Composition, ceramide liposome freeze-dried powder as well as preparation method and application of ceramide liposome freeze-dried powder

Abstract

The invention relates to the technical field of cosmetics, in particular to a composition, ceramide liposome freeze-dried powder and a preparation method and application of the ceramide liposome freeze-dried powder. The invention provides a composition containing ceramide and low-molecular polysaccharide, and further provides ceramide liposome prepared from the composition, lecithin, cholesterol, buffer salt and a freeze-drying protective agent, ceramide liposome freeze-dried powder as well as a preparation method and application of the ceramide liposome freeze-dried powder. The compositions provided by the invention are mutually matched, strong in compatibility and synergistic, the ceramide content in the prepared ceramide liposome freeze-dried powder is high, a suspension and a transparent aqueous solution are more easily absorbed by skin, the skin barrier can be effectively maintained, the absorption efficiency of the product is improved, and water is replenished while the barrier is repaired. And compared with a suspension, a transparent aqueous solution and other liquids and solids, the prepared ceramide liposome freeze-dried powder is easier to store and transport, small in organic reagent residue and high in safety.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a composition, ceramide liposome freeze-dried powder, and a preparation method and application thereof. Background Art

[0002] The biological membrane is a supramolecular system composed of membrane lipids such as phospholipids, cholesterol, glycolipids and membrane proteins, which provides a functional physical barrier for cells and their organelles. Phospholipids as bilayer scaffolds can be divided into glycerophospholipids (Phosphoglyceride) and sphingomyelin (Sphingomyelin, SM). Glycerophospholipids are composed of two fatty acid chains and a phosphate group with glycerol as the backbone. Due to the different molecules bound by the phosphate group, they can be divided into phosphatidylcholine (also known as egg phosphatidylcholine, PC), phosphatidylethanolamine (also known as brain phosphatidylcholine, PE), phosphatidylserine PS, phosphatidylinositol PI, etc. Sphingomyelin is composed of sphingosine (Sph) as the backbone to bind fatty acid chains and phosphorylcholine. These phospholipids are asymmetrically distributed in the cell membrane of eukaryotic organisms. Phosphatidylcholine and sphingomyelin are mainly in the outer cell membrane, while phosphatidylserine, phosphatidylinositol and phosphatidylethanolamine are limited to the inner membrane. Some phospholipids and their metabolites are also involved in important signal transduction processes. For example, phosphatidylserine is transported to the cell surface by membrane proteins TMEM16F and Xkr8 by different mechanisms, which will stimulate coagulation and cell apoptosis.

[0003] In particular, sphingolipids with longer saturated fatty acid chains, including sphingomyelin, glycosphingolipid cerebrosides (GlcCer, GalCer), and gangliosides (GM), together with cholesterol and special proteins, will aggregate into functional microdomains of lipid raft structures on the cell membrane, which are closely related to membrane signal transduction and membrane protein sorting.

[0004] like Figure 1 As shown in the figure, ceramide (Cer), as the central molecule of sphingolipid metabolism, is the common precursor of sphingolipids such as sphingomyelin, cerebroside, ganglioside, 1-phosphoceramide (Cer-1-P). Among them, sphingomyelin synthase SMS first hydrolyzes phosphatidylcholine into diacylglycerol and choline phosphate, and ceramide replaces diacylglycerol and forms a bond with choline phosphate to produce widely distributed sphingomyelin. At the same time, a small amount of ceramide will be hydrolyzed by ceramidase CDase to produce sphingosine and free fatty acids.

[0005] The de novo synthesis of ceramide begins with the formation of 3-ketosphinganine (3KS) from serine and palmitoyl-CoA under the action of serine palmitoyltransferase SPT, and then is reduced to dihydrosphinganine (Sphinganine, Spa) under the action of enzyme KDSR and cofactor NADPH. Dihydrosphinganine is then condensed with fatty acyl-CoA amide under the action of synthetase CerS to form dihydroceramide (dhCer), and finally dehydrogenase DES1 introduces a double bond at position 4 to form ceramide. The second main synthesis pathway is the recovery and rescue of sphingosine, which is condensed with fatty acyl-CoA amide under the action of synthetase CerS to form ceramide. In addition, under stress conditions, sphingomyelin hydrolysis is also the main pathway for the production of ceramide. Sphingomyelinase SMase is divided into three families: acidic, alkaline and neutral due to the different optimal pH, cellular sublocalization and structure. Among them, neutral sphingomyelinase requires activation by phosphatidylserine.

