A liposome encapsulating hyaluronic acid, a moisturizing composition, and a preparation method and application thereof

Abstract

The present invention belongs to the technical field of skin care products, and specifically relates to a liposome encapsulating hyaluronic acid, a composition, and a preparation method and application thereof. The present invention provides a liposome encapsulating hyaluronic acid, comprising a hyaluronic acid compound and a choline phosphate lipid bilayer encapsulated outside the hyaluronic acid compound; the particle size of the liposome is 50 to 300 nm. Compared with the products containing hyaluronic acid on the market, the liposome provided by the present invention uses choline phosphate lipids to encapsulate hyaluronic acid. Compared with other carriers, choline phosphate lipids have excellent cell membrane fusion ability, which can effectively overcome the skin barrier and more effectively deliver molecules to the interior of the target cells, greatly improving the efficiency of hyaluronic acid through the skin, and for the first time realizing the non-invasive transdermal absorption of ultra-high molecular weight hyaluronic acid (~2,500KDa), which is conducive to exerting the best cosmetic effect of hyaluronic acid.

Description

Technical Field

[0001] The invention belongs to the technical field of skin care products, and specifically relates to a liposome encapsulating hyaluronic acid, a moisturizing composition, and a preparation method and application thereof. Background Art

[0002] Hyaluronic acid, also known as hyaluronic acid or uronic acid, is an acidic mucopolysaccharide and one of the important components of human dermis. As a moisturizing factor for the skin, hyaluronic acid has a strong ability to absorb water and can absorb and retain hundreds of times more water than its own weight. This property makes hyaluronic acid one of the most effective moisturizers discovered so far. It is widely used in cosmetics to increase the moisture content of the skin and make the skin look plump, soft and smooth. In addition, hyaluronic acid can fill the space between cells and collagen fibers, play a supporting role, and make the skin fuller and more elastic. Supplementing hyaluronic acid helps to restore and maintain the youthfulness of the skin and reduce the formation of wrinkles.

[0003] Currently, hyaluronic acid is widely used in the field of skin care products. However, due to the multiple barriers of the stratum corneum, hyaluronic acid is difficult to penetrate the skin, which affects the effect of use and makes it difficult to fully exert its efficacy. Summary of the invention

[0004] The purpose of the present invention is to provide a liposome encapsulating hyaluronic acid, a moisturizing composition, and a preparation method and application thereof. The liposome provided by the present invention greatly improves the efficiency of hyaluronic acid penetrating the skin.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The present invention provides a liposome encapsulating hyaluronic acid, comprising a hyaluronic acid compound and a choline phosphate lipid bilayer encapsulating the hyaluronic acid compound; the particle size of the liposome is 50 to 300 nm;

[0007] The inner and outer surfaces of the choline phospholipid bilayer are hydrophilic phosphocholine heads, and the inner layer is a hydrophobic fatty acid tail.

[0008] Preferably, the choline phospholipid in the choline phospholipid bilayer has a chemical structure shown in Formula I:

[0009]

[0010] In Formula I,

[0011] A1 and A2 are independently C x H 2x+y , x is an integer from 5 to 35, and y is 1, -1, -3, -5, -7, -9 or -11;

[0012] R is a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C3-C8 alkenyl group, a substituted or unsubstituted C3-C8 alkynyl group, a substituted or unsubstituted C3-C8 epoxy group, a C3-C8 azide group, an amino group, an amino group with a protecting group, a heteroalicyclic group, an aryl group or a heteroaryl group.

[0013] Preferably, the hyaluronic acid compound includes hyaluronic acid and / or sodium hyaluronate; the molecular weight of the hyaluronic acid compound is 2.5 to 2500 KDa;

[0014] The mass ratio of choline phosphate lipid to hyaluronic acid compound in the choline phosphate lipid bilayer is 0.1-9.2:0.1-6.1.

[0015] The present invention also provides a method for preparing the liposome described in the above technical solution, comprising the following steps:

[0016] The choline phosphate lipid, hyaluronic acid compound and water are mixed and homogenized to obtain the liposome.

