Emulsion and nanoliposome comprising ceramide

Abstract

One aspect relates to an emulsion comprising: a lipid core including ceramide and fatty acid; and a nonionic surfactant. The composition according to one aspect stably emulsifies ceramide, which is a poorly soluble substance, and can stably deliver a high content of ceramide without precipitation. In addition, the composition exhibits enhanced skin permeability, and thus can be advantageously applied to cosmetics, topical skin preparations, and the like comprising ceramide as an active ingredient. Another aspect relates to liposomes for simultaneous delivery of fat-soluble substances and water-soluble substances, comprising ceramides, fatty acids, amino acids, polyols, nonionic surfactants, and phospholipids. The liposome for simultaneous delivery of a fat-soluble substance and a water-soluble substance according to one aspect is prepared by first preparing an emulsion comprising ceramide and then adding a phospholipid, thereby significantly increasing storage stability of the prepared liposome and enabling stable delivery of fat-soluble substance and water-soluble substance to the skin without precipitation. Therefore, the liposome can simultaneously deliver ceramide, fat-soluble substances, and water-soluble substances to the skin, and thus can be advantageously applied to cosmetics, topical skin preparations, and the like.

Description

Emulsions and nanoliposomes containing ceramide

[0001] This relates to emulsions and nanoliposomes containing ceramide.

[0002] Liposomes are utilized as carriers for various drugs and are used to stably deliver water-soluble and fat-soluble active ingredients, which are used as raw materials in functional cosmetics, into the skin. However, liposomes have the disadvantage of high physical and chemical instability, which can lead to easy precipitation or the leakage of encapsulated active ingredients.

[0003] Ceramide is a type of sphingolipid with a structure in which fatty acids are linked to sphingosine or phytosphingosine. Ceramide accounts for more than 40% of the intercellular lipids in the stratum corneum of the skin and is a component of the skin's lipid membrane and the cell membrane of the skin's bilayer. In the skin, ceramide, along with cholesterol and free fatty acids, prevents water loss and forms a lipid mortar, an impermeable barrier that prevents evaporative water loss.

[0004] In addition, ceramides are essential components for the structural formation and function of the stratum corneum, and the ceramides present in the human body are classified into various types depending on their degree of polarity.

[0005] The diversity of ceramide structures plays an important role in the unique properties of the stratum corneum across different organs of the body. For example, the stratum corneum of the face is thin and flexible, allowing for various facial expressions, while the stratum corneum covering the heels is thick and hard, protecting against trauma; thus, depending on the unique properties of the stratum corneum, the structure of ceramides and the lipid composition of the skin epidermis are distributed differently depending on the body part.

[0006] Similar to the specific changes in ceramide abundance depending on the body part, there are also characteristic changes in epidermal ceramide expression in patients with inflammatory skin diseases. In patients with psoriasis, it has been reported that ceramide AS and ceramide NS are increased, while ceramide EOS, ceramide AP, and ceramide NP are decreased. Such changes in ceramide expression may contribute to defects in the impermeable barrier that prevents evaporative moisture loss from the skin. Furthermore, previous studies investigating ceramide expression in patients with atopic dermatitis and psoriasis reported that the length of ceramide sphingoid bases and fatty acid chains have the greatest influence on the potential for upregulation or downregulation of specific ceramide structures in inflamed skin. Ceramides possessing these characteristics can be used as ingredients in topical dermatological drugs to complement the treatment of skin diseases such as eczema, and are therefore actively utilized in related industries.

[0007] However, ceramides are insoluble in both water and oil, which limits their use, and they often exhibit low efficacy even when used in high concentrations. Therefore, research is actively underway to formulate them into microemulsions, liposomes, and cubosomes to address these issues.

[0008] However, when ceramide, which has the characteristics of a dielectric material, is formulated using the aforementioned methods, it is difficult to induce the dipole moment of the ceramide, resulting in low transdermal penetration. Furthermore, there is a problem in that the stability of the formulation containing the manufactured ceramide is very low, making it impossible to load high concentrations of ceramide.

[0009] Accordingly, the inventors of the present invention aim to provide a liposome capable of delivering ceramide to the skin and simultaneously encapsulating fat-soluble and water-soluble substances within the liposome by inducing a dipole moment in ceramide (delta positive) through fatty acids to form hydrogen bonds and positioning the ceramide between phospholipid bilayers to produce a liposome with a stable structure, and at the same time, by coating the outside of the ceramide particles with polar molecules to stably contain ceramide in a high amount, and an emulsion comprising ceramide, fatty acids, amino acids, polyols, and nonionic surfactants.

[0010] One aspect provides a lipid core comprising ceramide and fatty acid; and an emulsion comprising a nonionic surfactant.

[0011] Another aspect is to provide a cosmetic composition comprising the above-mentioned emulsion.

[0012] Another aspect is to provide a topical skin preparation containing the above-mentioned emulsion.

[0013] Another aspect is the step of obtaining a lipid mixture by mixing ceramide, fatty acids, and a nonionic surfactant; and

[0014] The present invention provides a method for preparing an emulsion comprising the step of mixing the above lipid mixture and a eutectic solvent to obtain an emulsion.

[0015] One aspect is a liposome for the simultaneous delivery of lipid-soluble and water-soluble substances, comprising ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid,

[0016] The above ceramide and fatty acid provide a liposome that forms a lipid core through hydrogen bonding.

[0017] Another aspect is to provide a cosmetic composition comprising the above-mentioned liposome, a fat-soluble substance, and a water-soluble substance.

[0018] Another aspect is to provide a topical skin preparation comprising the above-mentioned liposome, a fat-soluble substance, and a water-soluble substance.

[0019] Another aspect is the step of obtaining a lipid mixture by mixing ceramide, fatty acids, and a nonionic surfactant;

[0020] A step of obtaining an emulsion by mixing the above lipid mixture and a eutectic solvent; and

[0021] The present invention provides a method for preparing liposomes for the simultaneous delivery of lipid-soluble substances and water-soluble substances, comprising the step of mixing the above emulsion and phospholipid to obtain liposomes.

[0022] One aspect provides an emulsion comprising a lipid core comprising ceramide and fatty acids; and a nonionic surfactant. Specifically, the emulsion may comprise a lipid core comprising ceramide and fatty acids, wherein the ceramide is included in an amount of 1 to 15 weight% based on the total weight of the emulsion, and the lipid core is formed by hydrogen bonding between the ceramide and fatty acids; and a nonionic surfactant.

[0023] Specifically, the fatty acid included in the oil phase forms hydrogen bonds and van der Waals forces with the ceramide, thereby preventing the ceramide included in the oil phase from precipitating without being dispersed in the aqueous phase. In addition, since the fatty acid and the ceramide maintain a nano-sized average diameter by forming hydrogen bonds and van der Waals forces, the ceramide can be stably dispersed in the aqueous phase. The nonionic surfactant included in the oil phase acts as an auxiliary emulsifier and can significantly reduce the ceramide that precipitates and is released in the aqueous phase (see FIGS. 1 and 2).

[0024] In one embodiment, the nonionic surfactant may surround the surface of the lipid core, and the hydrophilic end of the surfactant may interact with the fatty acid of the lipid core, while the hydrophobic end may be arranged toward the interior of the lipid core surface.

[0025] In one embodiment, the ceramide may be included in an amount of 1 to 15 weight% relative to the total weight of the emulsion. For example, the ceramide may be included in an amount of 1 to 15 weight%, 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 15 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 15 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, 7.5 to 15 weight%, 7.5 to 10 weight%, or 10 to 15 weight% relative to the total weight of the emulsion. At this time, if the content of the ceramide is below the above range, there is a problem that it does not sufficiently exhibit skin moisturizing, skin barrier strengthening, skin aging improvement, skin wrinkle improvement, skin elasticity improvement, or skin regeneration effects, and if it exceeds the above range, there is a problem that ceramide crystals are formed.

[0026] The above "ceramide" is a type of sphingolipid, a molecule composed of sphingosine and fatty acids. Ceramide is present in large quantities in cell membranes and is involved in various cellular signaling processes, such as cell differentiation, growth, and apoptosis. Ceramide exists in various structures depending on the combination of sphingoids and fatty acids. The above ceramide may include natural ceramide, synthetic ceramide, pseudo-ceramide, or derivatives thereof. Natural ceramide refers to ceramide existing in nature and may be ceramide extracted from animals, plants, and microorganisms. Pseudo-ceramide is a general term for compounds having a double-chain lamellar structure similar to that of natural ceramide. Examples of the above-mentioned similar ceramides include 1,3-bis(N-2-(hydroxyethyl)palmitoylamino)-2-hydroxypropane, 1,3-bis(N-2-(hydroxyethyl)lauroylamino)-2-hydroxypropane, 1,3-bis(N-2-(hydroxyethyl)isostearoylamino)-2-hydroxypropane, and 1,3-bis(N-2-(hydroxyethyl)stearoylamino)-2-hydroxypropane.

[0027] In one embodiment, the ceramide may be one or more selected from the group consisting of ceramide NS, ceramide AS, ceramide EOS, ceramide NDS, ceramide ADS, ceramide EODS, ceramide NP, ceramide AP, ceramide EOP, ceramide NH, ceramide AH, and ceramide EOH, and specifically may be ceramide NP.

[0028] The above "ceramide NP" corresponds to a synthetic N-acylated sphingolipid in which a saturated or unsaturated fatty acid is bound to phytosphingosine having a D-erythro structure. Specifically, ceramide NP is represented by the following chemical formula 1, its IUPAC name is (Z)-N-[(2S,3S,4R)-1,3,4-trihydroxyoctadecan-2-yl]octadec-9-enamide, and its molecular weight is 582.0 g / mol:

[0029] [Chemical Formula 1]

[0030] .

[0031] In one embodiment, the fatty acid may be one or more selected from the group consisting of oleic acid, linoleic acid, linolenic acid, lauric acid, palmitic acid, stearic acid, elaidic acid, myristic acid, arachidonic acid, isopropyl myristate, and caprylic / capric triglycerides. Specifically, the fatty acid may be one or more selected from the group consisting of oleic acid, lauric acid, palmitic acid, stearic acid, isopropyl myristate, and caprylic / capric triglycerides, and more specifically, it may be oleic acid.

[0032] In one embodiment, the fatty acid may be included in an amount of 10 to 30 weight% relative to the total weight of the emulsion. For example, the fatty acid may be included in an amount of 10 to 30 weight%, 10 to 25 weight%, 10 to 20 weight%, 10 to 15 weight%, 15 to 30 weight%, 15 to 25 weight%, 15 to 20 weight%, 20 to 30 weight%, 20 to 25 weight%, or 25 to 30 weight% relative to the total weight of the emulsion.

[0033] In this specification, the term "lipid core" refers to a lipid complex in which ceramide and fatty acids are bonded through hydrogen bonds, which stabilizes the emulsion structure and inhibits the precipitation of active ingredients (e.g., ceramide) encapsulated within the emulsion structure.

[0034] In one embodiment, the ceramide and fatty acid may be included in a weight ratio of 3 to 5: 5 to 10 based on the weight of the amino acid included in the emulsion. Specifically, the ceramide and fatty acid may be included in a weight ratio of 3 to 5: 5 to 10, 3 to 4: 5 to 10, 3 to 5: 5 to 8, 3 to 5: 5 to 7, 3 to 4: 5 to 8, or 3 to 4: 5 to 7 based on the weight of the amino acid included in the emulsion. At this time, if the weight ratio of the ceramide and fatty acid is less than the above range, there is a problem of lipid leakage in the emulsion, and if it exceeds the above range, there is a problem of ceramide crystal formation.

[0035] In one embodiment, the weight ratio of the ceramide and the fatty acid may be 1:1 to 4. Specifically, the weight ratio of the ceramide and the fatty acid may be 1:1 to 4, 1:1 to 3.5, 1:1 to 3, 1:1 to 2.5, or 1:1 to 2.

[0036] Specifically, when the weight ratio of the ceramide and fatty acid contained in the emulsion is 1:1 to 4, or when the weight ratio of the ceramide and fatty acid is 3 to 5:5 to 10 based on the weight of the amino acid contained in the emulsion, the fatty acid induces a dipole moment in the ceramide, and the carboxyl group (-COOH) of the fatty acid forms a hydrogen bond with the amino group (-NH2) of the ceramide, thereby stabilizing the ceramide within the emulsion. Accordingly, the amount of precipitated ceramide in the emulsion is significantly reduced to less than 0.1% compared to conventional ceramide carriers, and it exhibits excellent storage stability even under harsh conditions (e.g., high temperature, low temperature, repeated freezing and thawing conditions, etc.).

[0037] Furthermore, when the weight ratio of the ceramide and fatty acid contained in the emulsion is 1:1 to 4, or when the weight ratio of the ceramide and fatty acid is 3 to 5:5 to 10 based on the weight of the amino acid contained in the emulsion, the amino group (-NH2) of the ceramide combines with the carboxyl group (-COOH) of the fatty acid and is arranged on the outermost surface of the emulsion, and the transdermal permeability of the emulsion containing the active ingredient is significantly increased by the arrangement of the emulsion surface.

[0038] The term "surfactant" above refers to a compound that relaxes the interface between two or more substances, liquids, liquid phases, or continuous phases to mix liquids that do not mix with each other, and is dispersed as one or more dispersed phases without being separated into or emulsified within one or more continuous phases.

[0039] The term "nonionic surfactant" above refers to a surfactant containing hydrophilic and hydrophobic functional groups that do not dissociate into ions within the molecule.

[0040] In one embodiment, the nonionic surfactant may be one or more selected from the group consisting of a polyglyceryl-based surfactant having 12 to 20 carbon atoms, a glyceryl-based surfactant having 12 to 20 carbon atoms, an alkyl glucoside-based surfactant having 12 to 20 carbon atoms, a polyethylene glycol sorbitan-based surfactant having 12 to 20 carbon atoms, and a polyethylene glycol-based surfactant having 12 to 20 carbon atoms. Specifically, it may be one or more selected from the group consisting of polyglyceryl-10 stearate, glyceryl stearate / PEG-100 stearate, polysorbate 80, polysorbate 20, decyl glucoside, polyglyceryl-10 laurate, and polyglyceryl-10 oleate. More specifically, it may be one or more selected from the group consisting of polyglyceryl-10 stearate, decyl glucoside, polyglyceryl-10 laurate, and polyglyceryl-10 oleate.

[0041] More specifically, the above nonionic surfactant may be polyglyceryl-10 stearate, decyl glucoside, polyglyceryl-10 laurate, or polyglyceryl-10 oleate.

[0042] In one embodiment, the nonionic surfactant may be included in an amount of 1 to 15 weight% relative to the total weight of the emulsion, and specifically, the nonionic surfactant may be included in an amount of 1 to 15 weight%, 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 15 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 15 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, 7.5 to 15 weight%, 7.5 to 10 weight%, or 10 to 15 weight% relative to the total weight of the emulsion.

[0043] Additionally, the term "emulsion" refers to a type of colloid formed when one liquid is dispersed in another of two immiscible liquids (e.g., an oil phase and a water phase), and can be used interchangeably with "emulsion."

[0044] Figure 1 shows a nanoemulsion captured by Cryo-EM (High Resolution Cryo-Transmission Electron Microscope packaging system), and Figures 2a and 2b show the structures of ceramide and fatty acids within the nanoemulsion.

[0045] As shown in FIGS. 1, 2a, and 2b, in one aspect, the lipid core can induce an induced dipole-dipole interaction between the ceramide and fatty acid molecules by forming hydrogen bonds between the amino group (-NH2) of the ceramide and the carboxyl group (-COOH) of the fatty acid. Additionally, a permanent dipole-dipole interaction can be induced between the nonionic surfactant and the fatty acid molecules. That is, in one aspect, the lipid core can form a spherical aggregate in which the fatty acid chain of the ceramide and the hydrophobic group of the nonionic surfactant face inward, and the hydrophilic group of the fatty acid and the nonionic surfactant faces outward.

[0046] Accordingly, in one embodiment, the emulsion can form a lamellar liquid crystal structure. In addition, the lipid core can form a multilamellar vesicle as a spherical structure surrounded by a multilayer ceramide and fatty acid bilayer.

[0047] In addition, in other embodiments, the emulsion may be of the water-in-water type (O / W), oil-in-water type (W / O), water-in-oil-in-water type (W / O / W), or oil-in-water type (O / W / O), and specifically, may be of the water-in-water type (O / W) or oil-in-water type (W / O).

