Stable inclusion composition containing retinoid family as well as preparation method and application of stable inclusion composition

Abstract

The invention discloses a stable inclusion composition containing a retinoid family as well as a preparation method and application of the stable inclusion composition, and the composition comprises the following components in percentage by mass: 0.02%-5% of retinoid family active ingredients, 0.1%-4% of a surfactant, 0.05%-10% of skin moistening lipid, 10%-35% of a humectant, 0.01%-3% of a light protective agent and 1%-16% of an antiseptic bacteriostatic agent, and 50%-82% of deionized water. The inclusion composition has high encapsulation efficiency and good stability, can maintain structural integrity and reduce skin irritation in the transdermal delivery process, is safe and mild, promotes rapid transdermal permeation of the retinoid family, improves the distribution condition of active ingredients in the skin, increases the proportion of the active ingredients in the active epidermis and corium layer, and improves the retinoid family transdermal delivery effect. The method has the application potential of accurately positioning and conveying, resisting basilar membrane damage caused by external factors, promoting metabolism of epidermis and dermis, and moistening and moisturizing.

Description

Technical Field

[0001] The present invention relates to the fields of cosmetics and biomedicine, and in particular to a stable inclusion composition containing a retinoid family, and a preparation method and application thereof. Background Art

[0002] As the largest organ of human beings, the skin is in direct contact with the external environment and maintains the stability of the internal environment of the human body. The skin is mainly composed of three parts: the epidermis, the dermis and the subcutaneous tissue. The stratum corneum located at the outermost part of the skin contains a large amount of lipids, forming a certain physical barrier function, which can resist the invasion of external bacteria and is called the "first line of defense" of the skin. In addition, the basement membrane located at the junction of the true epidermis has also been considered as another new barrier of the skin in recent years. It has selective permeability and can allow small molecules to pass through, while large molecules with a molecular weight higher than 40kD, such as inflammatory factors, are difficult to pass through, which ensures the health of the skin. The basement membrane is mainly composed of type IV collagen and laminin, which can play a comprehensive role in structural support, signal transduction and nutrient penetration metabolism for the skin. It is an important structure to maintain skin firmness and smooth the "golden cycle" of epidermis-basement membrane-dermis. When the skin is exposed to ultraviolet rays or other adverse factors for a long time, not only the physical barrier is damaged, but the basement membrane zone will also be damaged or broken, and the structural support of the epidermis and dermis and the transportation of nutrients are affected, thus causing a series of skin problems such as erythema, wrinkles, and pigmentation. Therefore, how to reasonably and effectively resist continuous damage to multiple skin barriers has gradually become a hot topic of concern.

[0003] The retinoid family is a classic anti-aging and acne treatment ingredient that is widely favored by consumers. It can stimulate the proliferation of keratinocytes and fibroblasts, promote the production of type I collagen, and regenerate the skin; in addition, the retinoid family has also been found to have the effect of inhibiting matrix metalloproteinases, so they can resist the continuous degradation of type IV collagen and repair damaged basement membranes.

[0004] However, there are still many problems in the practical application of the retinoid family. First, there are multiple conjugated double bonds in the molecular structure of this class of compounds, making them sensitive to light, temperature, oxygen, etc., and prone to inactivation and failure, which makes formulation processing and storage difficult. Second, the stratum corneum, which is located on the outermost part of the skin, is composed of 15-20 layers of flat dead keratinocytes, and it forms a firm "brick wall structure" with intercellular lipids, etc. It is generally considered to be the determining step of the percutaneous penetration rate of molecules. Only molecules with appropriate oil-water partition coefficients can penetrate. However, the retinoid family is extremely lipophilic and tends to remain in the stratum corneum during percutaneous penetration, making it difficult to be delivered to the basement membrane or deeper layers for efficient repair. Third, the retinoid family can bind to transient receptor potential vanilloid 1 (TRPV1) that is abundantly present in the viable epidermis, causing severe local irritation, and most users report difficulty in establishing skin tolerance. Some studies have used zebrafish experiments to verify that retinol can stimulate zebrafish to cause large-area and long-term inflammatory pain, resulting in an increase in the movement trajectory and an acceleration in speed. Sewon Kang et al. found that when subjects applied only 0.025% retinoic acid for 4 days, the erythema score was 3.7 times that of the control group, showing its extremely strong skin irritation. Fourth, the strong lipophilicity makes it necessary to add a certain amount of oil phase to meet the requirement of uniform dispersion in the formulation, which limits the addition amount in the product, resulting in the inability to fully exert the efficacy of the retinoid family. Excessive surfactants added to some products to increase the dosage may cause damage to the skin barrier function after long-term use, reducing the skin's resistance to external stimuli.

[0005] Nanocarriers have been widely used because they can improve drug stability and percutaneous absorption rate, control drug release rate, and reduce drug irritation to the skin. However, in the face of many problems coexisting in the skin structure characteristics and drug molecules themselves, there are still many problems and deficiencies in the design of existing nanocarriers. For example, the intercellular spaces of the skin are very narrow, and nanocarriers are extremely prone to disintegration and fragmentation when infiltrating. Currently, common methods such as adding surfactants like Tween 80 are used to adjust the fluidity of the carrier to reduce nanoscale fragmentation. However, the results are still not ideal. The enhanced permeability effect shown by such methods is mainly due to the disruption of the skin lipid arrangement by the surfactant, and the carrier itself still fails to effectively maintain its original structure during the transdermal process, resulting in premature drug release, reduced skin permeability, and the leaked drugs still showing significant skin irritation. Therefore, while ensuring that the nanocarrier has appropriate parameters to pass through the micropores of the skin, it is necessary to maintain the structural stability of the carrier during skin penetration, and be able to control the loaded drug so that it does not enter the blood, but can be stably and continuously delivered to the viable epidermis or dermis. On the one hand, this can improve the transdermal efficiency and drug efficacy, and on the other hand, it can also prevent the nanocarrier from prematurely rupturing and decomposing during transdermal delivery or storage, contacting the TRPV1 receptor and causing strong skin irritation, thereby improving the compliance of use.

