Vesicles encapsulating active substances, methods of preparation and use thereof

Abstract

The application provides a vesicle with a double-layer membrane structure, which comprises an outer membrane and an inner membrane, the outer membrane comprises a film forming agent and an inducer, the inner membrane comprises a film forming agent and an inducer, the vesicle further comprises an active substance, the active substance is located inside the inner membrane, and stable and efficient wrapping of the active substance can be realized through film forming agent and double-layer membrane inducer proportion selection.

Description

Technical Field

[0001] This application relates to the field of cosmetic technology, and more particularly to a vesicle, its preparation method and its application, especially to a vesicle when it encapsulates an active substance, its preparation method and its application. Background Technology

[0002] In the cosmetics industry, active ingredients are used to achieve various effects such as moisturizing, whitening, anti-aging, and anti-oxidation. A large portion of these active ingredients are water-soluble. It is well known that the stratum corneum allows lipid-soluble substances to penetrate more easily, while only a very small portion of water-soluble active ingredients can pass through the stratum corneum, thus failing to fully realize their functions. Transdermal absorption of water-soluble active ingredients is typically achieved using methods such as penetration enhancement, hydrophobic modification, and liposome encapsulation. However, penetration enhancement requires large amounts of penetration enhancers, severely interfering with the normal physiology of the stratum corneum and leading to skin redness and itching. Hydrophobic modification usually increases the lipid solubility of the active ingredient, but after absorption, it needs to undergo enzymatic or hydrolytic degradation in the body to exert its effects, thus significantly reducing its efficacy. Liposomes have complex manufacturing processes, are expensive, and are prone to oxidation and deterioration, greatly limiting their application in skincare products. Summary of the Invention

[0003] To address the problems of existing technologies, this application provides a vesicle that overcomes the disadvantages of liposomes, such as high cost and instability. Using the vesicles of this application to encapsulate active substances can effectively improve the encapsulation efficiency and transdermal absorption of water-soluble active ingredients.

[0004] Specifically, this application relates to the following aspects:

[0005] 1. A vesicle, wherein the vesicle has a bilayer membrane structure, comprising an outer membrane and an inner membrane, the outer membrane comprising a film-forming agent and an inducing agent, and the inner membrane comprising a film-forming agent and an inducing agent.

[0006] 2. The vesicle according to claim 1, wherein the vesicle further includes an active substance located inside the inner membrane.

[0007] 3. The vesicle according to item 1, wherein the film-forming agent is a fatty alcohol polyether emulsifier, preferably, the film-forming agent includes stearyl alcohol polyether-2 and / or oleyl alcohol polyether-3;

[0008] The inducer is cholesterol or a derivative thereof;

[0009] More preferably,

[0010] The mass ratio of the film-forming agent to the inducing agent is 1:2-2:1, preferably 1:1.5-1.5:1.

[0011] 4. The vesicle according to item 1, wherein,

[0012] The active substance is selected from one or more of the following: ectoine, ergothioneine, polypeptide, glutathione, ascorbic acid, nicotinamide, asiaticoside, and NMN.

[0013] 5. The vesicle according to item 1, wherein,

[0014] The film-forming agent is 1 wt% to 10 wt% of the vesicles by mass percentage, preferably 2 wt% to 8 wt%.

[0015] More preferably,

[0016] The inducing agent is 1 wt% to 10 wt% of the mass percentage of the vesicles; preferably, the inducing agent is 2 wt% to 8 wt%.

[0017] 6. A method for preparing vesicles, wherein,

[0018] Preparation of the oil phase: The film-forming agent and the inducing agent are dissolved in liquid alcohol and heated to dissolve, thus obtaining the oil phase;

[0019] Preparation of the aqueous phase: Water is used as the aqueous phase, or the active substance is dissolved in water and heated to obtain the aqueous phase;

[0020] Mixing: While homogenizing, the aqueous phase is added to the oil phase. After cooling, the mixture is stirred to obtain vesicles.

[0021] 7. A composition, characterized in that it comprises vesicles as described in any one of claims 1-4 or vesicles prepared by the method of claim 5.

