A collagen liposome and moisturizing anti-aging composition and preparation method and application thereof

Abstract

The present invention belongs to the technical field of skin care products, and provides a collagen liposome and a moisturizing and anti-aging composition, and a preparation method and application thereof. The collagen liposome of the present invention includes collagen, and a choline phosphate lipid layer that wraps the collagen; the choline phosphate lipid layer is a bilayer formed by choline phosphate lipids, and the choline phosphate lipids have a structure shown in the following formula. The choline phosphate lipid layer formed by the choline phosphate lipids shown in the following formula of the present invention wraps the collagen, and the formed choline phosphate lipid layer has a bilayer membrane structure similar to that of the cell membrane components, which can effectively overcome the skin barrier and the dense stratum corneum lipid layer protein, and realize the non-invasive transdermal absorption of high molecular weight (250-300KDa) collagen, and more effectively deliver the collagen to the inside of the target cells, thereby improving the effect and reducing the impact of side effects, and greatly improving the efficiency of collagen penetrating the skin. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the technical field of skin care products, and in particular to a collagen liposome and a moisturizing and anti-aging composition, and a preparation method and application thereof. Background Art

[0002] Currently, the four types of absorbable materials approved by the FDA for use in plastic surgery include: collagen, sodium hyaluronate, hydroxyapatite and polylactic acid. Among them, collagen is a type of natural protein. As the main component of skin tissue, it is an ideal material in plastic surgery.

[0003] Currently, collagen with a molecular weight less than 3000Da can be absorbed by the skin, while collagen with a molecular weight greater than 3000Da is difficult to penetrate the skin and be absorbed. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a collagen liposome and a moisturizing and anti-aging composition and a preparation method and application thereof. The collagen liposome provided by the present invention can improve the skin permeability of collagen with a molecular weight of 100KDa to 300KDa.

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

[0006] The present invention provides a collagen liposome, comprising collagen, and a choline phosphate lipid layer encapsulating the collagen;

[0007] The molecular weight of the collagen is 100KDa to 300KDa;

[0008] The choline phosphate lipid layer is a bilayer formed by choline phosphate lipids, and the choline phosphate lipids have a structure shown in Formula I:

[0009]

[0010] In Formula I, A1 and A2 are independently CH3(CH2) n - or CH3(CH2) m1 CH=CH(CH2) m2 -, n is an integer from 4 to 20, m1 is an integer from 5 to 7, and m2 is an integer from 7 to 13;

[0011] 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.

[0012] Preferably, the collagen is type III collagen;

[0013] The mass ratio of the choline phospholipid to the collagen is 1:0.8 to 1:3.

[0014] Preferably, the particle size of the collagen liposome is 50 to 300 nm.

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

[0016] The choline phospholipid, collagen and solvent are mixed and homogenized to obtain the collagen liposome.

[0017] Preferably, the mass ratio of the choline phospholipid to the collagen is 1:0.8 to 1:3; the dosage ratio of the collagen to the solvent is 0.1 to 5 g:100 mL.

[0018] Preferably, the homogenization treatment includes performing normal pressure homogenization and pressurized homogenization in sequence; the temperature of the normal pressure homogenization is 25-40°C, the pressure is 1 atmosphere, and the time of each normal pressure homogenization is 20-30 minutes; the temperature of the pressurized homogenization is 25-40°C, the pressure is 50-100MPa, and the time is 2-3 hours.

[0019] The present invention also provides a moisturizing and anti-aging composition, comprising liposomes, an emulsifier, a thickener, an emollient and a solvent;

[0020] Every 100 parts by weight of the solvent includes the following components in parts by weight:

[0021] 0.48 to 2.41 parts of the liposome, 6.6 to 7.7 parts of the emulsifier, 2 to 3.8 parts of the thickener, and 2.6 to 4.1 parts of the emollient;

[0022] The liposome is the collagen liposome described in the above technical solution or the collagen liposome prepared by the preparation method described in the above technical solution.

[0023] Preferably, the emulsifier includes one or more of glyceryl distearate, sorbitan stearate, cetearyl glucoside and glyceryl tristearate;

[0024] The thickener includes stearyl alcohol and / or beeswax;

[0025] The emollients include isohexadecane and / or ethylhexyl hydroxystearate.

