A composition for combating and repairing photo-damage and its use in skin care

Abstract

The present application relates to a kind of compositions for resisting and repairing photo-damage and its application in skin care, belong to the field of cosmetics.The present application first proposes to solve the problem of photo-damage from multiple dimensions, the composition described in the present application includes lactobacillus / grape juice extract, tetrahydro methyl pyrimidine carboxylic acid, glucosyl hesperidin, adenosine, helianthus spadiceus callus culture filtrate, each active ingredient complements each other, synergistic effect, so that it can resist photo-damage, improve skin repair ability, improve the problem of pigmentation caused by photo-damage, also can strengthen skin barrier, effectively resist external invasion, provide a mild solution for people with sensitive skin and troubled by photo-damage.

Description

Technical Field

[0001] This invention relates to a composition for resisting and repairing photodamage and its application in skincare, belonging to the field of cosmetic technology. Background Technology

[0002] The skin, the largest organ in the human body, is in direct contact with the external environment. Prolonged exposure to sunlight can have varying degrees of impact on the skin. Recent studies have shown that visible light and long-wave ultraviolet (UV) radiation have a synergistic effect on skin pigmentation and erythema. In addition to causing erythema, DNA damage, and free radical production, visible light can also cause skin darkening and persistent inflammatory responses. As the first interface between the human body and the external environment, the skin produces reactive oxygen species (ROS) induced by UV and visible light. The human body has various oxygen free radical scavengers, and under normal circumstances, oxidation and antioxidation are in a dynamic balance. When the production of ROS exceeds the processing capacity of the antioxidant defense system, excessive free radicals interact with DNA and proteins, leading to oxidative stress damage. Oxidative stress and the resulting oxidative damage exacerbate skin pigmentation.

[0003] The problem of sensitive skin is becoming increasingly prominent globally, making sensitive skin care a crucial skincare issue of public concern. The skin barrier is a vital barrier system in the human body, essential for maintaining overall health. Its homeostasis is strictly and precisely regulated, and damage to the skin barrier is a key factor in the development of sensitive skin. When the skin barrier is damaged, transepidermal water loss (TEWL) increases, easily leading to local inflammatory responses, causing vasodilation. Simultaneously, the destruction of sulfhydryl groups in the skin increases tyrosinase activity, inducing inflammatory pigmentation. Furthermore, a damaged skin barrier increases the skin's sensitivity to ultraviolet radiation and external microorganisms, disrupts keratinocyte function, and prevents melanocytes from being promptly expelled from the epidermis. This results in increased melanin retention time in the basal layer, leading to the production of inflammatory mediators, triggering inflammatory responses, and ultimately further causing pigmentation. This also exacerbates the damage to the skin barrier, worsening skin sensitivity.

[0004] Currently, existing technologies do not address the multi-dimensional approach to resisting and repairing photodamage; instead, they focus on either resisting or repairing photodamage in a single way. Therefore, it is essential to develop a composition that can target photodamage from multiple dimensions and multiple points and is suitable for people with sensitive skin. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a composition for resisting and repairing photodamage and its application in skincare. This composition can not only resist and repair photodamage, but also improve pigmentation problems caused by the damage, strengthen the skin barrier, and effectively resist external aggressors, providing a gentle solution for people with sensitive skin who are troubled by photodamage.

[0006] To solve the above-mentioned technical problems, the objective of this invention is achieved through the following technical solution:

[0007] A composition for resisting and repairing photodamage, comprising the following components: Lactobacillus / grape juice extract, tetrahydromethylpyrimidine carboxylic acid, glucosyl hesperidin, adenosine, and callus culture filtrate of Erycibe obtusifolia.

[0008] The above composition can not only resist and repair photodamage in multiple dimensions, but also the emulsion containing the composition has additional effects such as moisturizing, soothing and repairing redness.

[0009] In this invention, Lactobacillus / grape juice extract is an active ingredient that can soften keratinocytes, promote metabolism, and has moisturizing effects. Specifically, Lactobacillus refers to *Lactiplantibacillus plantarum*, whose fermentation filtrate mainly consists of lactic acid and amino acids, with a lactic acid content of 3.0-7.0 wt%.

[0010] According to a preferred embodiment of the present invention, the lactobacillus / grape juice extract is obtained by fermenting grape juice with Lactobacillus plantarum.

[0011] More preferably, the preparation method of the lactobacillus / grape juice extract is as follows: *Lactobacillus plantarum* is cultured to obtain a *Lactobacillus plantarum* bacterial suspension, wherein the bacterial concentration of the *Lactobacillus plantarum* bacterial suspension is 1–10 × 10⁻⁶. 7 cfu / mL; After pressing the grapes, add water at a material-to-liquid ratio of 1:(10-15), homogenize, sterilize, and then inoculate with a suspension of Lactobacillus plantarum at a volume percentage of 1-2%. Culture in anaerobic fermentation at 35-40℃ for 30-60 hours. After fermentation, centrifuge and collect the supernatant. After filtration and sterilization, the Lactobacillus / grape juice extract is obtained.

[0012] In this invention, tetrahydromethylpyrimidine carboxylic acid, also known as ectoine, is an amino acid derivative that not only balances cell osmotic pressure but also provides excellent protection for enzymes, DNA, proteins, nucleic acids, cell membranes, and the entire cell under adverse conditions such as high temperature, cold, drought, extreme pH, high pressure, high salinity, and high radiation. It constructs a strong protective layer composed of water molecules in the skin, strengthening and restoring cell function, stabilizing the skin barrier, and restoring and regulating moisture content. It also activates Langerhans cells in the skin, stimulating the skin's self-repair ability and enhancing the skin cells' immune defense capabilities. Data shows that 0.5% ectoine can completely protect Langerhans cells in the skin from UV radiation damage, while 1.0% ectoine provides a faster emergency response, preventing cell damage and increasing the self-healing and repair speed by 2-3 times.