[0006] Ceramides are most abundant in the epidermis and can be divided into twelve categories according to the differences in fatty acid chains (N, A, EO) and sphingosine (DS, S, P, H), such as Figure 2 and Figure 3 As shown. Ceramide in the interstitial matrix of stratum corneum cells can also form a lamella structure with cholesterol and fatty acids, playing multiple barrier protection roles: as a permeability barrier, it prevents excessive evaporation of water in the skin, inhibits the entry of exogenous small molecules, allergens and microorganisms into the skin, and is also a heat insulation barrier; as an oxidation barrier, it is rich in antioxidants such as glutathione and vitamin C to neutralize oxidative stress; it can also serve as a physical barrier to absorb UV radiation and buffer mechanical pressure damage.

[0007] Allergic dermatitis, psoriasis, dry skin, sensitive skin and other problems are often accompanied by damage to the skin barrier. Exogenous supplementation of ceramide is considered to be an effective means of repair, but its poor water solubility hinders its widespread application in skin care products.

[0008] The artificial phospholipid spherical bilayer liposome was invented in 1964 as a research model for in vitro biological membranes. In 1974, liposomes were shown to be able to be used for nucleic acid delivery, which set off a research boom. In recent years, liposome encapsulation and delivery technology has been extended to the fields of food, health care products and skin care products. It has many advantages: lipid components improve cell permeability, hydrophilic surface improves water solubility, encapsulation inhibits degradation and thus improves stability and reduces abnormal odor and color effects, such as Figure 4 shown.

[0009] The preparation method of ceramide liposomes prepared in the current prior art is complicated, and the prepared liposome suspension has the problems of large particle size, low ceramide concentration, poor skin permeability, easy oxidation discoloration, stratification, short shelf life, poor stability, high organic solvent residue, etc. Summary of the invention

[0010] In view of this, the technical problem to be solved by the present invention is to provide a composition, a ceramide liposome freeze-dried powder, and a preparation method and application thereof. The ceramide liposome prepared by the composition provided by the present invention has low organic solvent residue, is easy to penetrate and absorb, and can be stably stored.

[0011] The present invention provides a composition, comprising 2.4 to 20 parts by weight of ceramide and 2 to 2.5 parts by weight of low molecular weight polysaccharide, wherein:

[0012] The ceramide comprises at least one of ceramide NP stearic acid, ceramide NP oleic acid, ceramide AP, ceramide NS, ceramide AS and ceramide EOP;

[0013] The low molecular weight polysaccharide includes at least one of chitosan oligosaccharide, low molecular weight hyaluronic acid and low molecular weight heparin.

[0014] The composition provided by the present invention can effectively maintain the skin barrier, improve the absorption efficiency of the product, replenish water while repairing the barrier, and thus obtain a more accurate technical effect because the components and their concentrations cooperate with each other and enhance synergy.

[0015] In some embodiments, the ceramide is ceramide NP oleic acid, and the low molecular weight polysaccharide is chitosan oligosaccharide.

[0016] Compared with other components, the combination of the above components has a higher degree of mutual coordination and synergistic effect, thereby obtaining a more accurate technical effect.

[0017] In some specific embodiments, 20 parts by weight of ceramide NP oleic acid and 2 parts by weight of chitosan oligosaccharide are included.

[0018] Within the above concentration range, since the components and their concentrations cooperate with each other to the highest degree and enhance synergy, they can most effectively maintain the skin barrier, further improve the absorption efficiency of the product, replenish moisture while repairing the barrier, thereby obtaining a more accurate technical effect.

[0019] The present invention provides application of the composition in preparing ceramide liposomes and / or ceramide liposome freeze-dried powder.

[0020] The present invention provides ceramide liposome and / or ceramide liposome freeze-dried powder, the raw materials of which include the composition.

[0021] In some embodiments, the raw materials further include 10 to 22 parts by weight of lecithin, 6 to 7.3 parts by weight of cholesterol, 2 to 2.5 parts by weight of buffer salt, and 50 to 66 parts by weight of freeze-drying protective agent, wherein:

[0022] The lecithin comprises at least one of soybean lecithin, sunflower lecithin and egg yolk lecithin;

[0023] The buffer salt includes sodium dihydrogen phosphate and / or potassium dihydrogen phosphate;

[0024] The low molecular weight polysaccharide comprises at least one of chitosan oligosaccharide, low molecular weight hyaluronic acid and low molecular weight heparin;

[0025] The lyoprotectant includes at least one of mannitol, trehalose and sucrose.

