WO2016140473A1 - Multilamella nanoliposome which contains skin lipid components, and preparation method therefor - Google Patents

Abstract

The present invention relates to a multilamella nanoliposome which is biocompatible and in which a ceramide, a peptide derived from a living body, and hyaluronic acid are stably collected. The present invention provides a multilamella nanoliposome and a preparation method therefor, the multilamella nanoliposome being suitable for a cosmetic composition for overall skin improvements such as skin moisturizing enhancement, skin aging prevention, skin wrinkle improvement, etc. due to having excellent skin absorptivity and permeability.

Description

Nano multilayer liposomes containing skin lipid components and preparation method thereof

The present invention relates to a nano-layered liposome that is bio-friendly, low skin irritation, and effective substances such as bio-derived peptides are effectively stabilized, which can be applied to cosmetics for moisturizing effect of skin, anti-aging and the like, and a preparation method thereof.

The skin, which is the first contact with the outside in the human body and provides the first defense against the external environment, has aesthetically significant effects in addition to simply protective functions. The composition of the skin is divided into the epidermis and the dermis. Among them, the epidermis has been most clearly observed according to the skin aging and moisture content, and has been the main subject of research for skin beauty.

The main component of the skin is lipids, and many skin components, including lipids, form a solid skin barrier for maintaining the shape of the skin and defending it from foreign environments. Representative lipids that form the skin barrier are cholesterol, fatty acids, ceramides, it is known that it is very difficult to penetrate the water-soluble components into the skin by the lipid component of the skin barrier and the skin barrier containing them.

Among various methods for infiltrating water-soluble components into the skin, a method using liposomes is widely used for anti-aging, moisturizing, and wrinkle improvement of skin. The main reason for using liposomes is that they can be delivered stably without altering the active ingredients contained in liposomes and can be delivered into the skin through liposomes regardless of whether the active ingredient is fat-soluble or water-soluble.

Liposomes can be classified into multilamella or unilamella liposomes, and monolithic liposomes can be further divided into large-membrane liposomes and small monolithic liposomes according to their size. In general, multilayer membrane liposomes have a size of 400 to 3,500 nm and can contain about 5 to 15% of the components of the liposome, and the size of single-layer liposomes can contain about 100 to 1,000 nm and about 30 to 65% of the components of the large membrane. In the case of the small monolayer, it may contain about 0.5 to 1.0% of the liposome in a size of 20 to 50 nm.

Macromolecular liposomes can contain the highest amount of components but are unstable, and small membrane liposomes are stable but contain only a small amount of components. On the other hand, the multi-layered liposome has a sufficient amount of components that can be contained in the liposome, and has a good affinity for the skin forming the multi-layered structure. However, in the case of the multi-layered liposomes, there is a problem that the size thereof is too large to maintain stability and the penetration into the skin is not excellent.

In addition to liposomes, a method of infiltrating water-soluble components into the skin through niosomes is also used. Niosomes are 10 to 200 nm in size and contain nonionic surfactants and cholesterol as main components and have a closed double layer structure through high pressure emulsification, etc., and are relatively stable compared to liposomes carrying components by lipid particles such as phospholipids. It is easy to produce and inexpensive, but it is easy to aggregate or fuse, and it is not biocompatible.

Among the lipid components used in the preparation of liposomes, lecithin is known to be effective in reducing the size of liposomes and enhancing skin absorption in the case of non-hydrogenated lecithin, but due to the nature of the unsaturated group, liposomes have chemical instability due to unsaturated groups. This is accompanied by a change in color and odor due to forest litter, and has a long term stability. On the other hand, hydrogenated lecithin has excellent chemical stability as the unsaturated group is removed, and has excellent long-term stability, such as no abrasion or odor change even when it is in a dispersed state, while saturated hydrocarbon group has a large curvature radius due to its rigid characteristic. It is difficult to prepare liposomes having nanoscales, but rather have a problem of transferring to lamellar phase.

In order to solve these problems or shortcomings, it is necessary to develop a multi-layer liposome that is stable and small in size but can stably collect various materials and has excellent biocompatibility and long-term stability.

[Patent Document] Republic of Korea Patent Publication 2004-0078500

In order to solve the problem of technology using multilayer liposomes, the present invention has been completed by developing nano-layered liposomes including a skin lipid component and having excellent skin permeability and stably collecting various agonists.