[0013] In this invention, glucosylhesperidin is a structurally stable flavonoid. Hesperidin is a dihydroflavonoid compound unique to citrus peel, also known as vitamin P. This type of substance is widely found in plants in nature and is a product of natural plant metabolism. Flavonoid molecules have a conjugated structure, enabling them to strongly absorb ultraviolet and visible light. Furthermore, the phenolic hydroxyl groups in the structure are reducing and easily oxidized, thus having a certain scavenging effect on free radicals already generated in the skin. In addition, it also maintains normal vascular osmotic pressure, enhances capillary toughness, shortens bleeding time, and reduces vascular fragility. Its glycosylated product, glucosylhesperidin, has superior water solubility and bioactivity compared to hesperidin. Furthermore, glucosylhesperidin has skin-warming, brightening, and dark circle-improving effects, making it an important raw material in high-end cosmetics.

[0014] In this invention, adenosine is a compound formed by the N-9 of adenine and the C-1 of D-ribose linked by a β-glycosidic bond, scientifically known as 9-β-D-rifuranosyladenine (ADO). It plays a crucial role in the body's energy metabolism, transferring energy in the form of adenosine triphosphate (ATP) or adenosine diphosphate (ADP). It can replenish skin energy, significantly improve skin elasticity, promote skin cell metabolism, prevent skin tissue sagging, and restore skin elasticity by tightening and shrinking pores.

[0015] In this invention, the callus culture filtrate of *Eryngium sibiricum* is obtained through stem cell technology, containing complete plant components and rich in flavonoids, amino acids, and sugars. In medicine, *Eryngium sibiricum* is frequently used as an analgesic and anti-inflammatory drug. In the skincare field, it primarily acts on the healing, repair, and proliferation stages, promoting the proliferation and differentiation of keratinocytes and the production of integrin β1 and type IV collagen.

[0016] According to a preferred embodiment of the present invention, the mass fractions of each component in the composition are as follows:

[0017] 5-10 parts of Lactobacillus / grape juice extract, 0.01-0.5 parts of tetrahydromethylpyrimidine carboxylic acid, 0.01-1 part of glucosyl hesperidin, 0.01-0.5 parts of adenosine, and 0.01-0.5 parts of callus culture filtrate of Erycibe obtusifolia.

[0018] More preferably, the mass fractions of each component in the composition are as follows:

[0019] 6-8 parts of Lactobacillus / grape juice extract, 0.01-0.2 parts of tetrahydromethylpyrimidine carboxylic acid, 0.01-0.5 parts of glucosyl hesperidin, 0.01-0.2 parts of adenosine, and 0.01-0.2 parts of callus culture filtrate of Erycibe obtusifolia.

[0020] More preferably, the mass fractions of each component in the composition are as follows:

[0021] 7 parts of Lactobacillus / grape juice extract, 0.1 part of tetrahydromethylpyrimidine carboxylic acid, 0.2 parts of glucosyl hesperidin, 0.1 parts of adenosine, and 0.1 parts of callus culture filtrate of Erycibe obtusifolia.

[0022] The above composition is used in the preparation of skin care products that have the function of resisting and repairing photodamage.

[0023] An emulsion with properties that resist and repair photodamage, comprising the following components by weight percentage:

[0024] The composition for resisting and repairing photodamage consists of 1-3% emulsifier, 0.1-3% oil, 1-10% rheology modifier, 0.1-0.8% humectant, 2-11% preservative, 0.2-2% chelating agent, 0.01-0.1% pH adjuster, and water to make up to 100%.

[0025] According to a preferred embodiment of the present invention, the emulsifier comprises, by mass percentage: 0.01-0.2% sodium di(lauramide-glutamine)lysine, 0.2-0.5% C12-20 alkyl glucoside, and 0.5-2% C14-22 alcohol.

[0026] According to a preferred embodiment of the present invention, the oil comprises, by mass percentage: 0.05-2% polydimethylsiloxane, 0.1-0.2% tocopheryl acetate, 2-4% isononyl isononanoate, and 0.1-0.5% shea butter.

[0027] According to a preferred embodiment of the present invention, the rheology modifier comprises, by mass percentage: 0.1-0.5% carbomer and 0.01-0.2% hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer.

[0028] According to a preferred embodiment of the present invention, the moisturizer comprises, by weight percentage: 1-5% glycerin, 1-5% 1,3-butanediol, and 0.01-0.5% hydrolyzed sodium hyaluronate.

[0029] According to a preferred embodiment of the present invention, the preservative comprises, by mass percentage: 0.1-1% 1,2-hexanediol and 0.1-0.8% p-hydroxyacetophenone.

[0030] According to a preferred embodiment of the present invention, the chelating agent is disodium EDTA.

[0031] According to a preferred embodiment of the present invention, the pH adjuster is tromethamine.

[0032] The preparation method of the above-mentioned emulsion with the ability to resist and repair photodamage includes the following steps:

[0033] (1) Mix the humectant, rheology modifier, emulsifier, chelating agent and some water evenly, heat to 80-85℃, and stir to dissolve evenly;

[0034] (2) Stir the oil evenly, heat it to 75-80℃, and then add it to the mixture in step (1). Homogenize it at 3000-5500 rpm for 3-6 minutes, and then cool it down to 70-75℃.

[0035] (3) Add preservative to the homogenized mixture in step (2), and after it is completely dissolved, add pH adjuster. After it is dissolved evenly, cool it to 40-45℃, add the composition solution of resisting and repairing light damage prepared from the remaining water, mix evenly, and cool it to room temperature to obtain the final product.

