Composition with efficacy of enhancing extracellular matrix and barrier repair and application thereof

Abstract

The invention relates to a composition with an effect of enhancing extracellular matrix and barrier repair and application thereof. The composition with the effect of strengthening extracellular matrix and barrier repair comprises galactomannan and rice fermentation liquor, and the mass ratio of the galactomannan to the rice fermentation liquor is (0.5-2.5): (0.1-10). Compared with the prior art, by utilizing the synergistic effect of the galactomannan and the rice fermentation liquor, the function of an extracellular matrix (ECM) can be remarkably enhanced, and the composition has excellent barrier repairing capacity and shows excellent characteristics of resisting aging, preserving moisture and promoting skin micro-ecological balance. The active ingredients are clear, and the product quality is easy to control. A multi-target and multi-path intervention strategy is adopted, and comprehensive optimization and deep repair of the skin barrier function are achieved through the synergistic effect of multiple biological mechanisms such as regulation and control of intracellular and extracellular signal paths, promotion of gene expression, enhancement of protein synthesis and inhibition of degrading enzyme activity.

Description

Technical Field

[0001] The invention belongs to the technical field of daily chemicals, and specifically relates to a composition with the functions of strengthening extracellular matrix and barrier repairing and application thereof. Background Art

[0002] At present, the common barrier repair products on the market are mostly based on relatively superficial mechanisms of action. They mainly perform their functions by supplementing or simulating the core components of the skin's natural barrier (such as ceramides to strengthen intercellular lipid connections, and squalane to build a physical barrier to reduce transepidermal water loss). Although these strategies can optimize the initial function of the skin barrier to a certain extent and relieve superficial symptoms such as dryness and sensitivity, they are limited by the singleness and incompleteness of their mechanisms, making it difficult to touch and solve the deep and complex problems behind skin barrier damage.

[0003] Chinese patent CN111741743A discloses a cosmetic composition comprising a spinulose nut extract, a long-hearted Kappaphycus algae extract, at least one prebiotic and a probiotic, which is intended to maintain the ecological balance of the skin and the diversity of the microbiome, enhance the skin barrier and resistance, improve the uniformity of skin color, promote skin peeling and epidermal renewal, reduce the loss of firmness and pore elasticity, improve the uniformity of skin color, promote skin energy balance and the durability of makeup and the uniformity of its deposition. The main ingredients involved in this scheme are spinulose nut extract, long-hearted Kappaphycus algae extract, prebiotics and probiotics. This scheme also does not involve the relevant content of strengthening the extracellular matrix.

[0004] Chinese patent CN111920716A discloses a raw material capable of improving symptoms of skin aging, comprising a solvent, a macromolecular thickener and an active ingredient combination; the active ingredient combination comprises: active ingredient one, an active ingredient having the function of repairing the extracellular matrix; active ingredient two, an active ingredient having the function of promoting the increase of collagen at the junction of the epidermis and dermis; active ingredient three, an active ingredient having antioxidant properties; active ingredient four, an active ingredient having the function of repairing the skin barrier, using plant ceramide 3; and the mass ratio of active ingredient one, active ingredient two, active ingredient three and active ingredient four is 3:1:1:0.5 to 3:1:1:2. Starting from the mechanism of skin aging and taking into account the anti-aging needs of the epidermis and dermis, the raw material design is carried out from four directions: repairing the skin barrier, repairing the junction DEJ of the epidermis and dermis, repairing the degradation of the dermis ECM and anti-oxidation. The combination of the real and the surface, from the inside out, has a good effect of improving the aging phenomenon. However, the active ingredients in this patent scheme include hydroxypropyl tetrahydropyrantriol, glycosaminoglycan, collagen, African oak bark extract, jojoba seed extract, hydrolyzed flaxseed extract, carnosine, tea extract, decarboxylated carnosine and ergothioneine, etc., and its ingredients are too complicated.

[0005] Therefore, the current industry is in urgent need of an innovative solution that aims to achieve comprehensive and deep repair of the skin barrier through the synergistic effects of multiple mechanisms, deep into the cellular level, and strengthening of the extracellular matrix (ECM). Summary of the invention

[0006] In view of the current situation that skin care products in the prior art have a single mechanism of action and limited effects, the present invention provides a composition with the effects of strengthening the extracellular matrix and barrier repairing and its application.

[0007] The technical solution provided by the present invention scientifically mixes galactomannan and rice fermentation liquid and achieves synergistic effects, so as to solve the problems of single action mechanism and limited effect of existing skin care products.

[0008] The composition of the present invention with the efficacy of strengthening the extracellular matrix and barrier repairing significantly enhances the repair and protection functions of the skin barrier through multiple mechanisms, while exhibiting excellent anti-aging, moisturizing and skin microecological balance promoting properties.

[0009] The purpose of the present invention can be achieved by the following technical solutions:

[0010] The invention firstly provides a composition with the functions of strengthening extracellular matrix and barrier repairing, comprising galactomannan and rice fermentation broth, wherein the mass ratio of the galactomannan to the rice fermentation broth is (0.5-2.5): (0.1-10).

[0011] In one embodiment of the present invention, the mass ratio of the galactomannan to the rice fermentation broth is (0.5-1.5): (0.1-3).

[0012] In one embodiment of the present invention, the galactomannan can be obtained through commercial channels, or extracted from alfalfa.

[0013] In one embodiment of the present invention, the rice fermentation liquid is a relatively common additive in the field of cosmetics. It should be noted that the rice fermentation liquid can be purchased commercially or prepared by oneself.