[0006] In the prior art, a variety of encapsulation strategies have been proposed to solve some of the above-mentioned problems of the retinoid family when used externally. For example, Chinese patent CN111658632A discloses a calcium acetate gradient method for preparing retinoic acid liposomes, which can achieve the purpose of improving encapsulation rate and stability. However, its operation steps are complicated and difficult to achieve industrialization. Studies have shown that the shelf life of liposomes themselves is relatively short, and there is a problem of long-term drug leakage. At the same time, it is difficult to maintain structural integrity during the transdermal process, so the practical application of such nanocarriers is limited. Chinese patent CN108451787B also discloses a nanolipid carrier for embedding vitamin A alcohol and a preparation method thereof. The invention uses solid oil palm wax with a high melting point and surfactant decyl glucoside and other raw materials to embed vitamin A alcohol, but the preparation temperature is higher than 85°C, which easily causes the heat-sensitive retinoid family to be destroyed during the preparation process, reducing the actual acquisition rate.

[0007] In addition, some scholars have used microcapsules and microspheres to encapsulate retinoids (Chinese patent CN112741773A, US patent US20030232091A1), which stabilizes the active ingredients and can also achieve the purpose of slow release. However, in actual applications, microspheres or microcapsules are difficult to penetrate the stratum corneum due to their large particle size. They are usually broken on the skin surface under the pressure of hand application and then penetrate through the skin. Therefore, they only stabilize the active ingredients and facilitate storage, but do not significantly improve the penetration characteristics of the active ingredients in the skin. In addition, these patents add silicone analogs to the formula. Long-term use and contact with the skin will damage the skin barrier, thereby increasing the user's skin sensitivity to the retinoid family.

[0008] Therefore, designing and developing an inclusion body that does not damage the skin barrier function and has a strong and stable structure will help to achieve the delivery of active ingredients deep in the skin and reduce the skin irritation of retinoids, which is conducive to improving the utilization rate of active ingredients and consumer compliance. At the same time, it is also very important to develop a preparation method that is simple to operate and does not affect the stability of retinoids during the production process.

[0009] Based on this, the present invention proposes a new type of flexible and stable inclusion composition. By rationally matching surfactants, emollient lipids, photoprotectants, moisturizers and preservatives and antibacterial agents, the characteristics of the inclusion itself are regulated, so that it can improve the stability of the retinoid family while reducing skin irritation, maintaining a stable structure to facilitate deep penetration of ingredients, accurately delivering them to the site of action, repairing damaged basement membranes, and avoiding the risk of entering the blood, ensuring safe use and eliminating user concerns. Summary of the invention

[0010] The purpose of the present invention is to provide a stable inclusion composition containing a retinoid family, and a preparation method and application thereof. The method has mild conditions, can improve the stability and skin irritation of the retinoid family, and achieve precise transdermal delivery with a more complete structure. The inclusion composition can be used to rebuild the skin barrier.

[0011] The purpose of the present invention can be achieved through the following technical solutions:

[0012] In the first aspect, the present invention claims protection for a stable inclusion composition containing a retinoid family, wherein the composition comprises the following components in percentage by mass: 0.02%-5% of a retinoid family active ingredient, 0.1%-4% of a surfactant, 0.05%-10% of an emollient lipid, 10%-35% of a moisturizer, 0.01%-3% of a photoprotectant, 1%-16% of a preservative and an antibacterial agent, and 50%-82% of deionized water.

[0013] Furthermore, the retinoid family active ingredient is selected from one or more of vitamin A, vitamin A palmitate, retinoic acid, and vitamin A aldehyde.

[0014] Furthermore, the surfactant is selected from one or more of polysorbate-80, polysorbate-20, polyglyceryl-6 distearate, sodium stearoyl glutamate, sodium cocoyl glutamate, egg yolk lecithin, soybean lecithin, hydrogenated lecithin, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylethanolamine, and dioleoyl phosphatidylethanolamine. The present invention improves the elasticity of the inclusion body by rationally assembling and matching the surfactant and the emollient lipid, and forming a special structure through intermolecular forces such as hydrogen bonds and van der Waals forces, thereby achieving efficient encapsulation and stabilization of the retinoid family.

[0015] Furthermore, the emollient lipid is selected from one or more of cetyl alcohol, shea butter, polydimethylsiloxane, cyclopentasiloxane, squalane, jojoba oil, sweet almond oil, isononyl isononanoate, isostearyl isostearate, beeswax, ceramide I, ceramide II, ceramide III, ceramide IV and ceramide VI, cholesterol, stearic acid, glyceryl behenate, and myristic acid; the present invention selects lipids that are similar in composition to the skin and have an emollient effect, so that the ingredients can quickly penetrate the stratum corneum into the deep layer of the skin, further enhancing the skin's physical and basement membrane barrier functions, and is safe and non-irritating.

[0016] Furthermore, the moisturizing agent is selected from one or more of propylene glycol, glycerol, butylene glycol, pentylene glycol, allantoin, sodium pyrrolidone, trehalose, serum albumin, whey protein, hydrolyzed soy protein, hydrolyzed wheat protein, hydrolyzed rice protein, hyaluronic acid, sodium hyaluronate, and dipropylene glycol; the present invention improves skin hydration by adding a moisturizing agent, alleviates skin dryness, itching, and the like caused by retinoid family components, and can also assist the active ingredients in penetrating the skin and improve their skin absorption rate.

[0017] Furthermore, the photoprotectant is selected from one or more of tocopherol, troxerutin, titanium dioxide, zinc oxide, astaxanthin, resveratrol, ectoine, vitamin C ethyl ether, ferulic acid, soybean oil, and sunflower seed oil; the retinoid family components are highly photo-instable, and the present invention utilizes ingredients that are beneficial to the skin and have anti-ultraviolet or blue light effects to avoid degradation of retinoids during use, and synergizes with the inclusions to improve the photoinstability of the retinoid family. At the same time, the photoprotectant itself has the ability to repair the skin, and can work together with the retinoid to deeply repair the skin basement membrane barrier.