[0017] The present invention also provides a moisturizing composition, which comprises, by weight percentage, 0.2% to 15.3% of liposomes, 2.5% to 7.7% of emulsifiers, 2% to 3.8% of thickeners, 2.6% to 4.1% of emollients, and the balance of water;

[0018] The liposome is the liposome encapsulating hyaluronic acid described in the above technical solution or the liposome encapsulating hyaluronic acid obtained by the preparation method described in the above technical solution.

[0019] Preferably, the emulsifier is a nonionic emulsifier;

[0020] The thickener includes a fatty alcohol thickener and / or beeswax;

[0021] The emollients include isohexadecane and / or ester emollients.

[0022] Preferably, it also includes one or more of an antioxidant, a pH regulator, a stabilizer, a preservative, an emulsifier, a solubilizer, a surfactant, a film former and a buffer;

[0023] The antioxidant has a mass percentage of 0.6% to 3.3% in the moisturizing composition; the antioxidant comprises one or more of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol;

[0024] The mass percentage of the pH regulator in the moisturizing composition is 0.6% to 1.4%; the pH regulator includes citric acid and / or sodium dihydrogen phosphate;

[0025] The mass percentage of the stabilizer in the moisturizing composition is 1.2% to 4.1%; the stabilizer includes one or more of an inorganic salt stabilizer, cetearyl alcohol and carbomer;

[0026] The mass percentage of the preservative in the moisturizing composition is 0.2% to 1.0%; the preservative is an acidic preservative.

[0027] The present invention also provides a method for preparing the moisturizing composition described in the above technical solution, comprising the following steps:

[0028] The moisturizing composition is obtained by mixing liposomes, emulsifiers, thickeners, emollients and water.

[0029] Preferably, the mixed raw materials further include one or more of an antioxidant, a pH regulator, a stabilizer, a preservative, an emulsifier, a solubilizer, a surfactant, a film former and a buffer.

[0030] The present invention also provides the liposome encapsulating hyaluronic acid described in the above technical solution, the liposome encapsulating hyaluronic acid obtained by the preparation method described in the above technical solution, the moisturizing composition described in the above technical solution, or the use of the moisturizing composition obtained by the preparation method described in the above technical solution in skin care products.

[0031] The present invention provides a liposome encapsulating hyaluronic acid, including a hyaluronic acid compound and a choline phosphate lipid bilayer encapsulated outside the hyaluronic acid compound; the particle size of the liposome is 50 to 300 nm; the inner and outer surfaces of the choline phosphate lipid bilayer are hydrophilic phosphorylcholine heads, and the inner layer is a hydrophobic fatty acid tail. Compared with the products containing hyaluronic acid on the market, the liposome provided by the present invention uses choline phosphate lipids to form a bilayer to encapsulate hyaluronic acid compounds. Compared with other carriers, choline phosphate esters have excellent cell membrane fusion ability, can effectively overcome the skin barrier, are easily absorbed by cells, and more effectively deliver hyaluronic acid molecules to the interior and gaps of skin cells, thereby improving the therapeutic effect and reducing the effects of side effects such as skin discomfort, and greatly improving the efficiency of hyaluronic acid compounds through the skin. The present invention realizes the non-invasive transdermal absorption of ultra-high molecular weight hyaluronic acid (~2,500KDa) for the first time in the world. At the same time, choline phosphate lipids have strong scalability and stable physical and chemical properties, which is conducive to the best cosmetic effect of hyaluronic acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0033] Figure 1 is a schematic diagram of the structure of the liposome of the present invention;

[0034] Figure 2 This is a SEM image of the moisturizing composition obtained in Example 1;

[0035] Figure 3 It is a data graph showing the cytotoxicity evaluation results of the moisturizing compositions obtained in Example 2 and Comparative Examples 1-2;

[0036] Figure 4 This is a data graph showing the evaluation results of the hyaluronic acid transdermal ability of the moisturizing compositions obtained in Example 2 and Comparative Examples 1-2. DETAILED DESCRIPTION

[0037] The present invention provides a liposome encapsulating hyaluronic acid, comprising a hyaluronic acid compound and a choline phosphate lipid bilayer encapsulating the hyaluronic acid compound; the particle size of the liposome is 50 to 300 nm;

[0038] The inner and outer surfaces of the choline phospholipid bilayer are hydrophilic phosphocholine heads, and the inner layer is a hydrophobic fatty acid tail.