[0014] As a non-limiting example, yeast Saccharomyces veronae can be used to prepare rice fermentation broth through fermentation and enrichment processes, wherein Alpha-KG is enriched, and the mass proportion of Alpha-KG in the rice fermentation broth is 400ppm-1000ppm. The preparation method of rice fermentation broth includes:

[0015] Step a. Add water to the rice powder and heat and stir to form 30% rice milk;

[0016] Step b. adding β-glucanase to the rice milk, the enzyme added in an amount of 0.001%-0.5% by mass of the rice milk, the enzyme activity is 150-200U / g, and enzymolysis is performed to obtain a rice milk hydrolyzate;

[0017] Step c. The yeast strain is inoculated into the rice milk hydrolyzate, and the fermentation process conditions are: the fermentation temperature is 20-40 ° C, the fermentation time is 24-48 hours, the pH of the fermentation process is controlled between 6-6.5, and the rice fermentation liquid is obtained after fermentation;

[0018] Step d. The fermented rice fermentation liquid is concentrated by reverse osmosis technology to efficiently enrich the active substances, and the concentrated rice fermentation liquid is obtained by heating, sterilizing, cooling and filtering, i.e., the rice fermentation liquid containing the active substances.

[0019] The present invention further provides the use of a composition having the effects of strengthening the extracellular matrix and repairing the barrier in the preparation of a skin care product, wherein the skin care product may be any form of skin care product or cosmetic, including lotion, cream, freeze-dried powder, skin care water, etc. The skin care product has the effects of strengthening the extracellular matrix and repairing the barrier.

[0020] The present invention also provides a skin care product with the functions of strengthening the extracellular matrix and repairing the barrier, comprising a matrix auxiliary material and a functional component, wherein the functional component comprises a composition with the functions of strengthening the extracellular matrix and repairing the barrier.

[0021] In some preferred embodiments, based on 100 parts by weight of the total weight of the skin care product, the composition having the effects of strengthening the extracellular matrix and repairing the barrier is 1 to 10 parts, preferably 2 to 7.5 parts, and more preferably 3.5 to 7.5 parts.

[0022] In some embodiments of the present invention, the matrix excipient is selected from any one or more of a solvent, a skin conditioner, a thickener, a moisturizer, an emulsifier, a pH adjuster, a fragrance or a preservative.

[0023] In a specific embodiment of the present invention, the present invention provides an essence having the effects of strengthening the extracellular matrix and repairing the barrier, the essence comprising a phase A auxiliary material, a phase B auxiliary material, a phase C auxiliary material, a phase D functional ingredient and a phase E auxiliary material;

[0024] Based on 100 parts by weight of the essence, the phase A auxiliary material includes 0.03 to 10 parts of thickener A, 0.12 to 5 parts of skin conditioner A and 3 to 20 parts of moisturizer;

[0025] The phase B auxiliary material includes 0.3-15 parts of thickener B, 0.1-5 parts of emulsifier and 0.5-5 parts of skin conditioner B;

[0026] The phase C auxiliary material includes 0.1 to 5 parts of a pH adjuster;

[0027] The dosage of the D phase functional ingredient is 1 to 10 parts;

[0028] The E phase auxiliary material includes 0.01 to 2 parts of fragrance and 0.15 to 1 part of preservative;

[0029] Optionally, the phase A auxiliary material includes water, and the amount of water used is such that the weight parts of the essence reaches 100 parts;

[0030] The thickener A and the thickener B are independently selected from one or more of acrylic acid (esters) / C10-30 alkyl acrylate cross-linked polymer, polyacrylamide, xanthan gum, propylene glycol, behenyl alcohol, carbomer, laureth-7, cetyl alcohol, stearyl alcohol or hydroxyethyl cellulose;

[0031] The skin conditioning agent A and the skin conditioning agent B are each independently selected from one or more of glycerin, isohexadecane, olive oil, sodium hyaluronate, lauryl alcohol, avocado butter, allantoin, butylene glycol, squalane, panthenol, lactic acid, tocopherol or dipotassium glycyrrhizinate;

[0032] The moisturizing agent is selected from one or more of hyaluronic acid, allantoin, glycerin, ceramide, sodium lactate, butylene glycol, ethylhexylglycerin, hydrogenated grape seed oil, olive oil or shea butter;

[0033] The emulsifier is selected from one or more of glyceryl stearate citrate, cetearyl glucoside (and) cetearyl alcohol, behenyl trimethyl ammonium methyl sulfate, cocoglyceryl esters or sorbitan olivate;

[0034] The pH regulator is selected from one or more of arginine or lactic acid;

[0035] The aromatic agent is selected from essences;

[0036] The preservative is selected from one or more of phenoxyethanol, ethylhexylglycerin or pentylene glycol.