[0048] The above water-in-water type may contain a water-soluble component on the outside of the emulsion particles and a fat-soluble component on the inside, and the oil-in-water type may contain a fat-soluble component on the outside of the emulsion particles and a water-soluble component on the inside. Additionally, the water-in-oil-in-water type or the oil-in-water type may contain a water-soluble or fat-soluble component on the outside of the emulsion particles and a fat-soluble or water-soluble component on the inside.

[0049] The term "emulsion particle" refers to the oil phase or aqueous phase particle dispersed within the aqueous phase or oil phase. For example, if the emulsion according to one aspect is of the water-in-water type, the emulsion particle may refer to spherical oil phase particles dispersed like colloids within the aqueous phase. Additionally, if the emulsion according to one aspect is of the water-in-oil type, the emulsion particle may refer to spherical aqueous phase particles dispersed like colloids within the oil phase.

[0050] In one embodiment, the average particle diameter of the emulsion may be 10 nm to 250 nm, specifically 10 nm to 250 nm, 10 nm to 200 nm, 10 nm to 150 nm, 10 nm to 100 nm, 10 nm to 50 nm, 50 nm to 250 nm, 50 nm to 200 nm, 50 nm to 150 nm, 50 nm to 100 nm, 100 nm to 250 nm, 100 nm to 200 nm, 100 nm to 150 nm, 150 nm to 250 nm, 150 nm to 200 nm, or 200 nm to 250 nm.

[0051] According to one aspect, the particles of the emulsion are nano-sized particles with an average diameter of 10 nm to 250 nm. Accordingly, the emulsion particles can easily penetrate the stratum corneum of the skin and deliver ceramide into the skin. When the average diameter of the particles is larger than 250 nm, the penetration rate into the stratum corneum is significantly lower, resulting in low efficacy of ceramide.

[0052] In one embodiment, the average zeta potential of the particles of the emulsion may be -10 to -60 mV. Specifically, the average zeta potential of the particles of the emulsion may be -10 to -60 mV, -10 to -50 mV, -10 to -40 mV, -10 to -30 mV, -10 to -20 mV, -20 to -60 mV, -20 to -50 mV, -20 to -40 mV, -20 to -30 mV, -30 to -60 mV, -30 to -50 mV, -30 to -40 mV, -40 to -60 mV, -40 to -50 mV, or -50 to -60 mV.

[0053] The term "zeta potential" refers to the potential difference between a medium and a fixed fluid layer attached to dispersed particles, and zeta potential is utilized as a key indicator of the stability of colloidal dispersions.

[0054] When the average zeta potential of the particles of the above emulsion is -10 to -60 mV, the particles do not aggregate or precipitate due to electrostatic repulsion between the particles and can be stably dispersed in the solution. On the other hand, when the absolute value of the above zeta potential is less than 10, the repulsion between the particles is significantly reduced, causing aggregation and precipitation to occur easily, thereby reducing emulsion stability.

[0055] In one experimental example, when ceramide, fatty acid, and nonionic surfactant are included in the emulsion in a specific weight ratio of the experimental example, it was confirmed that the ceramide is stably dispersed without precipitating even under harsh conditions (e.g., high temperature, low temperature, repeated freezing and thawing conditions, etc.) (see Experimental Examples 1 to 3). Therefore, the emulsion according to one embodiment can exhibit significantly excellent storage stability.

[0056] Accordingly, the emulsion according to one aspect contains ceramide, fatty acid, and a nonionic surfactant in a specific weight ratio of the experimental example, thereby significantly reducing the amount of ceramide released during storage and exhibiting high storage stability, and can stably deliver a high content of ceramide into the skin without precipitation. In addition, since the emulsion has excellent skin permeability and high skin delivery efficiency of ceramide, it can be utilized in cosmetics, topical skin preparations, etc., containing ceramide as an active ingredient.

[0057] In one experimental example, when the emulsion contains a nonionic surfactant as polyglyceryl-10 stearate, polyglyceryl-10 oleate, polyglyceryl-10 laurate, or decyl glucoside (Examples 1 to 4), it was confirmed that the emulsion particles are stably dispersed nanoparticles with an average size of 180 nm or less and a PDI of 0.321 or less. In addition, it was confirmed that there were no precipitated ceramides when the emulsion solution was observed visually (see Experimental Examples 4 and 5).

[0058] In other experimental examples, the emulsion particles (Examples 1 to 4) exhibited a negative charge, and in particular, when decyl glucoside was used as a nonionic surfactant, a zeta potential of -45.64 mV was exhibited, confirming that the emulsion particles were stably dispersed by inter-particle repulsion (see Experimental Example 6).

[0059] In addition, in other experimental examples, the ceramide encapsulation rate in the emulsion was 99.9% (Examples 1 to 4), confirming that the emulsion according to one aspect can encapsulate a high content of ceramide without any released ceramide (see Experimental Example 7).

[0060] In one embodiment, the ceramide encapsulation rate of the emulsion may be 90 to 99.99%. Specifically, the ceramide encapsulation rate of the emulsion may be 90 to 99.99%, 90 to 99.90%, 90 to 99.00%, 90 to 95%, 95 to 99.99%, 95 to 99.90%, 95 to 99.00%, 99.00 to 99.99%, 99.00 to 99.90%, or 99.90 to 99.99%.

[0061] The above term "encapsulation rate" is a value calculated as the ratio of ceramide actually encapsulated in the emulsion among the ceramide actually added.

[0062] The above emulsion encapsulates a high concentration of ceramide within the emulsion by introducing a fatty acid having a carboxyl group (-COOH) capable of forming hydrogen bonds with the amino group (-NH2) of ceramide. In addition, the emulsion is stabilized to prevent the encapsulated ceramide from precipitating by including a nonionic surfactant.

[0063] The term "storage stability" means that when an emulsion and / or cosmetic composition is stored for a certain period (e.g., one week, one month, three months, etc.) at a specific temperature (e.g., -20 ℃, 4 ℃, 20 ℃, 37 ℃, etc.) or under conditions where the temperature changes (e.g., changing from -20 to 20 ℃), the particles within the emulsion containing ceramide remain stably dispersed without clumping among particles, sedimentation of particles, or separation of the oil and water phases occurring, even under conditions of low temperature, high temperature, or change from low temperature to high temperature. Additionally, the above storage stability may be used interchangeably with emulsion stability.

[0064] The term "skin permeability" refers to the ability of an emulsion and / or cosmetic composition to pass through the stratum corneum, which is the outermost layer of the skin's epidermis, and to deliver active ingredients into the stratum corneum; furthermore, it refers to the ability to deliver active ingredients into the epidermal layers (stratum lucidum, stratum granulosum, stratum spinosum, stratum basale), dermal layers, or into the body that exist within the stratum corneum. Additionally, the term "skin permeability" may be used interchangeably with "transdermal permeability."

[0065] In one embodiment, the polydispersity index (PDI) of the particles of the emulsion may be 0.001 to 0.325. Specifically, the polydispersity index (PDI) of the particles of the emulsion is 0.001 to 0.325, 0.001 to 0.250, 0.001 to 0.200, 0.001 to 0.150, 0.001 to 0.100, 0.001 to 0.050, 0.050 to 0.325, 0.050 to 0.250, 0.050 to 0.200, 0.050 to 0.150, 0.050 to 0.100, 0.100 to 0.325, 0.100 to 0.250, 0.100 to 0.200, 0.100 to 0.150, 0.150 to It may be 0.325, 0.150 to 0.250, 0.150 to 0.200, 0.200 to 0.325, 0.200 to 0.250, or 0.250 to 0.325.

[0066] The term "Polydispersity Index (PDI)" is an index representing the particle size distribution, and the closer the index is to 0, the more uniform the particle size, that is, the closer the particle distribution is to a stable monodispersity.

[0067] When the polydispersity index (PDI) of the particles of the above emulsion is between 0.001 and 0.325, it means that the emulsion has a uniform particle distribution and is in a stable state. As the PDI of the particles of the above emulsion approaches 1, the size of the emulsion particles varies, and the possibility of aggregation, precipitation, etc. becomes significantly higher.

[0068]

[0069] In one embodiment, the emulsion may further include an amino acid, a polyol, or a combination thereof. The amino acid may be one or more selected from the group consisting of arginine, histidine, and lysine, and specifically may be arginine.

[0070] In addition, the polyol may be a polyol having 3 to 7 carbon atoms, specifically a polyol having 3 to 5 carbon atoms, and more specifically a polyol having 3 or 4 carbon atoms. In addition, the polyol may be a diol or a triol. Specifically, the polyol may be a diol or a triol having 3 to 7 carbon atoms, more specifically a diol or a triol having 3 to 5 carbon atoms, and even more specifically a diol or a triol having 3 carbon atoms or a diol or a triol having 4 carbon atoms. For example, the polyol may be glycerin, propylene glycol, butylene glycol, diethylene glycol, etc.

[0071] Specifically, the amino acids, polyols, and nonionic surfactants included in the emulsion serve to stabilize the ceramide by surrounding it during delivery, thereby preventing the ceramide from precipitating during the skin delivery process.

[0072] In one embodiment, the emulsion may comprise an aqueous phase comprising an amino acid and a polyol; and an oil phase comprising a fatty acid and a nonionic surfactant. Specifically, the aqueous phase may comprise an amino acid, a polyol, and water.

[0073] Specifically, the amino acid and polyol included in the aqueous phase surround the outside of the ceramide together with the nonionic surfactant, thereby not only stabilizing the ceramide but also significantly reducing the ceramide that precipitates and is released in the aqueous phase.

[0074] In one embodiment, the emulsion may further include additional components. The "additional components" may include, for example, functional components such as stabilizers, thickeners, antioxidants, UV blockers, pigments, fragrances, and preservatives, or active components effective for skin moisturization, skin barrier strengthening, skin aging improvement, skin wrinkle improvement, skin elasticity improvement, and skin regeneration. For example, 1,2-hexanediol, ethylhexylglycerin, hydroxyacetophenone, vitamin A, vitamin B, vitamin C, vitamin E, vitamin derivatives, collagen, ceramide, peptide, adenosine, arbutin, hyaluronic acid, chondroitin sulfate, alpha-bisabolol, guaiazulene, coenzyme Q10, oil-soluble licorice (glycyrrhiza) extract, hydroxydecyl ubiquinone, betaine, allantoin, urea, It may further include niacinamide, adenosine, and tranexamic acid.

[0075] If the above additional component is a water-soluble component, it may be included in the aqueous phase of the emulsion, and if the above additional component is an oil-soluble component, it may be included in the oil phase of the emulsion.

[0076] In one experimental example, the emulsion contains ceramide, fatty acid, amino acid, polyol, and nonionic surfactant in a weight ratio of 3 to 4: 5 to 8: 1: 5 to 8: 3 to 4, and when the nonionic surfactant is polyglyceryl-10 stearate, decyl glucoside, polyglyceryl-10 laurate, or polyglyceryl-10 oleate, it was confirmed that the emulsion particles are stably dispersed nanoparticles with an average size of 180 nm or less and a PDI of 0.321 or less. In addition, it was confirmed that there was no precipitated ceramide when the emulsion solution was observed visually.

[0077] In one embodiment, the emulsion may comprise the ceramide, fatty acid, amino acid, polyol, and nonionic surfactant in a weight ratio of 3 to 5: 5 to 10: 1: 5 to 10: 3 to 5, specifically comprising the ceramide, fatty acid, amino acid, polyol, and nonionic surfactant in a weight ratio of 3 to 5: 5 to 10: 1: 5 to 10: 3 to 5, 3 to 5: 5 to 8: 1: 5 to 10: 3 to 5, 3 to 5: 5 to 10: 1: 5 to 8: 3 to 5, 3 to 5: 5 to 10: 1: 5 to 10: 3 to 4, 3 to 5: 5 to 8: 1: 5 to 8: 3 to 5, and 3 to 5: 5 to 10: 1: 5 to 8: 3 to 4, 3 to 5: 5 to 8: 1: 5 to 10: 3 to 4, 3 to 5: 5 to 8: 1: 5 to 8: 3 to 4, 3 to 4: 5 to 10: 1: 5 to 10: 3 to 5, 3 to 4: 5 to 8: 1: 5 to 10: 3 to 5, 3 to 4: 5 to 10: 1: 5 to 8: 3 to 5, 3 to 4: 5 to 10: 1: 5 to 8: 3 to 5, 3 to 4: 5 to 10: 1: 5 to 10: 3 to 4, 3 to 4: 5 to 8: 1: 5 to 8: 3 to 5, 3 to 4: 5 to 10: 1: 5 to 8: 3 to 4, 3 to 4: It may be included in a weight ratio of 5 to 8: 1: 5 to 10: 3 to 4 or 3 to 4: 5 to 8: 1: 5 to 8: 3 to 4.

[0078] In one embodiment, the amino acid may be included in an amount of 1 to 5 weight% relative to the total weight of the emulsion, and specifically, the amino acid may be included in an amount of 1 to 5 weight%, 1 to 4 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 4 weight%, 2 to 3 weight%, 3 to 5 weight%, 3 to 4 weight%, or 4 to 5 weight% relative to the total weight of the emulsion.

[0079] In one embodiment, the polyol may be included in an amount of 10 to 30 weight% relative to the total weight of the emulsion, and specifically, the polyol may be included in an amount of 10 to 30 weight%, 10 to 25 weight%, 10 to 20 weight%, 10 to 15 weight%, 15 to 30 weight%, 15 to 25 weight%, 15 to 20 weight%, 20 to 30 weight%, 20 to 25 weight%, or 25 to 30 weight% relative to the total weight of the emulsion.

[0080] When the above emulsion contains ceramide, fatty acid, amino acid, polyol, and nonionic surfactant in a weight ratio of 3 to 5: 5 to 10: 1: 5 to 10: 3 to 5, the ceramide does not precipitate even under long-term storage or harsh conditions (e.g., high temperature, low temperature, repeated freezing and thawing conditions, etc.), allowing for the loading of a high content of ceramide within the emulsion particles. In addition, skin permeability is significantly increased, enabling the delivery of ceramide to the skin with high efficiency.

[0081] Specifically, if some of the above components are excluded or included outside the weight ratio range, the ceramide may not be stably captured within the emulsion and may easily precipitate. In addition, the prepared emulsion may not be stably dispersed within the aqueous phase and may aggregate, and as the size of the emulsion particles increases, the skin permeability is significantly reduced, resulting in a lower skin delivery efficiency of the ceramide.

[0082] In one embodiment, the emulsion may further include water.

[0083] In one embodiment, the emulsion may contain the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and water in a weight ratio of 3 to 5: 5 to 10: 1: 5 to 10: 3 to 5: 10 to 15. Specifically, the above ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and water are 3 to 5: 5 to 10: 1: 5 to 10: 3 to 5: 10 to 15, 3 to 5: 5 to 8: 1: 5 to 10: 3 to 5: 10 to 13, 3 to 5: 5 to 10: 1: 5 to 8: 3 to 5: 10 to 13, 3 to 5: 5 to 10: 1: 5 to 10: 3 to 4: 10 to 13, 3 to 5: 5 to 8: 1: 5 to 8: 3 to 5: 10 to 13, 3 to 5: 5 to 10: 1: 5 to 8: 3 to 4: 10 to 13, 3 to 5: 5 to 10: 1: 5 to 8: 3 to 4: 10 to 13, 3 to 5: 5 to 8: 1: 5 to 10: 3 to 4: 10 to 13, 3 to 5: 5 to 8: 1: 5 to 8: 3 to 4: 10 to 13, 3 to 4: 5 to 10: 1: 5 to 10: 3 to 5: 10 to 13, 3 to 4: 5 to 8: 1: 5 to 10: 3 to 5: 10 to 13, 3 to 4: 5 to 10: 1: 5 to 8: 3 to 5: 10 to 13, 3 to 4: 5 to 10: 1: 5 to 8: 3 to 5: 10 to 13, 3 to 4: 5 to 10: 1: 5 to 10: 3 to 4: 10 to 13, 3 to 4: 5 to 8: 1: 5 to 8: 3 to 5: 10 to It may be included in a weight ratio of 13, 3 to 4: 5 to 10: 1: 5 to 8: 3 to 4: 10 to 13, 3 to 4: 5 to 8: 1: 5 to 10: 3 to 4: 10 to 13 or 3 to 4: 5 to 8: 1: 5 to 8: 3 to 4: 10 to 13.

[0084]

[0085] Another aspect provides a cosmetic composition comprising the above-mentioned emulsion. Additionally, another aspect provides a topical skin preparation comprising the above-mentioned emulsion.

[0086] The above terms "ceramide," "surfactant," "emulsion," etc. may be within the aforementioned range.

