Preparation method and application of coated glass pigment

Abstract

The invention discloses a preparation method and application of a wrapped glass pigment, belongs to the technical field of cosmetics, and provides the preparation method of the wrapped glass pigment. Comprising the following steps: weighing lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol, and dissolving in absolute ethyl alcohol to obtain a phospholipid-cholesterol ethanol solution; performing ultrasonic treatment until no particles or insoluble substances can be seen by naked eyes, dissolving hydroxypropyl tetrahydropyrantriol in a phosphate buffer solution, and stirring under a 35-45 DEG C water bath condition after fully dissolving; the preparation method comprises the following steps: sucking a phospholipid-cholesterol ethanol solution by using an injector, dropwise adding the phospholipid-cholesterol ethanol solution into a hydroxypropyl tetrahydropyrantriol solution, stirring to form a liposome suspension, carrying out rotary evaporation on the suspension to remove ethanol, then carrying out ultrasonic dispersion under the condition of ice-water bath, and finally extruding to pass through a film to obtain the liposome wrapping the glassine. The glassine is wrapped by the cyclic polyethylene glycol modified lipidosome, so that the transdermal rate of the glassine can be remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetics, and in particular to a preparation method and application of encapsulated Bossagra. Background Art

[0002] Hydroxypropyl tetrahydropyrantriol, commonly known as Bossain, is a xylose derivative with anti-aging active ingredients. It can stimulate the production of mucopolysaccharides, increase the water content between extracellular matrix, fill the gaps in extracellular matrix, reduce wrinkles on the skin and make it more delicate. At the same time, Bossain can also play a role in the junction of the true epidermis, promote the synthesis of collagen VII and collagen IV, make the epidermis and dermis link more closely, improve the firmness and elasticity of the skin, and long-term use can improve wrinkles and fine lines on the face and neck, and promote the regeneration of damaged tissues. However, due to the weak transdermal performance of Bossain raw materials and the high effective concentration, many brands actually add an astonishing 30% for product effects. However, high concentrations of Bossain have a strong greasy feeling and poor effect, and can also cause sensitive irritation, acne and other problems.

[0003] Liposomes refer to closed vesicles composed of a phospholipid bilayer with an aqueous phase inside. The basic components are usually amphiphilic phospholipids and cholesterol, among which amphiphilic phospholipids form a bilayer structure, and cholesterol supports and maintains the bilayer structure. In an aqueous environment, phospholipid molecules can spontaneously arrange into liposomes driven by hydrophobic interactions and other intermolecular interactions. Cholesterol can promote the accumulation of lipid chains and the formation of bilayers, and reduce the fluidity of the bilayer. Studies have shown that drugs with liposomes as carriers can easily enter the stratum corneum and form drug reservoirs in the epidermis and dermis. The drugs can continuously play a therapeutic role on diseased cells, greatly improve the therapeutic index, and reduce systemic side effects. This feature just meets the purpose of cosmetic applications. For cosmetics with liposomes as the encapsulation layer, liposomes can not only penetrate the epidermal barrier so that the encapsulated active ingredients can enter the dermis, but also the cell membrane-like structure of liposomes gives it some characteristics that other cosmetic carriers do not have. However, liposomes belong to a coarse dispersion system, and there are problems such as easy flocculation, easy leakage of encapsulated materials, and poor compatibility with dispersants that need to be solved. Summary of the invention

[0004] In order to overcome the shortcomings of the above-mentioned prior art, the purpose of the present invention is to provide a preparation method and application of encapsulated bosin, and the liposome encapsulation technology modified by cyclic polyethylene glycol is used to encapsulate hydroxypropyl tetrahydropyrantriol, which can significantly improve the skin penetration rate of bosin and improve the efficacy of bosin, thereby solving the problem that the skin feel and product efficacy of hydroxypropyl tetrahydropyrantriol products cannot be taken into account at the same time and it is not suitable for people with sensitive skin.

[0005] The present invention is achieved through the following technical solutions: A preparation method for encapsulating bosin comprises the following steps: weighing lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol and dissolving them in anhydrous ethanol to obtain a phospholipid-cholesterol ethanol solution; ultrasonically treating until no particles or insoluble matter are visible to the naked eye, dissolving hydroxypropyl tetrahydropyrantriol in a phosphate buffer, and stirring in a water bath at 35-45°C after fully dissolving; using a syringe to draw the phospholipid-cholesterol ethanol solution, adding it dropwise to the hydroxypropyl tetrahydropyrantriol solution, stirring to form a liposome suspension, rotary evaporating the suspension to remove ethanol, and then ultrasonically dispersing it in an ice-water bath, and finally extruding through a membrane to obtain a liposome encapsulating bosin, wherein the choline phospholipid-polyethylene glycol-choline phospholipid and the choline phospholipid-polyethylene glycol-cholesterol have structures shown in Formula 1 and Formula 2, respectively: Formula 1 Formula 2 Wherein, a and b are independently selected from any positive integer between 11 and 15.