[0026] When the raw materials provided by the present invention are used to prepare ceramide liposomes or freeze-dried powder thereof, the components are compatible with each other, have good synergistic effect, high drug loading capacity, high encapsulation rate, and the prepared liposomes have small particle size, low organic solvent residue, easy penetration and absorption, and stable storage, thereby obtaining more excellent technical effects.

[0027] In some embodiments, the raw materials further include 18 parts by weight of soy lecithin or egg yolk lecithin, 6 parts by weight of cholesterol, 2 parts by weight of sodium dihydrogen phosphate and 52 parts by weight of mannitol.

[0028] Within the above concentration range, the ceramide liposomes or lyophilized powder thereof prepared using the raw materials are compatible with each other, have better synergistic effects, higher drug loading capacity, higher encapsulation efficiency, smaller particle size of the prepared liposomes, lower organic solvent residue, easy penetration and absorption, and stable storage, thereby obtaining more excellent technical effects.

[0029] The method for preparing the ceramide liposome freeze-dried powder is characterized by comprising the following steps:

[0030] The ceramide, the lecithin and the cholesterol are dissolved in an organic solvent, and a lipid film is obtained by distillation under reduced pressure;

[0031] The lipid film, the buffer salt solution and the low molecular weight polysaccharide solution are mixed and homogenized to obtain ceramide liposomes;

[0032] The ceramide liposomes and the lyophilization protectant are mixed and lyophilized to obtain the ceramide liposome lyophilized powder.

[0033] In some embodiments, the organic solvent includes at least one of isopropanol, ethyl acetate, and n-butanol.

[0034] In some specific embodiments, the organic solvent is isopropanol.

[0035] In some embodiments, the method further includes dissolving and filtering the freeze-dried powder, wherein the filtration includes membrane filtration.

[0036] In some embodiments, the membrane filtration is performed using a polycarbonate membrane (PC membrane), and the machine used for the membrane filtration includes an Avanti microliposome extruder.

[0037] In some embodiments, the membrane filtration is performed 5 to 8 times.

[0038] The present invention provides application of the composition, the ceramide liposome or the ceramide liposome prepared by the preparation method in preparing skin care products.

[0039] The present invention provides the composition, the ceramide liposome or ceramide liposome freeze-dried powder, and the use of the ceramide liposome freeze-dried powder prepared by the preparation method in preparing skin care products.

[0040] The present invention provides a skin care product, comprising the composition, the ceramide liposome or ceramide liposome freeze-dried powder, and the ceramide liposome freeze-dried powder prepared by the preparation method.

[0041] The present invention provides a composition comprising ceramide and low molecular weight polysaccharides, further provides ceramide liposomes prepared by using the composition and lecithin, cholesterol, buffer salt and freeze-dried protective agent as raw materials and freeze-dried powder thereof, and also provides a preparation method and application of the ceramide liposomes. Compared with the prior art, the present invention has the following advantages:

[0042] 1. The composition provided by the present invention cooperates with each other, has strong compatibility and synergistic effect. The ceramide liposome freeze-dried powder, suspension and transparent aqueous solution prepared by the composition are more easily absorbed by the skin, can effectively maintain the skin barrier, improve the absorption efficiency of the product, and replenish water while repairing the barrier;

[0043] 2. Compared with liquids such as suspensions and transparent aqueous solutions, the ceramide liposome freeze-dried powder prepared by the present invention is easier to store and transport as a solid, and the ceramide content in the freeze-dried powder is as high as 19.5%;

[0044] 3. The ceramide liposome transparent aqueous solution prepared by the present invention has a small uniform particle size of liposomes, which is less likely to be stratified during storage and more stable than the suspension. The processing of the suspension into the transparent aqueous solution only requires a physical process of heating and extruding the film, without the need for other reagents to assist.

[0045] 4. The organic solvent added in the early processing of the ceramide liposome freeze-dried powder, suspension and transparent aqueous solution prepared by the present invention is subjected to reduced pressure distillation and freeze-drying treatment, and the residual content is less than 100 ppm. Other ingredients such as lecithin, cholesterol, low molecular weight polysaccharides, mannitol, etc. are edible and have high safety.