[0026] Preferably, the moisturizing and anti-aging composition further comprises one or more of an antioxidant, a pH adjuster, a stabilizer and a preservative;

[0027] Based on the weight of the liposome, the weight of the antioxidant is 2.4 to 3.3 parts per 100 parts by weight of the solvent, and the antioxidant includes one or more of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol;

[0028] Based on the weight of the liposome, the weight of the pH adjuster is 1.2 to 1.4 parts per 100 parts by weight of the solvent, and the pH adjuster includes citric acid and / or sodium dihydrogen phosphate;

[0029] Based on the weight of the liposome, the weight of the stabilizer is 3.4 to 4.1 parts per 100 parts by weight of the solvent, and the stabilizer includes one or more of sodium chloride, cetearyl alcohol and carbomer;

[0030] Based on the weight of the liposome, the weight of the preservative is 0.2 to 1.0 parts per 100 parts by weight of the solvent, and the preservative is potassium sorbate.

[0031] The present invention also provides the use of the collagen liposome described in the above technical solution, the collagen liposome prepared by the preparation method described in the above technical solution, or the moisturizing and anti-aging composition described in the above technical solution in the preparation of skin care products.

[0032] The present invention provides a collagen liposome, comprising collagen and a choline phosphate lipid layer encapsulating the collagen; the molecular weight of the collagen is 100KDa to 300KDa;

[0033] The choline phosphate lipid layer is a bilayer formed by choline phosphate lipids, and the choline phosphate lipids have a structure shown in Formula I:

[0034]

[0035] In Formula I, A1 and A2 are independently CH3(CH2) n - or CH3(CH2) m1 CH=CH(CH2) m2 -, n is an integer from 4 to 20, m1 is an integer from 5 to 7, and m2 is an integer from 7 to 13;

[0036] 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.

[0037] The present invention wraps collagen with a choline phosphate lipid layer formed by the choline phosphate lipid shown in Formula I, and the formed choline phosphate lipid layer has a double-layer membrane structure similar to the cell membrane component, which can effectively overcome the skin barrier and the dense stratum corneum lipid layer protein, and realize the non-invasive transdermal absorption of high molecular weight collagen (250-300KDa), and more effectively deliver collagen to the inside of the target cell, thereby improving the effect and reducing the impact of side effects, and greatly improving the efficiency of collagen through the skin. At the same time, the choline phosphate lipid shown in Formula I has strong scalability, stable physical and chemical properties, which is conducive to exerting the best beauty and anti-aging effect of collagen. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 A schematic diagram of the structure of the collagen liposome provided by the present invention;

[0039] Figure 2 This is a scanning electron microscope photograph of type III collagen liposomes in the type III collagen liposome suspension obtained in Example 1;

[0040] Figure 3 The particle size diagram of type III collagen liposome in the type III collagen liposome suspension obtained in Examples 1 to 5;

[0041] Figure 4 The cytotoxicity of type III collagen liposomes obtained in Example 1 and Comparative Examples 1-2 to CRL1474 cells;

[0042] Figure 5 The transdermal test results of type III collagen liposomes obtained in Examples 1 to 5 and Comparative Examples 1 to 2;

[0043] Figure 6 This is a flow cytometry result of human dermal fibroblasts endocytosing the incubated collagen liposomes obtained in Example 1. DETAILED DESCRIPTION

[0044] The present invention provides a collagen liposome, comprising collagen, and a choline phosphate lipid layer encapsulating the collagen;

[0045] The molecular weight of the collagen is 100KDa to 300KDa;

[0046] The choline phosphate lipid layer is a bilayer formed by choline phosphate lipids, and the choline phosphate lipids have a structure shown in Formula I:

[0047]

[0048] In Formula I, A1 and A2 are independently CH3(CH2) n - or CH3(CH2)m1 CH=CH(CH2) m2 -, n is an integer from 4 to 20, m1 is an integer from 5 to 7, and m2 is an integer from 7 to 13;

[0049] 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.

[0050] Unless otherwise specified, the raw materials used in the present invention are preferably commercially available products.

[0051] The collagen liposome provided by the present invention comprises collagen, and the collagen is preferably type III collagen. In the present invention, the molecular weight of the collagen is 100KDa to 300KDa, and is preferably 100KDa, 150KDa, 200KDa, 250KDa, 285KDa or 300KDa.