[0037] In one embodiment of the present invention, the essence having the effects of strengthening the extracellular matrix and repairing the barrier, based on 100 parts by weight of the essence, the phase A auxiliary material comprises 0.01 to 5 parts of xanthan gum, 0.02 to 10 parts of carbomer, 0.02 to 1 part of sodium hyaluronate, 0.1 to 5 parts of allantoin, 2 to 15 parts of glycerol and 1 to 10 parts of butylene glycol; the phase B auxiliary material comprises 0. 2-10 parts of cetyl alcohol, 0.1-10 parts of stearyl alcohol, 0.1-5 parts of cetearyl glucoside (and) cetearyl alcohol, 0.5-5 parts of squalane; the C phase auxiliary material includes 0.1-5 parts of arginine; the D phase functional ingredients are 1-10 parts; the E phase auxiliary material includes 0.1-1 part of phenoxyethanol, 0.05-1 part of ethylhexylglycerin and 0.01-2 parts of fragrance. The present invention also provides a preparation method of the above-mentioned specific essence with the effect of strengthening extracellular matrix and barrier repair, comprising the following steps:

[0038] a. Obtaining the aqueous phase: Place water in a container with a stirring device and stir, fully wet and mix the other parts of the phase A excipients, add them to the water, stir until completely dissolved, and heat to 75-80°C;

[0039] b. Obtain the oil phase; mix the B phase excipients and heat to 75-80°C, stirring until completely dissolved;

[0040] c. Emulsification: Add the mixture of step b to the aqueous phase of step a, stir at 250-350 rpm for 3-5 minutes and homogenize at 2500-3500 rpm for 3-10 minutes;

[0041] d. Add phase C auxiliary material, stir at 250-350 rpm for 3-5 minutes and homogenize at 2500-3500 rpm for 3-10 minutes.

[0042] Stir and cool to 45°C;

[0043] e. Add E phase auxiliary materials, stir at 250-350 rpm for 3-5 minutes and homogenize at 2500-3500 rpm for 3-10 minutes;

[0044] f. After the temperature drops to 35-38°C, add phase D, stir at 250-350 rpm and homogenize at 2500-3500 rpm for 3-10 minutes to obtain the desired essence.

[0045] It should be noted that the above steps all require strict control of sterility. The obtained essence is filtered and sterilized before filling, and the bacterial content, toxins, immunogenicity, etc. are tested for quality control.

[0046] The galactomannan in the present invention is a unique neutral heteropolysaccharide, the main chain of which is composed of d-mannose via β-1,4 glycosidic bonds, and the side chain of which is composed of d-galactose connected via α-1,6 glycosidic bonds. The galactomannan can be extracted from alfalfa, the oldest forage plant in the world.

[0047] Galactomannan can effectively enhance the skin barrier function, promote cell renewal and limit protein oxidation by activating and restoring key physical and moisture barrier proteins including Ki67, loricrin, tight junction protein-1, aquaporin-3, as well as neutral lipid BODIPY and epidermal-dermal anchoring proteins (such as integrin β4, laminin 332, collagen VII).

[0048] In addition, galactomannan can promote the synthesis of extracellular matrix (ECM) while inhibiting the degradation of its components, significantly improving fiber quality and dermal density, especially by promoting the synthesis of type I collagen and inhibiting the synthesis of MMP-1, thereby enhancing the quality of dermal matrix and showing excellent barrier repair and anti-aging properties.

[0049] ECM is a dynamic network composed of various biomacromolecules such as collagen, elastin, proteoglycans, glycosaminoglycans (such as hyaluronic acid), adhesion molecules and growth factors. It runs through the periphery of cells and provides structural support and biochemical signaling environment for cells. In the skin, ECM is a key factor in maintaining skin elasticity, firmness and regeneration ability.

[0050] Collagen is the most abundant protein in the ECM, especially type I and type III collagen, which plays a core role in the structure and elasticity of the skin. Galactomannan can upregulate the expression of genes related to collagen synthesis, such as activating the TGF-β (transforming growth factor-β) signaling pathway, thereby promoting the synthesis of type I collagen and enhancing the structural integrity and elasticity of the skin.

[0051] Proteoglycans and glycosaminoglycans such as hyaluronic acid are important water-retaining components of the ECM. They are able to attract and retain large amounts of water, giving the skin a hydrated feel and elasticity. Galactomannan increases the hydration of the ECM and improves the skin's moisturizing properties by stimulating cells to synthesize these molecules.

[0052] During skin aging, ECM degradation accelerates, mainly due to the increase in the activity of degradative enzymes such as matrix metalloproteinases (MMPs). Galactomannan can downregulate the expression of MMPs or inhibit their activity, thereby slowing down the degradation process of ECM and maintaining the integrity and stability of ECM.

[0053] Adhesion molecules in ECM, such as integrins and laminins, are not only involved in the adhesion between cells and ECM, but also in the signal transduction between cells. Galactomannan enhances the expression and function of these molecules, promotes communication and cooperation between cells, and thus optimizes the remodeling and repair process of ECM.

[0054] Galactomannan also has the physiological activity of promoting the growth of probiotics. In an in vitro fermentation environment, it can effectively promote the proliferation of potential beneficial bacteria such as Bifidobacterium, Lactobacillus, Bacteroidetes, and Porites, while inhibiting the growth of potential harmful bacteria such as Fusobacterium, Trichoderma, and spirochetes, providing a new way to promote skin healing.

[0055] Rice fermentation liquid is a product obtained by a certain period of fermentation process using special strains and high-quality rice based on biological fermentation technology. The rice fermentation filtrate significantly increases the expression of tight junction proteins (such as Claudin-1, Claudin-4 and Occludin) through precise regulation. Claudin-1, Claudin-4 and Occludin contribute to the formation of tight junction protein barriers. The lack of Claudin-4 in the tight junction protein structure will lead to impaired barrier function of keratinocytes and MDCK cells. These tight junction proteins are key components of the skin's physical barrier. Their enhancement not only improves the skin's resistance to external stimuli, but also effectively reduces the skin's water loss, thereby enhancing the skin's barrier function in all directions. In addition, when repairing damaged skin cells (such as human skin fibroblasts HES and HaCaT cells), rice fermentation liquid shows the characteristics of being effective at low concentrations and has no toxic side effects. This mechanism allows rice fermentation liquid to provide a long-term protective barrier for the skin and have anti-aging effects while instantly repairing the damaged barrier.