[0022] The beneficial effects of this application include at least the following aspects:

[0023] (1) The vesicles of this application not only have good stability, but also can achieve efficient encapsulation of active substances.

[0024] (2) The vesicles of this application are also stable at high temperatures.

[0025] (3) The vesicles of this application can promote the permeable absorption of active substances into the skin. Attached Figure Description

[0026] Figure 1 Microscopic photograph of the vesicles in Example 2;

[0027] Figure 2 This is a microscopic photograph of the vesicles in Comparative Example 6.

[0028] Figure 3 This is a microscopic photograph of Example 2 taken after being placed at 45°C for one month.

[0029] Figure 4 Microscopic images of liposomes

[0030] Figure 5 Microscopic images of liposomes after being stored at 45°C for one month. Detailed Implementation

[0031] The present application is further illustrated below with reference to embodiments. It should be understood that the embodiments are only used to further illustrate and explain the present application and are not intended to limit the present application.

[0032] Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. While similar or identical methods and materials may be applied in experimental or practical applications, materials and methods are described herein. In case of conflict, the definitions included herein shall prevail. Furthermore, materials, methods, and examples are for illustrative purposes only and are not intended to be limiting. The present application is further described below with reference to specific embodiments, but is not intended to limit the scope of the application.

[0033] This application provides a vesicle with a bilayer membrane structure, comprising an outer membrane and an inner membrane. The outer membrane includes a film-forming agent and an inducing agent, and the inner membrane includes both a film-forming agent and an inducing agent. The vesicle also includes an active substance located inside the inner membrane, encapsulated within the bilayer membrane structure. The inducing agent is embedded between the film-forming agents in a regular or irregular manner. The arrangement of the film-forming agent and the inducing agent in the outer membrane can be the same as or different from that in the inner membrane. The vesicle of this application forms a bilayer membrane structure based on the properties and structure of the hydrophilic and lipophilic groups of the film-forming agent and the inducing agent.

[0034] In some embodiments of this application, the film-forming agent is 1 wt%-10 wt% of the vesicles by mass percentage, preferably 2 wt%-8 wt%.

[0035] For example, the film-forming agent may be 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, or any range thereof, based on the mass percentage of the vesicles.

[0036] In some embodiments of this application, the inducing agent is 1 wt%-10 wt% of the mass percentage of the vesicles, preferably 2 wt%-8 wt%.

[0037] For example, the inducer may be 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, or any range thereof, as a percentage of the mass of the vesicle.

[0038] In some embodiments of this application, the mass ratio of the film-forming agent to the inducing agent is 1:2-2:1, more preferably 1:1.5-1.5:1. For example, the mass ratio of the film-forming agent to the inducing agent can be 1:2.5, 1:2, 1:1.5, 1:1, 2:1.5, 2:1, 2.5:1 or any range therebetween.

[0039] In some embodiments of this application, the film-forming agent is a fatty alcohol polyether emulsifier, preferably, the film-forming agent includes stearyl alcohol polyether-2 and / or oleyl alcohol polyether-3.

[0040] In some embodiments of this application, the inducing agent is cholesterol or a derivative thereof. The cholesterol derivatives include, but are not limited to, 7-dehydrocholesterol, C10-C40 isoalkyl sterol esters, cholesterol macadamia oleate, cholesterol butyrate, cholesterol dichlorobenzoate, cholesterol succinate, cholesterol hydroxystearate, cholesterol nonanoate, cholesterol lanolinate, cholesterol isostearyl carbonate, cholesterol isostearyl carbonate, cholesterol oleate, dihydrocholesterol, dihydrocholesterol butyrate, dihydrocholesterol oleate, and cholesterol chloride.

[0041] In some embodiments of this application, the active substance may be an active substance that needs to enter cells to exert its effects, and the active substance is selected from one or more of ectoine, ergothioneine, polypeptide, glutathione, ascorbic acid, nicotinamide, asiaticoside, and β-nicotinamide mononucleotide (NMN).

[0042] This application also provides a method for preparing the above-mentioned vesicles, comprising the following steps:

[0043] Preparation of the oil phase: The film-forming agent and the inducing agent are dissolved in liquid alcohol to obtain the oil phase;

[0044] Preparation of the aqueous phase: The active substance is dissolved in water to obtain the aqueous phase;

[0045] Mixing: While homogenizing, the aqueous phase is added to the oil phase. After cooling, water is added and stirred to obtain vesicles.