[0087] In one embodiment, the cosmetic composition or external skin preparation may have one or more activities selected from the group consisting of skin moisturization, skin barrier strengthening, skin wrinkle improvement, skin elasticity improvement, antioxidant, skin whitening, and hair loss improvement; specifically, the cosmetic composition or external skin preparation may have all of the activities of skin moisturization, skin barrier strengthening, skin wrinkle improvement, skin elasticity improvement, antioxidant, skin whitening, and hair loss improvement.

[0088] In one embodiment, the cosmetic composition may be prepared in any formulation conventionally manufactured in the art to which the present invention belongs. For example, the cosmetic composition may be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, spray, etc. Specifically, the cosmetic composition may be manufactured in the form of a lotion such as a softening lotion or a nourishing lotion, a fluid such as a facial lotion or a body lotion, a cream such as a nourishing cream, a moisturizing cream, or an eye cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a foundation in liquid, solid, or spray form, a powder, a makeup remover such as a cleansing cream, a cleansing lotion, or a cleansing oil, a cleanser such as a cleansing foam, a soap, or a body wash, a liquid or gel formulation of mesotherapy that can be injected into the skin layer, and a formulation mixed with hyaluronic acid gel. More specifically, the cosmetic composition may be manufactured in one or more formulations selected from the group consisting of a serum, lotion, essence, paste, mask pack, patch, gel, cream, lotion, nourishing lotion, nourishing cream, moisturizing cream, massage cream, powder, soap, cleanser, oil, foundation, makeup base, wax, and spray.

[0089] The above cosmetic composition may further comprise any conventional cosmetic ingredient selected from additional ingredients commonly used in cosmetics, such as excipients, thickeners, dispersants, fragrances, fillers, preservatives, antiseptics, neutralizing agents, sweeteners, vitamins, free radical scavengers, metal ion chelating agents, functional ingredients, and mixtures thereof. A person skilled in the art may select any additional ingredient and / or the amount thereof so that the advantageous properties of the composition according to one aspect are not adversely affected or are substantially affected by the expected addition.

[0090] For example, the above excipients may include surfactants, emulsifiers, saponic acid, solvents, coloring agents, preservatives, antioxidants, antifoaming agents, antibacterial agents, anti-redeposition agents, enzymes, plant or mineral oils, fats, fluorescent substances, fungicides, hydrotropy-inducing substances, humectants, fragrances, preservatives, proteins, silicones, solubilizers, sugar derivatives, sunblocks, vitamins, plant extracts, waxes, fatty acids, higher alcohols, hydrocarbon oils, ester oils, triglycerides, vegetable oils, etc.

[0091] According to one aspect, the cosmetic composition may additionally include ingredients typically added to cosmetic compositions, such as purified water, surfactants, moisturizers, lower alcohols, chelating agents, disinfectants, preservatives, antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0092] In addition, the above cosmetic composition may be appropriately formulated with ingredients commonly used in external skin preparations such as cosmetics or pharmaceuticals, for example, aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof, as needed. The above cosmetic composition may also appropriately incorporate metal chelating agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; caffeine, tannin, bellapamil, licorice extract, glablidin, hot water extract of the fruit of Carin, various herbal medicines, pharmaceuticals such as tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts thereof; and sugars such as vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, and trehalose.

[0093] When the above cosmetic composition is a surfactant-containing cleansing formulation, it may further include, as a carrier component, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative, or ethoxylated glycerol fatty acid ester, etc.

[0094] In addition, if the cosmetic composition according to one aspect is in the form of a cream or gel, it may further include animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide as a carrier component.

[0095] In addition, if the above cosmetic composition is in the form of a solution or emulsion, it may further include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, propylene glycol, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan as a solvent, solvating agent, or emulsifying agent.

[0096] In the case where the above cosmetic composition is in the form of a suspension, it may further include, as a carrier component, a liquid diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracanth.

[0097] In the case where the above cosmetic composition is in the form of a powder or spray, it may further include lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder as a carrier component, and in particular, in the case of a spray formulation, it may further include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0098] The above cosmetic composition may be used by applying it alone or in combination, or by applying it in combination with other cosmetic compositions other than the cosmetic composition according to one aspect. Furthermore, all ingredients included in the above cosmetic composition do not exceed the regulations set by each country. Additionally, the cosmetic composition according to one aspect may be used according to a standard method of use, and the frequency of use may be varied depending on the user's skin condition or preference.

[0099] In one experimental example, when the emulsion contained ceramide, oleic acid, and decyl glucoside (Example 4), approximately 16.82 ppm of ceramide was detected after 12 hours of penetration, confirming that it exhibited significantly superior skin penetration ability compared to Comparative Example 1 (fatty acid: MCT; nonionic surfactant: polyglyceryl-10 stearate) (see Experimental Example 8).

[0100] The above-mentioned topical skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, or a combination thereof. The above-mentioned topical skin preparation may be appropriately formulated as needed with ingredients commonly used in topical skin preparations such as cosmetics or pharmaceuticals, for example, aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof. The above topical preparation may also appropriately incorporate metal chelating agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; caffeine, tannin, bellapamil, licorice extract, glablidin, hot water extract of the fruit of Carin, various herbal medicines, pharmaceuticals such as tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts, and sugars such as vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, and trehalose.

[0101] In addition, the above-mentioned external skin agent may be included in a quasi-drug composition.

[0102] The term "quasi-drug" above refers to articles falling under one of the following categories: fibers, rubber products, or similar items used for the purpose of treating, alleviating, managing, or preventing diseases in humans or animals; items similar thereto that have a weak effect on the human body or do not act directly on the human body and are not instruments or machines; and preparations used for sterilization, insecticidal, and similar purposes for the prevention of infection; excluding articles used for the purpose of diagnosing, treating, alleviating, managing, or preventing conditions or diseases in humans or animals that are not instruments, machines, or devices, and articles used for the purpose of exerting pharmacological effects on the structure and function of humans or animals that are not instruments, machines, or devices; and may include not only topical skin preparations but also personal hygiene products.

[0103] Accordingly, a cosmetic composition or topical skin preparation according to one aspect includes an emulsion containing a high amount of ceramide, thereby significantly reducing the amount of ceramide released during storage, exhibiting high storage stability, and can stably deliver a high amount of ceramide into the skin without precipitation. In addition, the cosmetic composition has excellent skin permeability, which can significantly increase the effects of ceramide, such as skin moisturization and skin barrier improvement.

[0104]

[0105] Another aspect is a step of mixing a ceramide, a fatty acid, and a nonionic surfactant to obtain a lipid mixture, wherein the ceramide is included in an amount of 1 to 15 weight% based on the total weight of the emulsion; and

[0106] A method for preparing an emulsion is provided, comprising the step of mixing the above lipid mixture and a eutectic solvent to obtain an emulsion.

[0107] In one embodiment, the method may further include the step of mixing an amino acid and a polyol to obtain the eutectic solvent. Specifically, the step of obtaining the eutectic solvent may be performed before, simultaneously with, or after the step of obtaining the lipid mixture.

[0108] In one embodiment, the step of mixing the ceramide, fatty acid, and nonionic surfactant to obtain a lipid mixture may further include the step of heating the lipid mixture at 60 to 100°C.

[0109] Specifically, the heating step may be performed at 60 to 100 ℃, 60 to 90 ℃, 60 to 80 ℃, 60 to 70 ℃, 70 to 100 ℃, 70 to 90 ℃, 70 to 80 ℃, 80 to 100 ℃, 80 to 90 ℃, or 90 to 100 ℃.

[0110] In the step of obtaining the above lipid mixture and heating the mixture, hydrogen bonds are formed between the carboxyl group (-COOH) of the fatty acid and the amino group (-NH2) of the ceramide, and intermolecular forces are formed due to van der Waals forces. In addition, by mixing a nonionic surfactant together, the precipitation of ceramide is prevented.

[0111] In one embodiment, the step of obtaining the eutectic solvent may further include the step of heating the eutectic solvent at 60 to 100°C.

[0112] Specifically, the heating step may be performed at 60 to 100 ℃, 60 to 90 ℃, 60 to 80 ℃, 60 to 70 ℃, 70 to 100 ℃, 70 to 90 ℃, 70 to 80 ℃, 80 to 100 ℃, 80 to 90 ℃, or 90 to 100 ℃.

[0113] The above "eutectic solvent" is a substance made by mixing hydrogen bond acceptors such as choline chloride, betaine, proline, etc. and hydrogen bond donors such as amides, sugars, alcohols, organic acids, fatty acids, etc., and has the advantage of having high solubility, a low melting point, and low toxicity.

[0114] Specifically, when a eutectic solvent is used, the dissolution of the lipid mixture at a low temperature is enabled, allowing the emulsion to be prepared without using an organic solvent. In addition, when the eutectic solvent is used, intermolecular interactions (e.g., hydrogen bonding, van der Waals forces, etc.) between the amino acids and polyols in the eutectic solvent and the ceramide in the lipid mixture are maximized, thereby enabling the formation of a stable emulsion.

[0115] In one embodiment, the step of mixing the lipid mixture and the eutectic solvent to obtain an emulsion may further include the step of pressurizing to 500 to 1500 bar.

[0116] Specifically, the pressurizing step may be performed at 500 to 1500 bar, 500 to 1300 bar, 500 to 1100 bar, 500 to 900 bar, 500 to 700 bar, 700 to 1500 bar, 700 to 1300 bar, 700 to 1100 bar, 700 to 900 bar, 900 to 1500 bar, 900 to 1300 bar, 900 to 1100 bar, 1100 to 1500 bar, 1100 to 1300 bar, or 1300 to 1500 bar.

[0117] Additionally, the above-mentioned pressurizing step may be performed 1 to 10 times, and specifically, 1 to 10 times, 1 to 7 times, 1 to 5 times, 1 to 3 times, 3 to 10 times, 3 to 7 times, 3 to 5 times, 5 to 10 times, 5 to 7 times, or 7 to 10 times.

[0118] In one embodiment, the pressurizing step may be performed simultaneously with mixing the lipid mixture and the eutectic solvent or after mixing the lipid mixture and the eutectic solvent.

[0119] By performing the above pressurizing step, the size of the manufactured emulsion particles becomes uniform. Specifically, by performing the above pressurizing step 1 to 10 times at 500 to 1500 bar, a nano-sized emulsion with an average particle diameter of 10 nm to 250 nm can be obtained. Accordingly, since the manufactured emulsion exhibits a small size, its ability to penetrate the skin barrier is significantly increased.

[0120] The emulsion prepared by the manufacturing method according to one aspect is stabilized by the formation of hydrogen bonds and van der Waals forces between the ceramide and fatty acids, so it does not precipitate upon delivery to the skin and exists stably.

[0121]

[0122] One aspect provides a liposome for the simultaneous delivery of a lipid-soluble substance and a water-soluble substance comprising ceramide, a fatty acid, an amino acid, a polyol, a nonionic surfactant, and a phospholipid, wherein the ceramide and the fatty acid form a lipid core through hydrogen bonding.

[0123] The above "liposome" is a self-assembling spherical phospholipid vesicle composed of an amphiphilic molecule having a water-soluble head (hydrophilic group) and a lipid-soluble tail (hydrophobic group). The liposome has a double membrane (bilayer) structure similar to a cell membrane.

[0124] In one embodiment, the ceramide can induce a +δ charge distribution and a dipole moment within the ceramide molecule by forming hydrogen bonds between the polar group of the ceramide and the fatty acid. Specifically, the amino group (-NH2) of the ceramide and the carboxyl group (-COOH) of the fatty acid can bond to form hydrogen bonds. These electrical properties can promote the spontaneous arrangement of the lipid bilayer and the stabilization of the lamellar liquid crystal structure.

[0125] In another embodiment, the lipid core containing the ceramide may be coated with a deep eutectic solvent (DES). By coating the outer surface of the lipid core with a deep eutectic solvent, the structural stability of the liposome, the preservation of the active ingredient, and the persistence upon skin application can be improved. Specifically, the deep eutectic solvent may be selected from the group consisting of amino acids and polyols.

[0126] In one embodiment, the ceramide and fatty acid may be included in a weight ratio of 1 to 5: 3 to 10 based on the weight of the amino acid included in the liposome. Specifically, the ceramide and fatty acid may be included in a weight ratio of 1 to 5: 3 to 10, 1 to 3: 3 to 10, 3 to 5: 3 to 10, 1 to 5: 3 to 6, 1 to 5: 6 to 10, 1 to 3: 3 to 6, 1 to 3: 6 to 10, 3 to 5: 3 to 6, or 3 to 5: 6 to 10 based on the weight of the amino acid included in the liposome.

[0127] In one embodiment, the weight ratio of the ceramide and fatty acid may be 1:0.5 to 10. Specifically, the weight ratio of ceramide to fatty acid may be 1:0.5 to 10, 1:0.5 to 7, 1:0.5 to 4, 1:0.5 to 3, 1:0.5 to 2, 1:0.5 to 1, 1:1 to 10, 1:1 to 7, 1:1 to 4, 1:1 to 3, 1:1 to 2, 1:2 to 10, 1:2 to 7, 1:2 to 4, 1:2 to 3, 1:3 to 10, 1:3 to 7, 1:3 to 4, 1:4 to 10, 1:4 to 7, or 1:7 to 10.

[0128] In one embodiment, the ceramide may be included in an amount of 1 to 10 weight% relative to the total weight of the liposome, and specifically, the ceramide may be included in an amount of 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, or 7.5 to 10 weight% relative to the total weight of the liposome.

[0129] In one embodiment, the fatty acid may be included in an amount of 5 to 20 weight% relative to the total weight of the liposome, and specifically, the fatty acid may be included in an amount of 5 to 20 weight%, 5 to 15 weight%, 5 to 12 weight%, 5 to 8 weight%, 8 to 20 weight%, 8 to 15 weight%, 8 to 12 weight%, 12 to 20 weight%, 12 to 15 weight%, or 15 to 20 weight% relative to the total weight of the liposome.

[0130] Specifically, when the weight ratio of the ceramide and fatty acid contained in the liposome is 1:0.5 to 10, or when the weight ratio of the ceramide and fatty acid is 1 to 5:3 to 10 based on the weight of the amino acid contained in the liposome, the fatty acid induces a dipole moment in the ceramide, and the carboxyl group (-COOH) of the fatty acid forms a hydrogen bond with the amino group (-NH2) of the ceramide, thereby enabling the liposome to maintain its shape stably. Accordingly, the liposome exhibits excellent storage stability even under harsh conditions (high temperature, low temperature, high relative humidity, low relative humidity, repeated freezing and thawing conditions, etc.).

[0131] Furthermore, when the weight ratio of the ceramide and fatty acid contained in the liposome is 1:0.5 to 10, or when the weight ratio of the ceramide and fatty acid is 1 to 5:3 to 10 based on the weight of the amino acid contained in the liposome, the amino group (-NH2) of the ceramide combines with the carboxyl group (-COOH) of the fatty acid and is arranged on the outermost surface of the liposome, and the transdermal permeability of the emulsion containing the active ingredient is significantly increased by the above liposome surface arrangement.

[0132] The above "lipid core" is a lipid complex in which ceramide and fatty acids are bonded through hydrogen bonds, and it serves to stabilize the phospholipid bilayer structure and inhibit the precipitation of active ingredients encapsulated within the phospholipid bilayer structure.

[0133] In one embodiment, the phospholipid may be a natural phospholipid or a synthetic phospholipid. For example, the phospholipid may be a neutral lipid, a cationic lipid, anionic lipid, etc., and may be a lipid derived from soybeans, eggs, cured soybeans, or cured eggs.

[0134] Specifically, the phospholipids include lecithin, hydrogenated lecithin, phosphatidyl choline, sphingomyelin, cholesterol, phosphatidic acid, phosphatidyl serine, phosphatidyl glycerol, phosphatidyl inositol, phosphatidyl ethanolamine, egg phosphatidyl choline, egg phosphatidyl glycerol, egg phosphatidyl ethanolamine, egg phosphatidyl serine, egg phosphatidic acid, egg phosphatidylinositol, soybean phosphatidyl choline, soybean phosphatidyl glycerol, soybean phosphatidyl ethanolamine, soybean phosphatidyl serine, soybean phosphatidic acid, soybean phosphatidylinositol, Dipalmitoyl phosphatidylcholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC), dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidyl glycerol (DPPG), dioleoyl phosphatidyl glycerol (DOPG), dimyristoyl phosphatidyl glycerol (DMPG), hexadecyl phosphocholine (HEPC), hydrogenated soybean phosphatidyl choline (HSPC), distearoyl phosphatidylcholine (DSPC),distearoyl phosphatidyl glycerol (DSPG), dioleoyl phosphatidyl ethanolamine (DOPE), palmitoyl stearoyl phosphatidylcholine (PSPC), palmitoyl stearoyl phosphatidyl glycerol (PSPG), monooleoyl phosphatidyl ethanolamine (MOPE), 1-palmitoyl-2-oleoyl-glycero-3-phosphatidylcholine (POPC), polyethyleneglycol distearoyl phosphatidylcholine (PEG-DSPE), Dipalmitoyl phosphatidyl serine (DPPS), 1,2-dioleoyl-sn-glycero-3-phosphatidyl serine (DOPS), dimyristoyl phosphatidyl serine (DMPS), distearoyl phosphatidyl serine (DSPS), dipalmitoyl phosphatidic acid (DPPA), 1,2-dioleoyl-sn-glycero-3-phosphatidic acid (DOPA), dimyristoyl phosphatidic acid (DMPA), distearoyl phosphatidic acid (distearoyl phosphatidic acid, DSPA),It may be one or more selected from the group consisting of dipalmitoyl phosphatidyl inositol (DPPI), 1,2-dioleoyl-sn-glycero-3-phosphatidyl inositol (DOPI), dimyristoyl phosphatidyl inositol (DMPI), and distearoyl phosphatidyl inositol (DSPI).