[0006] Preferably, the lecithin is at least one of soybean lecithin or egg yolk lecithin.

[0007] Phosphatidylcholine, or phosphatidylcholine, has a zwitterionic head group and is the main component of all eukaryotic cell membranes. It has excellent biocompatibility. Liposome formulations formed with phosphatidylcholine have been approved by the FDA and account for more than 50% of the clinical nanomedicine market. However, phosphatidylcholine lacks a modifiable group in its structure, and phosphatidylethanolamine with an amine group has low biocompatibility due to the lack of a zwitterionic structure. Therefore, some studies have synthesized choline phosphate to meet the requirements of good biocompatibility and modifiability, and have confirmed that choline phosphate has excellent biocompatibility and low toxicity.

[0008] The present invention adds choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol to the traditional liposome formula of lecithin and cholesterol. The inventor has found through research that when two choline phospholipid hydrophilic heads are connected to the same polyethylene glycol, when these two choline phospholipids form liposomes, the polyethylene glycol segment becomes cyclic, and the relatively linear polyethylene glycol segment can effectively improve the dispersibility and stability of liposome vesicles. This may be because the cyclic structure of the cyclic polyethylene glycol limits the free movement of the polyethylene glycol chain, reduces the mutual entanglement between the chains, makes the integrity of the liposome better, and introduces a steric hindrance structure between the liposomes, reduces the aggregation and flocculation of the liposomes, and improves the dispersibility of the liposomes. Similarly, the structure of choline phospholipid-polyethylene glycol-cholesterol can make cholesterol link with the phospholipid layer through the polyethylene glycol segment, so that cholesterol is no longer in a free state, thereby further improving the firmness of the liposome. In addition, modifying the surface of liposomes with cyclic polyethylene glycol can also improve the compatibility of liposomes with dispersants or solvents, which is beneficial for the dilution application of encapsulated bosine in lotions or creams.

[0009] Preferably, the mass ratio of lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol is (10-15): (1-5): (3-5).

[0010] Preferably, the molecular weight of polyethylene glycol in the choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol is 600-2000, wherein, when the molecular weight of polyethylene glycol is too short, the steric hindrance effect on the liposome surface is affected, and when the molecular weight of polyethylene glycol is too long, the anchoring effect of cholesterol and phospholipid is not obvious.

[0011] Preferably, the mass ratio of hydroxypropyl tetrahydropyrantriol to the total mass of phospholipids in lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol is 1:(5-10).

[0012] Preferably, the temperature of the rotary evaporation is 40-60°C.

[0013] Preferably, the method for preparing the choline phospholipid-polyethylene glycol-choline phospholipid comprises the following steps: S1. A saturated fatty acid, 3-dimethylamino-1,2-propylene glycol, a catalyst and a dehydrating agent are dissolved in dichloromethane, and an esterification reaction is carried out at room temperature to obtain a reactant 1; S2. Under anhydrous and oxygen-free conditions, polyethylene glycol and triethylamine are added to tetrahydrofuran, stirred evenly, and then cooled in an ice-water bath, 2-chloro-2-oxo-1,3,2-dioxaphospholane is slowly added dropwise, the reaction is stirred in the ice-water bath for a period of time, and then the temperature is raised to room temperature and the reaction is continued for a period of time to obtain reactant 2; S3. Add reactant 1 and reactant 2 into acetonitrile and react at 60-80° C. to obtain choline phospholipid-polyethylene glycol-choline phospholipid.

[0014] Preferably, the molar ratio of the saturated fatty acid to 3-dimethylamino-1,2-propanediol is (2.1~2.5):1; the saturated fatty acid is selected from at least one of palmitic acid, myristic acid, and lauric acid; the catalyst is 4-dimethylaminopyridine; the dehydrating agent is at least one of N,N'-dicyclohexylcarbodiimide and N,N'-diisopropylcarbodiimide; the molar ratio of the polyethylene glycol to 2-chloro-2-oxo-1,3,2-dioxaphospholane is 1:(2.1~2.5); the molar ratio of the reactant 1 to the reactant 2 is (2.1~2.5):1.