[0052] The collagen liposome provided by the present invention comprises a choline phosphate lipid layer encapsulating the collagen. In the present invention, the choline phosphate lipid layer is a bilayer formed by choline phosphate lipids, and the choline phosphate lipids have a structure shown in Formula I:

[0053]

[0054] In the present invention, in Formula I, A1 and A2 are independently CH3(CH2) n - or CH3(CH2) m1 CH=CH(CH2) m2 -, preferably CH3(CH2) n -; n is an integer of 4 to 20, preferably 15 to 19, and more preferably 17; m1 is an integer of 5 to 7, and m2 is an integer of 7 to 13. In a specific embodiment of the present invention, in Formula I, A1 is CH3(CH2) n -, n is 16. In one embodiment of the present invention, in Formula I, A2 is CH3(CH2) n -, n is 16.

[0055] In the present invention, in Formula I, 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 azido group, an amino group, an amino group with a protecting group, a heteroalicyclic group, an aryl group or a heteroaryl group, preferably an unsubstituted C1-C8 alkyl group, and more preferably a C4 alkyl group.

[0056] In a specific embodiment of the present invention, the choline phospholipid preferably has the following structure:

[0057]

[0058] In the present invention, choline phospholipids are used to encapsulate collagen. Compared with other phospholipids, the choline phospholipids shown in Formula I have a double-layer membrane structure similar to cell membrane components, which can effectively overcome the skin barrier, can pass through the dense colloid layer protein, and can be easily absorbed by cells, and more effectively deliver collagen to the inside of the target cells, thereby improving the effect and reducing the impact of side effects, and greatly improving the efficiency of collagen permeating the skin. At the same time, the choline phospholipids shown in Formula I have strong scalability and stable physical and chemical properties, which is conducive to exerting the best anti-aging and wrinkle removal effect of collagen.

[0059] In the present invention, the mass ratio of the choline phospholipid to collagen is preferably 1:0.8 to 1:3, and specifically preferably 1:0.8, 1:0.9, 1:1, 1:1.33, 1:1.5, 1:1.67, 1:2, 1:2.5 or 1:3.

[0060] In the present invention, the particle size of the collagen liposome is preferably 50 to 300 nm.

[0061] In the present invention, the structural schematic diagram of the collagen liposome is as follows Figure 1 As shown, the collagen is wrapped by a choline phosphate lipid layer, and the choline phosphate lipid layer is a bilayer.

[0062] In the present invention, the collagen liposomes are preferably present in the form of a collagen liposome suspension, and the mass concentration of the collagen liposomes in the collagen liposome suspension is preferably 0.5% to 2.0%, specifically preferably 0.5%, 0.75%, 1%, 1.25%, 1.5%, 1.6%, 1.75% or 2%.

[0063] The present invention also provides a method for preparing the collagen liposome described in the above technical solution, comprising the following steps:

[0064] The choline phospholipid, collagen and solvent are mixed and homogenized to obtain the collagen liposome.

[0065] In the present invention, the solvent preferably includes water or phosphate buffer. In the present invention, the pH value of the phosphate buffer is preferably 7.0 to 7.4. In the present invention, the water is preferably water for injection.

[0066] In the present invention, the mass ratio of the choline phospholipid to the collagen is preferably 1:0.8 to 1:3, and specifically preferably 1:0.8, 1:0.9, 1:1, 1:1.33, 1:1.5, 1:1.67, 1:2, 1:2.5 or 1:3. In the present invention, the amount ratio of the collagen to the solvent is preferably 0.1 to 5 g:100 mL, and more preferably 0.1 g:100 mL, 0.5 g:100 mL, 1 g:100 mL, 2 g:100 mL, 3 g:100 mL, 4 g:100 mL or 5 g:100 mL.

[0067] In the present invention, the temperature for mixing the choline phospholipid, collagen and solvent is preferably 20-30°C, more preferably 25°C; the rotation speed is preferably 500-600rpm, more preferably 590rpm; the time is preferably 0.5-1.5h, more preferably 1h. In the present invention, the mixing is preferably carried out in a stirrer.

