Preparation and application of a composition containing camellia sinensis extract and human-like sebum membrane

Abstract

The present application relates to a kind of preparation and application of containing camellia sinensis extract, human-like sebaceous membrane composition.The composition includes epidermal intercellular lipid composition, human-like sebaceous membrane component;According to mass percentage, epidermal intercellular lipid composition includes: oat kernel oil 33-60%, phytosterols 20-35%, camellia sinensis extract 20-35%;Human-like sebaceous membrane component includes: camellia sinensis extract 52-60%, squalane 10-14%, jojoba seed oil 23-29%, phytosterols 1-10%;Wherein, the mass ratio of epidermal intercellular lipid composition and human-like sebaceous membrane component is 0.1-5:5-0.1.All ingredients in the composition are of natural origin, and can be added as cosmetic efficacy raw materials to different types of cosmetic products, such as essence oil, emulsion, cream and other products, from the dimension of sebaceous membrane+keratin layer+active epidermis layer, reach the effect of three-dimensional repair skin barrier.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology and relates to the preparation and application of a composition containing red camellia extract and human-like sebum membrane. Specifically, it relates to a three-dimensional skin barrier protection composition containing red camellia extract and human-like sebum membrane, its preparation method, and its application. Background Technology

[0002] The surface area of ​​an adult's skin is approximately 2m². 2 The average thickness of skin is approximately 2.5 mm. Skin accounts for about 6% of body mass. It provides a vast physical barrier designed to protect us from a range of injuries. Typically, skin comprises two extensive tissue types: the epidermis (70-150 μm, up to 600 μm in the palms / soles, epithelial tissue) and the dermis (up to 4 mm, for example, on the adult back, but mostly <2 mm, connective tissue). Anatomically, it also includes a third layer: the subcutaneous tissue. The dermis also includes sebaceous glands, sweat glands, keratinized structures, and nerve receptors.

[0003] The epidermis of the skin, mainly composed of keratinocytes, is tightly connected to the dermis via the basement membrane. From the inside out, it consists of the basal layer (SB), spinous layer (SS), granular layer (SG), and stratum corneum (SC). The basal layer is composed of undifferentiated keratinocytes, stem cells, melanocytes, and Merkel cells. The proliferative activity of basal keratinocytes ensures continuous tissue renewal. In the spinous layer, the production of numerous desmosomes enhances intercellular adhesion, thus stabilizing the tissue. In the granular layer, not only are tight junctions (TJs) found, but the main process of stratum corneum formation is also prepared in this layer. Due to this gradual differentiation, different layers can perform different functions, such as forming new cells (SB), providing mechanical strength (SS, SG), and forming an effective barrier (SG, SC).

[0004] Broadly speaking, the skin barrier is divided into physical, chemical, microbial, and immune barriers. In a narrower sense, the skin barrier usually refers to the physical or mechanical barrier structure of the epidermis, especially the stratum corneum, also known as the permeable barrier. These barriers are mainly composed of keratinocytes and intercellular lipids forming a brick-wall-like structure. Keratinocytes are the "bricks," tightly connected by intercellular keratinized desmosomes; intercellular lipids are the "mortar," anchored to keratinocytes by structural lipids such as ceramides. Outside the stratum corneum is a sebum film composed of skin surface lipids and sweat, acting as a "coating layer" to collectively form the skin's physical barrier. Skin surface lipids mainly originate from lipids secreted by sebaceous glands, and also contain a small amount of lipids from the breakdown of keratinocytes. The lipids secreted by sebaceous glands mainly contain triglycerides, free fatty acids, squalene, wax esters, cholesterol, and cholesterol esters (see Table 1). Freshly secreted sebum does not contain free fatty acids; it is formed by the hydrolysis of triglycerides by parasitic bacteria within the hair follicles, and can inhibit some bacteria and fungi. It is worth noting that the components of intercellular lipids differ from those secreted by sebaceous glands. Intercellular lipids contain ceramides and phospholipids, which are absent in sebaceous gland lipids, and contain almost no squalene, wax esters, or other components. Skin surface lipids play a role in protecting the skin barrier, nourishing the skin, fighting infection, neutralizing acid and alkali damage, and maintaining the ecological stability of resident flora on the human skin surface.