[0015] Preferably, the method for preparing choline phospholipid-polyethylene glycol-cholesterol comprises the following steps: S1. A saturated fatty acid, 3-dimethylamino-1,2-propylene glycol, a catalyst and a dehydrating agent are dissolved in dichloromethane, and an esterification reaction is carried out at room temperature to obtain a reactant 1; S2. Under anhydrous and oxygen-free conditions, hydroxypolyethylene glycol carboxyl and triethylamine were added to tetrahydrofuran, stirred evenly and cooled in an ice-water bath, 2-chloro-2-oxo-1,3,2-dioxaphospholane was slowly added dropwise, the reaction was stirred in the ice-water bath for a period of time, and then the temperature was raised to room temperature and the reaction was continued to obtain reactant 3; S3. adding reactant 1 and reactant 3 to acetonitrile, and reacting at 60-80° C. to obtain choline phospholipid-polyethylene glycol-carboxyl; S4. Dissolve choline phospholipid-polyethylene glycol-carboxyl, cholesterol, a catalyst and a dehydrating agent in dichloromethane, and under the protection of an inert gas, stir the mixture at room temperature to carry out an esterification reaction to obtain choline phospholipid-polyethylene glycol-cholesterol.

[0016] Preferably, the molar ratio of the saturated fatty acid to 3-dimethylamino-1,2-propylene glycol is (2.1-2.5):1; the saturated fatty acid is selected from at least one of palmitic acid, myristic acid and lauric acid; the catalyst is 4-dimethylaminopyridine; the dehydrating agent is at least one of N,N'-dicyclohexylcarbodiimide and N,N'-diisopropylcarbodiimide; the molar ratio of the hydroxypolyethylene glycol carboxyl group to 2-chloro-2-oxo-1,3,2-dioxaphospholane is (1.05-1.2):1; the molar ratio of the reactant 1 to the reactant 3 is (1.05-1.2):1, and the molar ratio of the choline phospholipid-polyethylene glycol-carboxyl group to cholesterol is 1:(1.05-1.2).

[0017] Preferably, the step of extruding the liposomes through the membrane is performed by stacking and extruding a plurality of membranes, and the particle size combinations of the multi-layer membranes are 400 nm, 200 nm and 80 nm, respectively.

[0018] Another object of the present invention is to protect a transdermal bosera, wherein the bosera prepared by the above-mentioned preparation method of encapsulated bosera is dispersed in a mixed solution of water and butanediol at a mass fraction of 10-50wt%.

[0019] Another object of the present invention is to protect the use of the encapsulated Bossagra prepared by the above-mentioned preparation method of encapsulated Bossagra in essence water, lotion, essence milk, face cream and eye cream.

[0020] Beneficial Effects

[0021] The present invention provides a preparation method and application of encapsulated bosine, by adding choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol to the traditional liposome formula of lecithin and cholesterol, accessing the same polyethylene glycol to the two choline phosphorylation hydrophilic heads to obtain a liposome structure modified with cyclic polyethylene glycol, and the relatively linear polyethylene glycol chain segment can effectively improve the dispersibility and stability of liposome vesicles. Among them, the ring structure of the cyclic polyethylene glycol limits the free movement of the polyethylene glycol chain, reduces the mutual entanglement between the chains, makes the integrity of the liposome better, and introduces a steric hindrance structure between the liposomes, reduces the aggregation and flocculation of the liposomes, and improves the dispersibility of the liposomes. At the same time, the structure of choline phospholipid-polyethylene glycol-cholesterol allows cholesterol to be linked to the phospholipid layer through the polyethylene glycol chain segment, so that cholesterol is no longer in a free state, thereby further improving the firmness of the liposome. In addition, modifying the surface of liposomes with cyclic polyethylene glycol can also improve the compatibility of liposomes with dispersants or solvents, which is beneficial for the dilution application of encapsulated bosine in lotions or creams.