[0056] Furthermore, rice fermentation filtrate can also promote the proliferation and differentiation of epidermal keratinocytes. Experiments have shown that rice fermentation filtrate at different concentrations can significantly promote the growth of keratinocytes. This cell-level promotion is essential for accelerating the repair process of skin damage. It not only helps to restore the normal structure of the skin, but also provides a solid foundation for the reconstruction of the skin barrier.

[0057] The present invention utilizes the unique biological activity of galactomannan to activate and restore key proteins and neutral lipids of the skin barrier, effectively enhancing the physical and moisture barrier functions of the skin barrier. At the same time, rice fermentation liquid precisely regulates the expression of tight junction proteins, further strengthening the skin barrier. The two work synergistically to achieve comprehensive repair and strengthening of the skin barrier.

[0058] In the present invention, galactomannan not only promotes the synthesis of ECM, but also maintains the integrity and stability of ECM by inhibiting the activity of degrading enzymes, especially enhancing the synthesis of type I collagen, significantly improving the structural integrity and elasticity of the skin. The rice fermented liquid accelerates the natural repair process of skin damage by promoting the proliferation and differentiation of epidermal keratinocytes. The two work together to effectively inhibit skin aging and keep the skin young.

[0059] Compared with the prior art, the present invention utilizes the synergistic effect of galactomannan and rice fermentation liquid, which can significantly enhance the function of the extracellular matrix (ECM), and has excellent barrier repair ability, while showing excellent anti-aging, moisturizing and skin microecological balance promoting properties. Its active ingredients are clear and the product quality is easy to control.

[0060] The present invention adopts a multi-target and multi-pathway intervention strategy, which achieves comprehensive optimization and deep repair of skin barrier function through the synergistic effects of multiple biological mechanisms such as regulating intracellular and extracellular signal pathways, promoting gene expression, enhancing protein synthesis and inhibiting the activity of degradative enzymes.

[0061] As an important component of skin tissue, the quality of ECM directly affects the function of the skin barrier. The present invention achieves comprehensive remodeling and strengthening of ECM by enhancing the synthesis and stability of the main components of ECM (such as collagen, proteoglycans, glycosaminoglycans, etc.) and inhibiting their degradation process. This deep repair of the ECM level provides a more solid and lasting support for the skin barrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 10D of collagen expression after cell culture with the essences of different examples. DETAILED DESCRIPTION

[0063] The invention discloses a composition with the effects of strengthening extracellular matrix and barrier repair. The composition comprises galactomannan and rice fermentation broth. The mass ratio of the galactomannan to the rice fermentation broth is (0.5-2.5):(0.1-10). Preferably, the mass ratio of the galactomannan to the rice fermentation broth is (0.5-1.5):(0.1-3).

[0064] The present invention also provides a skin care product with the effects of strengthening extracellular matrix and barrier repair, comprising a matrix auxiliary material and an effective component, wherein the effective component comprises a composition with the effects of strengthening extracellular matrix and barrier repair. Based on the total weight of the skin care product being 100 parts by weight, the composition with the effects of strengthening extracellular matrix and barrier repair is 1 to 10 parts, preferably 2 to 7.5 parts, and more preferably 3.5 to 7.5 parts. The matrix auxiliary material is selected from any one or more of a solvent, a skin conditioner, a thickener, a moisturizer, an emulsifier, a pH adjuster, a fragrance or a preservative.

[0065] The following are some specific embodiments to illustrate the present invention. These embodiments are only used to further illustrate the present invention, and are not used to limit the present invention, and do not represent the limitation of the protection scope of the present invention. Some non-essential modifications and adjustments made by others based on the present invention still fall within the protection scope of the present invention.

[0066] The present invention takes essence as an example to set various groups of embodiments and comparative examples, but is not limited to the use of essence. The composition can also be applied to various skin care products such as lotion, cream, freeze-dried powder, skin care water, etc. The essence is the above-mentioned key combination and skin care agent, thickener, skin conditioner and pH regulator. Preferably, the raw materials for the preparation of the essence that enhances the extracellular matrix (ECM) and has barrier repair effect also include any one or a combination of at least two of preservatives and flavors.

[0067] Unless otherwise specified, the percentages (%) below are all by mass. All raw materials of the present invention are commercially available products.

[0068] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0069] Example 1

[0070] This embodiment provides an essence having the effects of enhancing the extracellular matrix (ECM) and barrier repairing, wherein the essence comprises a key composition having the barrier repairing effect.

[0071] The raw material components of the essence provided in this embodiment are shown in Table 1.

[0072] Table 1 Components and dosage in Example 1

[0073]

[0074] The preparation method of the essence of this embodiment comprises the following steps:

[0075] a. Obtaining the aqueous phase: Place water in a container with a stirring device and stir, fully wet and mix the other parts of the phase A excipients, add them to the water, stir until completely dissolved, and heat to 75-80°C;

[0076] b. Obtain the oil phase; mix the B phase excipients and heat to 75-80°C, stirring until completely dissolved;

[0077] c. Emulsification: Add the mixture of step b to the aqueous phase of step a, stir at 300 rpm for 4 minutes and homogenize at 3000 rpm for 5 minutes;

[0078] d. Add phase C auxiliary material, stir at 300 rpm for 4 minutes and homogenize at 3000 rpm for 5 minutes, and cool to 45°C with stirring;

[0079] e. Add phase E auxiliary material, stir at 300 rpm for 4 minutes and homogenize at 3000 rpm for 5 minutes;

[0080] f. After the temperature drops to 35-38°C, add phase D, stir at 300 rpm and homogenize at 3000 rpm for 5 minutes to obtain the desired essence.