[0135] More specifically, the phospholipid may be one or more selected from the group consisting of lecithin, hydrogenated lecithin, phosphatidyl choline, sphingomyelin, cholesterol, phosphatidic acid, phosphatidyl serine, phosphatidyl glycerol, phosphatidyl inositol, and phosphatidyl ethanolamine. More specifically, it may be hydrogenated lecithin.

[0136] In one embodiment, the liposome may further include one or more active ingredients selected from the group consisting of lipid-soluble substances and water-soluble substances. Specifically, the liposome may include both lipid-soluble substances and water-soluble substances.

[0137] In one embodiment, the fat-soluble or water-soluble substance may be trapped between the phospholipid bilayers.

[0138] In one embodiment, the lipid-soluble substance or water-soluble substance may be captured in the inner or outermost membrane of the liposome.

[0139] The term "capture" encompasses all states in which substances are gathered within a separated space, specific substances are sandwiched between other substances, and substances are held together by interactions such as hydrogen bonding or van der Waals forces. The term "capture" may be used interchangeably with "encapsulation" or "loading."

[0140] Specifically, the phospholipids within the liposome form a bilayer to encapsulate lipid-soluble or water-soluble substances between the bilayers. Additionally, by encapsulating water-soluble or lipid-soluble substances in the spherical internal space (a space surrounded by the phospholipid bilayer) and the outermost layer of the bilayer created by the formation of the phospholipid bilayer, lipid-soluble and water-soluble substances can be delivered simultaneously.

[0141] More specifically, when the liposome particles are present in a water-soluble solvent, the liposome particles have a form in which phospholipids, ceramides, and fatty acids form an outer membrane (e.g., a double layer in which the hydrophobic groups of the phospholipids, ceramides, and fatty acids are in contact), encapsulate lipid-soluble substances between the double layers, and encapsulate or bind a water-soluble substance to the hydrophilic groups of the substances constituting the double layers and a spherical internal space formed by the double layers (see FIG. 1a and 1b). If the liposome particles are present in a water-soluble solvent, the liposome particles have a form in which phospholipids, ceramides, and fatty acids form an outer membrane (e.g., a double layer in which the hydrophilic groups of the phospholipids, ceramides, and fatty acids are in contact), encapsulate water-soluble substances between the double layers, and encapsulate or bind a water-soluble substance to the hydrophobic groups of the substances constituting the double layers and a spherical internal space formed by the phospholipid double layer.

[0142] For example, according to one aspect, the liposome may be utilized as a delivery vehicle to simultaneously deliver lipid-soluble and water-soluble substances to the skin by encapsulating tocopheryl acetate as a lipid-soluble substance between phospholipid bilayers and encapsulating niacinamide as a water-soluble substance in the spherical internal space formed by the phospholipid bilayers and in the outermost membrane of the bilayers.

[0143] In one embodiment, the fat-soluble substance may be one or more selected from the group consisting of tocopheryl acetate, retinol, calciferol, linolenic acid, biotin, menadione, bioflavonoids, astaxanthin, idebenone, bakuchiol, volufiline, bisabolol, and zeaxanthin. Specifically, the above-mentioned fat-soluble substances may all be ceramide, tocopheryl acetate, retinol, calciferol, linolenic acid, biotin, menadione, bioflavonoids, astaxanthin, idebenone, bakuchiol, volufiline, bisabolol, and zeaxanthin.

[0144] The above "bioflavonoids" are natural antioxidants of plant origin that have a basic structure of polyphenol rings and are mainly found in the roots, stems, and barks of citrus fruits, light-colored vegetables, and nuts, as well as in tea and wine. Examples include quercetin, hesperidin, naringenin, naringin, rutin, and kaempferol.

[0145] In one embodiment, the water-soluble substance may be one or more selected from the group consisting of niacinamide, niacin, thiamine, riboflavin, carnitine, pantothenic acid, panthenol, pyridoxine, ascorbic acid, peptide collagen, and cyanocobalamin. Specifically, the above water-soluble substances may all be niacinamide, niacin, thiamine, riboflavin, carnitine, pantothenic acid, panthenol, pyridoxine, ascorbic acid, peptide collagen, and cyanocobalamin.

[0146] The above "peptide collagen" refers to collagen in a high molecular weight form, which is difficult to ingest directly or absorb through the skin or body because its molecular weight is very large, approximately 300,000 Da. Therefore, collagen in a high molecular weight form is broken down by collagenases, etc., to a low molecular weight of 5,000 Da or less (e.g., 3,000 to 5,000 Da).

[0147] In one embodiment, the liposome may further include additional components. The "additional components" may include, for example, functional components such as stabilizers, thickeners, antioxidants, UV blockers, pigments, fragrances, and preservatives, or active components effective for skin moisturization, skin barrier strengthening, skin aging improvement, skin wrinkle improvement, skin elasticity improvement, and skin regeneration. For example, the liposome comprises 1,2-hexanediol, ethylhexylglycerin, hydroxyacetophenone, vitamin A, vitamin B, vitamin C, vitamin E, vitamin derivatives, collagen, ceramide, peptide, adenosine, arbutin, hyaluronic acid, chondroitin sulfate, alpha-bisabolol, guaiazulene, coenzyme Q10, oil-soluble licorice (glycyrrhiza) extract, hydroxydecyl ubiquinone, betaine, allantoin, urea, It may further include niacinamide, adenosine, and tranexamic acid.

[0148] If the liposome particle is present in a water-soluble solvent and the additional component is water-soluble, the additional component may be contained within the spherical internal space formed by the phospholipid bilayer of the liposome and / or bonded to the hydrophilic groups of the components constituting the bilayer. Additionally, if the additional component is lipid-soluble, the additional component may be contained between the phospholipid bilayers of the liposome. If the liposome particle is present in a lipid-soluble solvent and the additional component is lipid-soluble, the additional component may be contained within the spherical internal space formed by the phospholipid bilayer of the liposome and / or bonded to the hydrophobic groups of the components constituting the bilayer. Additionally, if the additional component is water-soluble, the additional component may be contained between the phospholipid bilayers of the liposome.

[0149] In one embodiment, the liposome may contain the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10. Specifically, the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid are 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 1 to 3: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 3 to 6: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 3 to 6: 1 to 5: 3 to 10, 3 to 5: It may be included in a weight ratio of 6 to 10: 1: 6 to 10: 1 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 6 to 10: 1 to 3: 3 to 10, 3 to 5: 6 to 10: 1: 6 to 10: 3 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 6 to 10: 3 to 5: 3 to 6, 3 to 5: 6 to 10: 1: 6 to 10: 3 to 5: 6 to 10 or 3 to 4: 6 to 7: 1: 6 to 7: 3 to 4: 6 to 7.

[0150] In one embodiment, the liposome may contain a lipid-soluble substance and a water-soluble substance in a weight ratio of 0.5 to 6: 0.5 to 6 based on the weight of the amino acid contained in the liposome. Specifically, the liposome may contain lipid-soluble substances and water-soluble substances in a weight ratio of 0.5 to 6: 0.5 to 6, 0.5 to 3: 0.5 to 6, 3 to 6: 0.5 to 6, 0.5 to 6: 0.5 to 3, 0.5 to 6: 3 to 6, 0.5 to 3: 0.5 to 3, 3 to 6: 0.5 to 3, 0.5 to 3: 3 to 6, 3 to 6: 3 to 6, 0.5 to 2: 0.5 to 2, or 0.5 to 1: 0.5 to 1, based on the weight of the amino acids contained in the liposome.

[0151] In one embodiment, the amino acid may be included in an amount of 1 to 5 weight% relative to the total weight of the liposome, and specifically, the amino acid may be included in an amount of 1 to 5 weight%, 1 to 4 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 4 weight%, 2 to 3 weight%, 3 to 5 weight%, 3 to 4 weight%, or 4 to 5 weight% relative to the total weight of the liposome.

[0152] In one embodiment, the polyol may be included in an amount of 5 to 20 weight% relative to the total weight of the liposome, and specifically, the polyol may be included in an amount of 5 to 20 weight%, 5 to 15 weight%, 5 to 12 weight%, 5 to 8 weight%, 8 to 20 weight%, 8 to 15 weight%, 8 to 12 weight%, 12 to 20 weight%, 12 to 15 weight%, or 15 to 20 weight% relative to the total weight of the liposome.

[0153] In one embodiment, the nonionic surfactant may be included in an amount of 1 to 10 weight% relative to the total weight of the liposome, and specifically, the nonionic surfactant may be included in an amount of 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, or 7.5 to 10 weight% relative to the total weight of the liposome.

[0154] In one embodiment, the phospholipid may be included in an amount of 1 to 10 weight% relative to the total weight of the liposome, and specifically, the phospholipid may be included in an amount of 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, or 7.5 to 10 weight% relative to the total weight of the liposome.

[0155] In one embodiment, the water-soluble material may be included in an amount of 0.5 to 5 weight% relative to the total weight of the liposome, and specifically, the water-soluble material may be included in an amount of 0.5 to 5 weight%, 0.5 to 3 weight%, 0.5 to 2 weight%, 0.5 to 1 weight%, 1 to 5 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 3 weight%, or 3 to 5 weight% relative to the total weight of the liposome.

[0156] In one embodiment, the lipid-soluble material may be included in an amount of 0.5 to 5 weight% relative to the total weight of the liposome, and specifically, the lipid-soluble material may be included in an amount of 0.5 to 5 weight%, 0.5 to 3 weight%, 0.5 to 2 weight%, 0.5 to 1 weight%, 1 to 5 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 3 weight%, or 3 to 5 weight% relative to the total weight of the liposome.

[0157] When the above liposomes contain the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, the ceramide, water-soluble, and fat-soluble substances do not precipitate even under long-term storage or harsh conditions (high temperature, low temperature, high relative humidity, low relative humidity, repeated freezing and thawing conditions, etc.), so it is possible to load a high amount of ceramide, water-soluble, and fat-soluble substances into the liposomes. In addition, skin permeability is significantly increased, allowing the ceramide, water-soluble, and fat-soluble substances to be delivered to the skin simultaneously with high efficiency.

[0158] The above "skin permeability" refers to the ability of liposomes for the simultaneous delivery of lipid-soluble and water-soluble substances to pass through the stratum corneum, which is the outermost layer of the skin's epidermis, and to deliver active ingredients into the stratum corneum. Furthermore, it refers to the ability to deliver active ingredients to the epidermal layers (stratum lucidum, stratum granulosum, stratum spinosum, stratum basale), dermal layers, or even into the body that exist within the stratum corneum. Additionally, the above skin permeability may be used interchangeably with transdermal permeability.

[0159] Specifically, if some of the above components are excluded or included outside the weight ratio range, for example, if some or all of the above components are less than the minimum value or greater than the maximum value of the weight ratio, such as when ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid are included in weight ratios of 8:3:1:3:1:3, 8:1:1:30:1:30, or 8:10:1:15:7:15, the ceramide, water-soluble substances, and fat-soluble substances may not be stably captured within the liposome and may easily precipitate. Furthermore, the manufactured liposome may not be stably dispersed and may aggregate, and the size of the liposome particles increases, significantly reducing skin permeability, thereby lowering the skin delivery efficiency of ceramide, water-soluble substances, and fat-soluble substances.

[0160] In one embodiment, the liposome may further contain water.

[0161] In one embodiment, the liposome may contain the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, phospholipid, and water in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10: 30 to 45. Specifically, the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, phospholipid, and water are 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10: 30 to 45, 1 to 3: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10: 30 to 45, 3 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10: 30 to 45, 3 to 5: 3 to 6: 1: 3 to 10: 1 to 5: 3 to 10: 30 to 45, 3 to 5: 6 to 10: 1: 3 to 10: 1 to 5: 3 to 10: 30 to 45 45, 3 to 5: 3 to 10: 1: 3 to 6: 1 to 5: 3 to 10: 30 to 45, 3 to 5: 3 to 10: 1: 6 to 10: 1 to 5: 3 to 10: 30 to 45, 3 to 5: 3 to 10: 1: 3 to 10: 1 to 3: 3 to 10: 30 to 45, 3 to 5: 3 to 10: 1: 3 to 10: 3 to 5: 3 to 10: 30 to 45, 3 to 5: 3 to 10: 1: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 6: 30 to 45, 3 to 5: 3 to 10: 1: 3 to 10: 1: 3 to 10: It may be included in a weight ratio of 1 to 5: 6 to 10: 30 to 45, 3 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 6 to 10: 30 to 40, or 3 to 4: 6 to 7: 1: 6 to 7: 3 to 4: 6 to 7: 30 to 40.

[0162] In one experimental example, when the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in the liposome are included in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, it was confirmed that the ceramide, lipid-soluble substances, and water-soluble substances are stably dispersed without precipitating even under harsh conditions (high temperature, low temperature, high relative humidity, low relative humidity, repeated freezing / thawing conditions, etc.), thereby confirming that the liposome exhibits significantly excellent storage stability (see Experimental Examples 1 and 4).

[0163] In one embodiment, the liposome may have a bilayer structure, a multilayer structure, or a mixture of a bilayer liposome and a multilayer liposome. Specifically, the liposome may have a bilayer structure.

[0164] In one embodiment, the average particle diameter of the liposome may be 10 nm to 100 nm, specifically 10 nm to 100 nm, 10 nm to 70 nm, 10 nm to 40 nm, 40 nm to 100 nm, 40 nm to 70 nm, or 70 nm to 100 nm.

[0165] According to one aspect, the liposome particles are nano-sized particles with an average diameter of 10 nm to 100 nm. Accordingly, the liposome particles can easily penetrate the stratum corneum of the skin to deliver ceramide and water-soluble and fat-soluble substances into the skin. When the average diameter of the particles is larger than 100 nm, the penetration rate into the stratum corneum is significantly lower, resulting in low efficacy of ceramide and water-soluble and fat-soluble substances.

[0166] According to one aspect, the liposome particles are nano-sized particles with an average diameter of 10 nm to 100 nm. Accordingly, the liposome particles can easily penetrate the stratum corneum of the skin to deliver ceramide, water-soluble substances, and fat-soluble substances into the skin. When the average diameter of the particles is larger than 100 nm, the penetration rate into the stratum corneum is significantly lower, resulting in low efficacy of ceramide, water-soluble substances, and fat-soluble substances.

[0167] In one embodiment, the polydispersity index (PDI) of the particles of the liposome may be 0.001 to 0.420. Specifically, the polydispersity index (PDI) of the particles of the liposomes may be 0.001 to 0.420, 0.001 to 0.300, 0.001 to 0.250, 0.001 to 0.100, 0.100 to 0.420, 0.100 to 0.300, 0.100 to 0.250, 0.100 to 0.200, 0.200 to 0.420, 0.200 to 0.300, 0.200 to 0.250, 0.250 to 0.420, 0.250 to 0.300, or 0.300 to 0.420.

[0168] The term "Polydispersity Index (PDI)" is an index representing the particle size distribution, and the closer the index is to 0, the more uniform the particle size, that is, the closer the particle distribution is to a stable monodispersity.

[0169] When the polydispersity index (PDI) of the above liposome particles is between 0.001 and 0.420, it means that the liposome has a uniform particle distribution and is in a stable state. As the PDI of the above liposome particles approaches 1, the possibility of aggregation, precipitation, etc., of the liposome particles varying in size becomes significantly higher.