[0046] Furthermore, the dissolution can be achieved by heating.

[0047] Preferably, the method for preparing vesicles according to this application includes the following steps:

[0048] Preparation of the oil phase: The film-forming agent and the inducing agent are dissolved in liquid alcohol and heated to dissolve, thus obtaining the oil phase;

[0049] Preparation of the aqueous phase: Water is used as the aqueous phase, or the active substance is dissolved in water to obtain the aqueous phase;

[0050] Mixing: While homogenizing, the aqueous phase is added to the oil phase. After cooling, water is added and stirred to obtain vesicles.

[0051] This application does not impose specific restrictions on the heating temperature in the oil phase preparation step, as long as the oil phase is in a dissolved state. Those skilled in the art can freely choose the temperature according to the properties of the oil phase. In some embodiments of this application, the heating temperature is 50-95℃, for example, it can be 50℃, 55℃, 60℃, 65℃, 70℃, 75℃, 80℃, 85℃, 90℃, 95℃, etc.

[0052] This application does not impose specific restrictions on the heating temperature in the aqueous phase preparation step, as long as the addition of the aqueous phase to the oil phase does not affect the dissolution state of the oil phase. Those skilled in the art can freely choose the temperature. In some embodiments of this application, the heating temperature is 50-95℃, for example, it can be 50℃, 55℃, 60℃, 65℃, 70℃, 75℃, 80℃, 85℃, 90℃, 95℃, etc.

[0053] In some embodiments of this application, the liquid alcohol is an alcohol that is liquid at room temperature. This application does not impose any restrictions, and those skilled in the art can select from existing liquid alcohols. Preferably, the liquid alcohol is a polyol, such as ethylene glycol, propylene glycol, butanediol, hexanediol, pentanediol, glycerol, etc.

[0054] This application also provides a cosmetic composition comprising the above-described vesicles.

[0055] This application also provides the use of the above-described vesicles or vesicles prepared by the above-described method or the above-described compositions in cosmetics.

[0056] This application also provides the use of the above-described vesicles or vesicles prepared by the above method for non-therapeutic purposes in enhancing the transdermal absorption of active substances.

[0057] Example

[0058] This application provides a general and / or specific description of the materials and test methods used in the experiments. In the following examples, unless otherwise specified, % represents wt%, i.e., weight percentage. Reagents or instruments used, unless otherwise specified, are all commercially available conventional reagent products.

[0059] Example 1

[0060] Dissolve 4g stearyl alcohol ether and 8g cholesterol in 4g polyol (dipropylene glycol), heat to 80℃ and mix evenly to obtain the oil phase;

[0061] Dissolve 4g of ergothioneine in 16g of water and heat to 80℃ to obtain an aqueous phase;

[0062] While homogenizing, the aqueous phase was added to the oil phase. After cooling, 64g of water was added, bringing the total mass to 100g. The mixture was stirred until homogeneous to obtain vesicles.

[0063] Example 2-11

[0064] Examples 2-11 were prepared using the same method as in Example 1.

[0065] Comparative Example 1

[0066] Dissolve 4g of cholesterol in 4g of polyol (dipropylene glycol), heat to 80℃ and mix thoroughly to obtain the oil phase;

[0067] Dissolve 4g of ergothioneine in 16g of water and heat to 80℃ to obtain an aqueous phase;

[0068] While homogenizing, the aqueous phase was added to the oil phase. After cooling, 72g of water was added, bringing the total mass to 100g. After stirring evenly, vesicles could not be obtained.

[0069] Comparative Example 2

[0070] Dissolve 4g of stearyl alcohol ether in 4g of polyol (dipropylene glycol), heat to 80℃ and mix evenly to obtain the oil phase;

[0071] Dissolve 4g of ergothioneine in 16g of water and heat to 80℃ to obtain an aqueous phase;

[0072] While homogenizing, the aqueous phase was added to the oil phase. After cooling, 72g of water was added, bringing the total mass to 100g. After stirring evenly, vesicles could not be obtained.

