Anti-aging, moisturizing and antibacterial vaginal gel

Abstract

The invention discloses anti-aging, moisturizing and antibacterial vaginal gel, and belongs to the field of daily necessities for health. The gel is composed of a solution A, a solution B and a solution C, wherein the solution A contains 15-25 parts of glycerol, 10-15 parts of sodium hyaluronate and 1-5 parts of xanthan gum; the solution B is prepared from 0.5 to 2 parts of hydroxyethyl cellulose and 1 to 2 parts of carboxymethyl chitosan; the solution C contains 0.5-3 parts of beta-nicotinamide mononucleotide, 0.5-1.5 parts of an antibacterial agent and 0.1-0.8 part of a transdermal absorption enhancer, and further contains coenzyme Q10 and a centella asiatica extract or a lactobacillus fermentation product on the basis. The antibacterial rate of the vaginal gel to pathogenic bacteria is greater than or equal to 99.5%, lactobacilli are not interfered, vaginal epithelial cell collagen can be improved by 35% or above, the mitochondrial activity can be improved by 40%, and the vaginal gel is suitable for preventing infection, delaying tissue aging and maintaining micro-ecological balance and can also be combined with clotrimazole to enhance the antifungal curative effect.

Description

Technical Field

[0001] This invention relates to the field of daily hygiene and health products, specifically to an anti-aging, moisturizing, and antibacterial vaginal gel. Background Technology

[0002] In recent years, with the increasing awareness of women's health, the market demand for vaginal care products has continued to grow. However, similar products on the market still have significant limitations in terms of functionality and safety: First, the problem of singular efficacy is prominent. Most products only focus on basic moisturizing or antibacterial functions, making it difficult to address multidimensional needs such as anti-aging and microecological regulation. For example, they lack active ingredients that promote collagen regeneration and improve mitochondrial activity. Second, the safety of the formula is insufficient. Some products, due to high concentrations of preservatives (such as benzalkonium chloride), irritating fragrances, or strong acid regulators, can easily damage the vaginal mucosal barrier, causing local irritation or allergic reactions. Third, there is a risk of microecological disruption. Although broad-spectrum antibacterial ingredients (such as chlorhexidine) can inhibit pathogenic bacteria, they can non-selectively kill beneficial lactobacilli in the vagina, leading to pH imbalance and secondary infections. Fourth, the technical verification is weak. The functions of "long-lasting moisturizing" and "anti-aging" claimed by products lack support from cell experiments or clinical data, and the actual effects are far from users' expectations.

[0003] Furthermore, existing products have significant flaws in dosage form design. Traditional suppositories or creams suffer from uneven application and a strong sense of residue, while ordinary hydrogels are prone to leakage or stickiness due to poor rheological properties. In the field of anti-aging, the application of active ingredients (such as NMN and coenzyme Q10) targeting collagen loss and decreased elasticity in vaginal tissue is almost non-existent. Summary of the Invention

[0004] In response to the shortcomings of existing technologies, this invention utilizes a multi-component, multi-target synergistic mechanism to produce a vaginal gel that is antibacterial, moisturizing, and anti-aging, while also being gentle and long-lasting, and can be applied evenly without leaving any residue.

[0005] To achieve the above objectives, the present invention provides the following technical solution to address the technical problem:

[0006] In a first aspect, the present invention provides an anti-aging, moisturizing, and antibacterial vaginal gel, comprising solution A, solution B, and solution C; by weight:

[0007] Solution A: 15-25 parts glycerin, 10-15 parts sodium hyaluronate, 1-5 parts xanthan gum, 50-100 parts water;

[0008] Solution B: 0.5-2 parts hydroxyethyl cellulose, 1-2 parts carboxymethyl chitosan, 10-20 parts water;

[0009] Solution C: 0.5-3 parts β-nicotinamide mononucleotide, 0.5-1.5 parts antibacterial agent, 0.1-0.8 parts PEG-40 hydrogenated castor oil, 0.1-0.3 parts fragrance, 30-60 parts water.

[0010] Specifically, the antibacterial agent is... PE 9010.