[0170] In one embodiment, the average zeta potential of the liposome particles may be -10 to -60 mV. Specifically, the average zeta potential of the liposome particles may be -10 to -60 mV, -10 to -50 mV, -10 to -40 mV, -10 to -30 mV, -10 to -20 mV, -20 to -60 mV, -20 to -50 mV, -20 to -40 mV, -20 to -30 mV, -30 to -60 mV, -30 to -50 mV, -30 to -40 mV, -40 to -60 mV, -40 to -50 mV, or -50 to -60 mV.

[0171] The term "zeta potential" refers to the potential difference between a medium and a fixed fluid layer attached to dispersed particles, and zeta potential is utilized as a key indicator of the stability of colloidal dispersions.

[0172] When the average zeta potential of the above liposome particles is -10 to -60 mV, the particles do not aggregate or precipitate due to electrostatic repulsion between the particles and can be stably dispersed in the solution. On the other hand, when the absolute value of the above zeta potential is less than 10, the repulsion between the particles is significantly reduced, causing aggregation and precipitation to occur easily, thereby reducing emulsion stability.

[0173] In one embodiment, the total encapsulation rate of the water-soluble material in the liposome may be 55 to 99.99%. Specifically, the total encapsulation rate of the water-soluble material in the liposome may be 55 to 99.99%, 55 to 90%, 55 to 80%, 55 to 70%, 55 to 60%, 60 to 99.99%, 60 to 90%, 60 to 80%, 60 to 70%, 70 to 99.99%, 70 to 90%, 70 to 80%, 80 to 99.99%, 80 to 90%, or 90 to 99.99%.

[0174] In one embodiment, the total encapsulation rate of the lipid-soluble material in the liposome may be 90 to 99.99%. Specifically, the total encapsulation rate of the lipid-soluble material in the liposome may be 90 to 99.99%, 90 to 99.90%, 90 to 99.00%, 90 to 95%, 95 to 99.99%, 95 to 99.90%, 95 to 99.00%, 99.00 to 99.99%, 99.00 to 99.90%, or 99.90 to 99.99%.

[0175] The term "total encapsulation rate" above is a value calculated as the ratio of the water-soluble and / or lipid-soluble substances bound to the liposome membrane as well as inside the actual liposome particles, out of the total amount of water-soluble and / or lipid-soluble substances actually introduced.

[0176] The above liposome was prepared by introducing a fatty acid having a carboxyl group (-COOH) capable of forming hydrogen bonds with the amino group (-NH2) of ceramide, thereby positioning a high concentration of ceramide between the phospholipid bilayers without precipitation, and as a result, the structure of the liposome was stabilized to encapsulate a high proportion of lipid-soluble substances within the phospholipid bilayers. In addition, water-soluble substances were also encapsulated in a high proportion within the internal space surrounded by the phospholipid bilayers, thereby producing a stabilized liposome capable of simultaneously delivering poorly soluble, lipid-soluble, and water-soluble substances.

[0177] In other experimental examples, the liposome particles (Examples 5 to 8) exhibited a negative charge (-42.88 mV to -45.86 mV), and it was confirmed that the liposome particles were stably dispersed by the repulsive force between the particles (see Experimental Example 6).

[0178] In addition, the encapsulation rates of ceramide and additional water-soluble and fat-soluble substances within the liposomes were found to be significantly higher compared to liposomes without ceramide, confirming that liposomes containing ceramide can also increase the encapsulation rates of other water-soluble and fat-soluble substances. In other words, it was confirmed that when liposomes containing ceramide are manufactured, not only ceramide but also other water-soluble and fat-soluble substances can be loaded in high concentrations and delivered to the skin without precipitation (see Experimental Example 7).

[0179] Furthermore, when water-soluble and lipid-soluble substances were encapsulated in the liposomes containing ceramide (Example 5) and applied to the skin, the transdermal permeability of the water-soluble (total 126.3 ppm) and lipid-soluble substances (total 21.2 ppm) was significantly superior compared to when delivered in a simple solution form, an emulsion form, or a liposome form not containing ceramide, and this was confirmed to be the same in clinical trial results (skin absorption amount: 92.7%, skin absorption rate: 107.7%, absorption depth: 101.7%) (see Experimental Example 8).

[0180] According to one aspect, the liposome contains ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, thereby significantly reducing the amount of water-soluble and fat-soluble substances released during storage, exhibiting high storage stability, and can simultaneously deliver high amounts of ceramide, water-soluble substances, and fat-soluble substances to the skin. In addition, since the liposome has excellent skin permeability, it can be utilized in cosmetics, topical skin preparations, etc., that simultaneously contain poorly soluble, water-soluble, and fat-soluble substances.

[0181]

[0182] Another aspect provides a cosmetic composition comprising the liposome, a fat-soluble substance, and a water-soluble substance. Another aspect provides a topical skin preparation comprising the liposome, a fat-soluble substance, and a water-soluble substance.

[0183] The above liposome contains ceramide, amino acid, polyol, nonionic surfactant, and phospholipid.

[0184] The above terms "ceramide," "surfactant," "liposome," "phospholipid," etc. may be within the aforementioned range.

[0185] In one embodiment, the cosmetic composition or external skin preparation may comprise the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10. Specifically, the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid are 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 1 to 3: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 3 to 6: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 3 to 10: 1 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 3 to 6: 1 to 5: 3 to 10, 3 to 5: It may be included in a weight ratio of 6 to 10: 1: 6 to 10: 1 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 6 to 10: 1 to 3: 3 to 10, 3 to 5: 6 to 10: 1: 6 to 10: 3 to 5: 3 to 10, 3 to 5: 6 to 10: 1: 6 to 10: 3 to 5: 3 to 6, 3 to 5: 6 to 10: 1: 6 to 10: 3 to 5: 6 to 10 or 3 to 4: 6 to 7: 1: 6 to 7: 3 to 4: 6 to 7.

[0186] In one embodiment, the cosmetic composition or external skin preparation may contain a fat-soluble substance and a water-soluble substance in a weight ratio of 0.5 to 6: 0.5 to 6 based on the weight of the amino acid included in the cosmetic composition or external skin preparation. Specifically, the cosmetic composition or external skin preparation may contain a fat-soluble substance and a water-soluble substance in a weight ratio of 0.5 to 6: 0.5 to 6, 0.5 to 3: 0.5 to 6, 3 to 6: 0.5 to 6, 0.5 to 6: 0.5 to 3, 0.5 to 6: 3 to 6, 0.5 to 3: 0.5 to 3, 3 to 6: 0.5 to 3, 0.5 to 3: 3 to 6, 3 to 6: 3 to 6, 0.5 to 2: 0.5 to 2, or 0.5 to 1: 0.5 to 1, based on the weight of the amino acid included in the cosmetic composition or external skin preparation.

[0187] A cosmetic composition or topical skin preparation according to one aspect comprises a liposome containing ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in a weight ratio of 1 to 5: 3 to 10: 1 to 3: 3 to 10: 1 to 5: 3 to 10, thereby significantly reducing the amount of ceramide, water-soluble substances, and fat-soluble substances released during storage, exhibiting high storage stability, and can stably deliver high concentrations of ceramide, water-soluble substances, and fat-soluble substances into the skin simultaneously without precipitation. In addition, the cosmetic composition or topical skin preparation has excellent skin permeability and can simultaneously deliver poorly soluble, water-soluble, and fat-soluble substances to the skin, thereby significantly increasing effects such as skin moisturization, improvement of the skin barrier, and antioxidant properties.

[0188]

[0189] Another aspect is the step of obtaining a lipid mixture by mixing ceramide, fatty acids, and a nonionic surfactant;

[0190] A step of obtaining an emulsion by mixing the above lipid mixture and a eutectic solvent; and

[0191] The present invention provides a method for preparing liposomes for the simultaneous delivery of lipid-soluble substances and water-soluble substances, comprising the step of mixing the above emulsion and phospholipid to obtain liposomes.

[0192] In one embodiment, the method may further include the step of mixing an amino acid and a polyol to obtain the eutectic solvent. Specifically, the step of obtaining the eutectic solvent may be performed before, simultaneously with, or after the step of obtaining the lipid mixture.

[0193] In one embodiment, in the step of obtaining the lipid mixture, the ceramide, fatty acid, and nonionic surfactant may be mixed in a weight ratio of 1 to 5: 3 to 10: 1 to 5 based on the weight of the amino acid contained in the eutectic solvent.

[0194] Specifically, in the step of obtaining the lipid mixture, the ceramide, fatty acid, and nonionic surfactant may be mixed in a weight ratio of 1 to 5: 3 to 10: 1 to 5, 1 to 3: 3 to 10: 1 to 5, 3 to 5: 3 to 10: 1 to 5, 3 to 5: 3 to 6: 1 to 5, 3 to 5: 6 to 10: 1 to 5, 3 to 5: 6 to 10: 1 to 3, or 3 to 5: 6 to 10: 3 to 5, based on the weight of the amino acids contained in the eutectic solvent.

[0195] In one embodiment, in the step of obtaining the lipid mixture, the ceramide and fatty acid can form a lipid core through hydrogen bonding.

[0196] In one embodiment, the hydrogen bond may be formed by the bonding of the amino group (-NH2) of the ceramide and the carboxyl group (-COOH) of the fatty acid.

[0197] In one embodiment, the ceramide and fatty acid may be mixed in a weight ratio of 1 to 5: 3 to 10 based on the weight of the amino acid contained in the liposome. Specifically, the ceramide and fatty acid may be mixed in a weight ratio of 1 to 5: 3 to 10, 1 to 3: 3 to 10, 3 to 5: 3 to 10, 1 to 5: 3 to 6, 1 to 5: 6 to 10, 1 to 3: 3 to 6, 1 to 3: 6 to 10, 3 to 5: 3 to 6, or 3 to 5: 6 to 10 based on the weight of the amino acid contained in the liposome.

[0198] In one embodiment, the weight ratio of the ceramide and the fatty acid in the step of obtaining the lipid mixture may be 1:0.5 to 10. Specifically, the weight ratio of the ceramide and fatty acid in the liposome may be 1:0.5 to 10, 1:0.5 to 7, 1:0.5 to 4, 1:0.5 to 3, 1:0.5 to 2, 1:0.5 to 1, 1:1 to 10, 1:1 to 7, 1:1 to 4, 1:1 to 3, 1:1 to 2, 1:2 to 10, 1:2 to 7, 1:2 to 4, 1:2 to 3, 1:3 to 10, 1:3 to 7, 1:3 to 4, 1:4 to 10, 1:4 to 7, or 1:7 to 10.

[0199] In one embodiment, in the step of obtaining the lipid mixture, the ceramide may be mixed in an amount of 1 to 10 weight% relative to the total weight of the liposome, and specifically, the ceramide may be mixed in an amount of 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, or 7.5 to 10 weight% relative to the total weight of the liposome.

[0200] In one embodiment, in the step of obtaining the lipid mixture, the fatty acid may be mixed in an amount of 5 to 20 weight% relative to the total weight of the liposome, and specifically, the fatty acid may be mixed in an amount of 5 to 20 weight%, 5 to 15 weight%, 5 to 12 weight%, 5 to 8 weight%, 8 to 20 weight%, 8 to 15 weight%, 8 to 12 weight%, 12 to 20 weight%, 12 to 15 weight%, or 15 to 20 weight% relative to the total weight of the liposome.

[0201] In one embodiment, in the step of obtaining the lipid mixture, the nonionic surfactant may be mixed in an amount of 1 to 10 weight% relative to the total weight of the liposome, and specifically, the nonionic surfactant may be mixed in an amount of 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, or 7.5 to 10 weight% relative to the total weight of the liposome.

[0202] In one embodiment, the amino acid and polyol may be mixed in a weight ratio of 1:3 to 10 in the step of obtaining the eutectic solvent.

[0203] Specifically, in the step of obtaining the eutectic solvent, the amino acid and the polyol may be mixed in a weight ratio of 1:3 to 10, 1:3 to 6, or 1:6 to 10.

[0204] In one embodiment, in the step of obtaining the eutectic solvent, the amino acid may be mixed in an amount of 1 to 5 weight% relative to the total weight of the liposome, and specifically, the amino acid may be mixed in an amount of 1 to 5 weight%, 1 to 4 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 4 weight%, 2 to 3 weight%, 3 to 5 weight%, 3 to 4 weight%, or 4 to 5 weight% relative to the total weight of the liposome.

[0205] In one embodiment, in the step of obtaining the eutectic solvent, the polyol may be mixed in an amount of 5 to 20 weight% relative to the total weight of the liposome, and specifically, the polyol may be mixed in an amount of 5 to 20 weight%, 5 to 15 weight%, 5 to 12 weight%, 5 to 8 weight%, 8 to 20 weight%, 8 to 15 weight%, 8 to 12 weight%, 12 to 20 weight%, 12 to 15 weight%, or 15 to 20 weight% relative to the total weight of the liposome.

[0206] In one embodiment, the step of mixing the ceramide, fatty acid, and nonionic surfactant to obtain a lipid mixture may further include the step of heating the lipid mixture at 60 to 100°C.

[0207] Specifically, the heating step may be performed at 60 to 100 ℃, 60 to 90 ℃, 60 to 80 ℃, 60 to 70 ℃, 70 to 100 ℃, 70 to 90 ℃, 70 to 80 ℃, 80 to 100 ℃, 80 to 90 ℃, or 90 to 100 ℃.

[0208] In the step of obtaining the above lipid mixture and heating the above lipid mixture, hydrogen bonds are formed between the carboxyl group (-COOH) of the fatty acid and the amino group (-NH2) of the ceramide, and intermolecular forces are formed according to van der Waals forces. In addition, by mixing a nonionic surfactant together, the precipitation of ceramide is prevented.

[0209] In one embodiment, the step of obtaining the eutectic solvent may further include the step of heating the eutectic solvent at 60 to 100°C.

[0210] Specifically, the heating step may be performed at 60 to 100 ℃, 60 to 90 ℃, 60 to 80 ℃, 60 to 70 ℃, 70 to 100 ℃, 70 to 90 ℃, 70 to 80 ℃, 80 to 100 ℃, 80 to 90 ℃, or 90 to 100 ℃.

[0211] The above "eutectic solvent" is a substance made by mixing hydrogen bond acceptors such as choline chloride, betaine, proline, etc. and hydrogen bond donors such as amides, sugars, alcohols, organic acids, fatty acids, etc., and has the advantage of having high solubility, a low melting point, and low toxicity.

[0212] Specifically, when a eutectic solvent is used, the dissolution of the lipid mixture at a low temperature is enabled, allowing the liposomes to be prepared without using organic solvents. Furthermore, when the eutectic solvent is used, intermolecular interactions (e.g., hydrogen bonding, van der Waals forces, etc.) between the amino acids and polyols in the eutectic solvent and the ceramide in the lipid mixture are maximized, thereby enabling the formation of stable liposomes.

[0213] A manufacturing method according to one aspect can produce a stable emulsion with a significantly reduced amount of precipitated ceramide by mixing the lipid mixture and a eutectic solvent to obtain an emulsion, and by using this for liposome manufacturing, it serves to provide structural stability to the liposome so that water-soluble and fat-soluble substances loaded inside the liposome do not precipitate and can be stably delivered to the skin.

[0214] In one embodiment, the step of obtaining the liposome may involve further mixing a water-soluble substance and a fat-soluble substance with the emulsion and phospholipid. Specifically, the water-soluble substance and the fat-soluble substance may be mixed after the emulsion and phospholipid are mixed; or they may be mixed simultaneously with the emulsion and phospholipid.

[0215] In one embodiment, in the step of obtaining the liposome, the phospholipid may be mixed in a weight ratio of 3 to 10 based on the weight of the amino acid. Specifically, in the step of obtaining the liposome, the phospholipid may be mixed in a weight ratio of 3 to 10, 3 to 8, 3 to 6, 6 to 10, 6 to 8, or 8 to 10 based on the weight of the amino acid.

[0216] In one embodiment, in the step of obtaining the liposome, the water-soluble material and the fat-soluble material may be mixed in a weight ratio of 0.5 to 6: 0.5 to 6 based on the weight of the amino acid. Specifically, in the step of obtaining the liposome, the water-soluble material and the fat-soluble material may be mixed in a weight ratio of 0.5 to 6: 0.5 to 6, 0.5 to 3: 0.5 to 6, 3 to 6: 0.5 to 6, 0.5 to 6: 0.5 to 3, 0.5 to 6: 3 to 6, 0.5 to 6: 0.5 to 6, or 0.5 to 2: 0.5 to 2 based on the weight of the amino acid.

[0217] In one embodiment, in the step of obtaining the liposome, the phospholipid may be mixed in an amount of 1 to 10 weight% relative to the total weight of the liposome, and specifically, the phospholipid may be mixed in an amount of 1 to 10 weight%, 1 to 7.5 weight%, 1 to 5.5 weight%, 1 to 3.5 weight%, 3.5 to 10 weight%, 3.5 to 7.5 weight%, 3.5 to 5.5 weight%, 5.5 to 10 weight%, 5.5 to 7.5 weight%, or 7.5 to 10 weight% relative to the total weight of the liposome.