[0073] Comparative Example 3

[0074] Prepared using the same method as Comparative Example 1.

[0075] Comparative Example 4

[0076] Prepared using the same method as Comparative Example 2.

[0077] Comparative Examples 5-8

[0078] Prepared using the same method as in Example 1.

[0079] The compositions of Examples 1-11 and Comparative Examples 1-8 are shown in Table 1:

[0080] Table 1

[0081]

[0082] Experimental Example

[0083] Experiment Example 1 Stability Test

[0084] Examples 1-11 and Comparative Examples 1-8 were diluted to 1% aqueous solution, and their particle size and zeta potential were measured using an Anton Paar Litesizer 500 instrument; at the same time, the appearance of the vesicles was observed under a microscope. The results are shown in Table 2.

[0085] Experiment Example 2 Encapsulation Efficiency Determination

[0086] The encapsulation efficiency of the vesicles encapsulated with ergothioneine or ectoine prepared in Examples 1-11 and Comparative Examples 1-8 was determined.

[0087] The encapsulation efficiency of the active ingredient was tested using ultrafiltration centrifugation: A certain proportion of the vesicle solution was diluted, and a precise amount of the solution was weighed and placed in an ultrafiltration centrifuge tube. Suitable rotation speed and time were set, and the samples from the examples and comparative examples were centrifuged. After centrifugation, the content of the active ingredient in the separated aqueous solution was detected and compared with the content of the active ingredient in the feed solution to determine the content of the encapsulated active ingredient. The test results are shown in Table 2.

[0088] Table 2

[0089]

[0090] Comparative Examples 5 and 7 show that when the cholesterol content is significantly lower than that of stearyl alcohol polyether, no vesicle structures are observed under the microscope. Comparative Examples 6 and 8 show that when the cholesterol content is significantly higher than that of stearyl alcohol polyether, no vesicle structures are observed under the microscope, and a large amount of crystals precipitate out. Therefore, stearyl alcohol polyether and cholesterol need to be within a certain ratio range to form structurally complete vesicles and obtain a high encapsulation efficiency.

[0091] As shown in Examples 1-11, if the amount of film-forming agent and inducer added is insufficient, the active substance cannot be effectively encapsulated.

[0092] As can be seen from Comparative Examples 1-4, the formation of vesicle structures requires the simultaneous presence of both film-forming agents and inducing agents; neither can be omitted.

[0093] A comparison of Examples 2 and 11 shows that the active substance has no effect on the stability of the vesicles of this application.

[0094] Experiment Example 3 High Temperature Stability Test

[0095] High-temperature stability tests were conducted on the samples from Example 2 and liposomes.

[0096] The preparation method of liposome samples is as follows: 4g of lecithin and 4g of cholesterol are dissolved in 4g of polyol (dipropylene glycol), heated to 80℃ and mixed evenly to obtain an oil phase; 4g of ergothioneine is dissolved in 16g of water and heated to 80℃ to obtain an aqueous phase; while homogenizing, the aqueous phase is added to the oil phase, and after cooling, 64g of water is added, with a total mass of 100g, and stirred evenly to obtain liposomes.

[0097] Figure 1 This is a microscope photograph of Example 2, showing a clear vesicle structure under the microscope.

[0098] Figure 3 This is a microscope image of Example 2 after being placed at 45°C for one month, compared with... Figure 1 In contrast, the vesicle structure did not show significant changes.

[0099] Figure 4 The liposome sample prepared by the above method shows a clear vesicle structure under a microscope.

[0100] Figure 5 These are microscope images of the liposome samples after they have been placed at 45°C for one month.

[0101] As shown in the figure, the structure of liposomes lost after being placed at high temperatures, and a large amount of crystals precipitated out. The vesicles of this application retained their vesicle structure after being placed at high temperatures, and no crystal precipitation was observed, demonstrating good high-temperature stability.

[0102] Experimental Example 4: In vitro transdermal test

[0103] Transdermal tests were performed on the sample from Example 2 and the sample in the unencapsulated state of the active ingredient (free active ingredient).