The present invention can be used as a cosmetic composition for improving skin, such as improving skin moisturizing power, preventing skin aging, and can be applied to the pharmaceutical and food fields in addition to the cosmetic field.

The present inventors are aware of the problems of known liposomes and can have chemical stability and long-term stability by using hydrogenated lecithin, as well as introducing a polyglyceryl-based surfactant and a fatty acid substituted with an acyl group, thereby introducing a multilayer liposome into a nano size. Invented is a multi-layered nano liposome that can be stably formed, biocompatible and capable of stably collecting agonists.

The present invention provides a nano-layered liposomes containing polyglyceryl-based surfactants, hydrogenated lecithins and fatty acids substituted with acyl groups of C ₁₆ to C ₂₂ and stably trapping agonists.

The present invention comprises the steps of: a) mixing an oil phase component comprising a polyglyceryl surfactant, hydrogenated lecithin, and a fatty acid substituted with an acyl group of C ₁₆ to C ₂₂ , b) emulsifying the oil phase component by adding an oil phase component to the aqueous phase component, c A high pressure emulsifying step to form a nano-sized liposomes, and d) stabilizes the liposomes by dispersing and cooling the thickener.

Nano-layered liposomes of the present invention are bio-friendly, low irritation to the skin, safe and efficacious materials are collected, and highly moisturizing and skin permeability, thereby improving skin moisturizing effect, preventing skin aging, and improving skin wrinkles. It can be used in various ways.

1 (a) ~ (d) shows the structure and size of the nano multilayer liposome according to the present invention.

Details of the present invention will be described. In the description and drawings of the present invention, descriptions of well-known contents that may obscure the gist of the present invention may be omitted, and terms not separately defined in the entire specification may be used by those skilled in the art. It can be interpreted in the usual sense.

The present invention relates to a nano-layered multilamella liposome containing polyglyceryl-based surfactants, hydrogenated lecithin and fatty acids substituted with acyl groups of C ₁₆ to C ₂₂ and stably trapping agonists, and a method for preparing the same. The present invention provides a nano-layered liposomes which are biocompatible and excellent in skin permeability and skin absorption, and are effective for skin improvement.

The content of the polyglyceryl-based surfactant substituted with the C ₁₆ to C ₂₂ acyl groups included in the nano multilayer liposome of the present invention is 1 to 15% by weight, preferably 3 to 12% by weight, and more preferably, relative to the nano multilayer liposome dispersion. Is 5 to 10% by weight. According to an embodiment of the present invention, when less than 3% by weight of the multi-layer liposomes are not produced firmly, the stability over time may worsen, and if more than 12% by weight of the emulsification by the surfactant is increased by the production of a fine emulsion (emulsion) Multi-layered liposomes may not be produced.

The number of glycerin added in the polyglyceryl-based surfactant is 6 to 20, preferably 8 to 12, but is not limited thereto, and may be effective in reducing the size of the multilayer liposome and improving stability in the above range. Specifically, the polyglyceryl-based surfactant substituted with the acyl group of C ₁₆ to C ₂₂ is at least one of polyglyceryl stearate, polyglyceryl palmitate and polyglyceryl behenate It may include.

In the present invention, lecithin, which is a major component of phospholipid, may be included as an auxiliary emulsifier to improve stability and biocompatibility of the nano multilayer liposomes, and to reduce the total amount of the surfactant. Lecithin may be classified into a lecithin having an unsaturated group and a hydrogenated lecithin from which an unsaturated group is removed by hydrogenating an unsaturated group. In the present invention, the lecithin is preferably hydrogenated lecithin.

Lecithin is produced by extracting from soybeans or eggs and purifying them.Phospholipids having fatty acid chains having 12 to 24 carbon atoms, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and phosphatidic acid Mixture comprising a. The fatty acid chain constituting the hydrophobic group of the extracted lecithin is an unsaturated lecithin having an average distribution of 0 to 3 double bonds, and a mixture having a phosphatidylcholine content of 70 to 95% may be used depending on the degree of purification. The double bond of the fatty acid chain in lecithin is easily oxidized by water or active oxygen, while hydrogenated lecithin has a high chemical stability by reducing the number of unsaturated groups by hydrogenation.