[0018] Furthermore, the preservative and antibacterial agent is selected from one or more of sodium chloride, phenoxyethanol, methylparaben, ethylparaben, propylparaben, ethanol, tert-butyl alcohol, benzoic acid, sodium benzoate, peppermint oil, potassium sorbate, benzalkonium chloride, p-hydroxyacetophenone, and ethylhexylglycerin; the aqueous solution is very likely to breed microorganisms such as bacteria and fungi during storage, resulting in a decrease in product quality or deterioration. Therefore, the present invention is unique in that the preservative and antibacterial agent is added to the inclusion structure, which can ensure the stability of the structure while extending the storage time of the inclusion and ensuring the quality of the inclusion.

[0019] In a second aspect, the present invention claims a method for preparing the above-mentioned stable inclusion composition containing the retinoid family, the method comprising the following steps:

[0020] S1. The retinoid family active ingredients, surfactants, emollients, photoprotectants and fat-soluble preservatives and antibacterial agents are mixed uniformly in ethanol and sonicated at 30-65° C. until completely dissolved to form phase A;

[0021] S2. Separately take a moisturizer and deionized water or a moisturizer, a water-soluble preservative and antibacterial agent and deionized water and mix well to form a phase B;

[0022] S3. After preheating the phase A and phase B to 30-65°C, continuously dropwise add phase A to phase B at a stirring speed of 500-2000 r / min, and stir and hydrate for 10-120 min at the temperature and speed to obtain phase C;

[0023] S4. Phase C obtained in S3 is added to a high-pressure homogenizer for homogenization at a homogenization pressure of 300-1500 Bar and the homogenization cycle is 9-250 times;

[0024] S5. The solution obtained in S4 is dialyzed, ultra-high-speed centrifuged or freeze-dried to obtain an inclusion composition, which can be in the form of a uniform fluid liquid, a viscous semi-solid or a powder.

[0025] In the third aspect, the present invention claims the use of the above-mentioned stable inclusion composition containing the retinoid family in the preparation of cosmetics or pharmaceutical preparations. The cosmetics or pharmaceutical preparations can be used for damaged skin repair. Furthermore, the cosmetics are lotions, essences, creams, lotions, masks, etc.; the pharmaceutical preparations are solutions, creams, ointments, gels, etc.

[0026] The inclusion composition of the present invention exhibits good structural and drug molecule stability, accurately repairs damaged skin basement membrane barriers and epidermal barriers, and stably and slowly releases active ingredients to exert a continuous repair effect, and can be applied to cosmetics or pharmaceutical preparations.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The present invention compounds surfactants, emollient lipids, moisturizers, light protectants and antiseptics in a certain proportion, synergistically, and self-assembles to form a wrapping structure after mixing, which can encapsulate the retinoid family inside, and obtain intermolecular forces with stronger binding force through internal and external stabilizing layers. This assembly structure and stable properties can significantly improve the solubility and stability while maintaining the activity of the active ingredients, thereby improving its applicability.

[0029] The present invention assists the inclusion composition in improving the light stability of the retinoid family components and reducing the degradation of the components by adding a unique light protectant. At the same time, the preservative and antibacterial agent in the formula can also prevent the growth and reproduction of microorganisms, thereby ensuring the long-term storage quality of the inclusion.

[0030] The inclusion composition provided by the present invention has the characteristic of slow release, which effectively reduces the skin irritation of the retinoid family, is conducive to the long-term and healthy repair of the skin basement membrane barrier and epidermal barrier, promotes skin microcirculation, improves skin color, and delays skin aging.

[0031] The present invention can change the penetration distribution of retinoids in various skin layers, increase the penetration rate of active substances, reduce stratum corneum accumulation while increasing the true epidermal retention, accelerate the combination of ingredients with targets, and shorten the onset time.

[0032] Through the combination of emollient lipids and moisturizers, a moisturizing film can be formed on the skin surface, which can repair the damaged physical barrier of the skin, increase the water content of the stratum corneum, improve skin wrinkles and roughness, promote penetration and enhance the repair effect of the retinoid family on the skin. Long-term use can produce beneficial effects on the skin.

[0033] The present invention provides a new retinoid encapsulation method, which is carried out under mild conditions and in a short time, thus avoiding the activity loss caused by high temperature and long preparation time during the preparation process, and is simple to operate and suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creativity and labor. Among them:

[0035] Figure 1 This is a graph showing the particle size of the inclusion composition prepared in the examples.

[0036] Figure 2 This is a diagram showing the encapsulation efficiency of the inclusion composition prepared in the examples.

[0037] Figure 3 Oxygen stability of the retinoid family in the inclusion composition prepared in the examples.

[0038] Figure 4 The photostability of the retinoid family in the inclusion composition prepared in the examples.

[0039] Figure 5 Transdermal structural integrity of the inclusion composition prepared in the examples.

[0040] Figure 6 This is the basement membrane repair ability of the inclusion composition prepared in the example.

[0041] Figure 7 The skin irritation of the inclusion composition prepared in the example. DETAILED DESCRIPTION

[0042] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments. These embodiments are provided in order to understand the present invention more thoroughly and completely, and they cannot be understood as limiting the scope of protection of the present invention. Professionals and technicians in this field make non-essential corrections and adjustments based on the content of the above invention, which still belong to the scope of protection of the present invention. Unless otherwise specified, the technical means used in the following embodiments are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following embodiments, unless otherwise specified, can be obtained from commercial channels.

[0043] Example 1

[0044] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 0.05% vitamin A, 1.5% polysorbate 80, 0.2% jojoba oil, 12% dipropylene glycol, 2.4% troxerutin, 5% peppermint oil, and 78.85% deionized water.