[0081] Examples 1, 2, 3 and Comparative Examples 1, 2, 3

[0082] The raw material composition ratios of Examples 1, 2, 3 and Comparative Examples 1, 2, 3 are shown in Table 2. The difference between Example 2 and Example 1 is only the increase of each effective ingredient, and the difference between Example 3 and Example 1 is only the reduction of each effective ingredient.

[0083] Comparative Examples 1-3 differ from Example 1 only in that one or two key composition ingredients are missing.

[0084] The specific ratios are shown in Table 2. The preparation method is the same as that in Example 1, except that in step f, the substances in phase D are adaptively added or not added (Comparative Example 3).

[0085] Table 2 Ratio of raw material components of Examples 1-3 and Comparative Examples 1-3

[0086]

[0087] Efficacy evaluation test of Examples 1, 2, 3 and Comparative Examples 1, 2, 3

[0088] 1. Cell proliferation effect test

[0089] Test method:

[0090] 1) Immortalized human epidermal cells (HaCat) were cultured at 10 3 -10 4 The cells were seeded at a density of 10 cells / well in a 96-well plate, 100 μL of DMEM medium was added to the wells, and the cells were cultured at 37° C. in a CO 2 incubator for 24 hours.

[0091] 2) Add samples of Examples 1-3 and Comparative Examples 1-3 to the plate and incubate in an incubator for 48 hours.

[0092] 3) Use a repeating pipette to add 10 μL of CCK8 solution to each well of the culture plate and incubate the culture plate in an incubator for 1-4 hours.

[0093] 4) Use an ELISA reader to measure the absorbance at 450 nm and create a standard curve for data analysis.

[0094] 5) Using Comparative Example 3 as a reference, calculate the cell proliferation change rate, cell proliferation change rate (%) = (absorbance of the test group - absorbance of Comparative Example 3) / absorbance of Comparative Example 3 * 100%. The test results are shown in Table 3 below.

[0095] Table 3 Cell proliferation rate change

[0096] sample Cell proliferation rate (%) Example 1 43.16 Example 2 47.52 Example 3 30.88 Comparative Example 1 17.39 Comparative Example 2 19.47 Comparative Example 3 0.00

[0097] As shown in Table 3, after cell culture using the essence of Examples 1-3 and Comparative Examples 1-2, the cell proliferation rate is significantly improved. However, compared with Comparative Examples 1-2, the improvement effect of Example 1-3 is more significant. From the comparison of the results, it can be seen that when galactomannan and rice fermentation broth are used at the same time, the cell proliferation rate is more significantly improved, compared with not using the essence composition with the effect of enhancing the extracellular matrix (ECM) and barrier repair, or only using one of galactomannan and rice fermentation broth. This shows that galactomannan and rice fermentation broth have a synergistic effect, which has the effect of improving cell viability and delaying cell aging.

[0098] 2. Promotion of epidermal tight junction proteins (Claudin-1 and Claudin-4)

[0099] Test method:

[0100] 1) Normal human epidermal keratinocytes (NHEK) were cultured at 75 cm 2 The cells were collected in the logarithmic phase and cultured at a cell density of 1×10 5 Cells / 500 μL were seeded in 8-well culture plates and cultured overnight.

[0101] 2) After the culture, 500 μL of fresh culture medium was taken and added to Examples 1-3 and Comparative Examples 1-3, respectively. The cells were then cultured for 24 hours and then fixed on each well.

[0102] 3) Claudin-1 was immunofluorescently stained with rabbit anti-Claudin-1 polyclonal antibody, and the cell nuclei were fluorescently stained with DAPI. The promotion of Claudin-1 production was observed using a fluorescence microscope. Claudin-4 was immunofluorescently stained with mouse anti-Claudin-4 monoclonal antibody, and the antibody and cell nuclei were fluorescently stained with DAPI. The promotion of Claudin-4 was observed using a fluorescence microscope.

[0103] Taking comparative example 3 as a reference, the Claudin-1 promotion rate was calculated, Claudin-1 promotion rate (%) = (Claudin-1 value of the test group - Claudin-1 value of comparative example 3) / Claudin-1 value of comparative example 3 * 100%. The test results are shown in Table 4 below.

[0104] Taking comparative example 3 as a reference, the Claudin-4 promotion rate was calculated, Claudin-4 promotion rate (%) = (Claudin-4 value of the test group - Claudin-4 value of comparative example 3) / Claudin-4 value of comparative example 3 * 100%. The test results are shown in Table 5 below.

[0105] Table 4 Claudin-1 promotion rate %

[0106] sample Claudin-1 promotion rate % Example 1 21.86 Example 2 24.31 Example 3 17.62 Comparative Example 1 7.13 Comparative Example 2 8.16 Comparative Example 3 0.00

[0107] Table 5 Claudin-4 promotion rate %

[0108] sample Claudin-4 promotion rate % Example 1 13.65 Example 2 15.42 Example 3 11.07 Comparative Example 1 5.39 Comparative Example 2 6.95 Comparative Example 3 0.00

[0109] As shown in Tables 4 and 5, after cell culture using the essences of Examples 1-3 and Comparative Examples 1-2, the Claudin-1 promotion rate and the Claudin-4 promotion rate were significantly improved. However, compared with Comparative Examples 1-2, the improvement effect of Example 1-3 is more significant. From the comparison of the results, it can be seen that when galactomannan and rice fermentation broth are used at the same time, the Claudin-1 promotion rate and the Claudin-4 promotion rate are more significantly improved, compared with not using the essence composition with the effect of enhancing the extracellular matrix (ECM) and barrier repair, or only using one of galactomannan and rice fermentation broth. This shows that galactomannan and rice fermentation broth have a synergistic effect, can enhance the formation of tight junction protein barriers, and have a barrier repair effect.