[0036] According to a preferred embodiment of the present invention, the composition solution for resisting and repairing photodamage is prepared according to the following method:

[0037] 1) Heat the water to 90-95℃ and keep it at that temperature for 30-50 minutes to sterilize it, then cool it down to 80-85℃;

[0038] 2) Add tetrahydromethylpyrimidine carboxylic acid and adenosine to the water prepared in step 1), stir until completely dissolved, and then cool to 40-45℃;

[0039] 3) Add the Lactobacillus / grape juice extract, glucosyl hesperidin, and filtrate of callus culture from Erycibe obtusifolia to the solution in step 2), and stir to mix evenly.

[0040] Beneficial effects:

[0041] 1. This invention is the first to propose a multi-dimensional approach to solving the problem of photodamage. The composition of this invention includes Lactobacillus / grape juice extract, tetrahydromethylpyrimidine carboxylic acid, glucosyl hesperidin, adenosine, and callus culture filtrate of Erycibe obtusifolia. The active ingredients complement each other and work synergistically to enhance the skin's repair ability and improve pigmentation caused by photodamage while resisting photodamage. It can also strengthen the skin barrier and effectively resist external aggressors, providing a gentle solution for people with sensitive skin who are troubled by photodamage.

[0042] 2. Literature reports that researchers used B16 melanocytes in related studies, and the results showed that adenosine has a regulatory effect on pigmentation in B16 cells, and only at a certain concentration does it have an inhibitory effect on melanin. In this invention, the combination of five components improves the phenomenon that low concentrations of adenosine cannot improve pigmentation. Attached Figure Description

[0043] Figure 1 Images showing the irritation evaluation of the chorioallantoic membrane vessels of chicken embryos in the sample solution of the composition of Example 1;

[0044] Figure 2 Bar chart showing the test results of DPPH free radical scavenging rate under the action of different functional compositions;

[0045] Figure 3 Bar chart showing the relative cell viability of fibroblasts under UVA irradiation under the action of different functional compositions;

[0046] Figure 4 Bar chart showing the skin ITA° values ​​after using the emulsions in Application Examples 1, 2, and Comparative Example 12; Statistical analysis results: "-": no statistically significant difference (P≥0.05); "*": statistically significant difference (0.01≤P<0.05); "**": statistically significant difference (0.001≤P<0.01); "***": statistically significant difference (P<0.001);

[0047] Figure 5 A bar chart showing the results of skin moisture content testing after using the lotion in Application Example 1;

[0048] Figure 6 A bar chart showing the results of the transdermal water loss (TEWL) test after using the emulsion in Application Example 1;

[0049] Figure 7 A bar chart showing the rate of change of the red area a* value relative to the initial value after applying the emulsion in Example 1;

[0050] Figure 8 Images of the facial red area before and after applying the lotion in Example 1. Detailed Implementation

[0051] The technical solution of the present invention will be further described in detail below with reference to embodiments. It should be noted that the following detailed descriptions are exemplary and intended to provide further explanation of the present invention, and do not constitute any limitation on the scope of protection of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Unless otherwise specified, the experimental methods used in the specific embodiments are conventional methods; the materials, reagents, etc., used are commercially available unless otherwise specified; and the percentages mentioned in the specific embodiments are mass percentages unless otherwise specified.

[0052] The sources of the main raw materials in this invention are shown in Table 1.

[0053] Table 1. Sources of the main raw materials in this invention

[0054] Main raw material components source Tetrahydromethylpyrimidine carboxylic acid Shandong Freda Biotechnology Co., Ltd. Glucosyl hesperidin INNOVATION LABO adenosine Jiangsu Suryu Biotechnology Co., Ltd. Filtrate of callus culture from sea celery SEPPIC SA

[0055] The preparation method of Lactobacillus / grape juice extract in the example is as follows: Lactobacillus plantarum (strain number: CICC 25125, sourced from the China Industrial Microbial Culture Collection Center) was inoculated and streaked on an MRS solid medium slant tube, and anaerobically cultured at 37℃ for 48 h. Single colonies were picked and anaerobically activated in MRS liquid medium at 37℃ for 24 h. The activated bacterial solution was then centrifuged at 6000 r / min for 5 min, and the lower layer of cells was collected. Sterile water was added to prepare 1×10⁻⁶ saturates. 7 A CFU / mL bacterial suspension was prepared for use. After washing the grapes, the juice was extracted using a juicer (without adding water during the juicing process) to obtain freshly pressed grape juice. Water was added at a material-to-liquid ratio of 1:10, and the fresh product was homogenized, sealed, and sterilized (autoclaved at 121℃ for 30 min). This was then used as the substrate. 5 mL of the bacterial suspension was added to 250 mL of the substrate, and the mixture was anaerobically fermented at 37℃ for 48 h. After fermentation, the supernatant was collected by centrifugation at 8000-10000 rpm and filtered through a 0.22 μm filter membrane to obtain the Lactobacillus / grape juice extract.

[0056] Examples 1-4: Composition and preparation of the efficacy composition

[0057] The compositions of the efficacy combination solutions in Examples 1, 2, 3, and 4 are shown in Table 2, and their preparation methods are as follows:

[0058] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0059] (2) Add tetrahydromethylpyrimidine carboxylic acid and adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0060] (3) Add the lactobacillus / grape juice extract, glucosyl hesperidin, and filtrate of callus culture from sea celery to the solution in step (2) and stir to mix evenly.

[0061] Table 2. Composition of the functional composition solution (mass percentage)

[0062]

[0063] Comparative Example 1:

[0064] The composition of the efficacy composition solution of Comparative Example 1 is shown in Table 2, and its preparation method is as follows:

[0065] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0066] (2) Add adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0067] (3) Add the lactobacillus / grape juice extract, glucosyl hesperidin, and filtrate of callus culture from sea celery to the solution in step (2) and stir to mix evenly.