[0104] The preparation method for samples with unencapsulated active ingredients is as follows:

[0105] Dissolve 4g stearyl alcohol ether and 4g cholesterol in 4g polyol (dipropylene glycol), heat to 80℃ and mix evenly to form the oil phase;

[0106] Dissolve 4g of ergothioneine in 83.9g of water and heat to 80℃ to obtain the aqueous phase.

[0107] The aqueous phase is added to the oil phase and emulsified directly to obtain the final product.

[0108] Test Method: Pigskin was fixed between the supply and receiving tanks of a Franz apparatus, with the cuticle facing the supply tank. Equal masses of emulsion were applied to the surface of the pigskin, using phosphate buffer solution as the receiving solution. The receiving system was placed in a constant temperature system at 32℃±2℃, with magnetic stirring at 300 rpm. At 2, 4, 8, and 24 hours, 1 mL of the receiving solution was collected (filtered through a 0.45 μm membrane and analyzed by HPLC), and the same volume of receiving solution was added. After 24 hours, the experiment was terminated, and the pigskin was immediately removed. After cleaning off any remaining sample, the pigskin was cut into small pieces, mixed with 5 mL of the receiving solution, and sonicated for 30 min. After centrifugation for 10 min, the supernatant was filtered through a 0.45 μm membrane, and the content was analyzed by HPLC. The test results are shown in Table 3.

[0109] Table 3. Transdermal Transfusion Rate Test Results

[0110] time Example 2: Permeation rate (ug) after encapsulating ergothioneine Unencapsulated ergothioneine permeation / ug 2 hours 77 57 4 hours 158 105 8 hours 219 173 24 hours 4797 2751 Total throughput 5252 3086

[0111] As shown in Experiment 4, when the active substance is encapsulated on the inner side of the vesicle membrane, it is easier for the active substance to penetrate into the skin.

[0112] In summary, the vesicles of this application exhibit good stability and high encapsulation efficiency of active substances. Furthermore, the vesicles of this application demonstrate good dispersibility in aqueous solutions, reduced viscosity, and increased fluidity, thereby improving the ease of use and making them readily incorporateable into various dosage forms.

[0113] Although the present invention has been disclosed above with reference to embodiments, it is not intended to limit the present invention. Anyone skilled in the art may make some modifications and refinements without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be determined by the scope of the appended patent application.

Claims

1. A vesicle, characterized in that, The vesicle has a bilayer membrane structure, including an outer membrane and an inner membrane. The outer membrane includes a film-forming agent and an inducing agent, and the inner membrane includes a film-forming agent and an inducing agent. The vesicles also include active substances located inside the inner membrane. The active substance is selected from one or two of ectoine and ergothioneine, and the inducer is cholesterol. The film-forming agent includes stearyl alcohol polyether-2 and / or oleyl alcohol polyether-3; The mass ratio of the film-forming agent to the inducing agent is 1:2-2:

1.

2. The vesicle according to claim 1, characterized in that, The film-forming agent is 1 wt% to 10 wt% of the mass percentage of the vesicles.

3. The vesicle according to claim 2, characterized in that, The film-forming agent is 2 wt%-8 wt% based on the mass percentage of the vesicles.

4. The vesicle according to claim 1, characterized in that, The inducing agent is 1 wt% to 10 wt% of the mass percentage of the vesicles.

5. The vesicle according to claim 4, characterized in that, The inducing agent is 2 wt% to 8 wt% of the mass percentage of the vesicles.

6. A method for preparing vesicles according to any one of claims 1-5, characterized in that, Preparation of the oil phase: The film-forming agent and the inducing agent are dissolved in liquid alcohol and heated to dissolve, thus obtaining the oil phase; Preparation of the aqueous phase: Water is used as the aqueous phase, or the active substance is dissolved in water to obtain the aqueous phase; Mixing: While homogenizing, the aqueous phase is added to the oil phase. After cooling, water is added and stirred to obtain vesicles.

7. A cosmetic composition, characterized in that, The vesicles include any one of claims 1-5 or vesicles prepared by the method of claim 6.

8. Use of the vesicles according to any one of claims 1-5 or the vesicles prepared by the method according to claim 6 in the preparation of cosmetics.

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