[0022] Using liposomes to encapsulate the raw material hydroxypropyl tetrahydropyrantriol can significantly increase the skin penetration rate of benzoyl peroxide and improve the efficacy of benzoyl peroxide, thereby solving the problem that the skin feel and efficacy of hydroxypropyl tetrahydropyrantriol products cannot be taken into account at the same time and that it is not suitable for people with sensitive skin. It can also achieve the sustained-release effect of the product, prolong the action time of the product, further reduce the adverse reactions of the product, and improve the overall anti-aging effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is the H NMR spectrum of reactant 2; Figure 2 This is a physical picture of the encapsulated Bose with a mass fraction of 30%; Figure 3 This is the effect of improving eye wrinkles based on human evaluation of 10% Bossagra. DETAILED DESCRIPTION

[0024] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] In the examples, the experimental methods used are conventional methods unless otherwise specified, and the materials, reagents, etc. used are all commercially available unless otherwise specified.

[0026] The raw materials used in the embodiments and comparative examples are described as follows: Palmitic acid: CAS: 57-10-3, purity above 99%, impurity limit 0.05%, purchased from Nanjing Fangshenghe Pharmaceutical Technology Co., Ltd.; Myristic acid: CAS: 544-63-8, purity 99% HPLC, purchased from Changzhou Zhichao Pharmaceutical Technology Co., Ltd.; Lauric acid: CAS: 143-07-7, purity: 99%, purchased from Jinan World Chemical Co., Ltd.; 3-Dimethylamino-1,2-propanediol: CAS: 623-57-4, purity: 98%, purchased from Wuhan Yuqing Jiaheng Pharmaceutical Co., Ltd.; 4-Dimethylaminopyridine (DMAP): CAS: 1122-58-3, purity: 99%, purchased from Shanghai Kaisai Chemical Co., Ltd.; N,N'-Diisopropylcarbodiimide (DIC): CAS: 693-13-0, purity: 99% GC, purchased from Suzhou Haofan Biotechnology Co., Ltd.; Dichloromethane: CAS: 75-09-2, purity: 99%, purchased from Jiangsu Runfeng Synthetic Technology Co., Ltd., dried with CaH; Hydroxypolyethylene glycol carboxyl 1: molecular weight 1000, purchased from Zhengzhou Huiju Chemical Co., Ltd.; Hydroxypolyethylene glycol carboxyl 2: molecular weight 2000, purchased from Zhengzhou Huiju Chemical Co., Ltd.; Hydroxypolyethylene glycol carboxyl 3: molecular weight 600, purchased from Zhengzhou Huiju Chemical Co., Ltd.; Polyethylene glycol 1: molecular weight 1000, purchased from Shandong Leli New Materials Co., Ltd.; Polyethylene glycol 2: molecular weight 2000, purchased from Shandong Leli New Materials Co., Ltd.; Polyethylene glycol 3: molecular weight 600, purchased from Shandong Leli New Materials Co., Ltd.; 2-Chloro-2-oxo-1,3,2-dioxaphospholane: CAS: 6609-64-9, purity: 98%, purchased from Henan Weitixi Chemical Technology Co., Ltd.; Triethylamine: CAS: 121-44-8, purity: 99%, purchased from Jiangsu Runfeng Synthetic Technology Co., Ltd.; Dicyclohexylcarbodiimide (DCC): CAS: 538-75-0, purity: 99.5%, purchased from Xinjiang Dajiang Runyang Chemical Co., Ltd.; Lecithin: Egg yolk lecithin (high purity), CAS: 8002-43-5, purity above 98%, purchased from Dalian Meilun Biotechnology Co., Ltd.; Cholesterol: CAS: 57-88-5, purity 99%, purchased from Wuxi Jingyao Biotechnology Co., Ltd.; Hydroxypropyl tetrahydropyrantriol (Bosine): CAS: 439685-79-7, purity more than 99%, purchased from Shandong Mayo Huawei Technology Co., Ltd.; Phosphate buffer: prepare 1 L of 0.2 mol / L NaH2PO4 solution and 1 L of 0.2 mol / L Na2HPO4 solution, and then prepare a buffer solution with a pH value of 7.4 at a NaH2PO4 / Na2HPO4 volume ratio of 19:81.

[0027] Unless otherwise specified, the components and raw materials used in the embodiments and comparative examples of the present invention are all commercially available raw materials, and the components and raw materials used in each parallel experiment are all of the same kind.