[0068] Comparative Example 2:

[0069] The composition of the efficacy composition solution of Comparative Example 2 is shown in Table 2, and its preparation method is as follows:

[0070] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0071] (2) Add tetrahydromethylpyrimidine carboxylic acid to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0072] (3) Add the lactobacillus / grape juice extract, glucosyl hesperidin, and filtrate of callus culture from sea celery to the solution in step (2) and stir to mix evenly.

[0073] Comparative Example 3:

[0074] The composition of the efficacy composition solution of Comparative Example 3 is shown in Table 2, and its preparation method is as follows:

[0075] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0076] (2) Add tetrahydromethylpyrimidine carboxylic acid and adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0077] (3) Add glucosyl hesperidin and the filtrate of callus culture of sea celery to the solution in step (2) and stir to mix evenly.

[0078] Comparative Example 4:

[0079] The composition of the efficacy composition solution of Comparative Example 4 is shown in Table 2, and its preparation method is as follows:

[0080] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0081] (2) Add tetrahydromethylpyrimidine carboxylic acid and adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0082] (3) Add Lactobacillus / grape juice extract and glucosyl hesperidin to the solution in step (2) and stir to mix evenly.

[0083] Comparative Example 5:

[0084] The composition of the efficacy composition solution of Comparative Example 5 is shown in Table 2, and its preparation method is as follows:

[0085] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0086] (2) Add tetrahydromethylpyrimidine carboxylic acid and adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0087] (3) Add the Lactobacillus / grape juice extract and the filtrate of callus culture of sea celery to the solution in step (2) and stir to mix evenly.

[0088] Comparative Example 6:

[0089] The composition of the efficacy composition solution of Comparative Example 6 is shown in Table 2, and its preparation method is as follows:

[0090] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0091] (2) Add adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0092] (3) Add the Lactobacillus / grape juice extract and the filtrate of callus culture of sea celery to the solution in step (2) and stir to mix evenly.

[0093] Comparative Example 7:

[0094] The composition of the efficacy composition solution of Comparative Example 7 is shown in Table 2, and its preparation method is as follows:

[0095] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0096] (2) Add tetrahydromethylpyrimidine carboxylic acid to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0097] (3) Add Lactobacillus / grape juice extract and glucosyl hesperidin to the solution in step (2) and stir to mix evenly.

[0098] Comparative Example 8:

[0099] The composition of the efficacy composition solution of Comparative Example 8 is shown in Table 2, and its preparation method is as follows:

[0100] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0101] (2) Add tetrahydromethylpyrimidine carboxylic acid and adenosine to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0102] (3) Add glucosyl hesperidin to the solution in step (2) and stir to mix evenly.

[0103] Comparative Example 9:

[0104] The composition of the efficacy composition solution of Comparative Example 9 is shown in Table 2, and its preparation method is as follows:

[0105] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 85℃;

[0106] (2) Add tetrahydromethylpyrimidine carboxylic acid to the water prepared in step (1), stir until completely dissolved, and then cool to 45°C;

[0107] (3) Add glucosyl hesperidin to the solution in step (2) and stir to mix evenly.

[0108] Comparative Example 10:

[0109] The composition of the efficacy composition solution of Comparative Example 10 is shown in Table 2, and its preparation method is as follows:

[0110] (1) Heat the water to 90-95℃ and keep it at that temperature for 30 minutes to sterilize it, then cool it down to 45℃;

[0111] (2) Add the Lactobacillus / grape juice extract and the filtrate of callus culture of sea celery to the water prepared in step (1) and stir to mix evenly.

[0112] Comparative Example 11:

[0113] The composition of the efficacy composition solution in Comparative Example 11 is shown in Table 2. Its preparation method is the same as in Example 1, except that the Lactobacillus / grape juice extract is different. The preparation method of the Lactobacillus / grape juice extract in Comparative Example 11 involves inoculating a grape juice culture medium with a concentration of 1×10⁻⁶. 7 The cfu / mL saliva-based Lactobacillus salivarius (strain number: CICC 23174, sourced from the China Industrial Microbial Culture Collection Center) suspension was fermented and cultured under the same conditions of inoculum size, fermentation temperature, and fermentation time. The resulting Lactobacillus / grape juice extract was then added to the efficacy composition solution of Comparative Example 11.

[0114] Test Example 1: Safety Evaluation

[0115] The test was conducted according to the "Clonal Allantoic Membrane Test for Eye Irritation / Corrosivity of Cosmetics" (SN / T2329-2009), using the reaction time method. Nine-day-old SPF-grade chicken embryos were candled to expose the allantoic membrane (CAM) by removing the top of the air cell. 0.3 mL of the test sample was directly applied to the CAM surface, and the time for the onset of bleeding, vascular lysis, and clotting within 5 minutes was observed and recorded. Six parallel experiments were performed for each test sample. A negative control (0.9% sodium chloride injection) and a positive control (0.1 mol / L sodium hydroxide solution) were also included.

[0116] The stimulus score is calculated as follows: IS = (301-sec H) × 5 / 300 + (301-sec L) × 7 / 300 + (301-sec C) × 9 / 300.

[0117] In the formula: sec H is the average time for the onset of bleeding observed on the CAM membrane, in seconds (s); sec L is the average time for the onset of vascular dissolution observed on the CAM membrane, in seconds (s); sec C is the average time for the onset of coagulation observed on the CAM membrane, in seconds (s).