[0110] 3. Extracellular matrix protein synthesis effect test

[0111] Test method:

[0112] 1) Immortalized human epidermal cells (HaCat) were cultured at 1x10 410 cells / well were seeded in an 8-well plate, and an appropriate amount of complete culture medium (DMEM or RPMI containing 10% FBS) was added to each well to ensure that the cells were evenly distributed.

[0113] 2) Place the culture plate in a CO2 incubator and culture at 37°C and 5% CO2 for 24 hours to allow the cells to adhere to the wall and reach a suitable growth state.

[0114] 3) After the culture, 500 μL of fresh culture medium was taken and added to Examples 1-3 respectively, and then the cells were cultured for 24 hours and then fixed on each well.

[0115] 4) The expression of collagen was measured by immunofluorescence staining.

[0116] The cell color development method is as follows:

[0117] a. Pour off the culture medium, remove the culture medium, and wash the cells three times with PBS, 5 minutes each time.

[0118] b. Fix the cells with 4% paraformaldehyde solution at 4°C for 20 minutes. Make sure the fixative covers the entire cell layer; after fixation, wash the cells 3 times with pre-cooled PBS for 5 minutes each time to remove residual fixative.

[0119] c. Use a tissue pen to draw circles to prevent the incubation solution from flowing away in the subsequent process, and then rinse with PBS.

[0120] d. Dilute the antibody (COL1A1) with 5% BSA at a ratio of 1:150, add an appropriate amount of primary antibody (COL1A1) working solution, and incubate at 4°C overnight.

[0121] e. Remove the culture dish from 4°C and rewarm it at room temperature for about 30 minutes. Wash the cells three times with pre-cooled PBS for 5 minutes each time to remove unbound primary antibody.

[0122] f. Dilute the antibody (CY3-labeled goat anti-mouse) with 5% BSA at a ratio of 1:100, add the secondary antibody (CY3-labeled goat anti-mouse) working solution, and incubate in a 37°C water bath in the dark for 40 minutes. After incubation, wash the cells 3 times with pre-cooled PBS for 5 minutes each time to remove unbound secondary antibody.

[0123] g. Add DAPI dye to stain the cell nucleus and incubate at room temperature in the dark for 20-30 minutes. After staining, wash the cells 3 times with pre-cooled PBS for 5 minutes each time to remove excess DAPI dye.

[0124] h. Seal the cells with anti-fluorescence quenching mounting medium to ensure that no bubbles are generated.

[0125] i. Place the sealed glass slide under a fluorescence microscope, observe and take pictures, and record the experimental results.

[0126] The results are as follows Figure 1 As shown, I1 is embodiment 1; J1 is embodiment 2; K1 is embodiment 3.

[0127] Depend on Figure 1 It can be seen that after using the essence of Examples 1-3 for cell culture, the collagen content is significantly increased, indicating that galactomannan and rice fermentation liquid can effectively enhance the construction of extracellular matrix and have a barrier repair effect.

[0128] Characterization data and effect data of the products of the embodiments and comparative examples

[0129] 1. Security Testing

[0130] 1) Test sample: Example 1.

[0131] 2) Test method: Use a third-party testing agency to test safety. The selected area should not exceed 50mm 2 , a suitable patch with a depth of about 1mm, add about 0.02-0.025ml (g) of the test substance into the patch tester by the closed patch test method. The patch tester is attached to the curved side of the subject's forearm, and the test substance is removed after 24 hours. The skin reaction is observed at 0.5h, 24h, and 48h after removal: the results are recorded according to the skin reaction grading standards in the "Cosmetic Safety Technical Regulations 2015".

[0132] Table 6 Patch test results

[0133]

[0134]

[0135] Note: Skin reaction grading standards:

[0136] Grade 0: negative reaction;

[0137] Grade 1: Suspicious reaction; only slight erythema;

[0138] Grade 2: weak positive reaction (erythema reaction); erythema, infiltration, edema, and papules may be present;

[0139] Grade 3: Strong positive reaction (herpes reaction); erythema, infiltration, edema, papules, herpes: the reaction may extend beyond the tested area;

[0140] Grade 4: Extremely strong reaction (confluent herpes reaction): obvious erythema, severe infiltration, edema, confluent herpes, and the reaction extends beyond the test area.

[0141] 3) Test results: The results in Table 6 show no irritation and are considered safe.

[0142] 2. Stability test

[0143] 1) Test sample: Example 1.

[0144] 2) Test method: Internal control standard, observe the appearance color, viscosity and pH for three months.

[0145] Table 7 Stability test results

[0146] time 45℃ 40℃ 37℃ 25℃ 5℃ -15℃ CYCLE illumination 1W √ √ √ √ √ √ √ √ 2W √ √ √ √ √ √ √ √ 3W √ √ √ √ √ √ √ √ 4W √ √ √ √ √ √ √ √ 2M √ √ √ √ √ √ √ √ 3M √ √ √ √ √ √ √ √

[0147] Among them, "CYCLE" refers to the cycle, and the cycle setting is: heating from -15℃ to 45℃, cooling from 45℃ to -15℃ for five times.