[0068] In the present invention, the homogenization treatment preferably includes performing normal pressure homogenization and pressurized homogenization in sequence, and the temperature of the normal pressure homogenization is preferably 25-40°C, and specifically preferably 37°C; the pressure is preferably 1 atmosphere; the time is preferably 20-30min, and specifically preferably 20min, 25min or 30min. In the present invention, the temperature of the pressurized homogenization is preferably 25-40°C, and specifically preferably 37°C; the pressure is preferably 50-100MPa, and specifically preferably 50MPa, 55MPa, 60MPa, 65MPa, 70MPa, 75MPa, 80MPa, 85MPa, 90MPa, 95MPa or 100MPa; the time is preferably 2-3h, and more preferably 2.5h. In the present invention, the combination of normal pressure homogenization and pressurized homogenization, and reasonable pressure and time control, make the sample more delicate; during the entire homogenization process, the temperature is controlled throughout the process, and the activity and structure of collagen will not be destroyed.

[0069] After the homogenization treatment, the present invention directly obtains the collagen liposome, and the collagen liposome is preferably dispersed in a solvent and exists in the form of a collagen liposome suspension.

[0070] The present invention also provides a moisturizing and anti-aging composition, comprising liposomes, an emulsifier, a thickener, an emollient and a solvent;

[0071] Every 100 parts by weight of the solvent includes the following components in parts by weight:

[0072] 0.48 to 2.41 parts of the liposome, 6.6 to 7.7 parts of the emulsifier, 2 to 3.8 parts of the thickener, and 2.6 to 4.1 parts of the emollient;

[0073] The liposome is the collagen liposome described in the above technical solution or the collagen liposome prepared by the preparation method described in the above technical solution.

[0074] The moisturizing and anti-aging composition provided by the present invention includes liposomes. In every 100 parts by weight of the solvent, the weight portion of the liposomes is preferably 0.48 to 2.41 parts, specifically preferably 0.48, 0.5, 0.55, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.75, 1.8, 1.9, 2, 2.1, 2.2, 2.3 or 2.41 parts.

[0075] The moisturizing and anti-aging composition provided by the present invention includes an emulsifier, and the weight parts of the emulsifier are preferably 6.6 to 7.7 parts per 100 parts by weight of the solvent, and are specifically preferably 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6 or 7.7 parts. In the present invention, the emulsifier preferably includes one or more of glyceryl distearate, sorbitan stearate, cetearyl glucoside and tristearin, and is further preferably glyceryl distearate. In the present invention, the emulsifier specifically has the function of making each component dispersed evenly.

[0076] The moisturizing and anti-aging composition provided by the present invention includes a thickener, and the weight parts of the thickener are preferably 2 to 3.8 parts per 100 parts by weight of the solvent, and are specifically preferably 2 parts, 2.1 parts, 2.2 parts, 2.3 parts, 2.4 parts, 2.5 parts, 2.6 parts, 2.7 parts, 2.8 parts, 2.9 parts, 3 parts, 3.1 parts, 3.2 parts, 3.3 parts, 3.4 parts, 3.5 parts, 3.6 parts, 3.7 parts or 3.8 parts. In the present invention, the thickener preferably includes stearyl alcohol and / or beeswax, and is more preferably stearyl alcohol. In the present invention, the thickener can improve the skin feel of the composition.

[0077] The moisturizing and anti-aging composition provided by the present invention includes an emollient, and the weight parts of the emollient are preferably 2.6 to 4.1 parts per 100 parts by weight of the solvent, and are specifically preferably 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4 or 4.1 parts. In the present invention, the emollient preferably includes isohexadecane and / or ethylhexyl hydroxystearate, and is further preferably isohexadecane. In the present invention, the emollient can enhance the skin feel of the composition.

[0078] The moisturizing and anti-aging composition provided by the present invention comprises a solvent, and the solvent is preferably water or a phosphate buffer. In the present invention, the pH value of the phosphate buffer is preferably 7.0 to 7.4. In the present invention, the water is preferably water for injection.

[0079] The moisturizing and anti-aging composition provided by the present invention preferably further comprises one or more of an antioxidant, a pH adjuster, a stabilizer and a preservative.

[0080] The moisturizing and anti-aging composition provided by the present invention preferably includes an antioxidant, and the weight portion of the antioxidant is preferably 2.4 to 3.3 parts per 100 parts by weight of the solvent, and is specifically preferably 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2 or 3.3 parts. In the present invention, the antioxidant preferably includes one or more of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol, and is further preferably decarboxylated carnosine hydrochloride. In the present invention, the antioxidant can increase the shelf life of the composition.