[0045] Example 1 was prepared by the following preparation method, comprising the following steps:

[0046] (1) jojoba oil, troxerutin, vitamin A, polysorbate 80 and peppermint oil are mixed in ethanol and heated at 50° C. and ultrasonicated until completely dissolved to form phase A;

[0047] (2) taking another dipropylene glycol and mixing it with deionized water to form phase B;

[0048] (3) After preheating the phase A and phase B to 50° C., phase A was continuously added dropwise to phase B while stirring at a speed of 600 r / min, and the mixture was stirred and hydrated at the same temperature for 35 min to obtain phase C;

[0049] (4) Add phase C into a high-pressure homogenizer for homogenization at a homogenization pressure of 1000 Bar and cycle homogenization 72 times;

[0050] (5) The solution obtained in step 4 is dialyzed to obtain the inclusion composition.

[0051] Example 2

[0052] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 1% vitamin A, 1.5% polysorbate-20, 0.2% squalane, 12% pentylene glycol, 2.4% tocopherol, 5% ethylparaben, and 77.9% deionized water.

[0053] Example 2 was prepared by the following preparation method, comprising the following steps:

[0054] (1) Squalane, tocopherol, vitamin A, polysorbate 20 and ethylparaben are mixed in ethanol and heated at 50° C. and ultrasonicated until completely dissolved to form phase A;

[0055] (2) taking another pentanediol and mixing it with deionized water to form phase B;

[0056] (3) After preheating the phase A and phase B to 50° C., phase A is continuously added dropwise to phase B while stirring at a speed of 600 r / min, and the mixture is stirred and hydrated at the same temperature for 50 min to obtain phase C;

[0057] (4) Add phase C into a high-pressure homogenizer for homogenization at a homogenization pressure of 1000 Bar and cycle homogenization 72 times;

[0058] (5) The solution obtained in step 4 is dialyzed to obtain the inclusion composition.

[0059] Example 3

[0060] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 0.05% retinoic acid, 1.5% distearoylphosphatidylethanolamine, 0.2% shea butter, 12% glycerol, 2% resveratrol, 5% phenoxyethanol, and 79.25% deionized water.

[0061] Example 3 was prepared by the following preparation method, comprising the following steps:

[0062] (1) distearoylphosphatidylethanolamine, shea butter, resveratrol, retinoic acid and phenoxyethanol are mixed in ethanol, heated and ultrasonicated at 55° C. until completely dissolved to form phase A;

[0063] (2) taking another glycerol and mixing it with deionized water to form phase B;

[0064] (3) After preheating the phase A and phase B to 55° C., phase A was continuously added dropwise to phase B while stirring at a speed of 600 r / min, and the mixture was stirred and hydrated at the same temperature for 35 min to obtain phase C;

[0065] (4) adding phase C into a high-pressure homogenizer for high-pressure homogenization at a homogenization pressure of 1000 Bar and cyclic homogenization for 144 times;

[0066] (5) The solution obtained in step 4 is subjected to ultra-high speed centrifugation to obtain the inclusion composition.

[0067] Example 4

[0068] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 1.5% vitamin A palmitate, 3% soybean lecithin, 8% cholesterol, 12% trehalose, 2% astaxanthin, 2% sodium chloride, and 71.5% deionized water.

[0069] Example 4 was prepared by the following preparation method, comprising the following steps:

[0070] (1) Soy lecithin, cholesterol, astaxanthin, and vitamin A palmitate are mixed in ethanol and heated and ultrasonicated at 50° C. until completely dissolved to form phase A;

[0071] (2) taking trehalose, sodium chloride and deionized water and mixing them evenly to form phase B;

[0072] (3) After preheating the phase A and phase B to 50° C., phase A was continuously added dropwise to phase B while stirring at a speed of 600 r / min, and the mixture was stirred and hydrated at the same temperature for 35 min to obtain phase C;

[0073] (4) Add phase C into a high-pressure homogenizer for high-pressure homogenization at a homogenization pressure of 1000 Bar and cycle homogenization 72 times;

[0074] (5) The solution obtained in step 4 is freeze-dried to obtain the inclusion composition.

[0075] Example 5

[0076] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 0.05% vitamin A aldehyde, 1.5% polyglycerol-6 distearate, 0.2% sweet almond oil, 15% hydrolyzed rice protein, 1.3% wild soybean oil, 14% tert-butyl alcohol, and 67.95% deionized water.

[0077] The preparation method of Example 5 is the same as that of Example 3.

[0078] Example 6

[0079] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 1% retinoic acid, 1.5% dipalmitoylphosphatidylcholine, 2.3% ceramide I, 12% butylene glycol, 1.3% sunflower seed oil, 3.2% ethylparaben, and 78.7% deionized water.

[0080] The preparation method of Example 6 is the same as that of Example 1.

[0081] Comparative Example 1

[0082] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 0.05% vitamin A, 1.5% polysorbate-80, 0.2% jojoba oil, 12% dipropylene glycol, 5% peppermint oil, and 81.25% deionized water.

[0083] The preparation method of Comparative Example 1 is the same as that of Example 1.

[0084] Comparative Example 2

[0085] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 1% vitamin A, 1.5% sodium lauryl sulfate, 0.2% squalane, 12% pentylene glycol, 2.4% tocopherol, 5% ethylparaben, and 77.9% deionized water.

[0086] Comparative Example 2 Preparation method is the same as Example 2

[0087] Comparative Example 3

[0088] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 0.05% retinoic acid, 3.5% distearoylphosphatidylethanolamine, 0.03% shea butter, 12% glycerol, 2% resveratrol, 5% phenoxyethanol, and 77.42% deionized water.

[0089] The preparation method of Comparative Example 3 is the same as that of Example 3.