The content of hydrogenated lecithin is 0.05 to 10% by weight, 0.5 to 6% by weight, preferably 1 to 5% by weight, more preferably 4 to 5% by weight relative to the nano multilayer liposome dispersion. The preferred content of hydrogenated lecithin may be important in order to maintain particularly long term stability in the stability of the nano multilayer liposomes. In addition, when the hydrogenated lecithin satisfying the specific content range is used, while maintaining the emulsification, low-temperature stability, high-temperature stability, discoloration prevention, and satisfactory effects on use on the skin may be exhibited. According to an embodiment of the present invention, when the hydrogenated lecithin content is less than 1%, the stability of the nano multilayer liposomes may be very low, and when the hydrogenated lecithin content exceeds 5%, discoloration may occur too easily.

According to the present invention, it may further include a fatty acid to more stabilize the multi-layered structure of the nano-layered liposomes. Fatty acids are fatty acids having 14 to 24 carbon atoms, preferably 16 to 22 carbon atoms, but are not limited thereto. For example, the fatty acid may include any one or more of lauric acid, myristic acid, palmitic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, arachidonic acid, and behenic acid. The fatty acid content is 0.1 to 5% by weight, 1 to 5% by weight, 1 to 2% by weight, 1.5 to 2.5% by weight relative to the nano multilayer liposome dispersion, but is not limited thereto.

The agonist to be collected and stabilized in the present invention is not limited as long as it can be stably collected in the nano-layered liposome of the present invention and is a material exhibiting water-soluble or fat-soluble properties. Preferably any one or more of the bio-derived peptide, ceramide, tocopherol and hyaluronic acid (hyaluronic acid) is trapped and stabilized as an agonist.

In the present invention, the bio-derived peptide refers to a protein present in vivo found in human blood, saliva, etc., and includes any one or more of water-soluble copper peptide, azireline, palmitoyl oligopeptide, and palmitoyl tetrapeptide. The content of the bio-derived peptide is 0.001 to 1% by weight, 0.1 to 0.5% by weight, preferably 0.01 to 0.1% by weight relative to the nano multilayer liposome dispersion, but is not limited thereto.

In the present invention, the hyaluronic acid may be used to supply high moisture to the skin, and may use high molecular weight (HMW) hyaluronic acid, and use low molecular weight (LMW) hyaluronic acid to absorb the skin. Can be. Preferably, the high molecular weight hyaluronic acid has a molecular weight of 30,000 to 100,000 Daltons, and the low molecular weight hyaluronic acid preferably has a molecular weight of 10,000 Daltons or less. The content of hyaluronic acid is preferably different in the case of high molecular weight and low molecular weight, and in the case of high molecular weight hyaluronic acid, 0.001 to 0.1% by weight compared to the nano multilayer liposome dispersion, and in the case of low molecular weight hyaluronic acid compared to the nano multilayer liposome dispersion It is preferably from 0.05 to 1% by weight, but is not limited thereto.

Tocopherol (tocopherol) in the present invention can further improve the skin absorption, it may be given an antioxidant effect can further improve the skin improvement effect. The content of tocopherol is 0.001 to 0.01% by weight, 0.01 to 5% by weight, preferably 0.1 to 3% by weight relative to the nano multilayer liposome dispersion, but is not limited thereto.

In the present invention, ceramide may improve skin affinity as one of the skin lipid components and may further improve the stability of the multilayer liposome by combining with fatty acids. Ceramide may be included as phytosphingosine in precursor form, and may include both ceramides and phytosphingosine. The content of ceramide is 0.1 to 10% by weight, preferably 1 to 5% by weight relative to the nano multilayer liposome dispersion, but is not limited thereto.

The nano-multilayer liposomes of the present invention have a size of 50 to 500 nm, which is smaller than the conventional multilayer membrane liposomes, and has excellent stability and excellent content of components that can be contained. According to an embodiment of the present invention, the multilayer liposome has 5 to 7 layers, and has a multilayer structure including a lamellar structure of 20 to 50 nm between layers.

The nano multilayer liposomes of the present invention have a smaller size than the general multilayer liposomes, have high skin absorbency and permeability, and maintain a stable multilayer structure and have excellent skin affinity. In addition, according to an embodiment of the present invention, the amount that can be contained in the nano multilayer liposomes can be delivered by trapping an agonist such as sufficient bioactive components such as 5 to 15%.