[0081] The moisturizing and anti-aging composition provided by the present invention preferably includes a pH regulator, and the weight portion of the pH regulator is preferably 1.2 to 1.4 parts per 100 parts by weight of the solvent, and is specifically preferably 1.2 parts, 1.3 parts or 1.4 parts. In the present invention, the pH regulator preferably includes citric acid and / or sodium dihydrogen phosphate, and is more preferably citric acid.

[0082] The moisturizing and anti-aging composition provided by the present invention preferably includes a stabilizer, and the weight parts of the stabilizer are preferably 3.4 to 4.1 parts per 100 parts by weight of the solvent, and are specifically preferably 3.4 parts, 3.5 parts, 3.6 parts, 3.7 parts, 3.8 parts, 3.9 parts, 4 parts or 4.1 parts. In the present invention, the stabilizer preferably includes one or more of sodium chloride, cetearyl alcohol and carbomer, and is more preferably sodium chloride.

[0083] The moisturizing and anti-aging composition provided by the present invention preferably includes a preservative, and the weight portion of the preservative is preferably 0.2 to 1.0 parts per 100 parts by weight of the solvent, and is specifically preferably 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 or 1.0 parts. In the present invention, the preservative is preferably potassium sorbate.

[0084] In the present invention, the preparation method of the moisturizing anti-aging composition preferably comprises the following steps: mixing liposomes, emulsifiers, thickeners, emollients and solvents to obtain the moisturizing anti-aging composition. In the present invention, the mixing temperature is preferably 37°C, the pressure is preferably 0.3-50MPa, and the time is preferably 1h.

[0085] The present invention also provides the use of the collagen liposome described in the above technical solution, the collagen liposome prepared by the preparation method described in the above technical solution, or the moisturizing and anti-aging composition described in the above technical solution in the preparation of skin care products.

[0086] In the present invention, the skin care product preferably includes lotion, essence, cream, mask, freeze-dried powder or facial cleanser.

[0087] The collagen liposome and moisturizing anti-aging composition provided by the present invention and their preparation methods and applications are described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0088] The structural formula of the choline phospholipid used in the examples is as follows:

[0089]

[0090] Example 1

[0091] 1 g of choline phosphate lipid and 2 g of type III collagen, wherein the molecular weight of type III collagen is 285 KDa, are mixed, 200 mL of phosphate buffer (pH 7.4) is added, and mixed at 25° C. and 590 rpm for 1 hour; then, homogenized at 37° C. under normal pressure for 30 minutes; then, homogenized at 37° C. and 50 MPa under high pressure for 150 minutes to obtain a type III collagen liposome suspension, wherein the concentration of type III collagen liposome is 1.5%.

[0092] To the type III collagen liposome suspension obtained in Example 1, 14.0 g of glyceryl distearate as an emulsifier, 4.0 g of stearyl alcohol as a thickener, 6.0 g of isohexadecane as an emollient, 6.0 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.4 g of citric acid as a pH adjuster, 8.0 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added, and the mixture was thoroughly mixed at 37° C. and 50 MPa for 1 h to obtain a moisturizing and anti-aging composition. The samples were collected and sealed for storage.

[0093] Example 2

[0094] 1.2 g of choline phosphate lipid and 2 g of type III collagen, wherein the molecular weight of type III collagen is 285 KDa, are mixed, 200 mL of phosphate buffer (pH 7.4) is added, and mixed at 25° C. and 590 rpm for 1 hour; then, homogenized at 37° C. under normal pressure for 30 minutes; then, homogenized at 37° C. and 50 MPa under high pressure for 150 minutes to obtain a type III collagen liposome suspension, wherein the concentration of type III collagen liposome is 1.6%.

[0095] To the type III collagen liposome suspension obtained in Example 2, 14.0 g of glyceryl distearate as an emulsifier, 4.0 g of stearyl alcohol as a thickener, 6.0 g of isohexadecane as an emollient, 6.0 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.4 g of citric acid as a pH adjuster, 8.0 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added, and the mixture was thoroughly mixed at 37° C. and 50 MPa for 1 h to obtain a moisturizing and anti-aging composition. The samples were collected and sealed for storage.