[0090] Comparative Example 4

[0091] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 1% retinoic acid, 1.5% dipalmitoylphosphatidylcholine, 2.3% ceramide I, 3% butylene glycol, 1.3% sunflower seed oil, 3.2% ethylparaben, and 87.7% deionized water.

[0092] The preparation method of Comparative Example 4 is the same as that of Experimental Example 1.

[0093] Comparative Example 5

[0094] A stable inclusion composition containing a retinoid family is composed of the following raw materials in percentage by mass: 1.5% of vitamin A palmitate, 3% of soy lecithin, 8% of cholesterol, 12% of trehalose, 2% of astaxanthin, 18% of sodium chloride, and 55.5% of deionized water.

[0095] The preparation method of Comparative Example 5 is the same as that of Example 4.

[0096] Effect experiment:

[0097] Test Example 1 Particle Size Distribution and Encapsulation Efficiency Determination of Inclusion Composition

[0098] 1 mL of the inclusions obtained in Experimental Examples 1-6 and Comparative Examples 1-5 were accurately measured and placed in a 10 mL volumetric flask, and the volume was fixed to the mark with ultrapure water. The particle size of the inclusions was measured using a Malvern laser particle size analyzer.

[0099] The encapsulation efficiency of the inclusion composition was determined by microcolumn centrifugation. A dextran gel microcolumn was constructed, and 200 μl of Experimental Example 1-6 and Comparative Example 1-5 were slowly dripped onto the top of the microcolumn, respectively. PBS solution was used as the eluent, and the mixture was centrifuged at 1000 rpm for 2 min. The centrifugation was repeated 5 times, and the centrifuge was collected. 9 times the volume of methanol was added to the centrifuge, and the mixture was vortexed for 2 min to break the emulsion. The drug concentration C was determined at 325 nm by high performance liquid chromatography. 1 200 μl of the inclusion composition solution that has not been separated by microcolumn centrifugation was taken and the drug concentration C was obtained after demulsification in the same way. 2 The encapsulation efficiency (EE%) of the inclusion composition was calculated.

[0100]

[0101] Depend on Figure 1 and 2 It can be seen that the particle size range of the inclusion composition obtained in Examples 1-6 is between 100 and 210 nm, and the encapsulation rate is higher than 80%, which can well encapsulate the active ingredients, reduce the contact probability of the ingredients with the external environment, and thus improve the stability. However, the particle sizes of Comparative Examples 1-3 and Comparative Example 5 are all above 350 nm, especially Comparative Example 1, Comparative Example 3 and Comparative Example 5 are as high as 500 nm. The obtained particle size is too large and difficult to penetrate the stratum corneum, which does not meet the required conditions for transdermal delivery of topical preparations. Among the five groups of comparative examples, the encapsulation rate of Comparative Example 1 and Comparative Example 4 is only about 50%, and the encapsulation rate of Comparative Example 5 is further reduced to 40.32±2.23%. More than half of the active ingredients are exposed to the solution, which is not conducive to the storage of sensitive ingredients.

[0102] Comparative Example 1 and Comparative Example 5 are compared with Example 1 and Example 4, and the samples are prepared mainly by not adding a light protectant and changing the ratio of the preservative and antibacterial agent, respectively. The results show that the particle size of the system of Comparative Example 1 without adding a light protectant and Comparative Example 5 with a changed ratio of the preservative and antibacterial agent increases, and the encapsulation rate decreases significantly. After reducing the ratio of the moisturizer in Comparative Example 4, there is no obvious difference in the particle size, but the encapsulation effect of the ingredients can be significantly reduced.

[0103] Test Example 2: Stability Study of Active Ingredients

[0104] Thermal stability test: The samples of Examples 1-6 and Comparative Examples 1-5 were subjected to thermal stability test. The test conditions were heating at 45°C and 60°C for 2 hours, respectively. After heating was completed and the mixture was restored to room temperature, 0.2 mL of the inclusion composition was added with 9 times the volume of methanol, vortexed to break the emulsion, and centrifuged to determine the active ingredient content by high performance liquid chromatography.

[0105] Oxygen stability study: 1 mL of samples of Examples 1-6 and Comparative Examples 1-5 were placed in a 2 mL centrifuge tube, and air was pumped into the centrifuge tube at a constant speed about 3 cm from the tube mouth using an air pump for air oxidation. All air oxidation experiments were conducted at 22-26°C in the dark, and 0.1 mL was sampled at 0, 2, and 4 h, respectively, and 9 times the volume of methanol was added for vortex demulsification. After centrifugation, the active ingredient content was determined by high performance liquid chromatography.

[0106] Light stability test: 1 mL of samples from Examples 1-6 and Comparative Examples 1-5 were placed in vials, and the vials were placed upside down in a drug stability test chamber. 2 The samples were irradiated at room temperature (25°C, RH 60%). The active ingredient content was detected by high performance liquid chromatography after demulsification at 0, 2, 6 and 8 hours.

[0107] Table 1 Thermal stability test results

[0108]

[0109] As can be seen from the above table, under the same heating conditions, Examples 1-6, whether at a relatively low temperature (45°C) or a high temperature (60°C), all exhibited excellent component stabilization capabilities, wherein Examples 1 and 3 could retain more than 90% of the active ingredients, greatly improving the availability. However, the thermal stability results of Comparative Example 3 decreased compared to Example 3, with the component retention rate being less than 65%, and the decrease in the component content of Comparative Example 1 and Comparative Example 2 was more obvious, especially after incubation at 60°C for 2 hours, almost no active ingredients were retained.