The nano multilayer liposomes of the present invention can improve biocompatibility and long-term stability by using polyglyceryl-based surfactants, hydrogenated lecithins and fatty acids substituted with C ₁₆ to C ₂₂ acyl groups, and increase the size of the multilayer liposomes from 50 to 500 nm. It can be prepared as and can be stabilized while sufficient content of the agonist can be collected. In addition, the content of the polyglyceryl-based surfactant can be controlled by adjusting the content of hydrogenated lecithin, thereby making it possible to prepare liposomes which are more bio-friendly and have good usability and have excellent long-term stability and discoloration prevention.

Method for producing a nano-layered liposome of the present invention comprises the steps of a) mixing an oily component comprising a polyglyceryl surfactant, hydrogenated lecithin and a fatty acid substituted with an acyl group of C ₁₆ to C ₂₂ , b) the oily component to the aqueous component Emulsifying by adding, c) a high pressure emulsifying step to form nano-sized liposomes, and d) stabilizing the liposomes by dispersing and cooling the thickener is a method for producing a nano-layered liposomes.

Surfactant substituted with C ₁₆ to C ₂₂ acyl groups in step a) of the method for preparing a nano multilayer liposome is 1 to 15% by weight, preferably 3 to 12% by weight, more preferably 5 to 10, based on the nano multilayer liposome dispersion. Weight percent. According to an embodiment of the present invention, when less than 3% by weight of the multi-layer liposomes are not produced firmly, the stability over time may worsen, and if more than 12% by weight of the emulsification by the surfactant is increased by the production of a fine emulsion (emulsion) Multi-layered liposomes may not be produced.

The content of hydrogenated lecithin in step a) of the method for preparing a nano multilayer liposome is 0.05 to 10% by weight, 0.5 to 6% by weight, preferably 1 to 5% by weight, more preferably 4 to 5% by weight, based on the nano multilayer liposome dispersion. %to be. The preferred content of hydrogenated lecithin may be important in order to maintain particularly long term stability in the stability of the nano multilayer liposomes. In addition, when the hydrogenated lecithin satisfying the specific content range is used, while maintaining the emulsification, low-temperature stability, high-temperature stability, discoloration prevention, and satisfactory effects on use on the skin may be exhibited. According to an embodiment of the present invention, when the hydrogenated lecithin content is less than 1%, the stability of the nano multilayer liposomes may be very low, and when the hydrogenated lecithin content exceeds 5%, discoloration may occur too easily.

In step a) of the method for preparing a nano-layered liposome, an agonist such as ceramide and tocopherol may be added to the oil phase component, and the mixture is dissolved and mixed at 60 to 80 ° C. The emulsification step b) can be carried out by a general emulsification method, and the aqueous phase component may be added with water-soluble peptides, agonists such as hyaluronic acid and preservative components, and the aqueous phase component is also dissolved in water at 60 to 80 ° C.

The high pressure emulsification of c) is a process for making a nanoemulsion state, and a multi-sized nano liposome form having a stable size at a micro size is formed, using a high pressure emulsifier at 50 to 60 ° C. under a pressure of 500 to 1500 bar. . The thickener of step d) is guar gum, gellan gum, cellulose, agar, pectin, carbomer, poloxamer, xanthan gum, carrageenan gum, locust bean gum, plant polymer, microbial polymer, animal polymer, starch polymer, ar It may include one or more of a long acid polymer, a vinyl polymer, a polyoxyethylene polymer, a polyoxyethylene polyoxypropylene copolymer system polymer, an acrylic polymer and an inorganic water-soluble polymer, the cooling temperature is 25 to 35 ℃ to cool Preferred for stabilization of multilayer liposomes.

In the preparation method, when the thickener is added before the high pressure emulsification step of c), the efficiency of the high pressure emulsification is low, and thus, a stable nano multilayer liposome of the present invention cannot be prepared. In addition, in order to satisfy the desired stability in the present invention, it is preferable that each step of the manufacturing method be performed by a continuous process.

The nano multilayer liposome of the present invention may further include vegetable oil, vegetable sterol or skin softener as an oily component. The nano multilayer liposome of the present invention may further include an alcohol, a skin conditioning agent, a dispersant, an emulsifier, a humectant, a moisturizer or a whitening agent as an aqueous phase component. In addition, the nano multilayer liposome of the present invention may further include a preservative component.

Hereinafter, specific examples for carrying out the present invention will be described, and the present invention is not limited or limited to the following examples.

[Example 1-8] Production of multi-layered nano-liposomes

Component contents of Examples 1 to 8 and Comparative Examples 1 to 6 are as shown in Table 1 and Table 2.