[0046] 5. The ceramide liposome freeze-dried powder, suspension and transparent aqueous solution prepared by the present invention are compounded with low molecular weight polysaccharide components such as chitosan oligosaccharide, low molecular weight hyaluronic acid or low molecular weight heparin. The low molecular weight polysaccharide is more easily absorbed by the skin, and synergistically replenishes water while repairing the barrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 Figure 2 shows the ceramide metabolic network diagram;

[0048] Figure 2 Show different types of fatty acid chains and sphingosine;

[0049] Figure 3 Representative ceramide structures are shown;

[0050] Figure 4 Schematic diagram of liposomes is shown;

[0051] Figure 5 2 shows the appearance of ceramide NP liposome freeze-dried powder, wherein the left picture is the appearance of ceramide NP stearic acid liposome freeze-dried powder in Example 2, and the right picture is the appearance of ceramide NP oleic acid liposome freeze-dried powder in Example 3;

[0052] Figure 6 The results show the dissolution effect of ceramide NP oleic acid in solvents, where the solvent in the left figure is isopropanol and the solvent in the right figure is ethanol;

[0053] Figure 7 The appearance of ceramide NP stearic acid liposome suspensions with different concentrations is shown, wherein the concentration of ceramide NP stearic acid liposomes in the left picture is 3%, and the concentration of ceramide NP stearic acid in the right picture is 0.6%;

[0054] Figure 8 The appearance of the transparent aqueous solution of ceramide NP stearic acid liposomes in Example 14 is shown;

[0055] Fig. 9 The appearance of the transparent aqueous solution of ceramide NP oleic acid liposomes in Example 15 is shown;

[0056] Fig.10 An electron microscopic image of the transparent aqueous solution of ceramide liposomes in Example 16 is shown;

[0057] Fig.11 The particle size distribution of the liposomes in the transparent aqueous solution of the ceramide liposomes of Examples 2 to 6 is shown;

[0058] Fig.12 An electron microscopic image of the transparent aqueous solution of ceramide liposomes in Example 4 is shown;

[0059] Fig.13 An electron microscopic image of the transparent aqueous solution of ceramide liposomes in Example 5 is shown;

[0060] Fig.14 The figure shows the moisturizing effect test results of the liposome transparent aqueous solution of Example 18. DETAILED DESCRIPTION

[0061] The present invention provides compositions, ceramide liposome freeze-dried powders, and preparation methods and applications thereof. Those skilled in the art can refer to the contents of this article and appropriately improve the process parameters to achieve the above. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0062] The preparation method of ceramide liposomes provided by the present invention comprises the following steps:

[0063] Step 1: Preparation of lipid film

[0064] Weigh 2.4-20 wt% ceramide (the total mass is the designed mass of liposome freeze-dried powder, the same below), 18-22 wt% lecithin, and 6-7.3 wt% cholesterol and dissolve them in an organic solvent, and then distill under reduced pressure until it becomes viscous to form a lipid film. Wherein, ceramide is one or more of ceramide NP stearic acid, NP oleic acid, AP, NS, AS, and EOP, lecithin is one of soybean lecithin, sunflower lecithin, and egg yolk lecithin, preferably egg yolk lecithin and soybean lecithin, and the organic solvent is one of isopropanol, ethyl acetate, and n-butanol, preferably isopropanol.

[0065] Step 2: Hydration

[0066] Inject an aqueous solution containing 2-2.5 wt% buffer salt and 2-2.5 wt% low molecular weight polysaccharide into a container containing a lipid film, stir evenly, and then use an IKA T18 disperser to shear at high speed to improve uniformity. The buffer salt is one of sodium dihydrogen phosphate and potassium dihydrogen phosphate, and the low molecular weight polysaccharide is one of chitosan oligosaccharide, low molecular weight hyaluronic acid, and low molecular weight heparin.

[0067] Step 3: Freeze Drying

[0068] Continue to add 50-66 wt% of mannitol, stir and transfer to freeze-drying bottle / freeze-drying plate, and start freeze-drying. After freeze-drying, the sample is tested for content by high performance liquid chromatography (HPLC) external standard method, water content by Karl Fischer method, and solvent residue by gas chromatography (GC) external standard method.

[0069] Freeze-drying procedure: pre-freeze at -40℃ for 60min, annealing temperature at -22℃ for 60min, and re-freeze at -35℃ for 60min; sublimation vacuum at 18Pa, -25℃ for 14h until sublimation of frozen ice is complete; desorption vacuum at 0Pa, 25℃ for 6h.