[0218] In one embodiment, in the step of obtaining the liposome, the water-soluble material may be mixed in an amount of 0.5 to 5 weight% relative to the total weight of the liposome, and specifically, the water-soluble material may be mixed in an amount of 0.5 to 5 weight%, 0.5 to 3 weight%, 0.5 to 2 weight%, 0.5 to 1 weight%, 1 to 5 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 3 weight%, or 3 to 5 weight% relative to the total weight of the liposome.

[0219] In one embodiment, the lipid-soluble material may be included in an amount of 0.5 to 5 weight% relative to the total weight of the liposome, and specifically, the lipid-soluble material may be included in an amount of 0.5 to 5 weight%, 0.5 to 3 weight%, 0.5 to 2 weight%, 0.5 to 1 weight%, 1 to 5 weight%, 1 to 3 weight%, 1 to 2 weight%, 2 to 5 weight%, 2 to 3 weight%, or 3 to 5 weight% relative to the total weight of the liposome.

[0220] In one embodiment, the step of obtaining the liposome may further include the step of applying pressure at 500 to 1500 bar.

[0221] Specifically, the pressurizing step may be performed at 500 to 1500 bar, 500 to 1300 bar, 500 to 1100 bar, 500 to 900 bar, 500 to 700 bar, 700 to 1500 bar, 700 to 1300 bar, 700 to 1100 bar, 700 to 900 bar, 900 to 1500 bar, 900 to 1300 bar, 900 to 1100 bar, 1100 to 1500 bar, 1100 to 1300 bar, or 1300 to 1500 bar.

[0222] Additionally, the above-mentioned pressurizing step may be performed 1 to 10 times, and specifically, 1 to 10 times, 1 to 7 times, 1 to 5 times, 1 to 3 times, 3 to 10 times, 3 to 7 times, 3 to 5 times, 5 to 10 times, 5 to 7 times, or 7 to 10 times.

[0223] In one embodiment, the pressurizing step may be performed simultaneously with the mixing of the emulsion, phospholipid, water-soluble substance and fat-soluble substance, or after the mixing of the emulsion, phospholipid, water-soluble substance and fat-soluble substance.

[0224] By performing the above pressurizing step, the size of the manufactured liposome particles becomes uniform. Specifically, by performing the above pressurizing step 1 to 10 times at 500 to 1500 bar, nano-sized liposomes with an average particle diameter of 10 nm to 100 nm can be obtained. Accordingly, since the manufactured liposomes exhibit a small size, their ability to penetrate the skin barrier is significantly increased.

[0225] Liposomes manufactured by a method according to one aspect are prepared by mixing phospholipids after preparing an emulsion containing ceramide, thereby possessing a phospholipid bilayer that exhibits significantly superior stability, as ceramide stabilized by hydrogen bonding and van der Waals forces exists between the phospholipids. Furthermore, both lipid-soluble and water-soluble substances loaded between the phospholipid bilayers and in the internal space surrounded by the bilayers do not precipitate upon skin delivery and exist stably.

[0226]

[0227] An emulsion according to one aspect stably emulsifies ceramide, which is a poorly soluble substance, and can stably deliver a high amount of ceramide without precipitation. In addition, the emulsion exhibits improved skin permeability, so it can be usefully utilized in cosmetics, topical skin preparations, etc. containing ceramide as an active ingredient.

[0228] A liposome for the simultaneous delivery of lipid-soluble and water-soluble substances, comprising ceramide, fatty acids, amino acids, polyols, nonionic surfactants, and phospholipids according to one aspect, stably disperses ceramide, which is a poorly soluble substance, and can stably deliver not only ceramide but also water-soluble and lipid-soluble substances to the skin simultaneously without precipitation. Furthermore, the liposome exhibits enhanced skin permeability of ceramide and water-soluble and lipid-soluble substances, making it useful for applications such as cosmetics and topical skin preparations containing ceramide and water-soluble and lipid-soluble substances simultaneously.

[0229] Figure 1 is a figure of a nanoemulsion captured using Cryo-EM (High Resolution Cryo-Transmission Electron Microscope packaging system).

[0230] Figure 2 is a figure showing the structure of ceramide and fatty acid in a nanoemulsion. Figure 2a is a figure showing the overall structure of ceramide and fatty acid in an emulsion. Figure 2b is a figure showing an enlarged view of a part of the structure in Figure 2a.

[0231] Figure 3 is a figure confirming the external characteristics of the nanoemulsion.

[0232] Figure 4 is a figure showing the measured average diameter of nanoemulsion particles.

[0233] Figure 5 shows the measured zeta potential of nanoemulsion particles.

[0234] Figure 6 is a figure confirming whether ceramide precipitates in the nanoemulsion.

[0235] Figure 7 is a graph showing the results of a transdermal penetration test of ceramide in a nanoemulsion.

[0236] Figure 8a is a figure showing the morphology of nanoliposomes confirmed using a Cryo-EM (High Resolution Cryo-Transmission Electron Microscope packaging system) instrument.

[0237] Figure 8b is a figure confirming the morphology of nanoliposomes using Cryo-TEM equipment.

[0238] Figure 8c is a schematic diagram of a nanoliposome encapsulating water-soluble and fat-soluble substances.

[0239] Figure 9 is a figure confirming the external appearance of the nanoliposome.

[0240] Figure 10 is a figure showing the measured average diameter of nanoliposome particles.

[0241] Figure 11 shows the measured zeta potential of nanoliposome particles.

[0242] Figure 12 shows the transdermal permeability of niacinamide contained in nanoliposome particles over time.

[0243] Figure 13 shows the transdermal permeability of tocopheryl acetate contained in nanoliposome particles over time.

[0244] Figure 14 shows the results of a clinical trial showing the skin absorption of water-soluble and fat-soluble components when nano-liposomes are applied to the skin.

[0245] Figures 15a and 15b show the results of checking the depth of wrinkles after 4 weeks of use compared to before using the eye cream.

[0246] Figures 16a and 16b show the results confirming a reduction in melanin area after 4 weeks of use compared to before using the eye cream.

[0247] The present invention will be explained in more detail below through examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0248]

[0249] <Experimental Example 1> Selection of Excellent Ceramide Stabilizer

[0250] In order to select a stabilizer capable of strongly forming hydrogen bonds and van der Waals forces with ceramide, each lipid was mixed with ceramide in the ratios shown in Table 1 below, and the change in the lipid transition temperature of the ceramide was measured, and a ceramide stabilizer exhibiting a low lipid transition temperature was selected.

[0251] Specifically, to determine the lipid transition temperature, lipid cores, i.e., ceramide and lipids, were mixed according to the composition of Table 1 below and then dissolved in a water bath at 95°C for 10 minutes. Subsequently, the mixed lipid cores were cooled by repeatedly lowering the temperature of the water bath by 5°C increments and maintaining it for 15 minutes, and changes in the external appearance of samples 1 to 4 and comparison samples 1 and 2 were observed during this process.

[0252] As a result, as shown in Table 1 below, ceramide exhibited a low lipid transition temperature of 80°C when mixed with fatty acids, particularly among the lipids mentioned above.

[0253] When mixed with fatty acids, it exhibited a low lipid transition temperature of 80°C.

[0254] INCI Name Sample 1 Sample 2 Sample 3 Sample 4 Comparison Sample 1 Comparison Sample 2 Ceramide NP (CAS NO. 34354-88-6 / 100403-19-8) 10 10 10 10 10 10 oleic acid 10 Lauric acid 10 Palmitic acid 10 Stearic acid 10 Caprylic / Capric Triglyceride 10 Isopropyl Myristate 10 Curing Temperature (Lipid Transition Temperature) 70 70 80 80 Insoluble 90

[0255] The content of each ingredient is based on weight %.

[0256] In other words, since ceramide can be mixed with fatty acids to regulate lipid transition temperature, a balance between fluidity and stability can be secured.

[0257]

[0258] <Experimental Example 2> Evaluation of the Stability of Ceramide Emulsion According to the Type of Fatty Acid

[0259] The presence or absence of ceramide precipitation in the emulsion according to the type of fatty acid was confirmed. An emulsion containing ceramide (hereinafter referred to as the ceramide emulsion) was prepared through the following process.

[0260] Specifically, ceramide and lipids were mixed according to the composition of Table 2 below, heated to 80°C and stirred for 5 minutes (1,000 rpm), and then cooled to prepare a lipid core. Subsequently, amino acids and water were mixed according to the composition of Table 2 below, heated to 80°C and stirred for 10 minutes (1,000 rpm), and then cooled to prepare a deep eutectic solvent (DES). Next, the lipid core and the deep eutectic solvent were mixed, and a homogenized ceramide emulsion was prepared by circulating a high-pressure homogenizer 5 times at 1,000 bar. Afterward, a preservative was mixed, cooled to 10°C, and stirred at 1,000 rpm for 10 minutes.

[0261] INCI Name Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Lipid Core Ceramide NP (CAS NO. 34354-88-6 / 100403-19-8) 10 10 10 10 10 oleic acid 20 Caprylic / Capric Triglyceride 20 Lauric acid 20 Palmitic acid 20 Stearic acid 20 Polyglyceryl-10 Stearate 10 10 10 10 10 Deep eutectic solvent arginine 33 33 Glycerin 20 20 20 20 Water 35 35 35 35 35 Preservative 1,2-hexandiol 22 22 2 Total Content 100 100 100 100 100

[0262] The content of each ingredient is based on weight %.

[0263] As a result, as shown in Table 3 below, in the case of Example 1, in which oleic acid is included in the lipid core, the precipitation of ceramide is suppressed under all temperature conditions even after 12 weeks have passed since the preparation of the emulsion, and it was confirmed that an emulsion containing ceramide is formed stably and exhibits excellent emulsion stability.

[0264] Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 1 2nd Week 4 No precipitation Gel Fossil Precipitation Precipitation Precipitation 20 No precipitation Gel Fossil No precipitation Precipitation No Precipitation Precipitation 37 No precipitation Gel Fossil No precipitation Precipitation No Precipitation Cycle No precipitation Gel Fossil No precipitation Precipitation Precipitation

[0265] In the case of the above cycle conditions, heating and cooling are repeated within a temperature range of -20 ℃ to 20 ℃, and the phenomena occurring when the samples of the examples and comparative examples are frozen and thawed under these conditions are confirmed.

[0266] <Experimental Example 3> Evaluation of the Stability of Ceramide Emulsion According to the Type of Nonionic Surfactant

[0267] An auxiliary emulsifier for a ceramide emulsion containing oleic acid, which can form the most stable emulsion at all temperatures confirmed in Experimental Example 2, was selected. Specifically, a ceramide emulsion was prepared using the same method as in Experimental Example 2, except that a nonionic surfactant was added to prepare the ceramide emulsion with the composition shown in Table 4 below. Subsequently, the presence or absence of ceramide precipitation in the emulsion at each temperature was checked, and the results are shown in Table 5.

[0268] INCI Name Example 2 Example 3 Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Lipid Core Ceramide NP (CAS NO. 34354-88-6 / 100403-19-8) 10 10 10 10 10 oleic acid 20 20 20 20 20 Polyglyceryl-10 oleate (CAS NO. 79665-93-3) 10 Polyglyceryl-10 laurate (CAS NO. 34406-66-1) 10 Decyl Glucoside (CAS NO. 68515-73-1) 10 PolySorbate 20 (CAS NO. 9005-64-5) 10 PolySorbate 80 (CAS NO. 9005-65-6) 10 Glyceryl Stearate / PEG-100 Stearate(CAS NO. 31566-31-1 / 9004-99-3)10Deep eutectic solventarginine333333Glycerin202020202020Water353535353535Preservative1,2 hexandiol222222Total content100100100100100100

[0269] The content of each ingredient is based on weight %.

[0270] Example 2 Example 3 Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 12 Week 4 No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation 20 No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation 37 No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation Cycle No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation No precipitation

[0271] *In the case of the above cycle conditions, heating and cooling were repeated within a temperature range of -20 ℃ to 20 ℃, and the phenomena occurring when the samples of the examples and comparative examples were frozen and thawed under these conditions were confirmed. As a result, as shown in Table 5 above, it was confirmed that a high-content ceramide emulsion containing oleic acid in the lipid core formed a ceramide emulsion stably under all temperature conditions and exhibited excellent emulsion stability when 12 weeks had elapsed from the preparation of the ceramide emulsion, regardless of the type of auxiliary emulsifier. Although the viscosity of the ceramide emulsions prepared varied depending on the type and molecular weight of the nonionic surfactant, no precipitation of ceramide was confirmed in all of Examples 2 to 4 and Comparative Examples 5 to 7.

[0272]

[0273] <Experimental Example 4> Analysis of the physical properties of ceramide emulsion

[0274] To evaluate whether a ceramide-containing emulsion was stably formed, a characterization experiment was performed as follows.

[0275] Specifically, Examples 1 to 4 were classified into cases where they were not diluted and cases where they were diluted to a concentration of 1 v / v% using purified water, and the transparency was checked visually and the formation of suspended solids and precipitates was checked.

[0276] As a result, as shown in Figure 3, no precipitates or suspended particles were detected in either the undiluted ceramide emulsion or the ceramide emulsion diluted to a concentration of 1 v / v%.

[0277] Based on the above results, it was confirmed that the ceramide emulsions of Examples 1 to 4 maintain a stably dispersed form in both diluted and undiluted cases, where a large amount of ceramide emulsion particles are present in the solution.

[0278] This is the result of confirming that a ceramide emulsion containing specific fatty acids, ceramide, and nonionic surfactants can exhibit significantly superior stability.

[0279]

[0280] <Experimental Example 5> Particle Size Analysis of Ceramide Emulsion

[0281] The size and homogeneity of the ceramide emulsions of Examples 1 to 4 and Comparative Examples 5 to 7 were measured using a particle size analyzer, Zetasizer PRO, from Malvern.

[0282] The above particle size analyzer analyzes particle size by calculating the Brownian motion of nanoparticles using a formula, and the results are shown in Table 6 and Figure 4 below.

[0283] Particle Size (z-average) PDI Example 1 96.56 nm 0.321 Example 2 179.3 nm 0.006 Example 3 114.5 nm 0.160 Example 4 170.8 nm 0.070 Comparative Example 5 283.6 nm 0.457 Comparative Example 6 278.6 nm 0.496 Comparative Example 7 505.8 nm 0.327

[0284] As shown in Table 6 and Figure 4 above, the ceramide emulsions of Examples 1 to 4 had an average particle size of 96.56 nm to 179.3 nm and a PDI of 0.006 to 0.321. That is, it was confirmed that the ceramide emulsions of Examples 1 to 4 all consisted of nano-sized particles and that the ceramide emulsions all had uniform particle sizes.

[0285] On the other hand, the average particle size of the ceramide emulsions of Comparative Examples 5 to 7 was found to be 278.6 nm to 505.8 nm, which is an increase of at least 1.6 times compared to Examples 1 to 3 containing polyglyceryl (PGL)-based nonionic surfactants and Example 4 containing the glycolipid decyl glucoside. In addition, the PDI was found to be 0.327 to 0.496. That is, it was confirmed that Comparative Examples 5 to 7 are not suitable for skin penetration because the particle size increased and the particle size distribution was non-uniform.

[0286] Accordingly, the ceramide emulsions of Examples 1 to 4 have a uniform distribution of nano-sized particles dispersed in a solution, which not only provides excellent stability but also exhibits properties suitable for skin penetration due to the uniform distribution of nano-sized particles, so they can be effectively applied to cosmetic compositions, topical skin preparations, etc.

[0287]

[0288] <Experimental Example 6> Measurement of Zeta Potential of Ceramide Emulsion

[0289] Zeta potential is the potential difference between a medium and a fixed fluid layer attached to dispersed particles, and it is used as a key indicator of the stability of a colloidal dispersion. The magnitude of the zeta potential indicates the degree of electrostatic repulsion between adjacent charged particles in the dispersion. Accordingly, the stability of the ceramide emulsions of Examples 1 to 4 was evaluated according to the criteria in Table 7 below by measuring the zeta potential using Malvern’s particle size analyzer Zetasizer PRO, and the results are shown in Table 8 below.