[0028] Example 1

[0029] Preparation of choline phospholipid-polyethylene glycol-choline phospholipid: S1. 2.3 mmol of palmitic acid, 1.0 mmol of 3-dimethylamino-1,2-propanediol, 0.02 mmol of DMAP and 2.3 mmol of DIC were completely dissolved in 20 ml of dichloromethane, reacted at room temperature for 24 h, filtered under reduced pressure to remove the precipitate, the filtrate was dried by rotary evaporation, purified by silica gel column chromatography (the eluent was a mixed solvent of petroleum and ethyl acetate in a volume ratio of 3:1), and the solvent was removed by vacuum to obtain reactant 1, with a yield of 92.6%; the structural formula of the reactant 1 is shown in Formula 3; Formula 3 S2. Under an argon atmosphere, 0.1 mol of polyethylene glycol 1 and 0.21 mol of triethylamine were added to 150 ml of anhydrous tetrahydrofuran, stirred evenly, and then placed in an ice-water bath for cooling for 30 min. Within 1 hour, 0.21 mol of 2-chloro-2-oxo-1,3,2-dioxaphospholane was slowly added dropwise, and the reaction was kept stirring in the ice-water bath for 2 hours. The temperature was then raised to room temperature and the reaction was continued for 2 hours. After filtering, the filtrate was distilled under an argon atmosphere, and finally purified by vacuum distillation to obtain reactant 2 with a yield of 66.4%; the structural formula of the reactant 2 is shown in Formula 4, and the nuclear magnetic hydrogen spectrum 1 H-NMR (500MHz, CDCl3) Figure 1 As shown; Formula 4 S3. 0.11 mol of reactant 1 and 0.05 mol of reactant 2 were added to 150 ml of anhydrous acetonitrile, reacted at 70°C for 48 hours, and then vacuum dried to remove the solvent. The mixture was washed three times with ether, filtered, and separated by column chromatography to obtain choline phospholipid-polyethylene glycol-choline phospholipid with a yield of 69.7%.

[0030] Preparation of choline phospholipid-polyethylene glycol-cholesterol: S1. Under an argon atmosphere, 0.11 mol of hydroxypolyethylene glycol carboxyl 1 and 0.11 mol of triethylamine were added to 150 ml of anhydrous tetrahydrofuran, stirred evenly and placed in an ice-water bath for 30 min, 0.1 mol of 2-chloro-2-oxo-1,3,2-dioxaphospholane was slowly added dropwise within 1 hour, the reaction was stirred in an ice-water bath for 2 hours, and then the temperature was raised to room temperature and the reaction was continued for 2 hours. After filtering, the filtrate was distilled under an argon atmosphere, and finally purified by vacuum distillation to obtain reactant 3 with a yield of 86.4%; the structural formula of the reactant 3 is shown in Formula 5; Formula 5 S2. 0.11 mol of reactant 1 and 0.1 mol of reactant 3 were added to 150 ml of anhydrous acetonitrile, reacted at 70°C for 48 h, vacuum dried to remove the solvent, washed three times with ether, filtered, and separated by column chromatography to obtain choline phospholipid-polyethylene glycol-carboxyl, with a yield of 75.4%; S3. 2 mmol of choline phospholipid-polyethylene glycol-carboxyl, 2.1 mmol of cholesterol, 0.016 mmol of DMAP and 0.016 mmol of DCC were dissolved in 60 ml of anhydrous dichloromethane. Under argon protection, the mixture was stirred and reacted at room temperature for 24 hours, filtered, and the filtrate was vacuum dried to remove the solvent, washed three times with ether, filtered, and separated by column chromatography to obtain choline phospholipid-polyethylene glycol-cholesterol with a yield of 71.2%.

[0031] Preparation of liposomes encapsulating bosone: 15 parts by weight of lecithin, 1 part by weight of choline phospholipid-polyethylene glycol-choline phospholipid and 3 parts by weight of choline phospholipid-polyethylene glycol-cholesterol are weighed and dissolved in 50 parts by weight of anhydrous ethanol, and ultrasonicated for 5 minutes until no particles or insoluble matter are visible to the naked eye; 2 parts by weight of hydroxypropyl tetrahydropyrantriol are dissolved in 600 parts by weight of phosphate buffer, and stirred in a water bath at 40° C. after being fully dissolved; the phospholipid-cholesterol ethanol solution is drawn with a syringe, and quickly added dropwise to the hydroxypropyl tetrahydropyrantriol solution, and stirred for 30 minutes to form a liposome suspension; the suspension is subjected to rotary evaporation at 40° C. for 2 hours to remove ethanol; the liposome suspension is ultrasonicated for 10 minutes in an ice water bath, ultrasonicated for 5 seconds and then stopped for 5 seconds; finally, it is extruded through polycarbonate filter membranes (Whatman) with particle sizes of 400 nm, 200 nm and 80 nm at 40° C., respectively, to obtain liposomes encapsulating bosone, i.e., encapsulating bosone.