[0039] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0040] In the present invention, the choline phospholipid in the choline phospholipid bilayer has a chemical structure shown in Formula I:

[0041]

[0042] In Formula I,

[0043] A1 and A2 are independently C x H 2x+y , x is an integer of 5 to 35, y is 1, -1, -3, -5, -7, -9 or -11; A1 and A2 are independently CH3(CH2) n -、CH3(CH2) m1 CH=CH(CH2) m2-, n is an integer of 4 to 20, m1 is an integer of 5 to 7, and m2 is an integer of 7 to 13. In a specific embodiment, CH3(CH2)4-, CH3(CH2)5-, CH3(CH2)6-, CH3(CH2)7-, CH3(CH2)8-, CH3(CH2)9-, CH3(CH2) 10 -、CH3(CH2) 11 -、CH3(CH2) 12 -、CH3(CH2) 13 -、CH3(CH2) 14 -、CH3(CH2) 15 -、CH3(CH2) 16 -、CH3(CH2) 17 -、CH3(CH2) 18 -、CH3(CH2) 19 -、CH3(CH2) 20 -, CH3(CH2)5CH=CH(CH2)7-, CH3(CH2)5CH=CH(CH2)9-, CH3(CH2)7CH=CH(CH2)7-, CH3(CH2)5CH=CH(CH2) 11 -、CH3(CH2)7CH=CH(CH2) 11 - or CH3(CH2)7CH=CH(CH2) 13 -.

[0044] R is a substituted or unsubstituted C1-C8 alkyl, a substituted or unsubstituted C3-C8 alkenyl, a substituted or unsubstituted C3-C8 alkynyl, a substituted or unsubstituted C3-C8 epoxy, a C3-C8 azido, an amino, an amino with a protecting group, a heteroalicyclic group, an aryl or a heteroaryl group; the substituent of the substituted alkyl is a halogen, and the halogen is an F element; in a specific embodiment, R can be CH3-, CH3CH2-, CH3(CH2)2-, (CH3)2CHCH2-, CH3(CH2)3-, CH2=CH(CH2)2-, CH2=CHCH2-, CH≡C(CH2)2-, CH≡CCH2, N3-(CH 2)2-, N3-(CH2)3-, N3-(CH2)4-, NH2CH2-, NH2(CH2)2-, NH2(CH2)3-, BOC-NH(CH2)2-, BOC-NH(CH2)3-, C6H5-CH2-, C6H5-(CH2)2-, C6H5-(CH2)3 -, C6H5-(CH2)4-, CH3-C6H5-CH2-, CF3-, CF3CF2-, CF3CH2-, CHF2CH2-, CF3(CF2)2-, CF3CH(CF3)2-, (CF3)3CCH2-, (CF3)2CHCH2- or CF3(CF2)3-.

[0045] In the present invention, the hyaluronic acid compound includes hyaluronic acid and / or sodium hyaluronate, and in a specific embodiment, it can be hyaluronic acid; the molecular weight of the hyaluronic acid compound is 2.5-2500KDa, and in a specific embodiment, it can be 30-45KDa, 200-400KDa, 700-1,400KDa or 1,500-2,500KDa. The low molecular weight hyaluronic acid of the present invention has a small molecular size and is easy to penetrate into the skin layer and tissue cells; the medium molecular weight hyaluronic acid can form a colloid substance to achieve a good moisturizing effect.