[0011] In some embodiments, the anti-aging, moisturizing, and antibacterial vaginal gel further comprises at least one ingredient selected from the following:

[0012] 0.5-2 parts Coenzyme Q10;

[0013] 0.5-1.5 parts Centella Asiatica extract;

[0014] 1-5 portions of Lactobacillus fermentation products.

[0015] In some embodiments, the preparation of the lactobacillus fermentation product includes the following steps:

[0016] S1. Inoculate Lactobacillus plantarum into MRS medium containing 1-2 wt% inulin, 1-3 wt% soybean peptide and 0.1-0.3 wt% glutamine, and culture anaerobically at 35-40℃ until OD600 = 3.0-3.5;

[0017] S2. Fermentation medium was loaded into the bioreactor, and the glucose flow rate was controlled by dissolved oxygen feedback to maintain the DO value at 10%-15%. The cells were collected after 48 hours of fermentation.

[0018] S3. After the bacterial suspension is homogenized under high pressure, it is sequentially passed through a 0.22μm ceramic membrane for ultrafiltration and a 10kDa tangential flow ultrafiltration membrane for fractionation, and the <10kDa active small molecules are collected.

[0019] S4. Add 3-8% trehalose and 0.5-1.5% mannitol to the filtrate, and freeze-dry under vacuum to obtain a powdered fermentation product.

[0020] In some embodiments, the fermentation medium comprises 20-30 g / L trehalose, 15-20 g / L whey protein hydrolysate, 0.05-0.1 g / L MnSO4·H2O, 2 g / L potassium dihydrogen phosphate, 0.2 g / L magnesium sulfate, 0.01 g / L vitamin B1, and 10 g / L initial glucose.

[0021] The inoculum size of *Lactobacillus plantarum* is 5%-10% (v / v); the homogenization pressure is 800-1000 bar.

[0022] Secondly, this invention discloses a method for preparing the above-mentioned anti-aging, moisturizing, and antibacterial vaginal gel, comprising the following steps:

[0023] (1) Add glycerin, sodium hyaluronate and xanthan gum to water for injection at 60±2℃ in sequence, stir at 500-600rpm for 15-20min, cool to 30℃ and let stand to remove bubbles to obtain solution A;

[0024] (2) Disperse hydroxyethyl cellulose and carboxymethyl chitosan in water for injection at 40±2℃, heat to 37±1℃ and stir at 200-300 rpm for 10-20 min, cool to 20-25℃ to form a pregel, and obtain solution B;

[0025] (3) Slowly add solution A to solution B, stir at 100-150 rpm for 10-15 min, and adjust the pH to 4.8-5.2 with lactic acid to obtain solution AB;

[0026] (4) Dissolve the antibacterial agent, PEG-40 hydrogenated castor oil, β-nicotinamide mononucleotide and cucumber flavoring in water for injection at 30±2℃ and disperse by ultrasonication for 5-10 min to obtain solution C;

[0027] (5) Add solution C dropwise to the AB mixed gel, homogenize under vacuum for 10-15 minutes, and then fill the gel.

[0028] Thirdly, this invention discloses a method for preparing the above-mentioned anti-aging, moisturizing, and antibacterial vaginal gel containing at least one of coenzyme Q10, Centella asiatica extract, and Lactobacillus fermentation product, comprising the following steps:

[0029] (1) Add glycerin, sodium hyaluronate and xanthan gum to water for injection at 60±2℃ in sequence, stir at 500-600rpm for 15-20min, cool to 30℃ and let stand to remove bubbles to obtain solution A;

[0030] (2) Disperse hydroxyethyl cellulose and carboxymethyl chitosan in water for injection at 40±2℃, heat to 37±1℃ and stir at 200-300rpm for 10-20min, cool to 20-25℃ to form a pregel, i.e. solution B;

[0031] (3) Slowly add solution A to solution B, stir at 100-150 rpm for 10-15 min, and adjust the pH to 4.8-5.2 with lactic acid to obtain solution AB;

[0032] (4) Dissolve the antibacterial agent, PEG-40 hydrogenated castor oil, β-nicotinamide mononucleotide and cucumber flavoring in water for injection at 30±2℃ and disperse by ultrasonication for 5-10 min to obtain solution C;

[0033] (5) Final product mixing: Add solution C dropwise to the AB mixed gel, and simultaneously add coenzyme Q10, Centella asiatica extract or lactobacillus fermentation product. After vacuum homogenization for 10-15 minutes, fill the gel.

