Female private care gel with multiple effects and preparation method thereof

Abstract

The invention discloses a female private care gel with multiple effects and a preparation method thereof. The gel is prepared from the following components: carbomer 980, palmitoyl 3 peptide, phenethyl caffeate, glycerol, pentanediol, sodium hyaluronate, hydrolyzed sodium hyaluronate, panthenol, glycerol glucoside, crithmum maritimum callus fermentation filtrate, lysozyme, an ecological nutrient solution and the balance of water. According to the female private care gel disclosed by the invention, through a scientific formula and a reasonable preparation process, the synergistic effect of all the components is fully exerted, the use limitation of palmitoyl 3 peptide in vaginal gel is effectively solved, and the gel can better adapt to the vaginal environment, promote the production of collagen and realize the effects of tightening and moisturizing. Meanwhile, the product has multiple effects of repairing in vitro, moisturizing, relieving, resisting inflammation, regulating flora balance and the like, comprehensively cares female private health, and has a wide market application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of female reproductive health care, in particular to a vaginal gel composition with firming, moisturizing and repairing functions and a preparation method thereof. BACKGROUND

[0002] With the enhancement of modern women's health consciousness, the demand for maintaining the microecological environment of the female reproductive system is increasing. The skin of the female private parts is delicate, and is easily affected by physiological cycles, fertility and other factors, leading to problems such as dryness, relaxation and imbalance of flora. The existing private care products have single functions and are difficult to fully meet the needs of women for private part care. Some products add ingredients with firming effect, but the stability and effectiveness in the complex environment of the vagina are insufficient, and they cannot fully play their roles. Therefore, it is of great significance to develop a private care gel that can effectively adapt to the vaginal environment and has multiple functions. SUMMARY

[0003] The present application provides a vaginal gel composition with firming, moisturizing and repairing functions and a preparation method thereof.

[0004] The vaginal gel composition comprises the following components: Carbomer 980 (0.5-1.5 wt%): as the main component of the gel matrix, it has good thickening, suspending and stabilizing effects, can make the gel maintain appropriate viscosity and texture, and is convenient for application and use, and at the same time provides a stable dispersion environment for other active ingredients.

[0005] Palmitoyl 3 peptide (0.01-0.3 wt%): a signal peptide that can stimulate fibroblast activity and promote the synthesis of extracellular matrix such as collagen, elastin and fibronectin, thereby increasing skin elasticity and playing a role in firming the skin. However, there are certain limitations in using palmitoyl 3 peptide in vaginal gel. The vaginal environment is complex and contains various enzymes and microorganisms. When used alone, palmitoyl 3 peptide is easily degraded by enzymes, and its activity is greatly affected by the pH value of the vagina, and its stability is poor in the acidic environment of the vagina, making it difficult to fully exert the function of promoting the production of collagen.

[0006] Caffeic acid phenethyl ester (0.01-0.1 wt%): It has multiple biological activities such as antioxidant, anti-inflammatory, and antibacterial. In the present application, caffeic acid phenethyl ester is used in combination with palmitoyl 3 peptide, and the two components can exert synergistic effects. On the one hand, the antioxidant and anti-inflammatory properties of caffeic acid phenethyl ester can protect palmitoyl 3 peptide from damage by free radicals and inflammatory factors in the vagina, thereby improving the stability of palmitoyl 3 peptide in the vaginal environment. On the other hand, the antibacterial effect of caffeic acid phenethyl ester helps to maintain the balance of vaginal flora, creating a good microenvironment for palmitoyl 3 peptide to exert its effects. At the same time, caffeic acid phenethyl ester itself can also promote the synthesis of collagen, and together with palmitoyl 3 peptide, it can enhance the tightening and moisturizing effects. To improve the solubility of caffeic acid phenethyl ester, it is pre-dissolved in propylene glycol, and the amount of propylene glycol used is 5-10 times the weight of caffeic acid phenethyl ester.

[0007] Glycerin (6-8 wt%): It is a commonly used moisturizer that can absorb moisture from the air, keeping the skin in the private parts moist and preventing dryness and itching.

[0008] Pentylene glycol (2-4 wt%): It has moisturizing and antibacterial effects, and can synergistically enhance the moisturizing effect with glycerin while inhibiting the growth of harmful microorganisms, thereby maintaining vaginal health.

[0009] Sodium hyaluronate (0.05-0.5 wt%) and hydrolyzed sodium hyaluronate (0.01-0.02 wt%): Sodium hyaluronate is a natural moisturizing factor that can form a moisturizing film on the skin surface, locking in moisture and increasing the moisture level of the skin. Hydrolyzed sodium hyaluronate has a smaller molecular weight and can penetrate deeper into the skin, providing moisture and nutrients to the skin and enhancing the elasticity and luster of the skin.

[0010] Panthenol (0.5-1.5 wt%): Also known as vitamin B5, it has moisturizing, soothing, and repairing effects. It can penetrate deep into the skin, promote skin cell metabolism, enhance the skin barrier function, and relieve discomfort in the private parts.

[0011] Glycerin grape glycoside (0.1-0.5 wt%): It has good moisturizing and antioxidant properties, can increase the water content of the skin, reduce water loss, and at the same time scavenge free radicals to delay skin aging.

[0012] Sea fennel callus fermentation filtrate (0.01-0.1 wt%): It is rich in various nutrients such as amino acids, vitamins, and minerals, and has antioxidant, anti-inflammatory, and repairing effects, which can promote the repair and regeneration of damaged skin and enhance the resistance of the skin.

[0013] Lysozyme (0.05-0.3 wt%): can dissolve bacterial cell wall, destroy the structure of bacteria, thus playing the role of antibacterial and anti-inflammatory, effectively preventing and improving vaginal infection. To improve the stability and targeting of lysozyme, it can be added after being wrapped by liposomes.