[0005] Table 1 Summary of Relative Contents of Sebum Components

[0006] lipid types Content range (mass fraction) / % Average content (mass fraction) / % Triglycerides 20～60 45 Wax esters 23～29 25 Squalene 10～14 12 Free fatty acids 5～40 10 Cholesterol and cholesterol esters 1～5 4 diglyceride 1～2 2

[0007] Tight junctions are a class of cell-cell junctions first discovered in various cell and tissue types. They are complex structures composed of many types of tight junction transmembrane proteins that tightly connect adjacent cells. However, it wasn't until this year that the expression of these proteins in the epidermis of mammals was confirmed. Tight junction proteins include: the Claudin (Cldns) family of proteins (also known as clasping proteins), occludin, the junctional adhesion molecule family of proteins, and tight junction plaque proteins. The expression levels and sites of different tight junction proteins vary in the epidermis. Besides keratinocytes, Cldn-1 is also expressed in fixed and migrating Langerhans cells. However, although several tight junction proteins are expressed throughout the entire epidermal layer, typical tight junction structures are only found in the granular layer. Claudins are an important protein family that makes up keratinocyte tight junctions; 24 types have been identified so far, and they work with keratinocytes to help maintain the skin barrier function. A deficiency of slugin can cause mice to die within 24 hours of birth due to dry, wrinkled skin, severe dehydration, and increased epidermal permeability and TEWL values. In atopic dermatitis, the expression of both slugin-1 and slugin-23 is significantly reduced.

[0008] CN 111557874 B discloses a biomimetic skin barrier repair composition and its application containing camellia extract. It comprises the following components: a sebum film repair component: squalene, fatty acids, triglycerides, and cholesterol analogs; an intercellular matrix component: free fatty acids, ceramides, and cholesterol analogs; and synergistic components: soothing and anti-inflammatory ingredients, anti-aging ingredients, whitening ingredients, and skin penetrants; wherein the sebum film repair and intercellular matrix components are composed of camellia extract, flaxseed oil, oat kernel oil, squalane, phytosterols, and ceramide 3. CN 113041172 A discloses a biomimetic composition for skin barrier repair, its application, and cosmetics. The composition includes: 1-2 parts acetyl dipeptide-1 cetyl ester, 1-2 parts tea polyphenols, 3-5 parts squalane, 2-4 parts niacinamide, 8-12 parts glycerin, 12-4 parts bio-enamel-1, 4-8 parts PEG-100 stearate, 212-5 parts stearyl alcohol polyether-2, 1-2 parts vitamin E, and 46-80 parts water. However, both of these patents essentially only simulate the different proportions of individual sebum or individual intercellular lipids, without considering the ratio of sebum membrane to intercellular lipids. Summary of the Invention

[0009] The purpose of this invention is to address the limitations of existing technologies by providing a preparation and application of a composition containing red camellia extract and a human-like sebum membrane, which primarily functions to repair the skin barrier. This invention not only simulates the ratio of the sebum membrane and intercellular lipids, but also simulates an optimal ratio of sebum membrane to intercellular lipids to achieve the best barrier repair effect. Furthermore, the invention tests the level of CLAUDIN-1, a tight junction protein, secreted by keratinocytes, aiming to enable the composition to repair the skin barrier from three dimensions: sebum membrane, stratum corneum, and active epidermal layer.

[0010] The objective of this invention can be achieved through the following methods:

[0011] In a first aspect, the present invention provides a composition containing camellia extract and a human-like sebum membrane, the composition comprising intercellular lipid components of epidermal cells and human-like sebum membrane components;

[0012] Based on the percentage by mass of the total weight of intercellular lipids in epidermal cells, the intercellular lipid composition includes:

[0013] Oat kernel oil 33-60%,

[0014] Phytosterols 20-35%,

[0015] 20-35% red camellia extract;

[0016] Based on the percentage by mass of the total weight of human-like sebum membrane components, the human-like sebum membrane components include:

[0017]

[0018] In one embodiment of the present invention, the mass ratio of the intercellular lipid composition of the epidermal cells to the human-like sebum membrane component is 0.1-5:5-0.1. If the ratio is lower or higher than this range, the skin barrier will be damaged, such as a decrease in the skin's water-locking function and abnormal skin moisture content; it may also lead to an imbalance of the skin surface flora or cause various types of dermatitis.

[0019] Furthermore, the mass ratio of the intercellular lipid composition of the epidermal cells to the human-like sebum membrane component is 1-3:3-1.