[0032] Example 2

[0033] The preparation method is different from that in Example 1, in the preparation of choline phospholipid-polyethylene glycol-choline phospholipid, polyethylene glycol 1 is replaced by polyethylene glycol 2, and in the preparation of choline phospholipid-polyethylene glycol-cholesterol, hydroxy polyethylene glycol carboxyl group 1 is replaced by hydroxy polyethylene glycol carboxyl group 2; Example 3

[0034] The preparation method is different from that in Example 1, in the preparation of choline phospholipid-polyethylene glycol-choline phospholipid, polyethylene glycol 1 is replaced by polyethylene glycol 3, and in the preparation of choline phospholipid-polyethylene glycol-cholesterol, hydroxy polyethylene glycol carboxyl group 1 is replaced by hydroxy polyethylene glycol carboxyl group 3; Example 4

[0035] The preparation method is different from that in Example 1, the palmitic acid in reactant 1 is replaced by myristic acid; Example 5

[0036] The preparation method is different from that in Example 1, in that the palmitic acid in reactant 1 is replaced by lauric acid; Example 6

[0037] The preparation method is different from that in Example 1, the mass ratio of lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol in the preparation of liposomes encapsulating bosone is 10:1:3; Example 7

[0038] The preparation method is different from that in Example 1, the mass ratio of lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol in the preparation of liposomes encapsulating bosone is 15:5:3; Example 8

[0039] The preparation method is different from that in Example 1, the mass ratio of lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol in the preparation of liposomes encapsulating bosone is 15:1:5; Comparative Example 1 The preparation method is compared with Example 1, except that choline phospholipid-polyethylene glycol-choline phospholipid is replaced by choline phospholipid-polyethylene glycol, and the specific operation difference is that in the preparation step S2 of choline phospholipid-polyethylene glycol-choline phospholipid, 0.21 mol of 2-chloro-2-oxy-1,3,2-dioxaphospholane is replaced by 0.11 mol of 2-chloro-2-oxy-1,3,2-dioxaphospholane; Comparative Example 2 The preparation method is different from that in Example 1, in the preparation of the liposomes encapsulating bosone, choline phospholipid-polyethylene glycol-choline phospholipid is not added, and choline phospholipid-polyethylene glycol-cholesterol replaces cholesterol, that is, 20 parts by weight of lecithin and 3 cholesterol are used as raw materials of the liposomes; Comparative Example 3 Unwrapped Bosera.

[0040] Performance Testing The liposomes encapsulating bosone prepared in Examples 1-8 and Comparative Examples 1-3 were dispersed at a mass fraction of 30 wt % in a mixed solution of purified water and butanediol in a mass ratio of 10:1, and the following performance tests were performed. The results are shown in Table 1.

[0041] (1) Test of bosine content: The bosine content in the bosine encapsulated solution was tested by high performance liquid chromatography according to the standard GB 16631-2008 to determine the encapsulation rate. The instrument was LC-MS: Waters Acquity-SQ Detector 2; (2) Particle size test: Refer to the dynamic light scattering method recommended by GB / T 44223-2024 to test the particle size of the encapsulated boson. The instrument is: Dynamic Light Scattering (DLS): Nano ZS90; (3) Transdermal absorption test: The in vivo tape tearing method was used to determine the content of phosphene in the stratum corneum and calculate the permeability to compare the permeability difference of phosphene in the embodiment and the comparative example. The specific steps are as follows: 1) The inner side of the left and right front arms of the subject were cleaned and dried, and the subject sat in a constant temperature and humidity room for 1 hour; 2) The inner side of the left and right front arms of the subject were divided into sections, and each sample was tested in three areas. The area of ​​each area was 3 cm × 3 cm. Multiple areas on the same arm were at least 1 cm apart. 200 μL of the sample was used in each area and applied evenly; 3) After the sample was used, the subject sat for 2 hours, and then a D-Squame sampling patch was used to sample each area. A total of 11 layers were taken (the first layer was discarded), placed in a centrifuge tube, and every two layers were combined into one centrifuge tube. After ultrasonic extraction with methanol solution for 30 minutes, the sample was filtered through a 0.22 μm filter membrane, and the phosphene sample concentration was tested using high performance liquid chromatography. The permeability = sample concentration × fixed volume.