[0110] like Figure 3 and 4 As shown, the comparative example 1 without adding a photoprotectant showed extremely poor stability retention ability in the oxygen and light stability results. The retention rate of the ingredients was less than 40% after 4 hours of oxygen blowing, and almost no ingredients remained after 8 hours of light exposure. As shown in comparative examples 2 and 3, when the combination and ratio of surfactants and emollient lipids are changed, the oxygen and light stability of the active ingredients can also be adjusted. The poor stability shown in comparative examples 4 and 5 may be due to the moisturizers and preservatives and antibacterial agents in the prescription affecting the structure of the inclusion composition, causing the inclusions to be more prone to drug leakage, and the drugs are difficult to resist severe conditions after becoming free forms. In contrast, Examples 1-6 have improved the stability of the ingredients to varying degrees. Among them, Example 2 can still detect 95.75±0.81% of the active ingredients after 4 hours of oxygen stability test, and Example 3 can still retain 67.61±2.92% after 8 hours of light exposure, which has a strong ability to stabilize the ingredients.

[0111] Therefore, the encapsulation technology provided by the present invention can help make the structure of the inclusion composition more compact, stabilize the ingredients in the inclusion body, maintain the stability of the structure, improve the protection ability of the retinoid family, and improve its thermal, oxygen and light stability by selecting a suitable combination of surfactants and emollient lipids and an appropriate ratio of light protectants, moisturizers and preservatives and antibacterial agents.

[0112] Test Example 3 In vitro transdermal test

[0113] Use a stainless steel horseshoe clamp to clamp the pig ear back skin with an appropriate size between the supply tank and the receiving tank compartment of the Franz diffusion cell, with the skin stratum corneum facing up in contact with the solution to be tested, and preheat at 32°C for about 15 minutes to a hydrated state and temperature. Under the premise of meeting the conditions of the drain tank, a certain volume of Examples 1-6 and Comparative Examples 1-5 is transferred and evenly added to the surface of the pig skin for transdermal 12 hours. The receiving medium is 10% ethanol water, the temperature is 32±1°C, and the speed is 400r / min. Absorb all the receiving medium at 0.5h, 1h, 2h, 4h, 6h, 8h, and 12h, respectively, and supplement with an equal volume of insulated fresh medium. High performance liquid chromatography is used to determine the amount of active ingredients in the receiving medium at different time points.

[0114] After transdermal permeation, the stratum corneum and epidermis were separated by tape stripping. The stripping tape was placed in a centrifuge tube, 2 ml of methanol was added, and the tube was ultrasonically treated for 30 minutes to completely extract the drug. The tube was centrifuged at 12000 rpm and 4°C for 15 minutes. After drying and re-dissolving, the drug content C in the stratum corneum was measured by high performance liquid chromatography. 1 ; Cut the exfoliated skin into small particles with scissors, and mix methanol and exfoliated skin at a volume-to-mass ratio of 9:1 for extraction. Use a handheld high-speed homogenizer F6 / 10 to homogenize the skin in an ice bath at 10,000 rpm for 2.5 min, continue ultrasonication for 15 min to fully extract the drug, centrifuge at 12,000 rpm at 4°C for 15 min, blow dry and re-dissolve to measure the drug content C in the true epidermis 2 . Calculate the distribution of different groups in the skin.

[0115]

[0116] In all experimental groups, i.e., comparative examples 1-5 and examples 1-6, no active ingredient was detected in the receiving medium within 12 hours of in vitro transdermal permeation. This indicates that the technology provided by the present invention can regulate the transdermal penetration depth of the ingredients, avoid potential harm caused by the ingredients entering the blood, and reduce the user's concerns and risks of use.

[0117] Table 2 Distribution of active ingredients in each group in the skin after 12 hours of in vitro transdermal permeation

[0118]

[0119] Table 2 shows the distribution of active ingredients in the stratum corneum and epidermis of the skin after 12 hours of transdermal penetration under the same experimental conditions for different comparative examples and embodiments. The inclusion compositions obtained in Examples 1-6 have a lower stratum corneum accumulation rate and a higher epidermal retention rate, indicating that the inclusion compositions obtained in Examples 1-6 can flexibly pass through the stratum corneum barrier on the skin surface, and quickly and stably deliver the encapsulated active ingredients to the active epidermis or dermis. In contrast, Comparative Examples 1 to 5 do not have good stratum corneum penetration ability, especially when Comparative Example 2 adjusts the combination of surfactants and emollient lipids, it has the lowest epidermal retention rate, and the active ingredients are almost all accumulated in the stratum corneum, and it is difficult to reach the ideal action site such as the basement membrane to take effect. After long-term accumulation in the stratum corneum, penetration is also prone to cause skin irritation and sensitivity, and damage the skin barrier. This phenomenon may be because the selection of a suitable surfactant can affect the elasticity of the inclusions, thereby affecting its transdermal ability. Compared with Example 6, after Comparative Example 4 reduces the proportion of moisturizer, the deep retention of active ingredients is reduced due to decreased skin hydration. It can be concluded that the selection of appropriate surfactants, emollient lipids and moisturizers and their proportions can affect the transdermal performance of the inclusion composition, helping to accelerate the penetration of ingredients into the stratum corneum of the skin and reach deep layers to take effect.

[0120] Test Example 4: Investigation of Transdermal Integrity of Inclusion Composition

[0121] Fluorescence resonance energy transfer (FRET) is a method that can characterize the integrity of nanoformulations. It is very sensitive to changes in distance at the nanoscale. When two fluorescent molecules are close enough (<10nm), energy can be transferred from the donor to the acceptor. Therefore, by labeling two fluorescent dyes with FRET effect in the inclusion composition, the association or dissociation state of the inclusion composition can be reflected by monitoring the FRET signal before and after transdermal penetration. If the inclusion composition maintains structural integrity and the distance between the two dyes remains within a certain range, FRET can be maintained; if the preparation is disaggregated or destroyed, the change in distance leads to a decrease in FRET efficiency.

[0122] In this experiment, the stratum corneum of the pig ear dorsal skin was first successfully obtained by the trypsin method, and the stratum corneum was dried to constant weight for later use. A stratum corneum with an appropriate size was sandwiched between the supply pool and the receiving pool compartment of the Franz diffusion cell, and 0.4 mL of Examples 1-6 and Comparative Examples 1-5 labeled with two dyes, coumarin 6 and rhodamine isothiocyanate, were evenly coated on the surface of the stratum corneum and no bubbles were generated during the addition process. The preparation was allowed to naturally penetrate the stratum corneum into the receiving pool, and the solution in the receiving pool was aspirated for subsequent experiments.