[0020] As one embodiment of the present invention, the red camellia extract includes red camellia seed oil, red camellia flower extract, and red camellia leaf extract, with a mass percentage of 75-90%, 5-15%, and 5-15%, respectively.

[0021] In this invention, the preparation method of red camellia flower extract and red camellia leaf extract is described in patent CN109157454B. The method involves pulverizing and sieving red camellia flowers and red camellia leaves, followed by supercritical extraction and then two-stage separation. The parameters for supercritical extraction are: CO2 fluid flow rate of 15-30 ml / min; extraction pressure of 15-30 MPa; extraction temperature of 30-50℃; and extraction time of 1-2 h. The parameters for the two-stage separation are: stage I separation pressure of 8-12 MPa and temperature of 25-40℃; and stage II separation pressure of 7-8 MPa and temperature of 25-40℃.

[0022] In this invention, the intercellular lipids of the epidermal cells are composed of phase A, and the human-like sebum membrane components are composed of phase B.

[0023] Secondly, the present invention provides a method for preparing a composition containing red camellia extract and a human-like sebum membrane, the preparation method comprising the following steps:

[0024] S1. Mix and stir the intercellular lipid composition of epidermal cells;

[0025] S2. Mix and stir the human-like sebum membrane components;

[0026] S3. The epidermal intercellular lipid composition from step S1 is mixed with the human-like sebum membrane component from step S2 and stirred to obtain the composition.

[0027] As one embodiment of the present invention, in step S1, the stirring method includes water bath stirring, the temperature of the water bath stirring is 70-85℃, and the time is 20-40min.

[0028] In some embodiments, the mixing method in step S1 is as follows: phase A components are added sequentially to a beaker.

[0029] As one embodiment of the present invention, in step S2, the stirring method includes water bath stirring, the temperature of the water bath stirring is 70-85℃, and the time is 20-40min.

[0030] In some embodiments, the mixing method in step S2 is as follows: phase B components are added sequentially to the beaker.

[0031] In one embodiment of the present invention, in step S3, the mixture is stirred until it is cooled to room temperature.

[0032] Thirdly, this invention provides the use of a composition containing red camellia extract and human-like sebum membrane as an active ingredient in the preparation of cosmetics.

[0033] As one embodiment of the present invention, the cosmetic is a cosmetic used to repair the skin barrier.

[0034] As one embodiment of the present invention, the cosmetic type includes at least one of essential oil, lotion, and face cream.

[0035] Compared with the prior art, the present invention has the following beneficial effects:

[0036] 1. In addition to simulating the ratio of sebum membrane and intercellular lipids, this invention also explores a suitable ratio of sebum membrane and intercellular lipids to achieve the optimal barrier repair effect.

[0037] 2. The composition of the present invention significantly promotes the secretion of tight junction protein CLAUDIN-1 by keratinocytes through a human-like sebum film composition containing red camellia, thus achieving the effect of three-dimensional repair of the skin barrier from the dimensions of sebum film + stratum corneum + active epidermal layer.

[0038] 3. This invention uses jojoba seed oil to simulate the wax ester components in the sebum film, making the final composition more closely resemble the composition of the real skin sebum film, thereby enhancing the skin barrier's protective function.

[0039] 4. All ingredients in the composition of this invention are of natural origin and have the function of repairing the skin barrier. They can be added as cosmetic ingredients to different types of cosmetic products, such as essential oils, lotions, and face creams. Attached Figure Description

[0040] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0041] Figure 1This is an immunofluorescence assay of the model control group and the 1.0% human-like sebum membrane composition. Detailed Implementation

[0042] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. The following examples are implemented under the premise of the technical solution of the present invention, providing detailed implementation methods and specific operating procedures, which will help those skilled in the art to further understand the present invention. It should be noted that the scope of protection of the present invention is not limited to the following embodiments; any adjustments and improvements made under the concept of the present invention are all within the scope of protection of the present invention.

[0043] The present invention provides a method for preparing the compositions of the following embodiments or comparative examples, comprising the following steps:

[0044] S1: Add the A phase components sequentially to the beaker and stir in a water bath at 70-85℃ for 30 minutes;

[0045] S2: Add the B phase components sequentially to the beaker and stir in a water bath at 70-85℃ for 30 minutes;

[0046] S3: Then mix phase A and phase B together in a ratio of A:B = 0.1-5:5-0.1, stir and cool to room temperature.