[0118] Irritation classification: IS<1, non-irritating; 1≤IS<5, mildly irritating; 5≤IS<9, moderately irritating; IS≥10, strongly irritating / corrosive.

[0119] The comparison of the sample solution in Example 1 before and after its application to the chorioallantoic membrane is shown in the figure. Figure 1 The results of the stimulation scores for the sample solutions are shown in Table 3.

[0120] Table 3. Results of the chicken embryo chorioallantoic membrane test

[0121] experimental group sec H / s sec L / s sec C / s IS Stimulus Classification Positive control 15.0 31.0 301.0 11.06667 Strongly irritating / corrosive negative control 301.0 301.0 301.0 0.0 Non-irritating Example 1 Sample 301.0 301.0 301.0 0.0 Non-irritating

[0122] The results above show that, using 0.9% sodium chloride injection as a negative control, the irritation score was 0, indicating no irritation; using 0.1 mol / L sodium hydroxide solution as a positive control, the irritation score was 11.06667, and bleeding and coagulation could be observed under a stereomicroscope; the irritation score of the composition solution in Example 1 was 0, and there was no hemolysis in the blood vessels, indicating that the composition is non-irritating to the eyes. This indicates that the composition of the present invention has great potential to be used as a functional ingredient suitable for sensitive skin in cosmetics.

[0123] Experimental Example 2: In vitro antioxidant activity assay – DPPH free radical scavenging capacity assay

[0124] Take 2 mL of anhydrous ethanol and 2 mL of 0.3 mmol / L DPPH ethanol solution and place them in test tubes, mix well, let stand in the dark for 30 min, and measure the absorbance A0 at 517 nm using a UV spectrophotometer.

[0125] Samples from Examples 1-4 and Comparative Examples 1-11 were prepared into sample solutions with a mass concentration of 1.2 mg / mL. 2 mL of each sample solution with a mass concentration of 1.2 mg / mL was placed in a reaction tube, and 2 mL of 0.3 mmol / L DPPH ethanol solution was added. The mixture was stirred and each tube was allowed to stand in the dark for 30 min. The absorbance value A1 of each tube was measured at a wavelength of 517 nm using a UV spectrophotometer.

[0126] Take 2 mL of sample solution with a mass concentration of 1.2 mg / mL and place it in a reaction tube. Add 2 mL of anhydrous ethanol, mix well, and let each tube stand in the dark for 30 min. Use an ultraviolet spectrophotometer to measure the absorbance value A2 of each tube at a wavelength of 517 nm.

[0127] The formula for calculating the DPPH free radical scavenging rate is: Scavenging rate (%) = [A0 - (A1 - A2)] / A0 × 100%.

[0128] In the formula, A0 is the absorbance value of the DPPH ethanol solution; A1 is the absorbance value of the sample solution after scavenging DPPH free radicals; and A2 is the absorbance value of the sample solution.

[0129] The results of DPPH free radical scavenging are shown in Table 4 and Figure 2 As shown.

[0130] Table 4. Results of DPPH free radical scavenging rate

[0131] sample DPPH free radical scavenging rate (%) Example 1 95.96 Example 2 89.56 Example 3 93.57 Example 4 92.64 Comparative Example 1 62.41 Comparative Example 2 61.27 Comparative Example 3 65.29 Comparative Example 4 66.72 Comparative Example 5 62.98 Comparative Example 6 33.13 Comparative Example 7 30.25 Comparative Example 8 28.93 Comparative Example 9 33.3 Comparative Example 10 22.03 Comparative Example 11 70.92

[0132] From Table 3 and Figure 2 As can be seen, the DPPH free radical scavenging rates of Examples 1-4 are all above 90%, and the DPPH free radical scavenging ability of the compositions in Comparative Examples 1-10 is significantly less than that of Example 1. In particular, the DPPH free radical scavenging ability of the compositions in Comparative Examples 6 and 9, as well as the compositions in Comparative Examples 8 and 10, is significantly less than that of Example 1, which further illustrates the significant synergistic effect of the active ingredients in the compositions of the present invention.

[0133] Compared with Example 1, Comparative Example 11 shows that the free radical scavenging ability of the Lactobacillus / grape juice extract obtained by fermentation of Lactobacillus plantarum in the composition of the present invention is greater than that of Lactobacillus salivarii. Therefore, the composition obtained by fermentation of Lactobacillus plantarum in the composition of the present invention has better antioxidant capacity.

[0134] Experimental Example 3: Cellular UV Damage Repair Test

[0135] To test the effectiveness of the composition in repairing photodamage, this experiment used cell activity as a criterion; higher cell activity indicated better repair of UV damage. Succinate dehydrogenase in the mitochondria of living cells can reduce MTT to formazan, a function absent in dead cells. Dimethyl sulfoxide can dissolve formazan in cells. The absorbance (OD) value was measured at 490 nm using an ELISA reader. Within a certain cell count range, the more living cells present, the higher the OD value, indicating stronger cell activity.

[0136] Fibroblasts (HSF cells, purchased from Guangdong Boxi Biotechnology Co., Ltd.) were cultured in 10cm diameter culture dishes at 37℃ and 5% CO2 in a carbon dioxide incubator. The culture medium contained 10% fetal bovine serum. When the cell confluence reached 80-90%, the fibroblasts were digested, resuspended, and seeded into 96-well cell culture plates (cell density 1×10⁶). 4 The test can be performed with cells per well. The experiment is divided into control wells, experimental wells, and zeroing wells. The zeroing wells are not seeded with cells. The culture medium in the control wells is replaced with fresh culture medium. Using fresh culture medium as a solvent, samples from Examples 1-4 and Comparative Examples 1-11 are prepared into sample solutions with a mass concentration of 1.5 mg / mL for the experimental wells, replacing the old culture medium, and cultured for 24 hours. The control wells are further divided into non-irradiated and irradiated sections. The non-irradiated control wells are protected from light, while the irradiated control wells and experimental wells are continuously irradiated under a UVA lamp for 6 hours (irradiation dose of 9 J / cm²). 2 Discard the culture medium, then add 100 μL of MTT (1 mg / mL) solution to each well, and incubate the cells at 37°C and 5% CO2 for 3 h; then add 100 μL of DMSO to each well, shake to dissolve, and measure the OD value at 490 nm using a microplate reader.