[0070] The test materials used in the present invention are all common commercial products and can be purchased in the market. The present invention is further described below in conjunction with the embodiments.

[0071] Example 1 Screening of solvents with high solubility of ceramide NPs

[0072] Weigh different amounts of ceramide NPs in the organic reagents listed in the table below, shake in a 70°C water bath for 30 min, and observe whether they are completely dissolved after cooling. Record the maximum amount of ceramide NPs that remain clear in each solvent and calculate the solubility.

[0073] Table 1 Solubility of ceramide NPs

[0074]

[0075] The results showed that ceramide NPs were more soluble in isopropanol, ethyl acetate, tetrahydrofuran, and n-butanol, with isopropanol, ethyl acetate, and n-butanol being preferred.

[0076] Example 2 Preparation of liposome freeze-dried powder

[0077] Weigh 10 g of ceramide NP stearic acid, 9 g of soybean lecithin, and 3 g of cholesterol in 172 mL of isopropanol, dissolve completely at 70°C, and then distill at 60°C under reduced pressure until it becomes viscous and forms a lipid film.

[0078] Inject 200 mL of an aqueous solution containing 1 g of disodium hydrogen phosphate and 1 g of low-molecular hyaluronic acid into the rotary evaporator containing the lipid film, stir evenly, and then use an IKA T18 disperser to high-speed shear for 5 minutes at 19000 rpm to improve uniformity.

[0079] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. Freeze drying procedure: pre-freeze at -40℃ for 60min, annealing temperature at -22℃ for 60min, freeze again at -35℃ for 60min; sublimation at a vacuum of 18Pa, at -25℃ for 14h until the sublimation of frozen ice is complete; desorption at a vacuum of 0Pa, at 25℃ for 6h.

[0080] The liposome freeze-dried powder was tested to have a stearic acid content of 19.5% for ceramide NP, 1.3% for water, and 61 ppm for isopropanol residue.

[0081] Example 3: Preparation of liposome lyophilized powder

[0082] According to steps 2-4, 10 g of ceramide NP oleic acid, 9 g of soybean lecithin, and 3 g of cholesterol were weighed into 172 mL of isopropanol, dissolved completely at 70 °C, and then distilled under reduced pressure at 60 °C until a viscous lipid film was formed.

[0083] 200 mL of an aqueous solution containing 1 g of dipotassium hydrogen phosphate and 1 g of low molecular weight heparin was injected into the rotary evaporator containing the lipid film, stirred evenly, and then high-speed shearing was performed at 20,000 rpm for 5 minutes using an IKA T18 disperser to improve uniformity.

[0084] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. The liposome freeze-dried powder was tested to have a ceramide NP oleic acid content of 19.3%, a moisture content of 1.9%, and an isopropanol residue of 65ppm.

[0085] The lyophilized liposome powder prepared in Example 2 and Example 3 has the following appearance: Figure 5 As shown, the left picture is the appearance picture of ceramide NP stearic acid liposome freeze-dried powder in Example 2, and the right picture is the appearance picture of ceramide NP oleic acid liposome freeze-dried powder in Example 3.

[0086] Example 4: Preparation of liposome lyophilized powder

[0087] Weigh 4 g of ceramide NP stearic acid, 4 g of ceramide AP, 9 g of sunflower lecithin, and 3 g of cholesterol in 172 mL of ethyl acetate, dissolve completely at 70° C., and then distill at 60° C. under reduced pressure until a viscous lipid film is formed.

[0088] Into the rotary evaporator containing the lipid film, 200 mL of an aqueous solution containing 1 g of disodium hydrogen phosphate and 1 g of chitosan oligosaccharide was injected, stirred evenly, and then an IKA T18 disperser was used for high-speed shearing at 19000 rpm for 5 min to improve the uniformity.

[0089] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. The liposome freeze-dried powder was tested to have a ceramide NP stearic acid content of 8.1%, a ceramide AP content of 8.0%, a moisture content of 1.7%, and an ethyl acetate residue of 80ppm.

[0090] Example 5: Preparation of liposome lyophilized powder

[0091] Weigh 3 g of ceramide NP oleic acid, 2.6 g of ceramide NS, 9 g of sunflower lecithin, and 3 g of cholesterol in 172 mL of ethyl acetate, dissolve completely at 70° C., and then distill at 60° C. under reduced pressure until a viscous lipid film is formed.