[0014] Ecological nutrient solution (0.3-1.5 wt%): contains a variety of beneficial microorganisms and nutrients, which can regulate vaginal flora balance, enhance the defense ability of the vagina itself, and maintain the stability of the vaginal microecology.

[0015] Water: as a solvent, dissolving other components to form a uniform system for gel formation.

[0016] Ectoin (0.05-0.3 wt%, optional): has strong moisturizing and protective effects, can form a protective film on the skin surface to resist external environmental irritation, and enhance the tolerance of the skin.

[0017] Centella asiatica extract (0.1-0.5 wt%, optional): has anti-inflammatory, soothing and repairing effects, can reduce inflammation of the private parts, promote wound healing and relieve discomfort.

[0018] Rhamnose (0.1-0.8 wt%, optional): has moisturizing and immune regulating effects, can improve the moisturizing ability of the skin and enhance the immunity of the skin.

[0019] Collagen (0.01-0.1 wt%, optional): directly supplements collagen to the skin, increases the elasticity and firmness of the skin, and improves the problem of vaginal relaxation.

[0020] The present application has multiple functions by reasonably matching the components. Carbopol 980 and a variety of moisturizing ingredients synergistically provide long-lasting moisturizing for the private parts; palmitoyl 3 peptide and caffeic acid phenethyl ester cooperate to overcome the limitations of palmitoyl 3 peptide in vaginal gel, better adapt to the vaginal environment, effectively promote the production of collagen, and have the effect of tightening and moisturizing; a variety of antibacterial and anti-inflammatory ingredients maintain the balance of vaginal flora, prevent and improve vaginal infection; ecological nutrient solution regulates vaginal microecology and enhances the defense ability of the vagina itself.

[0021] The present application also provides a preparation method of the vaginal gel, which specifically comprises the following steps: Carbopol 980 is dissolved in water, and A phase is formed after repeated dissolution at 50-60℃.

[0022] Glycerol, sodium hyaluronate, panthenol and pentylene glycol are mixed to form B phase, which is added to A phase, and after fully stirring and complete dissolution, the temperature is reduced to 40℃.

[0023] Mix phenethyl caffeic acid with propylene glycol at 50-60°C until completely dissolved, cool, add the mixture from step (2), and stir at low speed to avoid direct contact between phenethyl caffeic acid and carbomer 980.

[0024] Add palmitoyl tripeptide, lysozyme (added after being encapsulated in liposomes), and ecological nutrient solution, and stir until fully dissolved.

[0025] Add the remaining components to the mixture from step (4) in sequence and stir until fully dissolved.

[0026] Adjust the pH to 4.5–5.5 with arginine, add water, and then sterilize and fill the product.

[0027] During the preparation process, each component is added stepwise to ensure its stability and effectiveness. First, Carbomer 980 is dissolved in water to form Phase A, providing the basic framework for the gel. Then, other moisturizing and auxiliary ingredients are mixed to form Phase B, which is then mixed with Phase A and cooled to create suitable conditions for the addition of active ingredients. Phenethyl caffeate is added after pre-dissolving in propylene glycol to avoid direct contact with Carbomer 980 and prevent adverse reactions. Palmitoyl tripeptide, lysozyme, and ecological nutrient solution are added at a suitable temperature to ensure their activity. Finally, the pH is adjusted and the product is sterilized and filled to ensure product quality and safety. Beneficial effects

[0028] This invention's feminine intimate care gel, through a scientific formula and rational preparation process, fully leverages the synergistic effects of its components, effectively overcoming the limitations of palmitoyl tripeptide in vaginal gels. It better adapts to the vaginal environment, promotes collagen production, and achieves a firming and moisturizing effect. Simultaneously, the product possesses multiple functions including external repair, moisturizing, soothing, anti-inflammatory, and regulation of vaginal flora balance, comprehensively protecting women's intimate health and possessing broad market application prospects. Attached Figure Description

[0029] Figure 1 This is a graph of Example 5; Figure 2 This is a diagram of the tissue morphology of Example 6; Figure 3 This is a bar chart showing the average dermal cell density results from Example 6; Figure 4 This is the Collagen I immunofluorescence image from Example 6; Figure 5 This is a bar chart of the average relative integrated optical density (IOD) of Collagen I in Example 6; Figure 6 This is the Collagen III immunofluorescence image from Example 6; Figure 7This is a bar chart of the average relative integrated optical density (IOD) of Collagen III in Example 6; Figure 8 This is an immunofluorescence image of α-SMA from Example 6; Figure 9 This is a bar chart of the average relative integrated optical density (IOD) of α-SMA in Example 6; Figure 10 This is a graph showing the collagen fiber content of Example 6; Figure 11 This is a bar chart showing the relative area of ​​collagen fibers in Example 6; Figure 12 This is the HA detection graph for Example 6; Figure 13 This is a bar chart of the average relative integrated optical density (IOD) of HA in Example 6; Figure 14 This is a bar chart showing the Elastin content detection results for Example 6; Figure 15 This is a skin moisture content detection graph from Example 7; Figure 16 This is the Claudin-1 immunofluorescence image from Example 7; Figure 17 This is a bar chart of the average relative integrated optical density (IOD) of Claudin-1 in Example 7; Figure 18 This is the AQP3 immunofluorescence image from Example 7; Figure 19 This is a bar chart of the average relative integrated optical density (IOD) of AQP3 in Example 7; Figure 20 This is a bar chart showing the PGE2 content detection results of Example 7; Figure 21 This is a bar chart showing the TNF-α content detection results of Example 7. Detailed Implementation

[0030] The present invention will be further described below with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and should not be used to limit the scope of protection of the present invention.