[0137] The formula for calculating relative cell viability is: Relative cell viability (%) = (OD of experimental wells - OD of zeroing wells) / (OD of control wells without irradiation - OD of zeroing wells) × 100%.

[0138] The results of the relative cell viability calculation are shown in Table 5 and Figure 3 As shown.

[0139] Table 5. Calculation results of relative cell viability

[0140]

[0141]

[0142] From Table 5 and Figure 3It can be seen that, compared with the unirradiated control wells, the relative cell activity of the UVA-irradiated control wells decreased to 64.47%, indicating that the photodamage cell model was successfully constructed. After UVA irradiation treatment in Examples 1-4 and after UVA irradiation treatment in the control wells, the relative cell activity was increased by 28.47%, 24.23%, 25.68%, and 23.85%, respectively. Example 1 showed the highest relative cell activity at 92.94%, indicating that the active composition of this invention can repair photodamage caused by ultraviolet irradiation. Comparative Examples 1-10, after UVA irradiation treatment and... Compared to the UVA irradiation treatment in Example 1, the relative cell activity decreased to some extent. This was especially evident when considering the increased relative cell activity after the UVA irradiation treatments in Comparative Examples 6 and 9, and in Comparative Examples 8 and 10. This demonstrates that the combined formulation of Lactobacillus / grape juice extract, tetrahydromethylpyrimidine carboxylic acid, glucosyl hesperidin, adenosine, and *Erycibe obtusifolia* callus culture filtrate, with all five components working synergistically, is essential; the absence of any one component leads to a decrease in cell activity. Furthermore, the comparison between the UVA irradiation treatment in Comparative Example 11 and Example 1 indicates that the combination of Lactobacillus / grape juice extract obtained from *Lactobacillus plantarum* fermentation with the other four components yielded a better effect.

[0143] Experimental Example 4: Melanin Inhibition Test

[0144] B16 melanocytes (purchased from Guangdong Boxi Biotechnology Co., Ltd.) were used at a rate of 1×10 5 Cells were seeded at a density of 2 mL / mL in 96-well plates and incubated at 37°C with 5% CO2 for 24 h. The supernatant was discarded, and control and experimental groups were established. The control group was supplemented with fresh RPMI 1640 medium, while the experimental groups were supplemented with RPMI 1640 medium containing 1.5 mg / mL of samples from Examples 1-4 and Comparative Examples 1-11, respectively. Cells were cultured for another 24 h, and the supernatant was discarded. Cells were washed twice with PBS buffer (pH 7.2-7.4). 0.5 mL of trypsin was added to each well for 3 min of digestion, followed by 2 mL of fresh RPMI 1640 medium containing serum to terminate the digestion. After refluxing, 0.5 mL of cell suspension from each group was taken for cell counting and density calculation. The cell suspension was then centrifuged at 2500 r / min for 5 min, the supernatant was discarded, and 200 μL of 1 mol / L sodium hydroxide solution was added to the precipitate to dissolve the melanin. The absorbance was measured at 490 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the melanin production inhibition rate was calculated.

[0145] The formula for calculating the melanin production inhibition rate is: Melanin production inhibition rate (%) = [1 - (sample well absorbance value ÷ sample well cell density) ÷ (control well absorbance value ÷ control well cell density)] × 100%

[0146] The calculated results of the melanin production inhibition rate are shown in Table 6.

[0147] Table 6. Calculation results of melanin production inhibition rate

[0148] experimental group Inhibition rate % Example 1 29.5 Example 2 28.2 Example 3 23.4 Example 4 23.8 Comparative Example 1 14.9 Comparative Example 2 12.1 Comparative Example 3 11.2 Comparative Example 4 13.2 Comparative Example 5 12.9 Comparative Example 6 9.2 Comparative Example 7 8.4 Comparative Example 8 7.8 Comparative Example 9 7.1 Comparative Example 10 6.2 Comparative Example 11 15.8

[0149] As shown in Table 6, the comparison revealed that the melanin production inhibition rate of samples from Examples 1-4 was significantly higher than that of Comparative Examples 1-10, indicating that the components of the composition of the present invention have a synergistic effect and a stronger ability to inhibit melanin production. Adenosine exhibits strong melanin production inhibition at high concentrations; however, a comparison of the melanin production inhibition rates of samples from Examples 1 and 2 showed that the inhibitory effect of the composition of Example 2 on melanin production was lower than that of Example 1. This indicates that the combination of the present invention can synergistically enhance the inhibitory effect of adenosine on melanin production, and can exert a strong inhibitory effect on melanin production even at low concentrations. A comparison of the melanin production inhibition rates of samples from Comparative Example 11 and Example 1 showed that the combination of Lactobacillus / grape juice extract obtained by fermentation of Lactobacillus plantarum with the other four components was more effective, and the resulting composition had a better effect on improving pigmentation after damage.

[0150] Application Example 1: Preparation of emulsions containing functional compositions

[0151] The formulation composition of emulsions containing functional compositions is shown in Table 7:

[0152] Table 7. Formulation composition of emulsions containing functional compositions

[0153]

[0154] The method for preparing the above-mentioned emulsion containing the functional composition includes the following steps:

[0155] (1) Mix the raw materials of phase A, including a portion of water, and heat to 80-85℃ and stir until dissolved.