[0046] In the present invention, the mass ratio of choline phosphate lipid to hyaluronic acid compound in the choline phosphate lipid bilayer is 0.1-9.2:0.1-6.1, and in specific embodiments, it can be 6.5:4.9, 8:5 or 7:6.

[0047] In the present invention, the particle size of the liposome is 50 to 300 nm, and in a specific embodiment, it can be 50 nm, 100 nm, 200 nm or 300 nm.

[0048] The liposomes provided by the present invention utilize choline phosphate lipids to encapsulate hyaluronic acid. Compared with products containing hyaluronic acid on the market, choline phosphate lipids have excellent cell membrane fusion ability, can effectively overcome the skin barrier, and are easily absorbed by cells, and more effectively deliver hyaluronic acid molecules to the interior and gaps of skin cells, thereby improving the therapeutic effect and reducing the impact of side effects such as skin discomfort, greatly improving the efficiency of hyaluronic acid penetrating the skin, and realizing the world's first non-invasive transdermal absorption of ultra-high molecular weight hyaluronic acid (~2,500KDa). At the same time, choline phosphate lipids have strong scalability and stable physical and chemical properties, which is conducive to exerting the best cosmetic effect of hyaluronic acid.

[0049] The present invention also provides a method for preparing the liposome described in the above technical solution, comprising the following steps:

[0050] The choline phosphate lipid, hyaluronic acid compound and water are mixed and homogenized to obtain the liposome.

[0051] In the present invention, the mixing can be: first mixing the choline phosphate lipid and the hyaluronic acid compound, and then adding water; the mixing temperature can be 50-60°C; the pressure of the homogenization treatment can be 50-100MPa, and in a specific embodiment, it can be 50MPa or 80MPa; the time can be 2-4h, and the temperature can be 50-60°C.

[0052] The liposome preparation method provided by the present invention has the advantages of high mixing efficiency, strong continuity and large preparation volume. The principle is to uniformly disperse the choline phosphate lipid and hyaluronic acid at the microscopic level through high-intensity shear force under high pressure, and automatically complete the self-assembly of the choline phosphate lipid and the double-layer membrane structure wrapping.

[0053] The present invention also provides a moisturizing composition, which comprises, by weight percentage, 0.2% to 15.3% of liposomes, 6.6% to 7.7% of emulsifiers, 2% to 3.8% of thickeners, 2.6% to 4.1% of emollients, and the balance of water;

[0054] The liposome is the liposome encapsulating hyaluronic acid described in the above technical solution or the liposome encapsulating hyaluronic acid obtained by the preparation method described in the above technical solution.

[0055] In the present invention, the moisturizing composition comprises 0.2% to 15.3% liposomes in terms of mass percentage, and in a specific embodiment, it may be 8%, 10% or 13%.

[0056] In the present invention, the moisturizing composition includes 2.5% to 7.7% of emulsifier in terms of mass percentage, and in a specific embodiment, it can be 3%, 4%, 5%, 6% or 7%; the emulsifier is a nonionic emulsifier; the nonionic emulsifier includes one or more of glyceryl distearate, sorbitan stearate, cetearyl glucoside and tristearic glyceryl; the mass ratio of glyceryl distearate, sorbitan stearate, cetearyl glucoside and tristearic glyceryl can be 2.5 to 2.6: 0 to 0.9: 0 to 0.9: 0 to 3.3, and in a specific embodiment, it can be 2.5% of glyceryl distearate. The role of the emulsifier in the present invention is to mix the ingredients evenly.

[0057] In the present invention, the moisturizing composition includes 2% to 3.8% of thickener in terms of mass percentage, and in a specific embodiment, it can be 2.5% or 3%; the thickener includes a fatty alcohol thickener and / or beeswax; the fatty alcohol thickener is stearyl alcohol; the mass ratio of the fatty alcohol thickener to beeswax can be 2 to 3.1:0 to 0.7, and in a specific embodiment, it can be 2% stearyl alcohol. The function of the thickener in the present invention is to make the skin feel of the skin care product better.

