F5P-F15-4COSM and F5P-F18COSM showed good delivery to the epidermis and upper skin layers, but not as deep. The addition (or increase) of isopropyl myristate to these formulations did not increase delivery, but rather seemed to decrease delivery in all three formulations. Here, F5P-F16COSM was selected as the lead formulation.

[0401] The lead formulation F5P2.6-F16COSM was further optimized: F5P2.6-F16STCOSM. Optimization was performed to increase the physical stability of the formulation. Optimization included the addition of one or more non-ionic surfactants as described herein. These two formulations were compared to confirm that optimization did not change the delivery efficiency. The two formulations were evaluated using confocal microscopy (Figure 12) and measurements of the mean fluorescence intensity of the epidermis and dermis of the two formulations (Table R). Both results showed that the final formulation was scalable, stable, and effective in delivering C7 peptide.

[0402] [Table 20]

[0403] While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the present invention. It is understood that various alternatives to the embodiments of the present invention described herein may be utilized in implementing the present invention. The following claims define the scope of the present invention, and it is intended that methods and structures within the scope of the claims, and equivalents thereof, be covered thereby.

Claims

1. A lipid vesicle composition comprising a plurality of lipid vesicles comprising diethylene glycol monooleyl ether, oleyl alcohol, and propylene glycol monocaprylate.

2. The lipid vesicle composition of claim 1, further comprising a lipid bilayer and an oil-in-water emulsion encapsulated within the lipid bilayer.

3. The lipid vesicle composition of claim 2, wherein the oil-in-water emulsion comprises one or more of the diethylene glycol monooleyl ether, the oleyl alcohol, and the propylene glycol monocaprylate.

4. A lipid vesicle composition described in any one of claims 1 to 3, further comprising a peptide antagonist of the muscle-type nicotinic acetylcholine receptor.

5. The lipid vesicle composition described in claim 4, wherein the peptide antagonist has an amino acid sequence identical to any one of the amino acid sequences of SEQ ID NOs: 1 to 52 or 60 to 99.

6. The lipid vesicle composition described in claim 4 or 5, wherein the peptide antagonist has an amino acid sequence identical to the amino acid sequence of SEQ ID NO: 60, 61, 73, 78, 82, 85, 91, or 95.

7. The lipid vesicle composition of claim 4, wherein the peptide antagonist is present at a concentration of about 0.1 mg / mL to about 10 mg / mL.

8. A lipid vesicle composition described in any one of claims 1 to 7, further comprising a nonionic surfactant or a combination of nonionic surfactants selected from polyethylene glycol ethers of aliphatic alcohols, sorbitan esters, polysorbates, sorbitan esters and polyethylene glycol fatty acid esters, and combinations thereof.

9. The lipid vesicle composition of claim 8, wherein the polyethylene glycol ether of an aliphatic alcohol comprises a C 8 -C 22 aliphatic alcohol and a polyethylene glycol group having from about 2 to about 8 ethylene glycol subunits.

10. The lipid vesicle composition of claim 8 or 9, wherein the polyethylene glycol ether of an aliphatic alcohol comprises diethylene glycol hexadecyl ether, 2-(2-octadecaoxyethoxy)ethanol, diethylene glycol monooleyl ether, polyoxyethylene (3) oleyl ether, or polyoxyethylene (5) oleyl ether, or any combination thereof.

11. The lipid vesicle composition of any one of claims 8 to 10, wherein the sorbitan ester comprises sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trioleate, sorbitan sesquioleate, or sorbitan isostearate, or any combination thereof.

12. The lipid vesicle composition of any one of claims 8 to 11, wherein the polyethylene glycol fatty acid ester comprises PEG-8 dilaurate, PEG-4 dilaurate, PEG-4 laurate, PEG-8 dioleate, PEG-8 distearate, PEG-8 distearate, PEG-7 glyceryl cocoate, and PEG-20 almond glyceride, or any combination thereof.

13. The lipid vesicle composition of any one of claims 8 to 12, wherein the polysorbate comprises polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, or any combination thereof.

14. A lipid vesicle composition according to any one of claims 8 to 13, wherein each of the non-ionic surfactants has a hydrophobic-lipophilic balance (HLB) of about 10 or less.

15. The lipid vesicle composition of any one of claims 1 to 14, further comprising monolauroyl lysine or dipalmitoyl lysine, or a combination thereof.

16. A lipid vesicle composition according to any one of claims 1 to 15, comprising a cationic surfactant.

17. The lipid vesicle composition of claim 16, wherein the cationic surfactant comprises a fatty acid amide-derived propylene glycol diammonium phosphate ester.

18. The lipid vesicle composition of claim 16, wherein the cationic surfactant comprises linoleamidopropyl PG-dimonium chloride phosphate.

19. A lipid vesicle composition according to any one of claims 1 to 18, comprising a sterol.

20. The lipid vesicle composition of claim 19, wherein the sterol comprises cholesterol or a phytosterol.

Images