[0156] (2) Mix the B phase raw materials and heat them to 75-80℃ and keep them warm for later use;

[0157] (3) Add the B phase raw material treated in step (2) to the A phase raw material treated in step (1), homogenize at 5000 rpm for 6 min, and cool down to 70-75℃ after homogenization.

[0158] (4) Then add raw materials 15 and 16, stir and dissolve evenly, then add raw material 17, stir and dissolve evenly, and then cool down to 40-45℃;

[0159] (5) Prepare the efficacy composition solution from the remaining water according to the mass ratio and method of Example 1, add it to the system of step (4), stir and mix evenly, and cool to room temperature to obtain the final product.

[0160] Application Example 2: Preparation of emulsions containing functional compositions

[0161] The formulation composition of emulsions containing functional compositions is shown in Table 8:

[0162] Table 8. Formulation composition of emulsions containing functional compositions

[0163]

[0164]

[0165] The method for preparing the above-mentioned emulsion containing the functional composition includes the following steps:

[0166] (1) Mix the raw materials of phase A, including a portion of water, and heat to 80-85℃ and stir until dissolved.

[0167] (2) Mix the B phase raw materials and heat them to 75-80℃ and keep them warm for later use;

[0168] (3) Add the B phase raw material treated in step (2) to the A phase raw material treated in step (1), homogenize at 5000 rpm for 6 min, and cool down to 70-75℃ after homogenization.

[0169] (4) Then add raw materials 15 and 16, stir and dissolve evenly, then add raw material 17, stir and dissolve evenly, and then cool down to 40-45℃;

[0170] (5) Prepare the efficacy composition solution from the remaining water according to the mass ratio and method of Example 1, add it to the system of step (4), stir and mix evenly, and cool to room temperature to obtain the final product.

[0171] Comparative Example 12: Preparation of an emulsion without functional composition

[0172] Compared to the emulsion composition of Application Example 1, the emulsion of Comparative Example 12 does not contain the functional composition, and its formulation composition is shown in Table 9:

[0173] Table 9. Formulation composition of emulsions without functional ingredients

[0174]

[0175]

[0176] The method for preparing the above-mentioned emulsion without functional composition includes the following steps:

[0177] (1) Mix the A phase raw materials and heat to 80-85℃, stirring until dissolved and homogeneous;

[0178] (2) Mix the B phase raw materials and heat them to 75-80℃ and keep them warm for later use;

[0179] (3) Add the B phase raw material treated in step (2) to the A phase raw material treated in step (1), homogenize at 5000 rpm for 6 min, and cool down to 70-75℃ after homogenization.

[0180] (4) Then add raw materials 15 and 16, stir and dissolve evenly, add raw material 17, stir and dissolve evenly, and then cool to room temperature to obtain the product.

[0181] Experimental Example 5: Skin ITA° Value Test

[0182] The emulsions prepared using Examples 1-2 and Comparative Example 12 were used for subject experience testing. Each group consisted of 10 female subjects aged 19-30 years, and the testing period was 28 days, with the emulsions applied twice daily, morning and evening.

[0183] Individual type angle (ITA°): Measured using a skin colorimeter (model: Colorimeter CL400, manufacturer: Courage and Khazaka GmbH, Germany) or a reflectance spectrophotometer. * a * b * The parameters used to characterize human skin color in color space are calculated using the following formula:

[0184]

[0185] Measurement method: At each visit time, L was measured in each test area using a skin colorimeter. * a * b * The ITA° value was measured three times for each area, and the value was recorded and calculated. The higher the ITA° value, the lighter the skin tone, and vice versa.

[0186] The ITA° test results are shown in Table 10 before sample use (T0), after 14 days of use (T14d), and after 28 days of use (T28d). Figure 4 The higher the ITA° value, the lighter the skin tone.

[0187] Table 10. Results of skin ITA° value detection (n=10)

[0188]

[0189] From Table 10 and Figure 4 It can be seen that in Application Examples 1 and 2, the functional composition of the present invention can effectively improve the problem of skin pigmentation, and the greater the dosage, the more obvious the improvement effect. The results of Comparative Example 12 show that without the addition of the functional composition, the number of people with effectively improved skin pigmentation is significantly reduced, indicating that the functional composition studied in this invention can effectively improve the problem of skin pigmentation.

[0190] Experimental Example 7: Application of the repairing, soothing, and moisturizing effects of the lotion in Example 1

[0191] 1. Experimental Methods:

[0192] Subject requirements: 18–41 years old, sensitive facial skin (screened via questionnaire), transepidermal water loss (TEWL) ≥ 20 g / m² before use. 2 h, Number of valid subjects: no less than 33, Test site: face.

[0193] How to use the lotion: After cleansing your face in the morning and evening, take an appropriate amount of the lotion from Example 1 into your palm, apply it evenly to your facial skin, and gently massage until absorbed. Apply once in the morning and once in the evening for 28 consecutive days.

[0194] 2. Test metrics:

[0195] Subjects were tested after 7 days, 14 days, and 28 days of use:

[0196] 2.1. Skin moisture content test (Corneometer), unilateral cheek;

[0197] 2.2. Transepidermal water loss rate (TEWL) test (Aqua Flux), unilateral cheek;

[0198] 2.3. Facial Imaging Test (VISIA-CR), Frontal View of the Face.

[0199] 3. Test Results:

[0200] 3.1 Skin Moisture Content Test Data

[0201] Skin moisture content test results as follows Figure 5 As shown. Figure 5 Data shows that after using the application example 1 lotion for 7, 14 and 28 days, the skin moisture content increased by 21.8%, 28.6% and 36.2% respectively, indicating that the lotion has a moisturizing effect.