[0058] In the present invention, the moisturizing composition includes 2.6% to 4.1% of the emollient in terms of mass percentage, and in a specific embodiment, it can be 3%; the emollient includes isohexadecane and / or an ester emollient; the ester emollient is ethylhexyl hydroxystearate; the mass ratio of isohexadecane to the ester emollient can be 2.6 to 3.1:0 to 1, and in a specific embodiment, it can be 2.6% of isohexadecane. The role of the emollient in the present invention is to enhance the skin feel of the skin care product.

[0059] In the present invention, the moisturizing composition further comprises one or more of an antioxidant, a pH adjuster, a stabilizer, a preservative, an emulsifier, a solubilizer, a surfactant, a film former and a buffer.

[0060] In the present invention, the moisturizing composition includes 0.6% to 3.3% of antioxidants in terms of mass percentage, and in a specific embodiment, it can be 1%, 2% or 2.8%; the antioxidant includes one or more of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol; the mass ratio of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol is 0.6 to 1: 0 to 0.7: 0 to 1.6, and in a specific embodiment, it can be 0.6% of decarboxylated carnosine hydrochloride. The function of the antioxidant in the present invention is to improve the stability of the skin care product.

[0061] In the present invention, the moisturizing composition includes 0.6% to 1.4% of pH regulator in terms of mass percentage, and in a specific embodiment, it can be 1%; the pH regulator includes citric acid and / or sodium dihydrogen phosphate; the mass ratio of citric acid to sodium dihydrogen phosphate is 0.6 to 0.7:0 to 0.7, and in a specific embodiment, it can be 0.6% of citric acid. The function of the pH regulator in the present invention is to maintain the stability of the skin care product.

[0062] In the present invention, the moisturizing composition includes 1.2% to 4.1% of a stabilizer in terms of mass percentage, and in a specific embodiment, it may be 2%, 3% or 3.8%; the stabilizer includes one or more of an inorganic salt stabilizer, cetearyl alcohol and carbomer; the inorganic salt stabilizer is sodium chloride; the mass ratio of the inorganic salt stabilizer, cetearyl alcohol and carbomer is 1.2 to 1.6: 0 to 1.8: 0 to 0.7, and in a specific embodiment, it may be 1.2% of sodium chloride. The function of the stabilizer in the present invention is to improve the storage stability of the skin care product.

[0063] In the present invention, the moisturizing composition includes 0.2% to 1.0% of preservatives by mass percentage, and in a specific embodiment, it can be 0.5%; the preservative is an acidic preservative, and in a specific embodiment, it can be potassium sorbate. The function of the preservative in the present invention is to prevent the skin care products from breeding bacteria.

[0064] The moisturizing composition provided by the present invention can be used as a skin care product with cosmetic effects that is easier to absorb, and has broad application prospects.

[0065] The present invention also provides a method for preparing the moisturizing composition described in the above technical solution, comprising the following steps:

[0066] The moisturizing composition is obtained by mixing liposomes, emulsifiers, thickeners, emollients and water.

[0067] In the present invention, the mixed raw materials further include one or more of an antioxidant, a pH regulator, a stabilizer, a preservative, an emulsifier, a solubilizer, a surfactant, a film former and a buffer.

[0068] In the present invention, the mixing pressure can be 50-100 MPa, and in a specific embodiment, it can be 50 MPa or 80 MPa; the temperature can be 50-60°C, and the time can be 1-4 hours, and in a specific embodiment, it can be 1 hour; after the mixing, the temperature is further reduced to 20-40°C, and in a specific embodiment, it can be 40°C.

[0069] The preparation method of the moisturizing composition provided by the invention has the advantages of high mixing efficiency, strong continuity and large preparation volume.

[0070] The present invention also provides the liposome encapsulating hyaluronic acid described in the above technical solution, the liposome encapsulating hyaluronic acid obtained by the preparation method described in the above technical solution, the moisturizing composition described in the above technical solution, or the use of the moisturizing composition obtained by the preparation method described in the above technical solution in skin care products.