[0031] Example 1: A vaginal gel composition with tightening, moisturizing and repairing functions This embodiment provides a vaginal gel composition, which, by weight percentage (wt%), comprises: Carbomer 980: 1.0% Palmitoyl tripeptide: 0.1% Phenyl ethyl caffeate: 0.05% Propylene glycol (used for predissolving phenethyl caffeate): 0.4% (8 times the weight of phenethyl caffeate) Glycerin: 7.0% Pentylene glycol: 3.0% Sodium hyaluronate: 0.2% Hydrolyzed sodium hyaluronate: 0.015% Panthenol: 1.0% Glyceryl glucoside: 0.3% Fermentation filtrate of sea fennel callus: 0.05% Lysozyme (liposome-encapsulated): 0.1% Ecological nutrient solution: 1.0% Ectocin: 0.2% Centella asiatica extract: 0.3% Rhamnose: 0.5% Collagen: 0.05% Arginine: As needed (for pH adjustment) Purified water: Add to 100% Its preparation method includes the following steps: Formation of Phase A: Carbomer 980 is slowly added to approximately 70% of the total volume of purified water (preheated to 55°C), and stirred for 15 minutes using a high-speed homogenizer (2000 rpm) to allow it to fully swell and disperse, forming a uniform and transparent gel matrix, which is denoted as Phase A.

[0032] Forming and adding phase B: In another container, mix glycerol, sodium hyaluronate, panthenol, and pentylene glycol, stirring until completely dissolved to form phase B. Add phase B to phase A, and stir at 500 rpm for 20 minutes at 45°C to ensure uniform mixing, then cool to 40°C.

[0033] Pre-dissolve and add phenethyl caffeate: Mix phenethyl caffeate with propylene glycol in a 55°C water bath and stir until completely dissolved to obtain a clear solution. Cool this solution to 40°C and slowly add it to the mixture from step (2) while stirring at a low speed (300 rpm) to avoid introducing too many air bubbles and to ensure uniform dispersion.

[0034] Add heat-sensitive active ingredients: Add palmitoyl 3-peptide, liposome-encapsulated lysozyme and ecological nutrient solution to the above system in sequence, and stir at low speed (200 rpm) for 15 minutes at 35-40℃ to ensure thorough mixing and maintain activity.

[0035] Add the remaining components: add hydrolyzed sodium hyaluronate, glyceryl glucoside, sea fennel callus fermentation filtrate, ectoine, centella asiatica extract, rhamnose and collagen in sequence, and stir until all components are completely dissolved and mixed evenly.

[0036] Adjust pH and bring to volume: Slowly adjust the pH of the system to 5.0 using a 10% arginine aqueous solution. Add the remaining purified water to bring the volume to 100%, and continue stirring for 10 minutes to ensure the entire system is homogeneous.

[0037] Filling and sterilization: After vacuum degassing the obtained gel, it is dispensed using an aseptic filling process to obtain the final product.

[0038] Example 2: A basic formulation of a vaginal gel composition This embodiment provides a relatively basic formula that focuses on core firming and moisturizing functions. Its composition, by weight percentage (wt%), is as follows: Carbomer 980: 0.8% Palmitoyl tripeptide: 0.05% Phenyl ethyl caffeate: 0.02% Propylene glycol: 0.2% (10 times the weight of phenethyl caffeate) Glycerin: 6.0% Pentylene glycol: 2.0% Sodium hyaluronate: 0.1% Hydrolyzed sodium hyaluronate: 0.01% Panthenol: 0.8% Glyceryl glucoside: 0.1% Lysozyme (liposome-encapsulated): 0.05% Ecological nutrient solution: 0.5% Arginine: Appropriate amount Purified water: Add to 100% The preparation method is the same as in Example 1.

[0039] Example 3: A high-efficacy vaginal gel composition This embodiment provides an enhanced formula with a higher content of active ingredients, designed to provide more significant firming and repairing effects. Its composition, by weight percentage (wt%), is as follows: Carbomer 980: 1.2% Palmitoyl tripeptide: 0.2% Phenyl caffeate: 0.08% Propylene glycol: 0.64% (8 times the weight of phenethyl caffeate) Glycerin: 8.0% Pentylene glycol: 4.0% Sodium hyaluronate: 0.4% Hydrolyzed sodium hyaluronate: 0.02% Panthenol: 1.5% Glyceryl glucoside: 0.5% Fermentation filtrate of sea fennel callus: 0.1% Lysozyme (liposome-encapsulated): 0.2% Ecological nutrient solution: 1.5% Ectocin: 0.3% Centella asiatica extract: 0.5% Arginine: Appropriate amount Purified water: Add to 100% The preparation method is the same as in Example 1.

[0040] Comparative Example 4: A gel composition without caffeic acid phenethyl ester has a formulation that is substantially the same as in Example 1, but omits caffeic acid phenethyl ester and propylene glycol used for pre-dissolving it, and replaces these portions with purified water.

[0041] Example of effect: To verify the beneficial effects of the present invention, relevant tests were conducted on the gels prepared in Examples 1-3 and the comparative examples.

[0042] Example 5: Stability Test 1. Experimental Objective Accelerated stability testing was used to quantitatively analyze and compare the changes in palmitoyl tripeptide content in the experimental formulation containing CAPE and the control formulation without CAPE, verifying that CAPE can significantly inhibit the degradation of palmitoyl tripeptide under high temperature and high humidity conditions and improve its stability.