[0202] 3.2 Transdermal Water Loss (TEWL) Test Data

[0203] Transdermal water loss rate (TEWL) test results are as follows Figure 6 As shown. Figure 6 Data shows that after using the application example 1 lotion for 7, 14 and 28 days, the transepidermal water loss of the skin decreased by 15.9%, 21.5% and 26.2% respectively, indicating that the lotion has a repairing effect.

[0204] 3.3 Soothing Efficacy Data

[0205] Facial red zone measurement results as follows Figure 7 and Figure 8 As shown. Figure 7 and Figure 8 The results showed that after using the application example 1 emulsion for 7, 14, and 28 days, the red area a... * The values ​​decreased by 9.1%, 11.7%, and 16.2% respectively, proving that the lotion has a soothing effect.

Claims

1. A composition for combating and repairing photo-damage, characterized in that it comprises: Lactobacillus / grape juice extract, tetrahydro methyl pyrimidine carboxylic acid, glucosyl hesperidin, adenosine, and Phoeniculeterum callus culture filtrate; The mass fraction of each component is as follows: lactobacillus / grape juice extract 5-10 parts, tetrahydro methyl pyrimidine carboxylic acid 0.01-0.5 parts, glucosyl hesperidin 0.01-1 part, adenosine 0.01-0.5 parts, and Phoeniculeterum callus culture filtrate 0.01-0.5 parts; The preparation method of the lactobacillus / grape juice extract is as follows: obtaining lactobacillus plantarum suspension after culturing lactobacillus plantarum, the bacterial concentration of the lactobacillus plantarum suspension is 1-10×10 7 After the grape fruit is squeezed, water is added according to the solid-liquid ratio 1:(10-15), homogenized, sterilized, inoculated with the lactobacillus plantarum suspension, the inoculation amount is 1-2% volume percentage, anaerobic fermentation culture is carried out at 35-40 DEG C for 30-60h, after the fermentation is finished, the supernatant is obtained by centrifugation, and the lactobacillus / grape juice extract is obtained after sterilization by filter membrane.

2. The composition of claim 1, wherein The mass fraction of each component is as follows: Lactobacillus / grape juice extract 6-8 parts, tetrahydro methyl pyrimidine carboxylic acid 0.01-0.2 parts, glucosyl hesperidin 0.01-0.5 parts, adenosine 0.01-0.2 parts, and Phoeniculeterum callus culture filtrate 0.01-0.2 parts.

3. The composition of claim 1, wherein The mass fraction of each component is as follows: Lactobacillus / grape juice extract 7 parts, tetrahydro methyl pyrimidine carboxylic acid 0.1 part, glucosyl hesperidin 0.2 part, adenosine 0.1 part, and Phoeniculeterum callus culture filtrate 0.1 part.

4. Use of the composition of claim 1 in the preparation of a skin care product having the effects of resisting and repairing photodamage.

5. An emulsion having a light damage resistant and light damage repairing effect, characterized by, According to the mass percentage, the following components are included: The composition for resisting and repairing photodamage of claim 1 1-3%, emulsifier 0.1-3%, oil 1-10%, rheological modifier 0.1-0.8%, humectant 2-11%, preservative 0.2-2%, chelating agent 0.01-0.1%, pH regulator 0.08-0.64%, and water is added to 100%.

6. The emulsion of claim 5, wherein, Satisfy one or more of the following conditions: i. The emulsifier is as follows according to the mass percentage: di(lauroylamido glutamoyl) lysine sodium 0.01-0.2%, C12-20 alkyl glucoside 0.2-0.5%, C14-22 alcohol 0.5-2%; ii. The oil is as follows according to the mass percentage: dimethicone 0.05-2%, tocopheryl acetate 0.1-0.2%, isononyl isononanoate 2-4%, shea butter 0.1-0.5%; iii. The rheological modifier is as follows according to the mass percentage: carbomer 0.1-0.5%, hydroxyethyl acrylate / sodium acryloyldimethyl taurate copolymer 0.01-0.2%; iv. The humectant is as follows according to the mass percentage: glycerin 1-5%, 1,3-butanediol 1-5%, sodium hyaluronate 0.01-0.5%; v. The preservative is as follows according to the mass percentage: 1,2-hexanediol 0.1-1%, p-hydroxyacetophenone 0.1-0.8%; vi. The chelating agent is disodium EDTA; vii. The pH regulator is tromethamine.

7. The method of claim 5, wherein the emulsion having resistance to and repair of photodamage is prepared by the steps of: The following steps are included: (1) Mix the humectant, rheological modifier, emulsifier, chelating agent, and part of the water uniformly, heat to 80-85℃, and stir to dissolve uniformly; (2) Stir the oil uniformly, heat to 75-80℃, then add to the mixture of step (1), homogenize at 3000-5500 rpm for 3-6 min, then cool to 70-75℃; (3) To the mixture after homogenization in step (2), add a preservative, after complete dissolution, add a pH regulator, after uniform dissolution, cool to 40-45℃, add the solution of the composition for resisting and repairing light damage prepared from the remaining water, mix uniformly, cool to room temperature to obtain.

8. The production method according to claim 7, wherein The solution of the composition for resisting and repairing light damage is prepared as follows: 1) Heat water to 90-95℃, sterilize for 30-50 min, then cool to 80-85℃; 2) Add tetrahydro-methyl pyrimidine carboxylic acid and adenosine to the water prepared in step 1), stir until completely dissolved, then cool to 40-45℃; 3) Add lactobacillus / grape juice extract, glucosyl hesperidin, and sea parsley callus culture filtrate to the solution in step 2), stir and mix uniformly to obtain.

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