[0092] Into the rotary evaporator containing the lipid film, 200 mL of an aqueous solution containing 1 g of disodium hydrogen phosphate and 1 g of chitosan oligosaccharide was injected, stirred evenly, and then an IKA T18 disperser was used for high-speed shearing at 18,000 rpm for 5 min to improve uniformity.

[0093] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. The liposome freeze-dried powder was tested to have a ceramide NP oleic acid content of 6.3%, a ceramide NS content of 5.5%, a moisture content of 2.6%, and an ethyl acetate residue of 88ppm.

[0094] Example 6: Preparation of liposome lyophilized powder

[0095] Weigh 1 g of ceramide NP stearic acid, 0.2 g of ceramide AS, 9 g of egg yolk phosphatidylcholine, and 3 g of cholesterol in 172 mL of n-butanol, dissolve completely at 70°C, and then distill at 60°C under reduced pressure until it becomes viscous to form a lipid film.

[0096] Inject 200 mL of an aqueous solution containing 1 g of dipotassium hydrogen phosphate and 1 g of low-molecular hyaluronic acid into the rotary evaporator containing the lipid film, stir evenly, and then use an IKA T18 disperser to high-speed shear for 5 minutes at 16,000 rpm to improve uniformity.

[0097] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. The liposome freeze-dried powder was tested to have a ceramide NP stearic acid content of 2.2%, a ceramide AS content of 4.7%, a moisture content of 2.1%, and a residual n-butanol content of 73ppm.

[0098] Example 7: Preparation of liposome lyophilized powder

[0099] Weigh 2.6 g of ceramide NP stearic acid, 1.1 g of ceramide AP, 1.2 g of ceramide NS, 0.1 g of ceramide EOP, 9 g of soybean lecithin, and 3 g of cholesterol in 172 mL of isopropanol, dissolve completely at 70°C, and then distill under reduced pressure at 60°C until the mixture becomes viscous to form a lipid film.

[0100] 200 mL of an aqueous solution containing 1 g of disodium hydrogen phosphate and 1 g of low molecular weight heparin was injected into the rotary evaporator containing the lipid film, stirred evenly, and then high-speed shearing was performed at 19000 rpm for 5 min using an IKA T18 disperser to improve uniformity.

[0101] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. The liposome freeze-dried powder was tested to have a ceramide NP stearic acid content of 5.6%, a ceramide AP content of 2.3%, a ceramide NS content of 2.6%, a ceramide EOP content of 0.2%, a moisture content of 1.7%, and an isopropanol residue of 69ppm.

[0102] Example 8: Preparation of liposome lyophilized powder

[0103] Weigh 10 g of ceramide NP oleic acid, 9 g of egg yolk phosphatidylcholine, and 3 g of cholesterol in 172 mL of isopropanol, dissolve completely at 70°C, and then distill at 60°C under reduced pressure until it becomes viscous and forms a lipid film.

[0104] Into the rotary evaporator containing the lipid film, 200 mL of an aqueous solution containing 1 g of dipotassium hydrogen phosphate and 1 g of chitosan oligosaccharide was injected, stirred evenly, and then an IKA T18 disperser was used for high-speed shearing at 16000 rpm for 5 min to improve uniformity.

[0105] Continue to add 26g of mannitol, stir and transfer to a plastic dish for freeze drying. The liposome freeze-dried powder was tested to have an oleic acid content of 19.2% for ceramide NP, a water content of 1.5%, and an isopropanol residue of 65ppm.

[0106] Example 9: Preparation of liposome lyophilized powder

[0107] Weigh 10 g of ceramide NP oleic acid, 9 g of egg yolk phosphatidylcholine, and 3 g of cholesterol in 172 mL of ethanol and dissolve them incompletely at 70 °C. Figure 6 As shown, the left figure shows the dissolution effect of ceramide NP oleic acid in Example 8 in isopropanol, and the right figure shows the dissolution effect of ceramide NP oleic acid in Example 9 in ethanol.

[0108] Example 10: Preparation of liposome lyophilized powder

[0109] Weigh 10 g of ceramide NP oleic acid, 9 g of egg yolk phosphatidylcholine, and 3 g of cholesterol in 172 ml of isopropanol, dissolve completely at 70°C, and then distill at 60°C under reduced pressure until it becomes viscous to form a lipid film.

[0110] 200 ml of an aqueous solution containing 1 g of dipotassium hydrogen phosphate was injected into the rotary evaporator containing the lipid film, stirred evenly, and then high-speed shearing was performed at 16,000 rpm for 5 minutes using an IKA T18 disperser to improve uniformity.