[0148] 3) Test results: The results in Table 7 show that the essence in Example 1 is stable in properties within 3 months.

[0149] 3. Cuticle moisture content test

[0150] Corneometer CM825 is used to measure the moisture content of the stratum corneum. The higher the measured value, the higher the moisture content of the stratum corneum of the skin.

[0151] 1) Test samples: Examples 1-3, Comparative Examples 1-3.

[0152] 2) Test method: Use the sample twice a day, morning and evening, for 13 weeks. Apply an appropriate amount of the test sample on one cheek, and do not use the test sample on the other cheek. Apply sunscreen products to the entire face every morning.

[0153] The test results were measured at the following four time periods: before using the sample (D0), 7 days after using the sample (D7), 14 days after using the sample (D14), and 91 days after using the sample (D91).

[0154] The test results can be calculated using the following formula:

[0155] Change rate = (analysis value after using the product - analysis value before using the product) ÷ analysis value before using the product × 100%

[0156] Table 8 Change rate of moisture content in stratum corneum

[0157] sample After 7 days of use % After 14 days of use % After 91 days of use % Example 1 14.58 18.75 30.27 Example 2 15.79 19.93 33.68 Example 3 12.31 17.26 27.51 Comparative Example 1 8.83 12.74 20.66 Comparative Example 2 9.35 13.42 22.73 Comparative Example 3 6.94 9.16 16.32

[0158] As shown in the results in Table 8, after using the essence, as the use time increases, the rate of change of the water content of the stratum corneum of the skin of Examples 1-3 and Comparative Examples 1-3 increases, indicating that the water content of the stratum corneum of the skin has improved. However, compared with Comparative Examples 1-3, when the same time is used, the rate of change of the water content of the stratum corneum of the skin of Examples 1-3 is greater, indicating that the improvement effect of the water content of the stratum corneum of the skin after the essence of Examples 1-3 is more significant. As shown in the comparison of the results of Examples 1-3 and Comparative Examples 1-3, compared with not using an essence composition having an enhanced extracellular matrix (ECM) and barrier repair effect, or only using one of galactomannan and rice fermentation liquid, when galactomannan and rice fermentation liquid are used simultaneously, the improvement effect of the water content of the stratum corneum of the skin is better. It shows that galactomannan and rice fermentation liquid have a synergistic effect, can effectively improve the water content of the stratum corneum of the skin, and have a barrier repair effect.

[0159] 4.TEWL(g / m 2 / h) Transepidermal water loss test

[0160] The transepidermal water loss (TEWL) is accurately measured using the non-invasive measurement device TEWL (Dermalab Combo). If the TEWL decreases significantly before and after the use of the test product, it means that the skin barrier has been restored and the product has a repairing effect.

[0161] 1) Test samples: Examples 1-3, Comparative Examples 1-3.

[0162] 2) Test method: Use the sample twice a day, morning and evening, for 13 weeks. Apply an appropriate amount of the test sample on one cheek, and do not use the test sample on the other cheek. Apply sunscreen products to the entire face every morning.

[0163] The test results were measured at the following four time periods: before using the sample (D0), 7 days after using the sample (D7), 14 days after using the sample (D14), and 91 days after using the sample (D91).

[0164] The test results can be calculated using the following formula:

[0165] Change rate = (analysis value after using the product - analysis value before using the product) ÷ analysis value before using the product × 100%

[0166] Table 9 Change rate of skin transepidermal water loss value

[0167] sample After 7 days of use % After 14 days of use % After 91 days of use % Example 1 -12.27 -16.53 -28.45 Example 2 -13.54 -17.49 -29.96 Example 3 -11.15 -14.24 -27.03 Comparative Example 1 -7.12 -9.33 -16.42 Comparative Example 2 -7.86 -10.71 -18.56 Comparative Example 3 -4.13 -6.05 -9.48

[0168] From the results in Table 9, it can be seen that after using the essence, as the use time increases, the change rate of the transepidermal water loss value of Examples 1-3 and Comparative Examples 1-3 decreases, indicating that the skin barrier is improved. However, compared with Comparative Examples 1-3, when used for the same time, the change rate of the transepidermal water loss value of Examples 1-3 is greater, indicating that the skin barrier is repaired more significantly after using the essence of Examples 1-3.

[0169] From the comparison of the results of Examples 1-3 and Comparative Examples 1-3, it can be seen that when galactomannan and rice fermentation broth are used together, the improvement effect of skin transepidermal water loss value is better than that of not using the essence composition with the effect of enhancing the extracellular matrix (ECM) and barrier repair, or using only one of galactomannan and rice fermentation broth. This shows that galactomannan and rice fermentation broth have a synergistic effect, can effectively improve the skin transepidermal water loss value, and have a barrier repair effect.

[0170] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. A composition having the effects of strengthening extracellular matrix and barrier repair, characterized in that: The invention comprises galactomannan and rice fermentation liquid, wherein the mass ratio of the galactomannan to the rice fermentation liquid is (0.5-2.5): (0.1-10).

2. A composition having the effects of strengthening extracellular matrix and barrier repair according to claim 1, characterized in that: The mass ratio of the galactomannan to the rice fermentation liquid is (0.5-1.5):(0.1-3).

3. Use of a composition having the effects of strengthening extracellular matrix and barrier repair according to any one of claims 1 to 2 in the preparation of skin care products.