[0043] 2. Experimental Principle Following the internationally accepted ICH accelerated stability testing guidelines, samples were placed under harsh environmental conditions (e.g., 40℃ ± 2℃, 75% ± 5% RH) to accelerate the chemical degradation reaction. Periodic sampling was performed, and the residual content of palmitoyl tripeptide in the samples was quantitatively determined using high-precision, high-specificity high-performance liquid chromatography (HPLC). The stabilization effect of CAPE could be evaluated by comparing the rate of degradation of the active ingredient in the two groups of samples.

[0044] 3. Experimental Materials and Equipment sample: Experimental groups: Example 1, Example 2, Example 3 Control group: Gel prepared according to the formulation of Comparative Example 4 (without CAPE, and the other components are exactly the same as the experimental group).

[0045] equipment: A constant temperature and humidity chamber (with precise temperature control of 40℃ ± 2℃ and humidity of 75% ± 5% RH), a high-performance liquid chromatograph (HPLC) equipped with a UV detector or mass spectrometer (MS) detector, an analytical balance, and a pH meter are also included. Rotational viscometers, centrifuges, ultrasonic water baths, micropipettes, volumetric flasks, sample vials, and other glassware.

[0046] Reagents: Palmitoyl tripeptide-1 standard (purity ≥98%), chromatographic grade acetonitrile, methanol, ultrapure water, and other chromatographic reagents (such as trifluoroacetic acid, TFA). 4. Experimental Procedure 4.1 Sample preparation and initial time point (T=0) detection The experimental and control group samples were prepared in parallel in the same batch, strictly following the preparation methods of Examples 1-3 and Comparative Example 4, to ensure that all conditions were completely consistent except for CAPE.

[0047] Before placing the two sets of samples into the stability test chamber, initial testing was immediately performed: Appearance, color, and odor: Visual inspection and olfactory recording.

[0048] pH value: Dissolve 1g of sample in 10mL of ultrapure water, stir well and then measure.

[0049] Viscosity: Measured using a rotational viscometer at a specified rotor and rotational speed.

[0050] Palmitoyl tripeptide content: Sample pretreatment: Accurately weigh approximately 1.0 g of gel and place it in a 10 mL volumetric flask. Add an appropriate amount of extraction solvent (such as acetonitrile-water mixture), vortex and sonicate for 20 minutes, cool to room temperature, and then make up to volume. Centrifuge a portion of the solution (12000 rpm, 10 min), filter the supernatant through a 0.22 μm organic filter membrane, and transfer it to an HPLC sample vial for analysis.

[0051] HPLC Analysis: Analytical methods were used. The initial concentration (denoted as C0) was accurately calculated by comparing the sample peak area with the palmitoyl tripeptide standard curve. This value served as the baseline for 100% retention.

[0052] 4.2 Accelerated stability test Place each group of samples (at least 3 independent packages) in an open or simulated actual package in a calibrated constant temperature and humidity chamber, with the following conditions: 40℃ ± 2℃, 75% ± 5% RH.

[0053] Samples from the experimental group and the control group were taken at preset time points (e.g., day 0, day 7, day 14, day 30).

[0054] Immediately after each sampling, the appearance, pH, and viscosity of the sample were evaluated according to the method in 4.1. Most importantly, the content of palmitoyl tripeptide was determined by HPLC, and its content (denoted as C) was recorded.t ).

[0055] 4.3 Data Analysis Content retention rate calculation: For each sample at each time point, calculate the content retention rate of palmitoyl 3-peptide.

[0056] Content retention rate (%) = (C t / C0) × 100% C t Content measured at time t C0: Content measured at the initial time point.

[0057] 5. Results and Judgments Appearance, pH, and viscosity: The experimental groups (Examples 1-3) and the control group showed no significant changes within 30 days, indicating that the basic formulation itself has good physical stability. The addition of CAPE did not negatively affect the physical stability of the system.

[0058] Palmitoyl tripeptide content retention rate: like Figure 1 As shown, Examples 1, 2, and 3: At all test time points (day 1, day 7, day 14, and day 30), the palmitoyl tripeptide content retention rates were relatively high in all three examples.

[0059] From day 1 to day 30, the content retention rate of all three examples showed a gradual decreasing trend, but the decrease was relatively small.

[0060] This indicates that, in these three embodiments, palmitoyl tripeptide exhibits relatively good stability and can maintain a high content over a longer period of time.

[0061] Comparative Example 4: Compared with Examples 1, 2 and 3, the palmitoyl 3-peptide content retention rate of Comparative Example 4 was significantly lower at all time points.

[0062] From day 1 to day 30, the content retention rate of Comparative Example 4 decreased significantly, especially between day 14 and day 30, where the decrease was particularly pronounced.

[0063] This indicates that, in Comparative Example 4, the stability of palmitoyl tripeptide was relatively poor, and its content decreased significantly in a short period of time.

[0064] Overall trend: The palmitoyl 3-peptide content in Examples 1, 2, and 3 remained relatively stable, with only a small decrease.

[0065] Compared with Example 4, the palmitoyl 3-peptide content retention rate decreased significantly, indicating poor stability.

[0066] Example 6: In vitro firming efficacy test 1. Test Basis The tests were conducted according to the "Methods for Determining the Content of Type I Collagen, Type III Collagen, α-Smooth Muscle Actin (α-SMA), Collagen Fibers, Elastin, and Hyaluronic Acid (HA) in a Skin Model Based on the Lamina Profunda of an Artificial Vagina".

[0067] 2. Test Objective This test uses an artificial vaginal lamina propria skin model stimulated with inflammatory factors (TNF-α). The tightening effect of the test sample is evaluated by detecting changes in tissue morphology, type I collagen, type III collagen, α-smooth muscle actin (α-SMA), collagen fibers, elastin, and hyaluronic acid (HA) content.