		Example 1	Example 2	Example 3	Example 4	Comparative Example 1	Comparative Example 2	Comparative Example 3
Oily ingredient	Meadownan Seed Oil (Limnanthes Alba Seed Oil)	5	5	5	5	5	5	5
	Octane Cetyl (CETYL OCTANOTE)	5	5	5	5	5	5	5
	Palmitic acid	2	2	2	2	2	2	2
	PEG-5 Rapeseed Sterol	2	2	2	2	2	2	2
	Polyglyceryl-10 Stearate	8	8	8	8	8	8	8
	Hydrogenated Lecithin	5	One	3	4	0.1	7	9
	Ceramide 3B	5	5	5	5	5	5	5
	Phytosphingosine	One	One	One	One	One	One	One
	Tocopherol	0.01	0.01	0.01	0.01	0.01	0.01	0.01
Water component 2	Propanediol	9	9	9	9	9	9	9
	Potassium Ceryl Phosphate (POTASSIUM CETYL PHOSPHATE)	0.3	0.3	0.3	0.3	0.3	0.3	0.3
	Acetyl Glucossamine	7	7	7	7	7	7	7
	Sodium Hyaluronate	0.1	0.1	0.1	0.1	0.1	0.1	0.1
	Low molecular weight sodium hyaluronate	0.2	0.2	0.2	0.2	0.2	0.2	0.2
	Peptide	0.5	0.5	0.5	0.5	0.5	0.5	0.5
	Niacinamide	5	5	5	5	5	5	5
Thickener	Polyacrylate-13 * Polyisobutene * Polysorbate 20	0.1	0.1	0.1	0.1	0.1	0.1	0.1
Preservative Ingredient	PhenoxyEthanol	0.4	0.4	0.4	0.4	0.4	0.4	0.4
	Ethylhexylglycerin	0.05	0.05	0.05	0.05	0.05	0.05	0.05
Water component 1	D.I Water	to 100	to 100	to 100	to 100	to 100	to 100	to 100
	Total	100	100	100	100	100	100	100

		Example 5	Example 6	Example 7	Example 8	Comparative Example 4	Comparative Example 5	Comparative Example 6
Oily ingredient	Meadownan Seed Oil (Limnanthes Alba Seed Oil)	5	5	5	5	5	5	5
	Octane Cetyl (CETYL OCTANOTE)	5	5	5	5	5	5	5
	Palmitic acid	2	2	2	2	2	2	2
	PEG-5 Rapeseed Sterol	2	2	2	2	2	2	2
	Polyglyceryl-10 Stearate	3	6	9	12	2	13	16
	Hydrogenated Lecithin	4	4	4	4	4	4	4
	Ceramide 3B	5	5	5	5	5	5	5
	Phytosphingosine	One	One	One	One	One	One	One
	Tocopherol	0.01	0.01	0.01	0.01	0.01	0.01	0.01
Water component 2	Propanediol	9	9	9	9	9	9	9
	Potassium Ceryl Phosphate (POTASSIUM CETYL PHOSPHATE)	0.3	0.3	0.3	0.3	0.3	0.3	0.3
	Acetyl Glucossamine	7	7	7	7	7	7	7
	Sodium Hyaluronate	0.1	0.1	0.1	0.1	0.1	0.1	0.1
	Low molecular weight sodium hyaluronate	0.2	0.2	0.2	0.2	0.2	0.2	0.2
	Peptide	0.5	0.5	0.5	0.5	0.5	0.5	0.5
	Niacinamide	5	5	5	5	5	5	5
Thickener	Polyacrylate-13 * Polyisobutene * Polysorbate 20	0.1	0.1	0.1	0.1	0.1	0.1	0.1
Preservative Ingredient	PhenoxyEthanol	0.4	0.4	0.4	0.4	0.4	0.4	0.4
	Ethylhexylglycerin	0.05	0.05	0.05	0.05	0.05	0.05	0.05
Water component 1	D.I Water	to 100	to 100	to 100	to 100	to 100	to 100	to 100
	Total	100	100	100	100	100	100	100

The nano multilayer liposomes of Examples 1 to 8 and Comparative Examples 1 to 6 were prepared according to the following methods. The oily ingredients were uniformly mixed at 70 ° C and dissolved. The aqueous phase component 2 and the antiseptic component were added to the aqueous phase component 1, and the mixture was dissolved at 70 ° C. After mixing and dissolving, the oil phase component was homogeneously mixed with the aqueous phase component and gradually added to emulsify. After 55 ℃, high pressure emulsified in a high pressure emulsifier of 1000 bar to make a nanoemulsion. After the high pressure emulsification was added to the nanoemulsion prepared by the thickening agent homo homomixed and completely dispersed, cooled to 30 ℃ to obtain a stabilized nano multilayer liposomes.