[0096] Example 3

[0097] 1.5 g of choline phosphate lipid and 2 g of type III collagen, wherein the molecular weight of type III collagen is 285 KDa, are mixed, 200 mL of injection water is added, and mixed at 25° C. and 590 rpm for 1 hour; then, normal pressure homogenization is performed at 37° C. for 30 minutes; then, high pressure homogenization is performed at 37° C. and 50 MPa for 150 minutes to obtain a type III collagen liposome suspension, wherein the concentration of type III collagen liposome is 1.75%.

[0098] To the type III collagen liposome suspension obtained in Example 3, 14.0 g of glyceryl distearate as an emulsifier, 4.0 g of stearyl alcohol as a thickener, 6.0 g of isohexadecane as an emollient, 6.0 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.4 g of citric acid as a pH adjuster, 8.0 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added, and the mixture was thoroughly mixed at 37° C. and 50 MPa for 1 h to obtain a moisturizing and anti-aging composition. The samples were collected and sealed for storage.

[0099] Example 4

[0100] 1.5 g of choline phosphate lipid and 2 g of type III collagen, wherein the molecular weight of type III collagen is 100 KDa, are mixed, 200 mL of injection water is added, and mixed at 25° C. and 590 rpm for 1 hour; then, normal pressure homogenization is performed at 37° C., and each homogenization time is 30 minutes; then, high pressure homogenization is performed at 37° C. and 50 MPa for 150 minutes to obtain a type III collagen liposome suspension, wherein the concentration of the type III collagen liposome is 1.75%.

[0101] To the type III collagen liposome suspension obtained in Example 4, 14.0 g of glyceryl distearate as an emulsifier, 4.0 g of stearyl alcohol as a thickener, 6.0 g of isohexadecane as an emollient, 6.0 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.4 g of citric acid as a pH adjuster, 8.0 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added, and the mixture was thoroughly mixed at 37° C. and 50 MPa for 1 h to obtain a moisturizing and anti-aging composition. The samples were collected and sealed for storage.

[0102] Example 5

[0103] 1.5 g of choline phosphate lipid and 2 g of type III collagen, wherein the molecular weight of type III collagen is 200 KDa, are mixed, 200 mL of injection water is added, and mixed at 25° C. and 590 rpm for 1 hour; then, normal pressure homogenization is performed at 37° C., and each homogenization time is 30 minutes; then, high pressure homogenization is performed at 37° C. and 50 MPa for 150 minutes to obtain a type III collagen liposome suspension, wherein the concentration of the type III collagen liposome is 1.75%.

[0104] To the type III collagen liposome suspension obtained in Example 5, 14.0 g of glyceryl distearate as an emulsifier, 4.0 g of stearyl alcohol as a thickener, 6.0 g of isohexadecane as an emollient, 6.0 g of decarboxylated carnosine hydrochloride as an antioxidant, 2.4 g of citric acid as a pH adjuster, 8.0 g of sodium chloride as a stabilizer, and 0.5 g of potassium sorbate as a preservative were added, and the mixture was thoroughly mixed at 37° C. and 50 MPa for 1 h to obtain a moisturizing and anti-aging composition. The samples were collected and sealed for storage.

[0105] Comparative Example 1

[0106] The only difference from the moisturizing and anti-aging composition of Example 1 is that hydrogenated lecithin is used instead of choline phosphate lipid.

[0107] Comparative Example 2

[0108] The only difference from the moisturizing and anti-aging composition of Example 1 is that choline phospholipid is not added.

[0109] Surface topography test

[0110] Figure 2 This is a scanning electron microscope photograph of type III collagen liposomes in the type III collagen liposome suspension obtained in Example 1. Figure 2 It can be seen that the particle size of the liposome formed by choline phosphate lipid encapsulating type III collagen is 50 to 300 nm.

[0111] The particle size of the type III collagen liposome in the type III collagen liposome suspension obtained in Examples 1 to 5 was measured by dynamic light scattering. Figure 3 As shown, Figure 3 The particle size diagram of type III collagen liposome in the type III collagen liposome suspension obtained in Examples 1 to 5 is shown in FIG. Figure 3 It can be seen that the particle size of the collagen liposome in Example 1 is stable at 162 nm, and the particle sizes of the collagen liposomes in Examples 2 to 5 are 151 nm, 148 nm, 146 nm, and 148 nm, respectively; while the particle sizes of the samples in Comparative Examples 1 and 2 are larger and cannot pass through the 0.45 μm filter membrane.