[0068] 3. Test materials 3.1 Test System The model used in this test was an artificial vaginal lamina propria skin model (batch number: FS250801), provided by Guangdong Boxi Shaanxi Branch.

[0069] 3.2 Sample Information Sample prepared in Example 1 3.3 Main Reagents Culture medium for the artificial vaginal lamina propria skin model, PBS, TGF-β1, TNF-α, paraformaldehyde, Collagen I antibody, Collagen III antibody, α-SMA antibody, Masson kit, Elastin ELISA kit, Hyaluronan Binding Protein (Biotin Conj.), xylene, hematoxylin, eosin.

[0070] 3.4 Main Equipment CO2 incubator, clean bench, ELISA reader, upright microscope, fluorescence microscope.

[0071] 4. Testing Methods Table 1 shows the test groups. Table 1

[0072] 2) Preparation before testing: Transfer the model to a 6-well plate and add 2 mL of culture medium to each well. At the same time, randomly divide the model into blank control group (BC), negative control group (NC), positive control group (PC), and sample group, with 3 replicates in each group.

[0073] 3) Model stimulation and drug administration: NC, PC, and sample groups were treated with culture medium containing TNF-α. Drug administration was performed according to the test groups, with no treatment for the BC and NC groups, and the corresponding positive control working solution added to each well of the PC group. Drug administration was applied to the surface of the sample groups.

[0074] 4) Model Culture: The model was transferred to a culture dish and incubated in a CO2 incubator (37℃, 5% CO2). The model was stimulated and treated once daily for a total of four times. After the last stimulation and treatment, the model was cultured for another 24 hours. After the culture, residual test material was washed with sterile PBS solution, and any remaining liquid was wiped away with sterile cotton swabs.

[0075] 5) Tissue morphology examination: The model used for examination was fixed with 4% paraformaldehyde for 24 hours, H&E staining was performed, and the images were taken and analyzed under a microscope.

[0076] 6) Immunofluorescence detection: The model used for detection was fixed with 4% paraformaldehyde. After 24 hours of fixation, immunofluorescence detection was performed. The images were photographed and analyzed under a fluorescence microscope.

[0077] 7) Collagen fiber detection: Skin models for testing were taken, fixed with 4% paraformaldehyde, and after 24 hours of fixation, Masson staining was performed. The images were then photographed and analyzed.

[0078] 8) ELISA detection: After the model culture is completed, collect the model culture medium into centrifuge tubes. After collection, place the samples for ELISA detection in a freezer at -80℃ and perform detection and analysis according to the ELISA kit instructions.

[0079] 9) Biotin colorimetric test: Take the model to be tested, fix it with 4% paraformaldehyde, and after 24 hours, take pictures under a microscope, collect images and analyze them.

[0080] 10) Calculation of improvement rate: Improvement rate (%) = Sample group − Negative control group × 100%.

[0081] 11) Statistical Analysis of Results: GraphPad Prism was used for plotting, and the results are expressed as Mean ± SD. Comparisons between groups were performed using... t-test statistical analysis. All statistical analyses are two-tailed. P A value <0.05 is considered statistically significant. P A value <0.01 is considered highly significant.

[0082] 5. Test Results 5.1 Results of Tissue Morphology Test Figure 2 For the tissue morphology test results, the images were taken using an upright microscope (Olympus, BX53) at a magnification of 200×.

[0083] Table 2 is a summary table of dermal cell density data.

[0084] Table 2

[0085] Note: When performing statistical analysis using the t-test method, significance compared to the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared to the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0086] like Figure 3 The image shows a bar chart of the average dermal cell density. The gel group is the sample from Example 1.

[0087] Compared with the BC group, the dermal cell density in the NC group was significantly reduced, indicating that the stimulation conditions in this test were effective.

[0088] Compared with the NC group, the dermal cell density in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0089] Compared with the NC group, the dermal cell density of the gel in Sample Example 1 increased significantly, with an increase rate of 55.56%.

[0090] 5.2 Results of Type I Collagen Test Figure 4 This is a summary image of the Collagen I immunofluorescence results. The images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 200×. Blue fluorescence represents cell nuclei, and green fluorescence represents Collagen I. The stronger the green fluorescence, the higher the Collagen I content.

[0091] Table 3 is a summary table of Collagen I immunofluorescence analysis results.

[0092] Table 3

[0093] Note: The average relative integrated optical density (IOD) reflects the content of Collagen I. t When performing statistical analysis using the -test method, significance compared to the BC group is indicated by #. P -Value < 0.05 is represented by #. P - Value < 0.01 is represented by ##; significance compared to the NC group is represented by *. P -value < 0.05 is represented as *. P -value < 0.01 is represented as **.

[0094] Figure 5 This is a bar chart showing the average relative integrated optical density (IOD) of Collagen I.

[0095] Compared with the BC group, the Collagen I content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0096] Compared with the NC group, the Collagen I content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0097] Compared with the NC group, the Collagen I content of the gel in Sample Example 1 increased significantly, with an increase rate of 122.73%.

[0098] 5.3 Results of Type III Collagen Test Figure 6 This is a summary of Collagen III immunofluorescence results. The above images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 200×. Blue fluorescence represents cell nuclei, and green fluorescence represents Collagen III. The stronger the green fluorescence, the higher the Collagen III content.

[0099] Table 4 is a summary table of Collagen III immunofluorescence analysis results.

[0100] Table 4

[0101] Note: The average relative integrated optical density (IOD) reflects the content of Collagen III. When performing statistical analysis using the t-test method, significance compared to the BC group is indicated by # (P-value < 0.05 is indicated by #, P-value < 0.01 is indicated by ##); significance compared to the NC group is indicated by * (P-value < 0.05 is indicated by *, P-value < 0.01 is indicated by **).