Comparison of Stability and Particle Size According to Incubation Time

The timing of adding the thickener should be added after high pressure emulsification, and when the timing of adding the thickener was before high pressure emulsification, nano multilayer liposomes having the desired stability and size could not be obtained. Only the point of introduction of the thickener was different from that before the high-pressure emulsification, and another manufacturing method measured stability and average size of the multilayer separator prepared in the same manner as in Examples 1-8. The size of each nano multilayer liposome obtained by varying the timing of adding the thickener and the results of stability experiments performed at 45 ° C. for 14 days are shown in Table 3. I also confirmed that the multilayer liposomes were formed uniformly and kept stable.

45 ℃ Stability and Particle Size According to Incubation Point

	Input before high pressure emulsification	Input after high pressure emulsification
45 ° C stability / 14 days	Upper part separation	Good
Average particle size	10 μm	100 nm

Comparison of stability by surfactant combination

In Examples 1 to 4, the amounts of hydrogenated lecithin were 1, 3, 4, and 5 wt%, respectively, and Comparative Examples 1 to 3 were the contents of hydrogenated lecithin, 0.01, 7, and 9 wt%, respectively. Examples 5 to 8 were 3, 6, 9, and 12% by weight, respectively, of polyglyceryl surfactants, and Comparative Examples 4 to 6 were polyglyceryl-10 stearates, respectively, which were polyglyceryl surfactants. It was set as 2, 13, and 16 weight%. Low temperature stability comparison of Examples 1 to 8 and Comparative Examples 1 to 6 was carried out by thawing for 1 day after storage for 1 day at minus 17 ℃, proceeded to check daily at 45 ℃ and 60 ℃ for high temperature stability experiments. It was. The freezing stability of Comparative Example 1 was excellent for one week, but the stability was decreased as the emulsification was broken when stored at 60 ° C. for 2 days or more during high temperature stability. In Comparative Examples 2 and 3, discoloration occurred at a high temperature, and the usability was not good due to an excess of lecithin content. As a result of stability evaluation of Examples 1 to 4 and Comparative Examples 1 to 3, the stability was excellent in the multilayer liposomes in which the lecithin content was in the range of 1 to 5 wt%, but the stability was not good in the case when the lecithin content was in the range of 1 to 5 wt%. . In Table 4, Examples 1 and 4 show that the results of evaluation of stability and discoloration are very good among the examples, and especially when the lecithin content is included in 4 to 5%, the stability and discoloration of the multilayer liposomes were excellent. Comparative Example 4 was excellent in freezing stability for one week, but when stored for more than two days at 45 ℃ and 60 ℃ and the stability was reduced as the emulsification is broken, in the case of Comparative Examples 5 and 6 the multi-layer nano liposome was not produced normally. As a result of stability evaluation of Examples 5 to 8 and Comparative Examples 4 to 6, when the content of the polyglyceryl surfactant is 3 to 12% by weight, it was confirmed that the stability was not good or the multilayer nano liposomes were not normally generated. Experimental results for stability and discoloration were as shown in Table 4 below.

	Stability	discoloration
Example 1	5	5
Example 2	4	5
Example 3	4	5
Example 4	5	5
Example 5	5	5
Example 6	5	5
Example 7	4	5
Example 8	5	5
Comparative Example 1	One	3
Comparative Example 2	2	One
Comparative Example 3	2	One
Comparative Example 4	One	4
Comparative Example 5	Not manufactured	4
Comparative Example 6	Not manufactured	4
1: bad, 2: a little bad, 3: moderate, 4: a little good, 5: very good

Confirmation of multilayer liposome formation

After preparing nano multilayer liposomes with the components according to Examples 1 to 8, the microstructure of the liposomes was confirmed. The lamellar structure of about 5-7 layers was formed in a multi-layered form and the length was about 200 nm, and the interlayer spacing was formed relatively regularly within 20-50 nm, and the size of liposomes corresponded to 50-500 nm (FIG. 1). When the nano-multilayer liposomes were prepared with the components according to Comparative Examples 5 and 6, a large number of micro emulsions of 20 to 40 nm were generated, and thus, nano-liposomes of the multi-layers were not produced.