[0111] Continue to add 26 g of mannitol, stir and transfer to a plastic dish for freeze drying.

[0112] Example 11: Preparation of liposome lyophilized powder

[0113] Weigh 9 g of egg yolk lecithin and 3 g of cholesterol into 172 ml of isopropanol, dissolve completely at 70°C, and then distill under reduced pressure at 60°C until it becomes viscous and forms a lipid film.

[0114] Into the rotary evaporator containing the lipid film, 200 ml of an aqueous solution containing 1 g of dipotassium hydrogen phosphate and 1 g of chitosan oligosaccharide was injected, stirred evenly, and then an IKA T18 disperser was used for high-speed shearing at 16000 rpm for 5 min to improve the uniformity.

[0115] Continue to add 26 g of mannitol, stir and transfer to a plastic dish for freeze drying.

[0116] The liposome freeze-dried powder formulas of Examples 2 to 11 are summarized in Table 2 below.

[0117] Table 2 Liposome freeze-dried powder formula of Examples 2 to 11

[0118]

[0119] Example 12: Preparation of liposome suspension

[0120] Weigh 3.3 g of the liposome freeze-dried powder prepared in Example 2, add 20 mL of pure water, and stir for 30-60 min to form a uniform yellow suspension. The stearic acid content of ceramide NP is 3%. Figure 7 Shown in the middle left picture.

[0121] Example 13: Preparation of liposome suspension

[0122] Weigh 0.66 g of the liposome freeze-dried powder prepared in Example 2, add 20 mL of pure water, and stir for 30-60 min to form a uniform white suspension. The stearic acid content of ceramide NP is 0.6%. Figure 7 As shown in the middle right picture.

[0123] Example 14: Preparation of liposome transparent aqueous solution

[0124] The liposome suspension containing 3% stearic acid of ceramide NP prepared in Example 12 was preheated to 70°C and then repeatedly passed through the membrane six times at 65°C using an Avanti microliposome extruder with a 100 nm polycarbonate membrane (PC membrane). The suspension gradually became transparent. Figure 8 shown.

[0125] Example 15: Preparation of liposome transparent aqueous solution

[0126] The ceramide NP oleic acid liposome freeze-dried powder obtained in Example 3 was taken, and a ceramide NP oleic acid content 3% liposome suspension was prepared according to the same process as in Example 12. The liposome suspension was preheated to 70° C., and then repeatedly passed through the membrane 6 times at 65° C. using an Avanti microliposome extruder with a 100 nm polycarbonate membrane (PC membrane). The suspension gradually became transparent, as shown in FIG. Fig. 9 shown.

[0127] Example 16: Transmission electron microscopy observation of liposomes

[0128] 5 μL of the transparent aqueous solution of ceramide NP oleic acid liposome prepared in Example 15 was adsorbed on a glow-discharged carbon-supported copper mesh for 1 min, and then 1% sodium phosphotungstate was added dropwise for staining for 30 s. The prepared liposome copper mesh was observed using a transmission electron microscope. Fig.10 As shown, the liposome particle size is uniformly less than 100 nm.

[0129] Example 17: Particle size distribution statistics of liposomes

[0130] The lyophilized liposome powders prepared in Examples 2 to 6 were prepared into suspensions of the same concentration and filtered through a membrane. The different liposome transparent aqueous solutions obtained were subjected to the same sample preparation process as in Example 16 and observed under a lens electron microscope. The particle sizes of the liposome particles were measured using Gatan software. Fig.11 As shown in the figure, the particle size distribution of liposomes is related to the source of lecithin. The particle size of egg yolk lecithin (ceramide NP stearic acid, AS) liposomes is mainly 20-60nM, the particle size of soybean lecithin (NP stearic acid, NP oleic acid) liposomes is mainly 40-80nM, and the particle size of sunflower lecithin (NP stearic acid, AP, NP oleic acid, NS) liposomes is more than 100nM (such as Fig.12 and Fig.13 as shown).

[0131] Example 18: Moisturizing efficacy test of liposome transparent aqueous solution

[0132] The liposome freeze-dried powders of Example 8, Example 10 and Example 11 were respectively prepared according to the same process as Example 12 and Example 14 to obtain liposome transparent aqueous solution sample 1 (containing ceramide NP oleic acid 3% + chitosan oligosaccharide 0.3%), liposome transparent aqueous solution sample 2 (containing ceramide NP oleic acid 3%) and liposome transparent aqueous solution sample 3 (containing chitosan oligosaccharide 0.3%).