[0071] In the present invention, the skin care products include lotions, essences, creams, masks, freeze-dried powders or facial cleansers; the present invention does not have any special restrictions on the application of the liposomes or moisturizing compositions in skin care products, and the process familiar to those skilled in the art can be used.

[0072] To further illustrate the present invention, the liposome, moisturizing composition, preparation method and application thereof provided by the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0073] Example 1

[0074] In this embodiment, the choline phospholipid has the following chemical structure:

[0075]

[0076] 5 g of choline phosphate lipid and 3 g of hyaluronic acid with a molecular weight of 400-800 KDa were mixed, 100 mL of injection water was added, the mixture was mixed at 50-60° C., and homogenized at a pressure of 50 MPa for 60 min to obtain a liposome suspension encapsulating hyaluronic acid. The concentration of the liposome encapsulating hyaluronic acid in the suspension was 80.0 mg / mL.

[0077] Example 2

[0078] To the liposome suspension obtained in Example 1, 2.5 g of glyceryl distearate as an emulsifier, 0.6 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.0 g of stearyl alcohol as a thickener, 0.6 g of citric acid as a pH adjuster, 2.6 g of hexadecane as an emollient, 1.2 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added. The mixture was fully mixed at 50 MPa and 50-60° C. for 1 h, and the mixture was cooled to 40° C. to obtain a moisturizing composition. The samples were collected and sealed for storage.

[0079] Example 3

[0080] 5 g of choline phosphate lipid and 3.25 g of hyaluronic acid, wherein the molecular weight of the hyaluronic acid is 400-800 KDa, are mixed, 100 mL of injection water is added, the mixture is mixed at 50-60° C., and the mixture is homogenized at a pressure of 50 MPa for 60 min to obtain a liposome suspension encapsulating hyaluronic acid, wherein the concentration of the liposome encapsulating hyaluronic acid in the suspension is 82.5 mg / mL.

[0081] Example 4

[0082] In the liposome suspension obtained in Example 3, 2.5 g of glyceryl distearate as an emulsifier, 0.6 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.0 g of stearyl alcohol as a thickener, 0.6 g of citric acid as a pH adjuster, 2.6 g of hexadecane as an emollient, 1.2 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added. The mixture was fully mixed at 50 MPa and 50-60° C. for 1 h, and the temperature was lowered to 40° C. to obtain a moisturizing composition. The samples were collected and sealed for storage.

[0083] Example 5

[0084] 5 g of choline phosphate lipid and 3.5 g of hyaluronic acid, wherein the molecular weight of the hyaluronic acid is 400-800 KDa, are mixed, 100 mL of injection water is added, the mixture is mixed at 50-60° C., and the mixture is homogenized at a pressure of 50 MPa for 60 min to obtain a liposome suspension encapsulating hyaluronic acid, wherein the concentration of the liposome encapsulating hyaluronic acid in the suspension is 85.0 mg / mL.

[0085] Example 6

[0086] In the liposome suspension obtained in Example 5, 2.5 g of glyceryl distearate as an emulsifier, 0.6 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.0 g of stearyl alcohol as a thickener, 0.6 g of citric acid as a pH adjuster, 2.6 g of hexadecane as an emollient, 1.2 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added. The mixture was fully mixed at 50 MPa and 50-60° C. for 1 h, and the mixture was cooled to 40° C. to obtain a moisturizing composition. The samples were collected and sealed for storage.

[0087] Comparative Example 1

[0088] The moisturizing composition was prepared according to the preparation method provided in Examples 1-2, except that hydrogenated lecithin was used instead of choline phosphate lipid.

[0089] Comparative Example 2

[0090] The moisturizing composition was prepared according to the preparation method provided in Examples 1 and 2, except that no choline phospholipid was added.