4. A skin care product having the effects of strengthening extracellular matrix and barrier repair, characterized in that: The invention comprises matrix auxiliary materials and functional components, wherein the functional components comprise the composition having the functions of strengthening the extracellular matrix and repairing the barrier as described in any one of claims 1 to 2.

5. The skin care product having the effects of strengthening the extracellular matrix and repairing the barrier according to claim 4, characterized in that: Based on 100 parts by weight of the total weight of the skin care product, the composition having the effects of strengthening the extracellular matrix and repairing the barrier is 1 to 10 parts.

6. The skin care product having the effects of strengthening the extracellular matrix and repairing the barrier according to claim 5, characterized in that: Based on 100 parts by weight of the total weight of the skin care product, the composition having the effects of strengthening the extracellular matrix and repairing the barrier is 2 to 7.5 parts.

7. The skin care product having the effects of strengthening the extracellular matrix and repairing the barrier according to claim 4, characterized in that: The matrix auxiliary material is selected from any one or more of solvents, skin conditioners, thickeners, moisturizers, emulsifiers, pH regulators, fragrances or preservatives.

8. The skin care product having the effects of strengthening the extracellular matrix and repairing the barrier according to claim 4, characterized in that: The skin care product is an essence, which includes phase A auxiliary materials, phase B auxiliary materials, phase C auxiliary materials, phase D functional ingredients and phase E auxiliary materials; based on 100 parts by weight of the essence, The phase A auxiliary material includes 0.03-10 parts of thickener A, 0.12-5 parts of skin conditioner A and 3-20 parts of moisturizer; The phase B auxiliary material includes 0.3-15 parts of thickener B, 0.1-5 parts of emulsifier and 0.5-5 parts of skin conditioner B; The phase C auxiliary material includes 0.1 to 5 parts of a pH adjuster; The dosage of the D phase functional ingredient is 1 to 10 parts; The E phase auxiliary material includes 0.01 to 2 parts of fragrance and 0.15 to 1 part of preservative; The phase A auxiliary material includes water, and the amount of water is such that the weight parts of the essence reaches 100 parts; The thickener A and the thickener B are independently selected from one or more of acrylic acid (esters) / C10-30 alkyl acrylate cross-linked polymer, polyacrylamide, xanthan gum, propylene glycol, behenyl alcohol, carbomer, laureth-7, cetyl alcohol, stearyl alcohol or hydroxyethyl cellulose; The skin conditioning agent A and the skin conditioning agent B are each independently selected from one or more of glycerin, isohexadecane, olive oil, sodium hyaluronate, lauryl alcohol, avocado butter, allantoin, butylene glycol, squalane, panthenol, lactic acid, tocopherol or dipotassium glycyrrhizinate; The moisturizing agent is selected from one or more of hyaluronic acid, allantoin, glycerin, ceramide, sodium lactate, butylene glycol, ethylhexylglycerin, hydrogenated grape seed oil, olive oil or shea butter; The emulsifier is selected from one or more of glyceryl stearate citrate, cetearyl glucoside (and) cetearyl alcohol, behenyl trimethyl ammonium methyl sulfate, cocoglyceryl esters or sorbitan olivate; The pH regulator is selected from one or more of arginine or lactic acid; The aromatic agent is selected from essences; The preservative is selected from one or more of phenoxyethanol, ethylhexylglycerin or pentylene glycol.

9. The skin care product having the effects of strengthening the extracellular matrix and repairing the barrier according to claim 8, characterized in that: Based on 100 parts by weight of the essence, the phase A auxiliary materials include 0.01-5 parts of xanthan gum, 0.02-10 parts of carbomer, 0.02-1 parts of sodium hyaluronate, 0.1-5 parts of allantoin, 2-15 parts of glycerin and 1-10 parts of butylene glycol; the phase B auxiliary materials include 0.2-10 parts of cetyl alcohol, 0.1-10 parts of stearyl alcohol, 0.1-5 parts of cetearyl glucoside (and) cetearyl alcohol, and 0.5-5 parts of squalane; the phase C auxiliary materials include 0.1-5 parts of arginine; the phase D effective ingredients are 1 to 10 parts; the phase E auxiliary materials include 0.1-1 parts of phenoxyethanol, 0.05-1 parts of ethylhexylglycerin and 0.01-2 parts of fragrance.

10. The method for preparing a skin care product having the functions of strengthening the extracellular matrix and repairing the barrier according to claim 8, characterized in that: The following steps are involved: a. Obtaining the aqueous phase: Place water in a container with a stirring device and stir, fully wet and mix the other parts of the phase A excipients, add them to the water, stir until completely dissolved, and heat to 75-80°C; b. Obtain the oil phase; mix the B phase excipients and heat to 75-80°C, stirring until completely dissolved; c. Emulsification: Add the mixture of step b to the aqueous phase of step a, stir at 250-350 rpm for 3-5 minutes and homogenize at 2500-3500 rpm for 3-10 minutes; d. Add phase C auxiliary material, stir at 250-350 rpm for 3-5 minutes and homogenize at 2500-3500 rpm for 3-10 minutes, and cool to 45°C with stirring; e. Add E phase auxiliary materials, stir at 250-350 rpm for 3-5 minutes and homogenize at 2500-3500 rpm for 3-10 minutes; f. After the temperature drops to 35-38°C, add phase D, stir at 250-350 rpm and homogenize at 2500-3500 rpm for 3-10 minutes to obtain the desired essence.

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