[0042] Table 1 Performance test results of embodiments and comparative examples

[0043] From the data of Comparative Example 3, it can be seen that making phosphene into liposomes can effectively improve the transdermal absorption effect of phosphene and improve the efficacy utilization rate of phosphene. From Comparative Example 2, it can be seen that conventional liposome vesicles are easily damaged, resulting in unstable phosphene content, and some phosphene that is not encapsulated exists in the system, affecting the transdermal absorption effect. From Comparative Example 1, it can be seen that the liposomes modified with cyclic polyethylene glycol are more stable than the linear modified liposomes. Figure 2 It can be seen that the encapsulated boson in the embodiment can exist as a clear and transparent homogeneous solution without flocculation or sedimentation, and has good compatibility with the dispersant.

[0044] Human efficacy testing The encapsulated bosine prepared in Example 1 was diluted to a bosine content of 10wt%, and a human efficacy evaluation test was conducted. 33 healthy Chinese male / female subjects with sagging facial skin, lack of elasticity, obvious facial lines, fine lines or wrinkles, and wrinkles at the corners of the eyes, which met the SKIN AGINGATLAS level 1-5, were selected. The age range was 25 to 60 years old. The skin elasticity and skin firmness of the subjects were measured by the self-before and after control method, and subjective evaluation was performed and local photos of the subjects were taken. The test results were compared by statistical test methods to determine whether there was a statistical difference.

[0045] The test sites are: 1) cheek skin elasticity, 2) cheek skin firmness, 3) number of eye corner wrinkles, 4) length of eye corner wrinkles, 5) area of ​​eye corner wrinkles, 6) full face self-assessment; the skin elasticity and skin firmness are measured by the skin elasticity tester Cutometer dualMPA580, and the number of eye corner wrinkles, eye corner wrinkle length, and eye corner wrinkle area are measured by the skin fast optical imaging system PRIMOS-CR; the higher the skin elasticity value, the better the skin elasticity; the lower the skin firmness value, the firmer the skin; The specific steps are as follows: 33 subjects were enrolled, and finally 33 subjects were counted, all of whom were female, aged 26 to 52 years old, with an average age of 42.06±1.20 years old. The test product: the encapsulated bosine of Example 1 was used. The method of use was: use in the morning and evening, after cleaning the face, take 2-3 pumps each time, and apply evenly on the face. Each subject used the test product as required within the specified time. According to the 2015 edition of the "Safety Technical Specifications for Cosmetics", according to the skin reaction record results, no adverse reactions occurred. The safety evaluation results are shown in Table 2: Table 2 Skin reactions in human trials

[0046] Table 3 Results of human efficacy evaluation test

[0047] Table 4 Self-assessment results of human efficacy evaluation

[0048] The results of human efficacy tests show that continuous use of products containing Bossagra has the effect of tightening the skin and anti-wrinkle, and the product is safe and non-irritating. Figure 3 The eye photos of the subjects show that after 4 weeks of use, the length, number and area of ​​the subjects' eye wrinkles have been significantly improved.

[0049] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A method for preparing encapsulated boson, characterized in that: The method comprises the following steps: weighing lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol and dissolving them in anhydrous ethanol to obtain a phospholipid-cholesterol ethanol solution; ultrasonically treating the mixture until no particles or insoluble matter are visible to the naked eye, dissolving hydroxypropyl tetrahydropyrantriol in a phosphate buffer, and stirring the mixture in a water bath at 35-45°C after the mixture is fully dissolved; using a syringe to draw the phospholipid-cholesterol ethanol solution, adding the solution dropwise to the hydroxypropyl tetrahydropyrantriol solution, stirring the mixture to form a liposome suspension, rotary evaporating the suspension to remove ethanol, and then ultrasonically dispersing the suspension in an ice-water bath, and finally extruding the mixture through a membrane to obtain liposomes encapsulating bosone, wherein the choline phospholipid-polyethylene glycol-choline phospholipid and the choline phospholipid-polyethylene glycol-cholesterol have structures shown in Formula 1 and Formula 2, respectively: Formula 1 Formula 2 Wherein, a and b are independently selected from any positive integer between 11 and 15.

2. The method for preparing the encapsulated boson as claimed in claim 1, characterized in that: The mass ratio of the lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol is (10-15): (1-5): (3-5).

3. The method for preparing the encapsulated boson as claimed in claim 1, characterized in that: The molecular weight of the polyethylene glycol in the choline phospholipid-polyethylene glycol-choline phospholipid and the choline phospholipid-polyethylene glycol-cholesterol is 600-2000.