[0102] Figure 7 This is a bar chart showing the average relative integrated optical density (IOD) of Collagen III.

[0103] Compared with the BC group, the Collagen III content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0104] Compared with the NC group, the Collagen III content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0105] Compared with the NC group, the Collagen III content of the gel in Sample Example 1 increased significantly, with an increase rate of 112.50%.

[0106] 5.4 Results of α-Smooth Muscle Actin (α-SMA) Test Figure 8 This is a summary table of α-SMA immunofluorescence results. The images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 200×. Blue fluorescence represents cell nuclei, and green fluorescence represents α-SMA. The stronger the green fluorescence, the higher the α-SMA content.

[0107] Table 5 is a summary table of α-SMA immunofluorescence analysis results.

[0108] Table 5

[0109] Note: The average relative integrated optical density (IOD) reflects the content of α-SMA. When performing statistical analysis using the t-test method, significance compared to the BC group is indicated by # (P-value < 0.05 is indicated by #, P-value < 0.01 is indicated by ##); significance compared to the NC group is indicated by * (P-value < 0.05 is indicated by *, P-value < 0.01 is indicated by **).

[0110] Figure 9 This is a bar chart showing the average relative integrated optical density (IOD) of α-SMA.

[0111] Compared with the BC group, the α-SMA content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0112] Compared with the NC group, the α-SMA content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0113] Compared with the NC group, the α-SMA content of the gel in Sample Example 1 increased significantly, with an increase rate of 54.17%.

[0114] 5.5 Collagen Fiber Test Results Figure 10 The table summarizes the collagen fiber content test results. The images were taken using an upright microscope (Olympus, BX53) at a magnification of 200×. The blue areas represent collagen fibers, and the more and darker the blue area, the higher the collagen fiber content.

[0115] Table 6 is a summary table of the relative area of ​​collagen fibers.

[0116] Table 6

[0117] Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0118] Figure 11 This is a bar chart showing the relative area of ​​collagen fibers.

[0119] Compared with the BC group, the collagen fiber content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0120] Compared with the NC group, the collagen fiber content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0121] Compared with the NC group, the collagen fiber content of the gel in Sample Example 1 increased significantly, with an increase rate of 86.49%.

[0122] 5.6 Hyaluronic Acid (HA) Test Results Figure 12 This is a summary table of HA detection results. The images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 200×. Blue fluorescence represents cell nuclei, and green fluorescence represents HA. The stronger the green fluorescence, the higher the HA content.

[0123] Table 7 is a summary table of HA analysis results.

[0124] Table 7

[0125] Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0126] Figure 13 This is a bar chart showing the average relative integrated optical density (IOD) of HA.

[0127] Compared with the BC group, the HA content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0128] Compared with the NC group, the HA content in the PC group was significantly increased, indicating that the positive control in this test was effective.

[0129] Compared with the NC group, the HA content of the gel in Sample Example 1 increased significantly, with an increase rate of 25.71%.

[0130] 5.7 Elastin Test Results Table 8 is a summary table of Elastin test results.

[0131] Table 8

[0132] Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0133] Figure 14 The bar chart shows the results of Elastin content detection.

[0134] Compared with the BC group, the NC group showed a significant decrease in elastin levels, indicating that the stimulation conditions in this test were effective.

[0135] Compared with the NC group, the PC group showed a significant increase in Elastin levels, indicating that the positive control in this test was effective.

[0136] Compared with the NC group, the gel of sample Example 1 showed a significant increase in Elastin content, with an increase rate of 53.60%.

[0137] in conclusion: Based on the artificial vaginal lamina propria skin model, compared with the control group, the dermal cell density of the antibacterial gel in Example 1 was significantly increased, with an increase rate of 55.56%. The contents of type I collagen, type III collagen, α-smooth muscle actin (α-SMA), collagen fibers, elastin, and hyaluronic acid (HA) were all significantly increased, with increase rates of 122.73%, 112.50%, 54.17%, 86.49%, 53.60%, and 25.71%, respectively. This indicates that the sample can improve tissue morphology and increase the contents of type I collagen, type III collagen, α-smooth muscle actin (α-SMA), collagen fibers, elastin, and hyaluronic acid (HA), thus having a firming effect.

[0138] Example 7: In vitro repair, moisturizing, and soothing efficacy test 1. Test Basis According to the study on skin water content, tight junction protein-1 (Claudin-1), and aquaporin 3 (AQP3) based on a vaginal mucosal epithelial model The detection method for the content of inflammatory factors (TNF-α) and inflammatory mediators (PGE2) was carried out.

[0139] 2. Test Objective This test uses a vaginal mucosal epithelial model stimulated by inflammatory factors (IL-1α) to detect the content of tight junction protein-1 (Claudin-1). The changes in quantity were used to evaluate the repair efficacy of the test samples. This was achieved by detecting changes in skin moisture content and aquaporin 3 (AQP3) content. The moisturizing efficacy of the test samples was evaluated by detecting changes in the levels of inflammatory factors (TNF-α) and inflammatory mediators (PGE2). The product has a soothing effect.

[0140] 3. Test Materials 3.1 Test System The model used in this test was a vaginal mucosal epithelial model (batch number: 250702), provided by Guangdong Boxi Shaanxi Branch.

[0141] 3.2 Sample Information Sample prepared in Example 1 3.3 Main Reagents EpiGrowth culture medium, PBS, glycerol, WY14643, CaCl2, dexamethasone, IL-1α, paraformaldehyde, AQP3 antibody, Claudin-1 antibody, TNF-α ELISA kit, PGE2 ELISA kit.