[0112] Cytotoxicity test

[0113] In a constant environment maintained at 37°C, while ensuring that the concentration of carbon dioxide was kept in the range of 4.0% to 6.0%, human skin dermal fibroblasts CRL1474 were cultured at a density of 1×10 5 / well, inoculated in a 96-well cell culture plate for incubation. After 24 hours of culture, the collagen liposomes of Examples 1 to 5 and Comparative Examples 1 to 2 were added to the 96-well plate, and a control group and a blank group were set up. After the cells were incubated for 48 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:

[0114] Cell viability (%) = [(control fluorescence intensity - experimental fluorescence intensity) / (control fluorescence intensity - blank fluorescence intensity)] × 100%.

[0115] The cell viability results were as follows Figure 4 As shown, Figure 4 The cytotoxicity of type III collagen liposomes obtained in Example 1 and Comparative Examples 1-2 to CRL1474 cells is shown in Figure 2. Figure 4 It can be seen that the cell survival rate of the type III collagen liposomes obtained in Example 1 is higher than 100%, which has a high biosafety; the cell survival rate of the type III collagen liposomes obtained in Comparative Examples 1-2 is also higher than 100%. After testing, the cell survival rate of the type III collagen liposomes obtained in Examples 2-5 is the same as that in Example 1, which is also higher than 100%.

[0116] Transdermal performance test

[0117] A 2cm×2cm pig skin was fixed on a Franz diffusion cell, and 100μL of type III collagen liposomes obtained in Examples 1 to 5 and Comparative Examples 1 to 2 were taken into the supply chamber, respectively, and phosphate buffer was used as the receiving solution. Stirring was performed at a constant temperature of 37°C, and 0.5mL of the receiving solution was taken at 0.5h, 1h, 2h, 3h, 5h, and 7h, respectively, and an equal amount of constant temperature fresh receiving solution was added in time. The ultraviolet spectrophotometer was used for analysis, and the transdermal conditions of the samples at different times were calculated. The results are shown in FIG. Figure 5 shown.

[0118] Figure 5 The results of the transdermal test of type III collagen liposomes obtained in Examples 1 to 5 and Comparative Examples 1 to 2 are shown in Table 1. Figure 5It can be seen that the type III collagen liposome obtained in Example 1 has the strongest ability to penetrate pig skin. After 7 hours of in vitro permeation, it was measured that 61.76% of the type III collagen liposomes penetrated pig skin. The permeation ability of the type III collagen liposomes obtained in Examples 2 to 5 is close to that of Example 1; the permeation rates of Comparative Example 1 are all lower than 13%, and Comparative Example 2 can hardly penetrate the skin. It can be seen that the permeation absorption of the collagen liposome of the present invention has a very excellent effect.

[0119] Human dermal fibroblast endocytosis assay

[0120] In a constant environment maintained at 37°C, while ensuring that the concentration of carbon dioxide is maintained in the range of 4.0% to 6.0%, human skin dermal fibroblasts CRL1474 were inoculated in a 24-well cell culture plate for incubation. After incubation for 24 hours, the incubated collagen liposomes obtained in Example 1 with different concentrations were added to the 24-well plate. After the cells were incubated for 24 hours, their endocytosis was detected by cell flow cytometry. The results are as follows: Figure 6 . Figure 6 The flow cytometry results of human dermal fibroblasts endocytosing the collagen liposomes obtained in Example 1 are shown in Figure 1. Figure 6 It can be seen that the cell endocytosis efficiency of the incubated collagen liposomes obtained in Example 1 can reach 36%.

[0121] In order to explore the irritation of the moisturizing anti-aging composition obtained in the present invention to the user's skin, the moisturizing anti-aging composition obtained in Examples 1 to 5 was selected for testing with reference to the human patch test in the "Technical Specification for Safety of Cosmetics" (2015 edition). The specific steps are as follows: 50 volunteers aged 20 to 55 years old, regardless of gender, were randomly selected, and a patch tester loaded with an appropriate amount of moisturizing anti-aging composition was covered on the curved side of the subject's forearm 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.