[0133] After uniform cleaning of the inner arms of 25 subjects, multiple test areas were marked, with an area of ​​3*3cm 2The interval between each area is at least 1 cm, and the sample application area and the blank control area are randomly distributed in the marked area. The subjects first sit quietly in a constant temperature and humidity environment of 21.0±1.0℃ and 50±10%RH for 30 minutes, and then use the Corneometer ® CM 825 Skin Moisture Tester measures the initial value in each area. The sample is measured at (2.0±1.0) mg / cm 2 Apply the sample evenly in the area with a latex fingertip, and measure the skin moisture content of the sample area and the blank area after a set time of 2 hours. Fig.14 As shown in Table 3.

[0134] Table 3

[0135]

[0136] like Fig.14 As shown in Table 3, the increase in stratum corneum moisture content in sample 1 area is significantly higher than that in sample 2 and sample 3 areas, and there is a synergistic effect.

[0137] The above are only preferred embodiments 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 principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composition, characterized in that It comprises 2.4-20 parts by weight of ceramide and 2-2.5 parts by weight of low molecular weight polysaccharide, wherein: The ceramide comprises at least one of ceramide NP stearic acid, ceramide NP oleic acid, ceramide AP, ceramide NS, ceramide AS and ceramide EOP; The low molecular weight polysaccharide includes at least one of chitosan oligosaccharide, low molecular weight hyaluronic acid and low molecular weight heparin.

2. The composition according to claim 1, characterized in that The ceramide is ceramide NP oleic acid, and the low molecular weight polysaccharide is chitosan oligosaccharide.

3. The composition according to claim 1 or 2, characterized in that The invention comprises 20 parts by weight of ceramide NP oleic acid and 2 parts by weight of chitosan oligosaccharide.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of ceramide liposomes and / or ceramide liposome lyophilized powder.

5. Ceramide liposomes and / or ceramide liposome freeze-dried powder, characterized in that: The raw materials include the composition according to any one of claims 1 to 3.

6. The ceramide liposome and / or ceramide liposome freeze-dried powder according to claim 5, characterized in that: The raw materials also include 10 to 22 parts by weight of lecithin, 6 to 7.3 parts by weight of cholesterol, 2 to 2.5 parts by weight of buffer salt and 50 to 66 parts by weight of freeze-drying protective agent, wherein: The lecithin comprises at least one of soybean lecithin, sunflower lecithin and egg yolk lecithin; The buffer salt includes sodium dihydrogen phosphate and / or potassium dihydrogen phosphate; The low molecular weight polysaccharide comprises at least one of chitosan oligosaccharide, low molecular weight hyaluronic acid and low molecular weight heparin; The lyoprotectant includes at least one of mannitol, trehalose and sucrose.

7. The ceramide liposome and / or ceramide liposome freeze-dried powder according to claim 5 or 6, characterized in that: The raw materials also include 18 parts by weight of soybean lecithin or egg yolk lecithin, 6 parts by weight of cholesterol, 2 parts by weight of sodium dihydrogen phosphate and 52 parts by weight of mannitol.

8. The method for preparing the lyophilized ceramide liposome powder according to any one of claims 5 to 7, characterized in that: The steps include: The ceramide, the lecithin and the cholesterol are dissolved in an organic solvent, and a lipid film is obtained by distillation under reduced pressure; The lipid film, the buffer salt solution and the low molecular weight polysaccharide solution are mixed and homogenized to obtain ceramide liposomes; The ceramide liposomes and the lyophilization protectant are mixed and lyophilized to obtain the ceramide liposome lyophilized powder.

9. The preparation method according to claim 8, characterized in that: The organic solvent includes at least one of isopropanol, ethyl acetate and n-butanol.

10. Use of the composition according to any one of claims 1 to 3, the ceramide liposome or ceramide liposome lyophilized powder according to any one of claims 5 to 7, or the ceramide liposome lyophilized powder prepared by the preparation method according to claim 8 or 9 in the preparation of skin care products.

11. A skin care product, characterized in that The invention comprises the composition according to any one of claims 1 to 3, the ceramide liposome or ceramide liposome lyophilized powder according to any one of claims 5 to 7, and the ceramide liposome lyophilized powder prepared by the preparation method according to claim 8 or 9.

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