[0142] 3.4 Main Equipment CO2 incubator, clean bench, ELISA reader, upright microscope, fluorescence microscope.

[0143] 4. Test Methods The test groups are shown in Table 9: Table 9

[0144] Note: Samples in orange-pink packaging were used to detect skin hydration, Claudin-1, and AQP3; samples in blue packaging were used to detect PGE2 and TNF-α. PC1 (glycerol) was a positive control for detecting skin hydration, PC2 (WY14643) was a positive control for detecting Claudin-1, PC3 (CaCl2) was a positive control for detecting AQP3, and PC4 (dexamethasone) was a positive control for detecting PGE2 and TNF-α.

[0145] 2) Preparation before testing: Prepare 6-well plates according to the test group, add 3.7 mL of model culture medium to each well, transfer the model received on the day to the marked 6-well plate, each test group needs 3 models, transfer all 6-well plates containing models to a CO2 incubator (37℃, 5% CO2) for incubation.

[0146] 3) Model stimulation and drug administration: NC, PC, and sample groups were added to culture medium containing IL-1α. Drug administration was performed according to the test groups, with no treatment for BC and NC groups, and the corresponding positive control working solution added to the PC group. Drug administration was applied to the surface of the sample groups.

[0147] 4) Model Culture: The model was transferred to a culture dish and incubated in a CO2 incubator (37℃, 5% CO2). The model was stimulated and treated once daily for a total of four times. After the last stimulation and treatment, the model was cultured for another 24 hours. After the culture, residual test material was washed with sterile PBS solution, and any remaining liquid was wiped away with sterile cotton swabs.

[0148] 5) Skin moisture content test: a. Cleaning: Prepare 24-well plates according to the number of models and label them accordingly. Add 0.3 mL of culture medium to each well. Place the 24-well plates containing the models in a clean bench, open the lids of the 24-well plates, and let them stand for 30 minutes before taking measurements.

[0149] b. Measurement: Wipe the bottom of the model dry, cut off the ring of the model, place it at the probe position of the tester, press the probe to measure, measure each model three times, and take the average value.

[0150] 6) Immunofluorescence detection: The model used for detection was fixed with 4% paraformaldehyde. After 24 hours of fixation, immunofluorescence detection was performed. The images were photographed and analyzed under a fluorescence microscope.

[0151] 7) ELISA detection: After the model culture is completed, collect the model culture medium into centrifuge tubes. After collection, place the samples for ELISA detection in a freezer at -80℃ and perform detection and analysis according to the ELISA kit instructions.

[0152] 8) Calculation of enhancement rate and inhibition rate: Improvement rate (%) = Sample group − Negative control group × 100%; Blank control group Inhibition rate (%) = Negative control group − Sample group × 100%.

[0153] Blank control group 9) Statistical Analysis of Results: GraphPad Prism was used for plotting, and the results are expressed as Mean ± SD. Comparisons between groups were performed using... t -test Statistical analysis. All statistical analyses are two-tailed. P A value <0.05 is considered statistically significant. P A value <0.01 is considered highly significant.

[0154] 5. Test Results 5.1 Skin moisture content test results Table 10 is a summary table of skin moisture content test results.

[0155] Table 10

[0156] Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, P-value < 0.05 is indicated by #, P-value < 0.01 is indicated by ##, and significance compared with the NC group is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0157] Figure 15 This is a bar chart showing the results of skin moisture content testing.

[0158] Compared with the BC group, the skin moisture content of the NC group was significantly reduced, indicating that the stimulation conditions in this test were effective.

[0159] Compared with the NC group, the skin moisture content of the PC1 group was significantly increased, indicating that the positive control in this test was effective.

[0160] Compared with the NC group, the skin moisture content of the gel in Sample Example 1 increased significantly, with an increase rate of 22.73%.

[0161] 5.2 Results of Claudin-1 assay Figure 16 This is a summary table of Claudin-1 immunofluorescence results. The images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 200×. Blue fluorescence represents cell nuclei, and green fluorescence represents Claudin-1. The stronger the green fluorescence, the higher the Claudin-1 content.

[0162] Table 11 is a summary table of Claudin-1 immunofluorescence analysis results.

[0163] Table 11

[0164] Note: The average relative integrated optical density (IOD) reflects the Claudin-1 content. When performing statistical analysis using the t-test method, significance compared to the BC group is indicated by # (P-value < 0.05 is indicated by #, P-value < 0.01 is indicated by ##); significance compared to the NC group is indicated by * (P-value < 0.05 is indicated by *, P-value < 0.01 is indicated by **).

[0165] Figure 17 This is a bar chart of the average relative integrated optical density (IOD) for Claudin-1.

[0166] Compared with the BC group, the Claudin-1 content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0167] Compared with the NC group, the Claudin-1 content in the PC2 group was significantly increased, indicating that the positive control in this test was effective.

[0168] Compared with the NC group, the Claudin-1 content of the gel in Sample Example 1 increased significantly, with an increase rate of 114.29%.

[0169] 5.3 Results of Aquaporin 3 (AQP3) assay Figure 18 This is a summary table of AQP3 immunofluorescence results. The images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 200×. Blue fluorescence represents cell nuclei, and green fluorescence represents AQP3. The stronger the green fluorescence, the higher the AQP3 content.

[0170] Table 12 is a summary table of AQP3 immunofluorescence analysis results: Table 12

[0171] Note: The average relative integrated optical density (IOD) reflects the AQP3 content. When performing statistical analysis using the t-test method, significance compared to the BC group is indicated by # (P-value < 0.05 is indicated by #, P-value < 0.01 is indicated by ##); significance compared to the NC group is indicated by * (P-value < 0.05 is indicated by *, P-value < 0.01 is indicated by **).