Stability test

The nano-layered liposomes prepared by the preparation method according to Examples 1 to 8 were subjected to in vitro patch test for stability evaluation in 10 subjects, and the results are shown in Table 5.

Test substance	Stimulation degree	Average stimulus score
(-)control	(-)	0.00
0.3% SDS	++	6.55
Examples 1-4	(-)	0.00
(-): No response / irritation score 0 ·: Faint mild erythema / irritation score 0.5 ±: Borderline weak erythema / irritation score 1+: Distinct erythema, papules and vesicles / stimulation score 2 ++: Severe erythema, alveolar vesicle Stimulus Score 6 +++: Crust Formation / Stimulus Score 12

The moisturizing test was conducted using a moisture measurement probe of ARAMO-TS (Aram Huvis Co., Ltd., Korea), and TEWL used the AquaFlux. (Biox, UK) device to measure the average moisturizing power of 10 subjects It measured, and the result is shown in Table 6 and Table 7.

Test substance	Before treatment	1 day	5 days	15th	20 days	30 days
Examples 1-4	33.40	+8.60	+5.90	+3.63	+5.73	+6.47

Moisturizing power was confirmed to be increased to more than 8 moisturizing power in one day using liposomes, and when used continuously it was confirmed that the improved moisturizing power of 6.47 compared to the existing day 30.

Test substance	Before treatment	1 day	5 days	15th	20 days	30 days
Examples 1-4	12.64	-7.13	-2.48	0.14	-1.8	-0.48

In the case of the average water loss (Transepidermal Water Loss, TEWL) value through the skin compared to before treatment it was confirmed that the loss amount is reduced by 7 or more in one day.

Claims (13)

1. Nano multilamella liposomes containing polyglyceryl surfactants, hydrogenated lecithin and fatty acids substituted with acyl groups of C ₁₆ to C ₂₂ and stably trapped agonists.
2. The method of claim 1,

   The agonist is a nano-layered liposome of any one or more of a bio-derived peptide, hyaluronic acid, ceramide, and tocopherol.
3. The method of claim 1,

   The polyglyceryl-based surfactant substituted with an acyl group of C ₁₆ to C ₂₂ is a nano multilayer liposome comprising at least one of polyglyceryl stearate, polyglyceryl palmitate and polyglyceryl behenate.
4. The method of claim 1,

   The hydrogenated lecithin content is nano multilayer liposomes of 1 to 5% by weight compared to the nano multilayer liposome dispersion.
5. The method of claim 1,

   The polyglyceryl-based surfactant content of the acyl group substituted C ₁₆ to C ₂₂ is 3 to 12% by weight compared to the nano multilayer liposome dispersion.
6. The method of claim 2,

   The bio-derived peptide is a nano-layered liposome comprising at least one of a copper peptide, azirelin, palmitoyl oligopeptide and palmitoyl tetrapeptide.
7. The method of claim 1,

   The nano multilayer liposomes have a size of 50 to 500 nm.
8. The method of claim 1,

   The lamellar structure of the nano multilayer liposome is 5 to 7 layers and the interlayer spacing is a multilayer structure of 20 to 50nm nano multilayer liposomes.
9. Cosmetic composition for skin moisturizing comprising the nano-multilayer liposome of any one of claims 1 to 8.
10. a) mixing an oily component comprising a polyglyceryl surfactant, hydrogenated lecithin, and a fatty acid substituted with an acyl group of C ₁₆ to C ₂₂ ;

    b) emulsifying by adding an oil phase component to the aqueous phase component;

    c) a high pressure emulsification step to form nano-sized liposomes; And

    d) stabilizing liposomes by dispersing and cooling the thickener;

    Method of producing a multi-layer liposomes comprising me.
11. The method of claim 10,

    The method of claim 1, wherein the hydrogenated lecithin content is 1 to 5 wt% based on the nano multilayer liposome dispersion in step a).
12. The method of claim 10,

    The method of preparing a nano-layered liposome having a polyglyceryl-based surfactant content substituted with the acyl group of C ₁₆ to C ₂₂ in step a) is 3 to 12% by weight relative to the nano-layered liposome dispersion.
13. The method of claim 10,

    The high pressure emulsifying step is a nano-layered liposome manufacturing method that proceeds at 50 to 60 ℃ and 500 to 1500bar.

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