[0122] The evaluation criteria are as follows:

[0123] Grade 0: negative reaction;

[0124] Grade 1: Suspicious reaction, only slight erythema;

[0125] Grade 2: weak positive reaction, erythema, infiltration, edema, and papules may be present;

[0126] Grade 3: Strong positive reaction, erythema, infiltration, edema, papules, and the reaction may extend beyond the test area;

[0127] Grade 4: Very strong positive reaction, obvious erythema, severe infiltration, edema, confluent herpes, and the reaction exceeds the test area.

[0128] 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.

[0129] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A collagen liposome, characterized in that: It comprises collagen, and a choline phospholipid layer encapsulating the collagen; The molecular weight of the collagen is 100KDa to 300KDa; The choline phosphate lipid layer is a bilayer formed by choline phosphate lipids, and the choline phosphate lipids have a structure shown in Formula I: In Formula I, A1 and A2 are independently CH3(CH2) n - or CH3(CH2) m1 CH=CH(CH2) m2 -, n is an integer from 4 to 20, m1 is an integer from 5 to 7, and m2 is an integer from 7 to 13; 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.

2. The collagen liposome according to claim 1, characterized in that The collagen is type III collagen; The mass ratio of the choline phospholipid to the collagen is 1:0.8 to 1:

3.

3. The collagen liposome according to claim 1 or 2, characterized in that: The particle size of the collagen liposome is 50 to 300 nm.

4. The method for preparing collagen liposomes according to any one of claims 1 to 3, characterized in that: The following steps are involved: The choline phospholipid, collagen and solvent are mixed and homogenized to obtain the collagen liposome.

5. The preparation method according to claim 4, characterized in that: The mass ratio of the choline phospholipid to the collagen is 1:0.8 to 1:3; the dosage ratio of the collagen to the solvent is 0.1 to 5 g:100 mL.

6. The preparation method according to claim 4, characterized in that: The homogenization treatment includes performing normal pressure homogenization and pressurized homogenization in sequence; the temperature of the normal pressure homogenization is 25-40°C, the pressure is 1 atmosphere, and the time of each normal pressure homogenization is 20-30 minutes; the temperature of the pressurized homogenization is 25-40°C, the pressure is 50-100MPa, and the time is 2-3 hours.

7. A moisturizing and anti-aging composition, characterized in that: These include liposomes, emulsifiers, thickeners, emollients, and solvents; Every 100 parts by weight of the solvent includes the following components in parts by weight: 0.48 to 2.41 parts of the liposome, 6.6 to 7.7 parts of the emulsifier, 2 to 3.8 parts of the thickener, and 2.6 to 4.1 parts of the emollient; The liposome is the collagen liposome described in any one of claims 1 to 3 or the collagen liposome prepared by the preparation method described in any one of claims 4 to 6.

8. The moisturizing and anti-aging composition according to claim 7, characterized in that: The emulsifier includes one or more of glyceryl distearate, sorbitan stearate, cetearyl glucoside and glyceryl tristearate; The thickener includes stearyl alcohol and / or beeswax; The emollients include isohexadecane and / or ethylhexyl hydroxystearate.

9. The moisturizing and anti-aging composition according to claim 7, characterized in that: The moisturizing and anti-aging composition further comprises one or more of an antioxidant, a pH adjuster, a stabilizer and a preservative; Based on the weight of the liposome, the weight of the antioxidant is 2.4 to 3.3 parts per 100 parts by weight of the solvent, and the antioxidant includes one or more of decarboxylated carnosine hydrochloride, 3-o-ethyl ascorbic acid and hydroxypropyl tetrahydropyrantriol; Based on the weight of the liposome, the weight of the pH adjuster is 1.2 to 1.4 parts per 100 parts by weight of the solvent, and the pH adjuster includes citric acid and / or sodium dihydrogen phosphate; Based on the weight of the liposome, the weight of the stabilizer is 3.4 to 4.1 parts per 100 parts by weight of the solvent, and the stabilizer includes one or more of sodium chloride, cetearyl alcohol and carbomer; Based on the weight of the liposome, the weight of the preservative is 0.2 to 1.0 parts per 100 parts by weight of the solvent, and the preservative is potassium sorbate.

10. Use of the collagen liposome according to any one of claims 1 to 3, the collagen liposome prepared by the preparation method according to any one of claims 4 to 6, or the moisturizing and anti-aging composition according to any one of claims 7 to 9 in the preparation of skin care products.

Images