[0172] Figure 19 This is a bar chart showing the average relative integrated optical density (IOD) of AQP3.

[0173] Compared with the BC group, the AQP3 content in the NC group was significantly lower, indicating that the stimulation conditions in this test were effective.

[0174] Compared with the NC group, the AQP3 content in the PC3 group was significantly increased, indicating that the positive control in this test was effective.

[0175] Compared with the NC group, the AQP3 content of the gel in Sample Example 1 increased significantly, with an increase rate of 31.71%.

[0176] 5.4 Results of Inflammatory Mediator (PGE2) Test Table 13 is a summary table of PGE2 test results: Table 13

[0177] Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0178] Figure 20 The bar chart shows the results of PGE2 content detection.

[0179] Compared with the BC group, the PGE2 content in the NC group was significantly increased, indicating that the stimulation conditions in this test were effective.

[0180] Compared with the NC group, the PGE2 content in the PC4 group was significantly lower, indicating that the positive control in this test was effective.

[0181] Compared with the NC group, the PGE2 content of the gel in Sample Example 1 was significantly reduced, with an inhibition rate of 19.30%.

[0182] 5.5 Results of TNF-α Inflammatory Factor Test Table 14 is a summary table of TNF-α detection results: Table 14

[0183] Note: When performing statistical analysis using the t-test method, significance compared with the BC group is indicated by #, with P-value < 0.05 indicated by # and P-value < 0.01 indicated by ##; significance compared with the NC group is indicated by *, with P-value < 0.05 indicated by * and P-value < 0.01 indicated by **.

[0184] Figure 21 The bar chart shows the results of TNF-α content detection.

[0185] Compared with the BC group, the TNF-α content in the NC group was significantly increased, indicating that the stimulation conditions in this test were effective.

[0186] Compared with the NC group, the TNF-α content in the PC4 group was significantly lower, indicating that the positive control in this test was effective.

[0187] Compared with the NC group, the TNF-α content of the gel in Example 1 was significantly reduced, with an inhibition rate of 31.49%.

[0188] in conclusion: Repairing effects: Based on the vaginal mucosal epithelial model, compared with the control group, the content of tight junction protein-1 (Claudin-1) in the gel of Example 1 was significantly increased, with an increase rate of 114.29%, indicating that the sample can increase the content of tight junction protein-1 (Claudin-1) and has a repair effect.

[0189] Moisturizing effect: Based on the vaginal mucosal epithelial model, compared with the control group, the skin moisture content of sample Example 1 gel increased significantly by 22.73%, and the content of aquaporin 3 (AQP3) increased significantly by 31.71%, indicating that the sample can increase skin moisture content and aquaporin 3 (AQP3) content, and has a moisturizing effect.

[0190] Soothing effect: Based on the vaginal mucosal epithelial model, compared with the control group, the content of inflammatory mediators (PGE2) and inflammatory factors (TNF-α) in the gel of Example 1 was significantly reduced, with inhibition rates of 19.30% and 31.49%, respectively. This indicates that the sample can inhibit the secretion of inflammatory mediators (PGE2) and inflammatory factors (TNF-α) and has a soothing effect.

[0191] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A feminine intimate care gel with multiple functions, characterized in that, It contains the following components: Carbomer 980 0.5–1.5 wt% Palmitoyl tripeptide 0.01–0.3 wt% Phenethyl caffeate 0.01–0.1 wt%. The caffeic acid phenethyl ester is pre-dissolved in propylene glycol, with the amount of propylene glycol being 5 to 10 times the weight of caffeic acid phenethyl ester. 6-8 wt% glycerol 2-4 wt% pentanediol Sodium hyaluronate 0.05–0.5 wt% Hydrolyzed sodium hyaluronate 0.01–0.02 wt% Panthenol 0.5–1.5 wt% Glyceryl glucoside 0.1–0.5 wt% 0.01–0.1 wt% of fermentation filtrate from sea fennel callus tissue; Lysozyme 0.05–0.3 wt% Ecological nutrient solution 0.3–1.5 wt%; The remainder is water; The composition is formulated with arginine to adjust the pH to 4.5–5.5, and phenethyl caffeate and carbomer 980 are added in steps to avoid direct contact.

2. The feminine intimate care gel with multiple functions according to claim 1, characterized in that, Also includes: Ectocin 0.05–0.3 wt%; Centella asiatica extract 0.1–0.5 wt% Rhamnose 0.1–0.8 wt% Collagen 0.01–0.1 wt%.

3. The feminine intimate care gel with multiple functions according to claim 1, characterized in that, The step of pre-dissolving phenyl ethyl caffeate in propylene glycol includes: mixing phenyl ethyl caffeate with propylene glycol at 50-60°C until completely dissolved, and then adding it to the gel matrix after cooling.

4. A method for preparing the nursing gel according to any one of claims 1 to 3, characterized in that, Includes the following steps: (1) Carbomer 980 is dissolved in water and repeatedly dissolved at 50℃~60℃ to form phase A; (2) Mix glycerol, sodium hyaluronate, panthenol and pentanediol to form phase B, add it to phase A, stir thoroughly until completely dissolved, and then cool to 40°C; (3) Add the pre-dissolved caffeic acid phenethyl ester and propylene glycol to the mixture in step (2) and stir at low speed; (4) Add palmitoyl tripeptide, lysozyme and ecological nutrient solution, and stir until fully dissolved; (5) Add the remaining components to the mixture from step (4) in sequence and stir until fully dissolved; (6) Adjust the pH to 4.5-5.5 with arginine, add water, sterilize and fill.

5. The method according to claim 4, characterized in that, The lysozyme was added after being encapsulated in liposomes.

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