[0123] The double-labeled drug delivery system before and after stratum corneum penetration and the single-labeled solution before transdermal delivery were respectively used. The fluorescence spectra in the range of 480 - 700 nm were scanned using a fluorescence spectrophotometer at the excitation wavelength of 465 nm for coumarin 6, and the FRET efficiency R and its percentage change X of each group were calculated.

[0124]

[0125] I c and I R are the fluorescence values of the emission peaks of the donor coumarin 6 and the acceptor rhodamine isothiocyanate, respectively.

[0126]

[0127] R 1 and R 2 are the FRET efficiencies before and after transdermal delivery of the double-labeled drug delivery system, respectively.

[0128] As Figure 5 shown, before and after stratum corneum penetration, the change in FRET efficiency of the inclusion complex compositions obtained in Examples 1 - 6 was relatively small, all remaining above 91%. Among them, Example 4 was 96.98 ± 2.38%, showing the structural stability of the inclusion complex, which was consistent with the in vitro transdermal results. The FRET effects of Comparative Example 1, Comparative Example 2, and Comparative Example 4 decreased significantly, indicating that the inclusion complex composition depolymerized and fragmented only after penetrating the stratum corneum, making it difficult to resist the layers of obstacles during transdermal delivery, resulting in premature release of the components and causing adverse reactions. The above data show that the photo-protective agent, surfactant, and humectant play an important role in the transdermal structural stability of the inclusion complex composition. Selecting appropriate photo-protective agent, surfactant, and humectant can maintain the structural stability, regulate the skin penetration depth of the preparation, and prolong the retention time of the components in the skin.

[0129] Test Example 5 Investigation of the ability to repair the basement membrane

[0130] Type IV collagen, as the main component of the skin basement membrane zone, accounts for more than 50% of the total components. The reticular structure formed by its cross-linking with each other has high stability and is an important supporting structure of the basement membrane zone. When the skin is damaged by external stimuli, type IV collagen is degraded by matrix metalloproteinase 9 (MMP9), the basement membrane loses its original structure, and the skin shows wrinkles and signs of aging. Therefore, the inclusion complex composition with the ability to inhibit MMP9 can reduce the continuous degradation of type IV collagen, stabilize the basement membrane, and rejuvenate the skin.

[0131] The Hacat cells in the logarithmic growth phase were diluted with complete medium and seeded at 1×10 per well 4Inoculate 200 μL of cells at a density into a 96-well plate, and fill the edges with an equal volume of PBS. After adaptively culturing until the cell density reaches about 50 - 60%, discard the supernatant medium, wash 3 times with PBS, add 100 μL of Examples 1 - 6 and Comparative Examples 1 - 5 diluted with blank medium to each well, and in the control group, only replace an equal volume of fresh blank medium. After continuing to culture for 24 h, aspirate and discard the supernatant medium, wash 3 times with PBS, and add 100 μl of PBS again. Irradiate the cells for 60 min under the condition of about 20 cm away from the ultraviolet lamp. After the irradiation ends, discard the PBS, add 100 μl of blank medium to each well and continue to culture for 24 h. After the culture ends, collect the supernatant medium, and detect the content of MMP9 in the supernatant of each group according to the ELISA kit method.

[0132] As Figure 6 shown, compared with the control group (Control), the MMP9 generation rates of the inclusion body compositions obtained in Example 1, Example 3, Example 4, Example 5, and Example 6 are all lower than 80%, which is superior to Comparative Examples 1 - 5. It can significantly inhibit the generation of MMP9 in Hacat cells after ultraviolet stimulation, can better resist the degradation of the basement membrane, and slow down skin aging. The above data show that after adding surfactants and emollient lipids in a specific proportion or selecting appropriate photoprotective agents in the inclusion body composition, it helps to improve the defense performance of the skin against external stimuli such as ultraviolet rays, and improve problems such as uneven skin color, skin relaxation, and skin spots.

[0133] Test Example 6 Skin irritation experiment

[0134] The skin irritation test is carried out according to the method of the human skin patch test in Chapter 7 of the "Cosmetics Technical Specifications" (2015 edition), specifically as follows:

[0135] Select 5 subjects. The subjects first clean the test area with warm water, and use a lint-free absorbent dry paper towel to blot the excess water on the test area. Sit quietly for 30 min in an environment with a humidity of 50 ± 5% and a temperature of 21 ± 1 °C in the efficacy evaluation laboratory. The test area is the flexor side of the forearm. Keep the test skin dry before the test and avoid contact with other topical preparations.

[0136] Select a qualified patch test device with an area not exceeding 50 mm 2 and a depth of about 1 mm. Put Examples 1 - 6 and Comparative Examples 1 - 5 into the small chamber of the patch test device, and the dosage is about 0.020 g - 0.025 g, and soak the filter paper. Stick the patch test device with the test substance on the flexor side of the subject's forearm with a low-sensitization tape, and gently press it with the finger pulp or palm to make it evenly adhere to the skin for 24 h. During the test, the volunteers need to keep the test area in a natural and relaxed state and do not squeeze or scratch.

[0137] After 24 hours, the volunteers returned to the laboratory, removed the patch device and sat quietly for 30 minutes. After the skin of the test area of ​​the volunteers returned to normal, the skin reaction was observed according to the standard 30 minutes after the test patch device was removed (after the indentation disappeared), and the observation results were recorded.

[0138] The skin red pigment value (EI) of the central area of ​​the test site and the surrounding uncoated area was measured using a Mexameter MX18 skin melanin hemoglobin tester. The measurement was performed three times and the average value was taken. The irritation of the inclusion composition to the skin was evaluated by comparing the change in the average value of skin EI before and after the use of the product. The higher the erythema index, the redder the skin in the test area, and the higher the skin irritation.