[0091] Test Example 1

[0092] Surface topography test

[0093] Take a small amount of sample prepared in Example 1 and test its surface morphology by scanning electron microscope. The result is as follows: Figure 2 As shown, the results showed that the particle size of the liposomes formed by choline phosphate lipid encapsulating hyaluronic acid was 50-200 nm.

[0094] Test Example 2

[0095] Cytotoxicity test

[0096] In a constant environment maintained at 36-38°C, while ensuring that the concentration of carbon dioxide was kept in the range of 4.0%-6.0%, human corneal epithelial cells (HCEC) were cultured at a density of 1×10 5 / well, inoculated in a 96-well cell culture plate for incubation. After 12 hours of culture, the moisturizing composition samples of Example 2, Comparative Example 1, and Comparative Example 2 were injected into the cell culture dish in a certain amount, and a 2% hyaluronic acid solution was injected into the cell culture dish as a control group. After the cells were incubated for 24 hours, 10 μL of Celltiter-Blue reagent was added to each well. After another 4 hours of incubation, the cell viability was detected by an ELISA instrument (λex=560nm, λem=590nm). The cell viability was calculated by the following formula:

[0097] Cell viability (%) = (sample fluorescence intensity / control fluorescence intensity) × 100%

[0098] The cell viability results were as follows Figure 3 As shown by Figure 3 It can be seen that the cell survival rate of the moisturizing composition obtained in Example 2 is 99.9%, which has high biosafety, while the cell survival rates of Comparative Examples 1 and 2 are 96.8% and 97.8%, respectively, which have a small amount of toxicity.

[0099] Test Example 3

[0100] Transdermal performance test

[0101] A 2cm×2cm pig skin was fixed on a Franz diffusion cell, and 0.05g of each moisturizing composition prepared in Example 2, Comparative Example 1, and Comparative Example 2 was taken into the supply chamber and evenly applied to the skin surface of the pig skin. Injection water was used as the receiving solution, and the mixture was stirred at a constant temperature of 37°C. 0.5mL of the receiving solution was taken at 0.5h, 1h, 2h, 4h, and 6h, and an equal amount of fresh receiving solution at a constant temperature was immediately added. The samples were analyzed using an ultraviolet spectrophotometer to calculate the transdermal conditions of the samples at different times.

[0102] Depend on Figure 4 It can be seen that the sample of Example 2 has the strongest ability to penetrate the pig skin. After 6 hours of in vitro permeation, it was measured that 90.8% of the sample penetrated the pig skin, and the permeability of the samples of Comparative Examples 1 and 2 was less than 0.2%. It can be seen that the present invention uses choline phospholipids to enhance the transdermal absorption of hyaluronic acid with very excellent effect.

[0103] Test Example 4

[0104] In order to explore the irritation of the invention to the user's skin, the human patch test in the "Technical Specification for Safety of Cosmetics" (2015 edition) was referred to, and the moisturizing compositions prepared in Examples 2, 4 and 6 were selected for testing. The specific steps are as follows: 50 volunteers aged 20 to 55 years old were randomly selected, and no gender requirements were required. The patch tester loaded with an appropriate amount of sample was covered on the curved side of the forearm of the subject with a non-irritating cloth-based tape, and the sample was evenly and tightly attached to the skin surface by lightly pressing with the palm of the hand. After about 12 hours, the patch tester was removed, and the skin reaction was observed at 30 minutes (after the indentation disappeared), 24 hours and 48 hours.

[0105] The evaluation criteria are as follows:

[0106] Grade 0: negative reaction;

[0107] Grade 1: Suspicious reaction, only slight erythema;

[0108] Grade 2: weak positive reaction, erythema, infiltration, edema, and papules may be present;

[0109] Grade 3: Strong positive reaction, erythema, infiltration, edema, papules, and the reaction may extend beyond the test area;

[0110] Grade 4: Very strong positive reaction, obvious erythema, severe infiltration, edema, confluent herpes, and the reaction exceeds the test area.

[0111] The results showed that the skin reactions of all 50 volunteers were level 0, with no irritation, allergies or other symptoms, and the product can be used safely.