[0047] It should be noted that in the following examples, the mass ratio of the intercellular lipids in the epidermal cells is oat kernel oil: phytosterols: camellia extract = 5:3:2; and the mass ratio of the human-like sebum membrane components is camellia extract: squalane: jojoba seed oil: phytosterols = 55:13:25:7.

[0048] In the following embodiments and comparative examples of this invention, the preparation methods of the camellia flower extract and the camellia leaf extract are as described in patent CN109157454B. Specifically, the camellia flowers and camellia leaves are pulverized and sieved, then subjected to supercritical extraction, followed by two-stage separation to obtain the extracts. The parameters for the supercritical extraction are: CO2 fluid flow rate of 20 ml / min; extraction pressure of 20 MPa; extraction temperature of 40°C; and extraction time of 2 h. The parameters for the two-stage separation are: stage I separation pressure of 10 MPa and temperature of 30°C; and stage II separation pressure of 8 MPa and temperature of 30°C.

[0049] Example 1

[0050] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0051]

[0052] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 1:1.

[0053] Example 2

[0054] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0055]

[0056] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 2:1.

[0057] Example 3

[0058] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0059]

[0060] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 3:1.

[0061] Example 4

[0062] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0063]

[0064] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 1:2.

[0065] Example 5

[0066] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0067]

[0068]

[0069] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 1:3.

[0070] Example 6

[0071] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0072]

[0073] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 1:4.

[0074] Example 7

[0075] This embodiment provides a composition containing camellia extract and a human-like sebum membrane, comprising the following components:

[0076]

[0077] The mass ratio of the intercellular lipid component to the human-like sebum membrane component is 4:1.

[0078] Comparative Example 1

[0079] This comparative example provides a composition containing red camellia extract, comprising the following components:

[0080]

[0081] Comparative Example 2

[0082] This comparative example provides a composition containing red camellia extract, comprising the following components:

[0083]

[0084]

[0085] Effect Verification 1: Security Test

[0086] a: Chicken embryo chorioallantoic membrane test

[0087] The irritant properties of the compositions were tested using a chicken embryo chorioallantoic membrane test. As can be seen from Tables 1 and 2, based on the irritant classification, all examples and comparative examples showed no irritation to the chicken embryo chorioallantoic membrane.

[0088] Table 1 Stimulus Rating Criteria

[0089] Finish line score Irritant Classification ES=0 Non-irritating 0 < ES ≤ 4 Mild irritation 4 < ES < 5.3 moderate irritation 5.3≤ES Strongly irritating / corrosive

[0090] Table 2 Results of Chicken Embryo Vulonallantoic Membrane Test

[0091]

[0092] b: Human patch test

[0093] Qualified patch testing equipment was selected, and a closed patch test method was used. Approximately 0.020g to 0.025g of the test substance was placed in the patch tester, and a hypoallergenic adhesive tape was applied to the inside of the subject's arm. The test substance was removed after 24 hours, and skin reactions were observed at 0.5, 24, and 48 hours after removal. The results were recorded according to the skin reaction grading standards in the "Cosmetic Safety Technical Specifications" (2015 edition). As shown in Table 3, no adverse reactions were observed in the 30 participants tested. Therefore, all examples and comparative examples passed the human skin patch test.

[0094] Table 3 Results of Human Patch Test

[0095]

[0096] Efficacy Verification 2: In Vitro Repair Test - Cell Scratch (Keratinocytes)

[0097] Epidermal keratinocytes are essential cells constituting the epidermal layer. When the skin surface is damaged, keratinocytes are stimulated to migrate and repair the damaged area. When cells grow to a monolayer in vitro, an artificially created blank area is formed on the monolayer. Cells at the edge of the scratch gradually enter the blank area, causing the scratch to heal, which to some extent simulates the in vivo cell migration process. The reparative efficacy of the test substance was evaluated by measuring the cell migration rate after sample treatment. The results are shown in Table 4. Different proportions of the A and B phase compositions showed different cell migration growth rates, and the differences were very significant. The cell migration growth rate of Example 2 was the highest, indicating that it had the best repair effect.