[0290] Zeta potential [mV] ±0~±5 Rapid coagulation or aggregation ±10~±30 Initial instability ±30~40 Moderate stability ±40~±60 Excellent stability ±61 Outstanding stability

[0291] Zeta potential [mV] Example 1-13.76 Example 2-27.29 Example 3-18.32 Example 4-45.64

[0292] As shown in Table 8 and Figure 5, Examples 1 to 4 all exhibited a negative charge. Specifically, in the case of Example 4, which used the glycolipid decyl glucoside as a nonionic surfactant, the zeta potential was found to be -45.64 mV, confirming excellent stability.

[0293] Meanwhile, Examples 1 to 3, which contain polyglyceryl (PGL)-based nonionic surfactants, exhibited relatively low absolute zeta potentials ranging from -13.76 mV to -27.29 mV. This is believed to be because the absolute value of the hydrophilic partial charge is low in the case of polyglyceryl-based nonionic surfactants. This result indicates that although the absolute value of the zeta potential in Examples 1 to 3 is relatively low, the particles can be stabilized by physically occupying a large space, as they consist of polyglyceryl-based nonionic surfactants with large volumes.

[0294] When the results of Experimental Examples 4 to 6 are combined, it can be seen that the ceramide emulsions of Examples 1 to 4 have a uniform nano-scale particle size and exhibit excellent dispersion stability. Specifically, Examples 1 to 3, which use polyglyceryl-based nonionic surfactants, exhibit dispersion stability based on the physical structure of the nonionic surfactants, and Example 4, which uses glycolipids, was confirmed to have excellent dispersion stability based on the electrostatic repulsion between charged ceramide emulsion particles.

[0295] That is, since the ceramide emulsion according to one aspect has significantly excellent dispersion stability, it can be applied and utilized in cosmetic compositions, external skin preparations, etc.

[0296]

[0297] <Experimental Example 7> Measurement of encapsulation efficiency of ceramide emulsion

[0298] To confirm the ceramide encapsulation rate of the ceramide emulsions of Examples 1 to 4, centrifugation was performed to separate the ceramide encapsulated and unencapsulated ceramide within the emulsion, and then quantified using UV and HPLC. HPLC analysis was performed under the following conditions.

[0299] -Method: ceramide_9_PDA set

[0300] -Column: YMC pack proC18 (250 x 4.6 mm, 5μm)

[0301] -Eluent: methanol

[0302] -Gradient: isocratic

[0303] -Flow: 0.5 mL / min

[0304] -Injection Volume: 50 µl

[0305] -Detector: PDA, 205 nm

[0306] -Blank: methanol

[0307] Specifically, 0.5 mL of the emulsion suspension was placed in an Ultra Centrifugal Filter (50 kDa), centrifuged at 4,000 rpm for 5 minutes using a microcentrifuge, and the lower layer was collected and analyzed by HPLC. The encapsulation rate was calculated using the following Equation 1.

[0308] [Mathematical Formula 1]

[0309] %EE(encapsulation efficiency) = (Total amount of ceramide - Amount of lower layer ceramide) / Total amount of ceramide) * 100

[0310] Lot number Sample (g) Solvent (mL) Area Results (ppm) Results (%) Encapsulation Rate (%) Example 1 1.00001 Not detected Not detected Not detected 99.9 Example 2 1.00001 Not detected Not detected Not detected 99.9 Example 3 1.00001 Not detected Not detected Not detected 99.9 Example 4 1.00001 Not detected Not detected Not detected 99.9

[0311] As a result, as shown in Table 9 above, all showed a high encapsulation rate of 99.9%.

[0312] Meanwhile, ceramide is one of the insoluble substances with high crystallinity and a specific gravity lower than that of water; therefore, since there are limitations to measuring the encapsulation rate solely through centrifugation and HPLC analysis, changes over time were also observed. Specifically, the particle characteristics of Examples 1 to 4 and Comparative Example 1 at 37°C during the 18th week of preparation were confirmed through optical and polarized light using a microscope.

[0313] As shown in Fig. 6, in the case of Examples 1 to 4 containing oleic acid, a fatty acid, no precipitation of ceramide was observed, whereas in the case of Comparative Example 1 containing Caprylic / Capric Triglyceride (MCT), which has relatively low polarity, it was confirmed that ceramide crystals were precipitated.

[0314] That is, it can be seen that the ceramide emulsion containing oleic acid with relatively high polarity (Examples 1 to 4) exhibits a significantly high encapsulation rate by stably encapsulating the ceramide within the emulsion, thereby preventing the precipitation of ceramide crystals.

[0315] Therefore, a ceramide emulsion according to one aspect can encapsulate ceramide in a high proportion within the emulsion, thereby overcoming the problem of existing emulsion formulations that showed low efficacy even with high-concentration ceramide administration, and enabling the desired efficacy of ceramide to be obtained.

[0316]

[0317] <Experimental Example 8> Evaluation of Transdermal Penetration Efficacy of Ceramide Emulsion

[0318] The transdermal penetration and absorption rates of the ceramide emulsion were measured in accordance with the Guidelines for In Vitro Skin Absorption Testing (Ministry of Food and Drug Safety). Specifically, in vitro skin absorption testing is a method to verify whether a test substance can penetrate the skin; it involves applying or injecting the test substance onto the skin surface and measuring the amount of substance that has migrated from the receptor site inside the skin after a certain period of time. In this experimental example, Millipore's Strat-M, an artificial membrane for transdermal absorption testing, was used. ® After mounting the membrane onto the Franz-Diffusion Cell and System (LOGAN), the experiment was performed according to the conditions in Table 10 below. The Franz-Diffusion Cell was maintained under non-occluded conditions, with the donor chamber not sealed with Parafilm, and the ceramide concentration in the sampled aliquot was measured after 12 hours and is shown in Table 10 below.

[0319] Artificial Skin Strat-M Membrane Skin Area: 4.90 cm 2 Volume of sample solution / Concentration: 0.2 mL / ceramide 1% Receptor medium: Purified water Volume of Receptor medium: 10 mL Temperature: 37 ℃ Stirbar speed: 500 rpm Sampling aliquot: 1 mL Sampling time: 12 h

[0320] Lot number Sample (g) Solvent (mL) Area Results (ppm) Results (%) Comparative Example 1 1.0000 12 39 24 27.03 60.00 1 Example 4 1.0000 15 40 18 216.8 270.00 2

[0321] As a result, as shown in Figure 7 and Table 11, the ceramide detection concentration over time was higher in Example 1, which contains a glycoside-based surfactant, compared to Comparative Example 1, which contains Caprylic / Capric Triglyceride (MCT). Specifically, in Comparative Example 1, ceramide was detected at 4.88 ppm and 7.03 ppm after 8 and 12 hours of transmission, respectively, whereas in Example 4, ceramide was detected at 10.01 ppm and 16.82 ppm after 8 and 12 hours of transmission, respectively, showing a transmission amount more than twice that of Comparative Example 1.

[0322] That is, since oleic acid stabilizes ceramide, the ceramide emulsion containing the oleic acid can have significantly increased skin permeability compared to a ceramide emulsion containing caprylic / capric triglyceride.

[0323] Therefore, a ceramide emulsion containing oleic acid and a nonionic surfactant can not only produce a stable formulation but also deliver ceramide into the skin with high efficiency.

[0324]

[0325] <Experimental Example 9> Preparation of an emulsion containing ceramide

[0326] An emulsion containing ceramide (hereinafter referred to as ceramide emulsion; emulsions 1 to 7) was prepared with the composition of Table 12 below.

[0327] Specifically, lipids were mixed according to the composition of Table 12 below, heated to 80°C and stirred for 5 minutes (1,000 rpm), and then cooled to prepare a lipid core. Subsequently, amino acids, polyols, and water were mixed according to the composition of Table 12 below, heated to 80°C and stirred for 10 minutes (1,000 rpm), and then cooled to prepare a deep eutectic solvent (DES). Next, the lipid core and the deep eutectic solvent were mixed, and a homogenized ceramide emulsion was prepared by circulating a high-pressure homogenizer 5 times at 1,000 bar. Afterward, a preservative was mixed, cooled to 10°C, and stirred at 1,000 rpm for 10 minutes.

[0328] INCI Name Emulsion 1 Emulsion 2 Emulsion 3 Emulsion 4 Emulsion 5 Emulsion 6 Emulsion 7 Lipid Core Ceramide NP(CAS NO. 34354-88-6 / 100403-19-8)10101010101010 Oleic acid20202020202020 Polyglyceryl-10 stearate(CAS NO. 79777-30-3)10 Polyglyceryl-10 oleate(CAS NO. 79665-93-3)10 Polyglyceryl-10 laurate(CAS NO. 34406-66-1)10 Decyl Glucoside(CAS NO. 68515-73-1)10 PolySorbate 20(CAS NO. 9005-64-5)10PolySorbate 80(CAS NO. 9005-65-6)10Glyceryl Stearate / PEG-100 Stearate(CAS NO. 31566-31-1 / 9004-99-3)10Deep eutectic solventarginine3333333Glycerin20202020202020Water35353535353535Preservative1,2 hexandiol2222222Total content100100100100100100100

[0329] *The content of each component is based on weight%. As a result, as shown in Table 13 below, emulsions 1 to 7 containing oleic acid in the lipid core formed a ceramide emulsion stably under all temperature conditions even after 12 weeks had passed since the preparation of the ceramide emulsion, regardless of the type of auxiliary emulsifier. In addition, although the viscosity of the ceramide emulsion varied depending on the type and molecular weight of the nonionic surfactant, no precipitation of ceramide was confirmed in any of emulsions 1 to 7.

[0330]

[0331] Emulsion 1 Emulsion 2 Emulsion 3 Emulsion 4 Emulsion 5 Emulsion 6 Emulsion 7 12th Week 4 Degree No precipitation

[0332] In the case of the above cycle conditions, heating and cooling are repeated within a temperature range of -20 ℃ to 20 ℃, and the phenomena occurring when the samples of the examples and comparative examples are frozen and thawed under these conditions are confirmed.

[0333] <Experimental Example 10> Preparation of Liposomes Containing Ceramide

[0334] Liposomes of Examples 5 to 8 and Comparative Examples 8 to 10 (hereinafter referred to as ceramide liposomes) were prepared using emulsions 1 to 7 prepared in Experimental Example 9 above without the use of an organic solvent.

[0335] Specifically, the lipid membrane component was mixed according to the composition of Table 14 below, heated to 80°C and stirred for 10 minutes (1,000 rpm), and then cooled. Subsequently, the lipid membrane component and the emulsion prepared in Experimental Example 9 (Emulsion 1 to 7) were mixed according to the composition of Table 14 below, and then homogenized ceramide liposomes were prepared by circulating a high-pressure homogenizer 5 times at 1,000 bar, and then a preservative was mixed, cooled to 10°C, and stirred for 10 minutes at 1,000 rpm.

[0336] INCI Name Example 5 Example 6 Example 7 Example 8 Comparative Example 8 Comparative Example 9 Comparative Example 10 Emulsion Emulsion 150 Emulsion 250 Emulsion 350 Emulsion 450 Emulsion 550 Emulsion 650 Emulsion 750 Lipid Membrane Hydrogenated Lecithin 10 10 10 10 10 10 Water 3 8 38 38 38 38 38 Preservative 1,2-hexandiol 2 2 2 2 2 2 Total Content 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 0

[0337] The content of each ingredient is based on weight %.

[0338] As a result, when liposomes were prepared with the compositions of Comparative Examples 8 to 10, no liposomes were formed, but when prepared with the compositions of Examples 5 to 8, stable liposomes were formed. That is, according to one aspect, liposomes can form a liposome structure by self-assembling a lipid core containing a specific non-phosphorous surfactant in contact with a phospholipid.

[0339]

[0340] <Experimental Example 11> Preparation of Ceramide Liposomes Containing Water-Soluble and Lipid-Soluble Substances

[0341] In order to stably load water-soluble and fat-soluble substances into the liposomes without precipitation and to deliver the water-soluble and fat-soluble substances to the skin with high efficiency, liposomes of Examples 9 to 12 with the compositions of Table 4 below were prepared using Examples 5 to 8 prepared in Experimental Example 10, and the morphology of the liposomes was confirmed using a Cryo-EM (High Resolution Cryo-Transmission Electron Microscope packaging system) device.

[0342] Specifically, the lipid core prepared in Experimental Examples 9 and 10 above, the eutectic solvent, the lipid membrane, and the water-soluble and fat-soluble substances of Table 15 below were mixed, and homogenized liposomes were prepared by circulating a high-pressure homogenizer five times at 1000 bar. Subsequently, a preservative was mixed, cooled to 10°C, and stirred at 1000 rpm for 10 minutes.

[0343] INCI Name Example 9 Example 10 Example 11 Example 12 Ceramide Liposome Example 198 Example 298 Example 398 Example 498 Active Product Water-soluble substance (niacinamide) 1111 Fat-soluble substance (tocopheryl acetate) 1111

[0344] *The content of each component is based on weight %. Figure 8a is a diagram confirming the morphology of nanoliposomes using a Cryo-EM (High Resolution Cryo-Transmission Electron Microscope packaging system) instrument.

[0345] As shown in Fig. 8a, it can be confirmed that the nanoliposome according to one aspect exhibits a spherical double-membrane structure.

[0346]

[0347] <Experimental Example 12> Evaluation of Ceramide Liposomes Containing Water-Soluble and Lipid-Soluble Substances (Analysis of Characteristics)

[0348] To evaluate whether ceramide liposomes containing water-soluble and fat-soluble substances were stably formed, a morphological analysis experiment was performed as follows.

[0349] Specifically, Examples 9 to 12 were classified into cases where they were not diluted and cases where they were diluted to a concentration of 1% using purified water, and the transparency was checked visually and the formation of suspended solids and precipitates was checked.

[0350] As shown in Fig. 9, no precipitates or suspended particles were detected in either the undiluted ceramide liposomes or the ceramide liposomes diluted to a concentration of 1%.

[0351] Based on the above results, it was confirmed that the ceramide liposomes of Examples 9 to 12 maintain a stably dispersed form in both diluted and undiluted cases, where a large amount of ceramide liposome particles are present in the solution.

[0352] It was confirmed that the above ceramide liposomes exhibit significantly higher storage stability compared to liposomes that do not contain ceramide, as they exhibit structural stability through hydrogen bonding between ceramide and phospholipids by first preparing an emulsion containing ceramide and then adding phospholipids.

[0353] In other words, it was confirmed that the ceramide liposome exhibits high stability as an active ingredient delivery system, as the ceramide and the water-soluble and fat-soluble substances are stably dispersed without precipitation even when loaded with water-soluble and fat-soluble substances.

[0354]

[0355] <Experimental Example 13> Particle Size Analysis of Ceramide Liposomes

[0356] The size and homogeneity of ceramide liposomes (Examples 9 to 12) containing water-soluble and fat-soluble substances were measured using Malvern’s particle size analyzer, Zetasizer PRO.

[0357] The above particle size analyzer analyzes particle size by calculating the Brownian motion of nanoparticles using a formula, and the results are shown in Table 16 and Figure 10 below.

[0358] Particle size (z-average) PDI Example 965.9 nm 0.285 Example 1062.5 nm 0.265 Example 1156.2 nm 0.304 Example 1247.77 nm 0.415

[0359] As shown in Table 16 and Figure 10 above, the ceramide liposomes of Examples 9 to 12 showed an average particle size of 47.77 nm to 65.9 nm, and the PDI was 0.265 to 0.415. That is, the liposomes of Examples 9 to 12 are all nano-sized particles, and it was confirmed that the ceramide liposomes all have uniform particle sizes.

[0360] Therefore, the ceramide liposomes according to one aspect have excellent stability as they are dispersed in a solution with a uniform distribution of nano-sized particles. In addition, since the ceramide liposomes have a uniform distribution of nano-sized particles and exhibit properties suitable for skin penetration, they can be applied to cosmetic compositions, topical skin preparations, etc.

[0361]

[0362] <Experimental Example 14> Measurement of Zeta Potential of Ceramide Liposomes

[0363] Zeta potential is the potential difference between a medium and a fixed fluid layer attached to dispersed particles, and it is used as a key indicator of the stability of a colloidal dispersion. The magnitude of the zeta potential indicates the degree of electrostatic repulsion between adjacent charged particles in the dispersion. Accordingly, to evaluate the stability of ceramide liposomes (Examples 9 to 12), the zeta potential was measured using the Zetasizer PRO particle size analyzer from Malvern. Specifically, the stability of the ceramide liposomes was evaluated according to the criteria in Table 17 below and is shown in Table 18 below.