[0034] In some embodiments, the vaginal gel exhibits a 24-hour inhibition rate of ≥99.5% against Candida albicans or Staphylococcus aureus, and has no significant inhibitory effect on vaginal lactobacilli.

[0035] In some embodiments, after the vaginal gel treated VK2 / E6E7 cells for 48 hours, the collagen content in the cells increased by ≥35%, and the mitochondrial activity increased by ≥40%.

[0036] Fourthly, the present invention also discloses the application of the above-mentioned anti-aging, moisturizing and antibacterial vaginal gel in the preparation of care products / medications for preventing vaginal infections, delaying vaginal tissue aging or maintaining vaginal microecological balance.

[0037] Fifthly, the present invention also discloses the application of the above-mentioned anti-aging, moisturizing and antibacterial vaginal gel in the preparation of clotrimazole vaginal gel in combination with clotrimazole, wherein the dose of clotrimazole is 50-100 mg and the mass ratio of clotrimazole to vaginal gel is 1:20 to 1:100.

[0038] Compared with the prior art, the present invention has the following beneficial effects:

[0039] This invention utilizes components such as glycerin, sodium hyaluronate, xanthan gum, hydroxyethyl cellulose, carboxymethyl chitosan, β-nicotinamide mononucleotide, antibacterial agents, and PEG-40 hydrogenated castor oil, and further includes coenzyme Q10, Centella asiatica extract, or lactobacillus fermentation products. The resulting vaginal gel not only possesses excellent antibacterial, anti-aging, and moisturizing properties, but also promotes the proliferation of beneficial vaginal bacteria. Furthermore, the trehalose / mannitol freeze-drying protection technology ensures the stability of the active ingredients.

[0040] The vaginal gel of this invention, with the proper formulation of its components, exhibits good fluidity and adhesion, allowing for even application without residue. This significantly enhances user safety and satisfaction, providing a new approach to the research and development of women's intimate health.

[0041] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0042] To more clearly illustrate the technical solution of the present invention, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0043] Figure 1 The graph shows the test results of the inhibition rate of elastase by gels in the embodiments and comparative examples of this invention. Detailed Implementation

[0044] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to specific examples. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0045] Unless otherwise specified, the experimental methods used in this invention are conventional methods; unless otherwise specified, the materials and reagents used are commercially available.

[0046] Table 1. Information on Main Raw Materials

[0047]

[0048]

[0049]

[0050] In this invention, the preparation of the lactobacillus fermentation product includes the following steps:

[0051] 1. Strain activation: Lactobacillus plantarum was inoculated into a modified MRS medium containing basic MRS components, 1.5 wt% inulin, 2 wt% soybean peptide, and 0.2 wt% glutamine, and cultured anaerobically at 37°C until OD600 = 3.0-3.5;

[0052] 2. High-density fermentation: A fermentation medium is packed into a bioreactor, the fermentation medium comprising trehalose 25 g / L, whey protein hydrolysate 18 g / L, MnSO4·H2O 0.075 g / L, potassium dihydrogen phosphate 2 g / L, magnesium sulfate 0.2 g / L, vitamin B1 0.01 g / L and initial glucose 10 g / L;

[0053] The inoculum was 7% (v / v), and the feeding was controlled by dissolved oxygen feedback to maintain the DO value at 10%-15%. The cells were harvested after 48 hours of fermentation.

[0054] 3. Product extraction: The bacterial suspension was homogenized under high pressure of 800-1000 bar to disrupt the cell walls, and then passed through a 0.22 μm ceramic membrane and a 10 kDa tangential flow ultrafiltration membrane in sequence to collect the active small molecule components with a molecular weight <10 kDa.

[0055] 4. Freeze-drying protection: Add 5% trehalose and 1% mannitol to the filtrate, and freeze-dry under vacuum. Pre-freeze at -50℃, main dry at -20℃ / 24h, and desorption dry at 25℃ / 12h to obtain powdered fermentation product.