[0098] Table 4. Effects of different samples on cell migration rate

[0099]

[0100]

[0101] Efficacy Verification 3: Repairing the skin barrier and soothing effects—Transepidermal water loss (TEWL) value and erythema index (EI) value

[0102] This protocol involved 33 healthy Chinese subjects who were stimulated on their backs with 1% SLS patches for 24 hours. The EI value was measured 10 minutes after application; TEWL and EI values ​​were measured before application (1 hour after patch removal), 3 days, and 7 days later. A blank control was used to evaluate the repair and soothing efficacy of the samples. The samples and blank control areas were treated with 2.00±0.05 μL / cm² of SLS patches after cleansing in the morning and evening. 2 Take a sample and massage for 5 circles. The TEWL and EI values ​​after 7 days are shown in Tables 5 and 6. The lower the TEWL value, the better the effect of repairing the skin barrier. There are significant differences between examples with different A / B values, with Example 2 showing the best effect. The lower the EI value, the better the soothing effect of the composition. As shown in Table 6, Example 2 is also the most effective in reducing skin redness.

[0103] Table 5 Results of TEWL Change Rate

[0104]

[0105] Table 6. Results of Erythema Index (EI) Tests

[0106]

[0107] Effect Validation 4: Test to Promote the Expression of Tight Junction Protein Claudin-1

[0108] When the skin barrier function is impaired, it accelerates the loss of moisture from the skin surface and skin aging. Due to the effects of natural aging and photoaging, skin problems such as dryness, wrinkles, and sagging occur. Claudin family proteins, belonging to the tight junction protein family, play an important role in the permeability specificity of the skin cell barrier and the transmembrane movement of substances. When the skin is exposed to ultraviolet radiation, Claudin-1 expression decreases, thereby causing impaired permeability barrier function in keratinocytes. Cellular immunofluorescence staining utilizes antigen-antibody specific binding and fluorescent labeling to display the expression and localization of target proteins within cells. The fluorescence intensity of the target protein can indicate its relative expression level. This experiment used HaCaT keratinocytes, which were cultured, seeded, modeled under ultraviolet light, and subjected to immunofluorescence testing. The results are shown in Table 7. The table shows that the human-like sebum membrane composition containing the red camellia extract significantly promoted Claudin-1 expression, with Example 2 showing the highest promotion rate. The immunofluorescence test results are shown in Table 7. Figure 1 The images are shown (representing the model control group and the 1.0% human-like sebum film composition, respectively; the model control group was modeled using ultraviolet light, and the human-like sebum film composition was 1.0 wt.%). Therefore, the human-like sebum film composition containing the camellia extract can significantly repair the skin barrier.

[0109] Table 7 Results of Claudin-1 relative expression promotion rate test

[0110]

[0111]

[0112] The specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A composition containing camellia extract and human-like sebum membrane, characterized in that, The composition includes intercellular lipid components of epidermal cells and human-like sebum membrane components; Based on the percentage by mass of the total weight of intercellular lipids in epidermal cells, the intercellular lipid composition includes: Oat kernel oil 33-60%, Phytosterols 20-35%, Red camellia extract 20-35%; Based on the percentage by mass of the total weight of human-like sebum membrane components, the human-like sebum membrane components include: Red camellia extract 52-60%, Squalane 10-14%, Jojoba seed oil 23-29%, Phytosterols 1-10%; The mass ratio of the intercellular lipid composition of the epidermal cells to the human-like sebum membrane component is 2:

1.

2. The composition according to claim 1, characterized in that, The red camellia extract includes red camellia seed oil, red camellia flower extract, and red camellia leaf extract, with a mass percentage of 75-90%, 5-15%, and 5-15%, respectively.

3. A method for preparing the composition according to any one of claims 1-2, characterized in that, The preparation method includes the following steps: S1. Mix and stir the intercellular lipid composition of epidermal cells; S2. Mix and stir the human-like sebum membrane components; S3. The epidermal intercellular lipid composition from step S1 is mixed with the human-like sebum membrane component from step S2 and stirred to obtain the composition.

4. The preparation method according to claim 3, characterized in that, In steps S1 and S2, the stirring method includes water bath stirring, with the temperature of the water bath being 70-85 ℃ and the time being 20-40 min.

5. The preparation method according to claim 3, characterized in that, In step S3, stir until cooled to room temperature.

6. Use of a composition according to any one of claims 1-2, or a composition obtained by any one of claims 3-5, as an active ingredient in the preparation of cosmetics.

7. The use according to claim 6, characterized in that, The cosmetic product in question is used to repair the skin barrier.

8. The use according to claim 6, characterized in that, The types of cosmetics include at least one of essential oils, lotions, and face creams.

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