[0364] Zeta potential [mV] ±0~±5 Rapid coagulation or aggregation ±10~±30 Initial instability ±30~40 Moderate stability ±40~±60 Excellent stability ±61 Outstanding stability

[0365] Zeta potential [mV] Example 9-45.86 Example 10-43.41 Example 11-42.88 Example 12-43.13

[0366] As shown in Table 18 and Figure 11, the zeta potentials of Examples 9 to 12 were measured, and all of Examples 9 to 12 exhibited a negative charge. Specifically, the zeta potentials of Examples 9 to 12 were measured in the range of -42.88 mV to -45.86 mV, confirming that they exist in a stable state in the solution due to electrostatic repulsion between the ceramide liposome particles.

[0367] Accordingly, it was confirmed that the ceramide liposomes of Examples 9 to 12 have a uniform nano-scale particle size and excellent dispersion stability due to electrostatic repulsion between charged ceramide liposome particles.

[0368] In other words, since ceramide liposomes according to one aspect have significantly superior dispersion stability, they can be applied and utilized in cosmetic compositions, topical skin preparations, etc.

[0369]

[0370] <Experimental Example 15> Measurement of Ceramide Liposome Encapsulation Efficiency

[0371] To determine the encapsulation rates of water-soluble (niacinamide) and fat-soluble (tocopheryl acetate) substances within the ceramide liposomes of Examples 9 to 12, centrifugation was performed to separate the substances encapsulated and unencapsulated within the liposomes, and then quantified using UV and HPLC. HPLC analysis was performed under the conditions of Table 19 below.

[0372] Niacinamide Tocopheryl Acetate Column YMC pack proC18 (250 x 4.6 mm) YMC pack proC18 (250 x 4.6 mm) Eluent Methanol (20): DW (80) methanol Detector 263 nm PDA or UVD (205 nm) Flow 1 mL / min 0.5 mL / min Injection volume 10 µl 50 µl Run time 30 min 40 min Temp 30 ℃-

[0373] Specifically, 0.5 mL of liposome suspension was placed in an Ultra Centrifugal Filter (50 kDa), centrifuged at 4,000 rpm for 5 minutes using a microcentrifuge, and the lower layer was collected and analyzed by HPLC. The encapsulation rate was calculated using the following Equation 2.

[0374] [Mathematical Formula 2]

[0375] %EE(encapsulation efficiency) = (Total amount of active ingredient - Amount of active ingredient in lower layer) / Total amount of active ingredient) * 100

[0376] Niacinamide Tocopheryl Acetate Total Encapsulation Rate Particle Internal Encapsulation Rate Total Encapsulation Rate Particle Internal Encapsulation Rate Control Group (Ceramide-free Liposomes) 54% 1% 62.9% 62% Example 9 64% 40% 99.9% 99.9% Example 10 80% 30% 99.9% 99.9% Example 11 63% 31% 99.9% 99.9% Example 12 60% 33% 99.9% 99.9%

[0377] As a result, as shown in Table 20 above, for lipid-soluble substances, both the total encapsulation rate and the particle internal encapsulation rate were 99.9%, showing a high encapsulation rate. On the other hand, for water-soluble substances, the total encapsulation rates were 64%, 80%, 63%, and 60% for the liposomes of Examples 9 to 12, respectively, showing the highest rate in Example 6, which used polyglyceryl-10 oleate as a nonionic surfactant. In addition, regarding the particle internal encapsulation rates, the liposomes of Examples 9 to 12 were 40%, 30%, 31%, and 33%, respectively, showing the highest rate in Example 9, which used polyglyceryl-10 stearate as a nonionic surfactant.

[0378] In addition, Examples 9 to 12 showed a high encapsulation rate compared to the control group, which is a liposome that does not contain ceramide, in both water-soluble and fat-soluble substances.

[0379] In other words, ceramide liposomes according to one aspect exhibit a significantly increased encapsulation rate of water-soluble and fat-soluble substances compared to liposomes that do not contain ceramide, thereby enabling the production of liposomes containing a high concentration of active ingredients. Consequently, compared to conventional liposomes composed solely of phospholipids, it is possible to deliver a high concentration of active ingredients to the target site.

[0380]

[0381] <Experimental Example 16> Evaluation of Transdermal Penetration Efficacy of Ceramide Liposomes Containing Water-Soluble and Lipid-Soluble Substances

[0382] The transdermal penetration and absorption rates of ceramide liposomes were measured in accordance with the guidelines for in vitro skin absorption testing (Ministry of Food and Drug Safety). Specifically, in vitro skin absorption testing is a method to determine whether a test substance can penetrate the skin; it involves applying or injecting the test substance onto the skin surface and measuring the amount transferred to a solution reservoir inside the skin after a certain period of time. In this experimental example, the Membrane Strat-M, an artificial membrane for transdermal absorption testing from Millipore, was mounted on the Franz-Diffusion Cell and System (LOGAN) and the experiment was conducted according to the conditions in Table 21 below. The Franz-Diffusion Cell was maintained under non-occluded conditions, with the donor chamber not sealed with Parafilm. As a control group, an aqueous solution of niacinamide, an emulsion without ceramide, and liposomes were used.

[0383] Artificial Skin Strat-M Membrane Skin Area: 4.90 cm 2 Volume of sample solution / Concentration: 0.2 mL / ceramide 1% Receptor medium: Purified water Volume of Receptor medium: 10 mL Temperature: 37 ℃ Stirbar speed: 500 rpm Sampling aliquot: 1 mL Sampling time: 12 h

[0384] Niacinamide Area 3h 6h 9h 12h total Solution (control) 5,597 5,421 5,786 5,584 22,388 Liposome (control) 284,563 247,157 292,546 277,482 1,101,748 Example 943 2,587 442,212 435,214 398,470 1,708,483

[0385] Tocopherol acetate Area3h6h9h12htotalEmulsion(control)194,373183,756175,642223,721777,492Liposome(control)235,484205,484247,854288,500977,322Example 92,684,7502,457,7603,452,1701,763,20110,357,881

[0386] As a result, as shown in Table 22 and Figure 12, it was confirmed that the liposome of Example 9 had a significantly wider permeation area of ​​the active ingredient over time compared to the control group. Specifically, the permeation amount of niacinamide after 12 hours of permeation in Example 9 was approximately 76 times higher and approximately 1.5 times higher than the control group niacinamide aqueous solution and control group liposome, respectively. In addition, the permeation amount of tocopherol acetate after 12 hours of permeation in Example 5 was approximately 133 times higher and approximately 10.5 times higher than the control group emulsion and liposome, respectively.

[0387] That is, liposomes according to one aspect exhibit significantly higher skin permeability than conventional cosmetic formulations such as aqueous solutions, emulsions, and liposome formulations, and can deliver water-soluble and fat-soluble substances into the skin with high efficiency.

[0388]

[0389] In addition, according to the conditions of Table 24 below, the amount, rate, and depth of absorption of water-soluble and oil-soluble substances into the skin were measured using a Raman spectroscope, and the results are shown in Tables 25 to 27 below, respectively. Ceramide-free liposomes were used as the control group.

[0390] Number of screened / Minimum valid evaluations 5 / 5 Skin type Dry (5) / Normal (0) / Oily (0) Average age 31 Gender Male (4) / Female (1)

[0391] Skin Absorption Amount Subject No. Before Use Example 5 Control Female 13 20 87 26 25 0 16 41 60 5 2 Female 23 27 84 7 60 7 7 9 860 32 8 8 Female 33 29 9 24 6 19 35 5 4 6 19 4 8 Female 43 24 19 4 6 36 14 4 4 8 12 5 3 Male 13 25 18 0 64 9 4 7 25 6 63 5 8 Improvement Rate 0% 92.7% 55.3%

[0392] Skin Absorption Rate Subject No. Before Use Example 5 Control Female 19 3 210.0 20 6.4 Female 210 5.6 210.0 16 7.4 Female 310 3.2 210.0 19 6.8 Female 410 4.4 210.0 17 1.6 Male 19 9.6 180.0 17 4.0 Improvement Rate 0% 107.7% 81.1%

[0393] Skin Absorption Depth Subject No. Before Use Example 5 Control Female 131.0 70.0 68.8 Female 235.2 70.0 55.8 Female 334.4 70.0 65.6 Female 434.8 70.0 57.2 Male 133.2 60.0 58.0 Improvement Rate 0% 101.7% 81.1%

[0394] As a result, as shown in Tables 25 to 27 and Figure 14, it was confirmed that Example 9 showed a significant increase in skin absorption amount, absorption rate, and absorption depth compared to the control group compared to before use. Specifically, compared to the control group, the skin absorption amount, absorption rate, and absorption depth of Example 9 increased by approximately 1.7 times, 1.3 times, and 1.3 times, respectively, showing excellent skin permeability.

[0395] In other words, liposomes according to one aspect have significantly increased skin penetration of active ingredients, including ceramide, and can effectively deliver water-soluble and fat-soluble substances.

[0396]

[0397] <Experimental Example 17> Evaluation of clinical efficacy of cosmetic formulations (1)

[0398] To confirm more directly, Tocopherol, which is effective for wrinkle improvement, was encapsulated within the ceramide liposomes of Example 9, and an eye cream using this was prepared and stored as shown in Table 28. The efficacy of the prepared eye cream was evaluated by conducting a human clinical trial in accordance with the relevant regulations of the Ministry of Food and Drug Safety (MFDS) and the Standard Operating Procedure (SOP) of the clinical laboratory.

[0399] Name: Anti-wrinkle Eye Cream Appearance: Lotion Cream / Opaque Storage Method: Store at room temperature between 5–25°C, away from high temperatures and direct sunlight Application Area: Face Liposome Content: Contains Tocopenol 50% and Liposome 2%

[0400] As a result, as shown in Figures 15a and 15b, it was confirmed that the depth of wrinkles was significantly reduced after 4 weeks of use compared to before using the eye cream.

[0401]

[0402] <Experimental Example 18> Evaluation of clinical efficacy of cosmetic formulations (2)

[0403] To confirm the whitening effect, the efficacy of the ceramide liposome was evaluated in the same manner as Experimental Example 9, except that niacinamide was encapsulated as the active ingredient as shown in Table 29 below.

[0404] Name: Whitening Essence Appearance: Eye Cream / Opaque Storage Method: Store at room temperature between 5–25°C, away from high temperatures and direct sunlight Application Area: Face Liposome Content: Contains Niacinamide 10% and Liposome 30%

[0405] As a result, as shown in Figures 16a and 16b, it was confirmed that the melanin area decreased significantly after 4 weeks of use compared to before using the eye cream.

Claims

1. An emulsion comprising a lipid core containing ceramide and fatty acids; and a nonionic surfactant, wherein An emulsion in which the above ceramide is included in an amount of 1 to 15 weight% based on the total weight of the emulsion, and the lipid core is formed through hydrogen bonding between the amino group (-NH2) of the ceramide and the carboxyl group (-COOH) of the fatty acid.

2. An emulsion according to claim 1, wherein the nonionic surfactant surrounds the surface of a lipid core, the hydrophilic terminals of the surfactant interact with the lipid core, and the hydrophobic terminals are arranged toward the outside of the surface of the lipid core.

3. An emulsion according to claim 1, wherein the ceramide is one or more selected from the group consisting of ceramide NS, ceramide AS, ceramide EOS, ceramide NDS, ceramide ADS, ceramide EODS, ceramide NP, ceramide AP, ceramide EOP, ceramide NH, ceramide AH, and ceramide EOH.

4. An emulsion according to claim 1, wherein the fatty acid is one or more selected from the group consisting of oleic acid, linoleic acid, linolenic acid, lauric acid, palmitic acid, stearic acid, elaidic acid, myristic acid, and arachidonic acid.

5. An emulsion according to claim 1, wherein the fatty acid is oleic acid.

6. An emulsion according to claim 1, wherein the lipid core comprises ceramide and fatty acid in a weight ratio of 1:1 to 1:

4.

7. An emulsion according to claim 1, wherein the nonionic surfactant is one or more selected from the group consisting of a polyglyceryl-based surfactant having 12 to 20 carbon atoms, a glyceryl-based surfactant having 12 to 20 carbon atoms, an alkyl glucoside-based surfactant having 12 to 20 carbon atoms, a polyethylene glycol sorbitan-based surfactant having 12 to 20 carbon atoms, and a polyethylene glycol-based surfactant having 12 to 20 carbon atoms.

8. An emulsion according to claim 7, wherein the nonionic surfactant is one or more selected from the group consisting of polyglyceryl-10 oleate, polyglyceryl-10 laurate, decyl glucoside, polysorbate 20, polysorbate 80, and glyceryl stearate / PEG-100 stearate.

9. An emulsion according to claim 1, wherein the ceramide and fatty acid form a lamellar liquid crystal structure.

10. The emulsion of Claim 1, wherein the emulsion is one or more selected from the group consisting of water-in-water type (O / W), oil-in-water type (W / O), water-in-oil-in-water type (W / O / W), and oil-in-water type (O / W / O).

11. The emulsion of claim 1, wherein the average particle diameter of the emulsion is 10 nm to 250 nm.

12. The emulsion of claim 1, wherein the zeta potential of the emulsion is -10 to -60 mV.

13. A cosmetic composition comprising the emulsion of Claim 1.

14. A cosmetic composition according to claim 13, having one or more activities selected from the group consisting of skin moisturization, skin barrier strengthening, skin aging improvement, skin wrinkle improvement, skin elasticity improvement, and skin regeneration.

15. A step of obtaining a lipid mixture by mixing a ceramide, a fatty acid, and a nonionic surfactant, wherein the ceramide is included in an amount of 5 to 15 weight% based on the total weight of the emulsion; A method for preparing an emulsion comprising the step of mixing the above lipid mixture and a eutectic solvent to obtain an emulsion.

16. A liposome having a lipid core comprising ceramide, fatty acid and nonionic surfactant; and a bilayer comprising phospholipids, A liposome having a lipid-soluble substance enclosed within the above double layer and a water-soluble substance captured inside and outside the above double layer.

17. The liposome of claim 16, wherein the ceramide is one or more selected from the group consisting of ceramide NS, ceramide AS, ceramide EOS, ceramide NDS, ceramide ADS, ceramide EODS, ceramide NP, ceramide AP, ceramide EOP, ceramide NH, ceramide AH, and ceramide EOH.

18. A liposome according to claim 16, wherein the fatty acid is one or more selected from the group consisting of oleic acid, linoleic acid, linolenic acid, lauric acid, palmitic acid, stearic acid, elaidic acid, myristic acid, arachidonic acid, isopropyl myristate, and caprylic / capric triglycerides.

19. The liposome according to claim 16, wherein the nonionic surfactant is one or more selected from the group consisting of a polyglyceryl-based surfactant having 12 to 20 carbon atoms, a glyceryl-based surfactant having 12 to 20 carbon atoms, an alkyl glucoside-based surfactant having 12 to 20 carbon atoms, a polyethylene glycol sorbitan-based surfactant having 12 to 20 carbon atoms, and a polyethylene glycol-based surfactant having 12 to 20 carbon atoms.

20. A liposome according to claim 16, wherein the lipid core has a surface coated with one or more selected from the group consisting of amino acids and polyols.

21. A liposome according to claim 16, wherein the phospholipid is one or more selected from the group consisting of lecithin, hydrogenated lecithin, phosphatidyl choline, sphingomyelin, cholesterol, phosphatidic acid, phosphatidyl serine, phosphatidyl glycerol, phosphatidyl inositol, and phosphatidyl ethanolamine.

22. A liposome according to claim 16, wherein the lipid-soluble substance is one or more selected from the group consisting of tocopheryl acetate, retinol, calciferol, linolenic acid, biotin, menadione, bioflavonoids, astaxanthin, idebenone, bakuchiol, volufiline, bisabolol, and zeaxanthin.

23. A liposome according to claim 16, wherein the water-soluble substance is one or more selected from the group consisting of niacinamide, niacin, thiamine, riboflavin, carnitine, pantothenic acid, panthenol, pyridoxine, ascorbic acid, peptide collagen, and cyanocobalamin.

24. The liposome of claim 16, comprising the ceramide, fatty acid, amino acid, polyol, nonionic surfactant, and phospholipid in a weight ratio of 1 to 5: 3 to 10: 1: 3 to 10: 1 to 5: 3 to 10.

25. The liposome of claim 16, wherein the liposome has an average particle diameter of 10 nm to 100 nm.

26. A cosmetic composition comprising the liposome of claim 16.

27. A cosmetic composition according to claim 26, having one or more activities selected from the group consisting of skin moisturization, skin barrier strengthening, skin wrinkle improvement, skin elasticity improvement, antioxidant, skin whitening, and hair loss improvement.

28. A step of obtaining a lipid core by mixing ceramide, fatty acid, and nonionic surfactant; A step of obtaining an emulsion by mixing the above lipid core and eutectic solvent; and A method for preparing liposomes comprising the step of mixing the above emulsion and phospholipid to obtain liposomes.

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