[0139] Erythema index (EI) = endpoint erythema difference - initial erythema difference

[0140] Table 3 Skin irritation reaction scores

[0141]

[0142]

[0143] Skin erythema index results are as follows Figure 7 The inclusion compositions obtained in Examples 1-6 had no adverse reactions on human skin, and there was no obvious redness on the skin after removing the spot tester. The objective test results of the skin melanin hemoglobin tester also verified this result. In contrast, the test areas of Comparative Examples 1 and 4 showed a higher erythema index and had a higher skin irritation.

[0144] Table 4 Skin irritation rating

[0145]

[0146] All subjects gave subjective scores for the skin reactions such as itching and tingling during the experiment, and the results are shown in the table above. The subjects reported that the control groups still had a certain degree of skin itching and tingling, especially control examples 1 and 4, while Examples 1-6 as a whole could alleviate this phenomenon. Even Example 2 containing a high concentration (1%) of active ingredients was reported to have low skin irritation and was acceptable. Example 3 had no subjective adverse skin reactions during the experiment and was very mild.

[0147] The above data show that the inclusion composition technology provided by the present invention can reduce the skin irritation of the retinoid family and avoid the irritation problems such as skin erythema, itching, burning, etc. caused by such compounds. Surfactants, emollient lipids, photoprotectants, moisturizers and antiseptics can stabilize the structure of the inclusion composition, allowing it to be delivered through the skin in its entirety, reducing the contact probability of the retinoid family with the TRPV1 receptor, slowly and continuously releasing the active ingredients, reducing skin irritation, and achieving a long-lasting and gentle repair effect.

Claims

1. A stable inclusion composition containing a retinoid family, characterized in that: The composition comprises the following components in percentage by mass: 0.02%-5% of a retinoid family active ingredient, 0.1%-4% of a surfactant, 0.05%-10% of an emollient lipid, 10%-35% of a moisturizer, 0.01%-3% of a light protectant, 1%-16% of an antiseptic and antibacterial agent, and 50%-82% of deionized water.

2. The stable inclusion composition containing the retinoid family according to claim 1, characterized in that: The retinoid family active ingredient is selected from one or more of vitamin A, vitamin A palmitate, retinoic acid, and vitamin A aldehyde.

3. The stable inclusion composition containing the retinoid family according to claim 1, characterized in that: The surfactant is selected from one or more of polysorbate 80, polysorbate 20, polyglycerol-6 distearate, sodium stearoyl glutamate, sodium cocoyl glutamate, egg yolk lecithin, soybean lecithin, hydrogenated lecithin, dipalmitoyl phosphatidylcholine, distearoyl phosphatidylethanolamine, and dioleoyl phosphatidylethanolamine.

4. The stable inclusion composition containing the retinoid family according to claim 1, characterized in that: The emollient lipid is selected from one or more of cetyl alcohol, shea butter, polydimethylsiloxane, cyclopentasiloxane, squalane, jojoba oil, sweet almond oil, isononyl isononanoate, isostearyl isostearate, beeswax, ceramide I, ceramide II, ceramide III, ceramide IV and ceramide VI, cholesterol, stearic acid, glyceryl behenate and myristic acid.

5. The stable inclusion composition containing the retinoid family according to claim 1, characterized in that: The moisturizing agent is selected from one or more of propylene glycol, glycerol, butylene glycol, pentylene glycol, allantoin, sodium pyrrolidone, trehalose, serum albumin, whey protein, hydrolyzed soy protein, hydrolyzed wheat protein, hydrolyzed rice protein, hyaluronic acid, sodium hyaluronate, and dipropylene glycol.

6. The stable inclusion composition containing the retinoid family according to claim 1, characterized in that: The photoprotective agent is selected from one or more of tocopherol, troxerutin, titanium dioxide, zinc oxide, astaxanthin, resveratrol, ectoine, vitamin C ethyl ether, ferulic acid, soybean oil, and sunflower seed oil.

7. The stable inclusion composition containing the retinoid family according to claim 1, characterized in that: The preservative and antibacterial agent is selected from one or more of sodium chloride, phenoxyethanol, methylparaben, ethylparaben, propylparaben, ethanol, tert-butyl alcohol, benzoic acid, sodium benzoate, peppermint oil, potassium sorbate, benzalkonium chloride, p-hydroxyacetophenone, and ethylhexylglycerin.

8. The method for preparing the stable inclusion composition containing retinoid family according to any one of claims 1 to 7, characterized in that: The method comprises the following steps: S1. The retinoid family active ingredients, surfactants, emollients, photoprotectants and fat-soluble preservatives and antibacterial agents are mixed uniformly in ethanol and sonicated at 30-65° C. until completely dissolved to form phase A; S2. Separately take a moisturizer and deionized water or a moisturizer, a water-soluble preservative and antibacterial agent and deionized water and mix well to form phase B; S3. After preheating the phase A and phase B to 30-65°C, continuously dropwise add phase A to phase B at a stirring speed of 500-2000 r / min, and stir and hydrate for 10-120 min at the temperature and speed to obtain phase C; S4. Phase C obtained in S3 is added to a high-pressure homogenizer for homogenization at a homogenization pressure of 300-1500 Bar and the homogenization cycle is 9-250 times; S5. The solution obtained in S4 is dialyzed, ultra-high-speed centrifuged or freeze-dried to obtain an inclusion composition, which is in the form of a uniform fluid liquid, a viscous semi-solid or a powder.

9. Use of the stable inclusion composition containing retinoid family as claimed in any one of claims 1 to 7 in the preparation of cosmetic or pharmaceutical preparations.

10. The use according to claim 9, characterized in that: The cosmetics are any one of lotion, essence, cream, toner and mask; the pharmaceutical preparations are any one of solution, cream, ointment and gel.

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