4. The method for preparing the encapsulated boson as claimed in claim 1, characterized in that: The mass ratio of hydroxypropyl tetrahydropyrantriol to the total mass of phospholipids in lecithin, choline phospholipid-polyethylene glycol-choline phospholipid and choline phospholipid-polyethylene glycol-cholesterol is 1:(5~10).

5. The method for preparing the encapsulated boson as claimed in claim 1, characterized in that: The preparation method of the choline phospholipid-polyethylene glycol-choline phospholipid comprises the following steps: S1. A saturated fatty acid, 3-dimethylamino-1,2-propylene glycol, a catalyst and a dehydrating agent are dissolved in dichloromethane, and an esterification reaction is carried out at room temperature to obtain a reactant 1; S2. Under anhydrous and oxygen-free conditions, polyethylene glycol and triethylamine are added to tetrahydrofuran, stirred evenly, and then cooled in an ice-water bath, 2-chloro-2-oxo-1,3,2-dioxaphospholane is slowly added dropwise, the reaction is stirred in the ice-water bath for a period of time, and then the temperature is raised to room temperature and the reaction is continued for a period of time to obtain reactant 2; S3. Add reactant 1 and reactant 2 into acetonitrile and react at 60-80° C. to obtain choline phospholipid-polyethylene glycol-choline phospholipid.

6. The method for preparing the encapsulated boson as claimed in claim 5, characterized in that: The molar ratio of the saturated fatty acid to 3-dimethylamino-1,2-propanediol is (2.1-2.5):1; the saturated fatty acid is selected from at least one of palmitic acid, myristic acid, and lauric acid; the catalyst is 4-dimethylaminopyridine; the dehydrating agent is at least one of N,N'-dicyclohexylcarbodiimide and N,N'-diisopropylcarbodiimide; the molar ratio of the polyethylene glycol to 2-chloro-2-oxo-1,3,2-dioxaphospholane is 1:(2.1-2.5); the molar ratio of the reactant 1 to the reactant 2 is (2.1-2.5):

1.

7. The method for preparing the encapsulated boson as claimed in claim 1, characterized in that: The preparation method of choline phospholipid-polyethylene glycol-cholesterol comprises the following steps: S1. A saturated fatty acid, 3-dimethylamino-1,2-propylene glycol, a catalyst and a dehydrating agent are dissolved in dichloromethane, and an esterification reaction is carried out at room temperature to obtain a reactant 1; S2. Under anhydrous and oxygen-free conditions, hydroxypolyethylene glycol carboxyl and triethylamine were added to tetrahydrofuran, stirred evenly and cooled in an ice-water bath, 2-chloro-2-oxo-1,3,2-dioxaphospholane was slowly added dropwise, the reaction was stirred in the ice-water bath for a period of time, and then the temperature was raised to room temperature and the reaction was continued to obtain reactant 3; S3. adding reactant 1 and reactant 3 to acetonitrile, and reacting at 60-80° C. to obtain choline phospholipid-polyethylene glycol-carboxyl; S4. Dissolve choline phospholipid-polyethylene glycol-carboxyl, cholesterol, a catalyst and a dehydrating agent in dichloromethane, and under the protection of an inert gas, stir the mixture at room temperature to carry out an esterification reaction to obtain choline phospholipid-polyethylene glycol-cholesterol.

8. The method for preparing the encapsulated boson as claimed in claim 7, characterized in that: The molar ratio of the saturated fatty acid to 3-dimethylamino-1,2-propylene glycol is (2.1-2.5):1; the saturated fatty acid is selected from at least one of palmitic acid, myristic acid and lauric acid; the catalyst is 4-dimethylaminopyridine; the dehydrating agent is at least one of N,N'-dicyclohexylcarbodiimide and N,N'-diisopropylcarbodiimide; the molar ratio of the hydroxypolyethylene glycol carboxyl group to 2-chloro-2-oxo-1,3,2-dioxaphospholane is (1.05-1.2):1; the molar ratio of the reactant 1 to the reactant 3 is (1.05-1.2):1, and the molar ratio of the choline phospholipid-polyethylene glycol-carboxyl group to cholesterol is 1:(1.05-1.2).

9. A transdermal Bossein, characterized in that: The encapsulated boson prepared by the preparation method of encapsulated boson according to any one of claims 1 to 8 is dispersed in a mixed solution of water and butanediol at a mass fraction of 10 to 50 wt%.

10. Use of the encapsulated Bossagra obtained by the preparation method of encapsulated Bossagra according to any one of claims 1 to 8 in essence water, lotion, essence milk, face cream and eye cream.

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