[0112] The test results of the liposome suspensions and moisturizing composition samples obtained in Examples 3 to 6 in Test Examples 1 to 4 are consistent with those in Examples 1 to 2, and are not described again herein.

[0113] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A liposome encapsulating hyaluronic acid, characterized in that: It comprises a hyaluronic acid compound and a choline phosphate lipid bilayer wrapped outside the hyaluronic acid compound; the particle size of the liposome is 50 to 300 nm; The inner and outer surfaces of the choline phospholipid bilayer are hydrophilic phosphocholine heads, and the inner layer is a hydrophobic fatty acid tail.

2. The liposome according to claim 1, characterized in that The choline phospholipid in the choline phospholipid bilayer has a chemical structure shown in Formula I: In Formula I, A1 and A2 are independently C x H 2x+y , x is an integer from 5 to 35, and y is 1, -1, -3, -5, -7, -9 or -11; R is a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C3-C8 alkenyl group, a substituted or unsubstituted C3-C8 alkynyl group, a substituted or unsubstituted C3-C8 epoxy group, a C3-C8 azide group, an amino group, an amino group with a protecting group, a heteroalicyclic group, an aryl group or a heteroaryl group.

3. The liposome according to claim 1 or 2, characterized in that The hyaluronic acid compound includes hyaluronic acid and / or sodium hyaluronate; the molecular weight of the hyaluronic acid compound is 2.5 to 2500 KDa; The mass ratio of the choline phosphate lipid bilayer to the hyaluronic acid compound is 0.1-9.2:0.1-6.

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4. The method for preparing liposomes according to any one of claims 1 to 3, characterized in that: The following steps are involved: The choline phosphate lipid, hyaluronic acid compound and water are mixed and homogenized to obtain the liposome.

5. A moisturizing composition, characterized in that: Calculated by mass percentage, it includes 0.2% to 15.3% of liposome, 2.5% to 7.7% of emulsifier, 2% to 3.8% of thickener, 2.6% to 4.1% of emollient and the balance of water; The liposome is the liposome encapsulating hyaluronic acid according to any one of claims 1 to 3 or the liposome encapsulating hyaluronic acid obtained by the preparation method according to claim 4.

6. The moisturizing composition according to claim 5, characterized in that The emulsifier is a nonionic emulsifier; The thickener includes a fatty alcohol thickener and / or beeswax; The emollients include isohexadecane and / or ester emollients.

7. The moisturizing composition according to claim 5, characterized in that It also includes one or more of an antioxidant, a pH adjuster, a stabilizer, a preservative, an emulsifier, a solubilizer, a surfactant, a film former, and a buffer; The antioxidant has a mass percentage of 0.6% to 3.3% in the moisturizing composition; the antioxidant comprises one or more of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol; The mass percentage of the pH regulator in the moisturizing composition is 0.6% to 1.4%; the pH regulator includes citric acid and / or sodium dihydrogen phosphate; The mass percentage of the stabilizer in the moisturizing composition is 1.2% to 4.1%; the stabilizer includes one or more of an inorganic salt stabilizer, cetearyl alcohol and carbomer; The mass percentage of the preservative in the moisturizing composition is 0.2% to 1.0%; the preservative is an acidic preservative.

8. The method for preparing the moisturizing composition according to claim 6, characterized in that: The following steps are involved: The moisturizing composition is obtained by mixing liposomes, emulsifiers, thickeners, emollients and water.

9. The preparation method according to claim 8, characterized in that: The mixed raw materials also include one or more of antioxidants, pH regulators, stabilizers, preservatives, emulsifiers, solubilizers, surfactants, film formers and buffers.

10. Use of the hyaluronic acid encapsulating liposome according to any one of claims 1 to 3, the hyaluronic acid encapsulating liposome obtained by the preparation method according to claim 4, the moisturizing composition according to claim 6 or 7, or the moisturizing composition obtained by the preparation method according to claim 8 or 9 in skin care products.

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