[0056] Example 1

[0057] I. An anti-aging, moisturizing, and antibacterial vaginal gel, composed of three solutions, A, B, and C, in parts by weight:

[0058] Solution A: 15 parts glycerin, 10 parts sodium hyaluronate, 1 part xanthan gum, 50 parts water for injection;

[0059] Solution B: 0.5 parts hydroxyethyl cellulose, 1 part carboxymethyl chitosan, 10 parts water for injection;

[0060] Solution C: 0.5 parts β-nicotinamide mononucleotide, 0.5 parts PE 9010 (antibacterial agent), 0.1 parts PEG-40 hydrogenated castor oil, 0.1 parts cucumber flavoring, 30 parts water for injection.

[0061] II. A method for preparing an anti-aging, moisturizing, and antibacterial vaginal gel, comprising the following steps:

[0062] 1. Add glycerin, sodium hyaluronate and xanthan gum sequentially to water for injection at 60±2℃, stir at 500-600rpm for 15-20min, then cool to 30℃ and let stand to remove bubbles to obtain solution A;

[0063] 2. Disperse hydroxyethyl cellulose and carboxymethyl chitosan in water for injection at 40±2℃, heat to 37±1℃ and stir at 200-300 rpm for 10-20 min, cool to 20-25℃ to form a pregel, and obtain solution B;

[0064] 3. Slowly add solution A to solution B, stir at 100-150 rpm for 10-15 min, and adjust the pH to 4.8-5.2 with lactic acid to obtain a mixed gel of A and B;

[0065] 4. PE 9010, PEG-40 hydrogenated castor oil, β-nicotinamide mononucleotide and cucumber flavoring were dissolved in water for injection at 30±2℃ and ultrasonically dispersed for 5-10 min to obtain solution C;

[0066] 5. Add solution C dropwise to the AB mixed gel, homogenize under vacuum for 10-15 minutes, and then fill the gel.

[0067] Example 2

[0068] I. An anti-aging, moisturizing, and antibacterial vaginal gel, composed of three solutions, A, B, and C, in parts by weight:

[0069] Solution A: 20 parts glycerin, 12.5 parts sodium hyaluronate, 3 parts xanthan gum, 75 parts water for injection;

[0070] Solution B: 1 part hydroxyethyl cellulose, 1.5 parts carboxymethyl chitosan, 15 parts water for injection;

[0071] Solution C: 1.5 parts β-nicotinamide mononucleotide, 1 part PE 9010, 0.5 parts PEG-40 hydrogenated castor oil, 0.2 parts cucumber flavoring, 45 parts water for injection.

[0072] II. The preparation method of an anti-aging, moisturizing, and antibacterial vaginal gel is the same as in Example 1.

[0073] Example 3

[0074] An anti-aging, moisturizing, and antibacterial vaginal gel, composed of three solutions, A, B, and C, in parts by weight:

[0075] Solution A: 25 parts glycerin, 15 parts sodium hyaluronate, 5 parts xanthan gum, 100 parts water for injection;

[0076] Solution B: 2 parts hydroxyethyl cellulose, 2 parts carboxymethyl chitosan, 20 parts water for injection;

[0077] Solution C: 3 parts β-nicotinamide mononucleotide, 1.5 parts PE 9010, 0.8 parts PEG-40 hydrogenated castor oil, 0.3 parts cucumber flavoring, 60 parts water for injection.

[0078] II. The preparation method of an anti-aging, moisturizing, and antibacterial vaginal gel is the same as in Example 1.

[0079] Example 4

[0080] Example 4 differs from Example 2 in that it also includes 3 portions of Lactobacillus fermentation products; otherwise, they are the same.

[0081] The preparation method of an anti-aging, moisturizing and antibacterial vaginal gel is the same as in Example 2, with the lactobacillus fermentation product added in step 5.

[0082] Example 5

[0083] Example 5 differs from Example 2 in that it also includes 3 portions of Lactobacillus fermentation product and 1 portion of Coenzyme Q10, while all other components are the same.

[0084] The preparation method of an anti-aging, moisturizing and antibacterial vaginal gel is the same as in Example 2, except that lactobacillus fermentation product and coenzyme Q10 are added in step 5.

[0085] Example 6

[0086] Example 6 differs from Example 2 in that it also includes 3 portions of Lactobacillus fermentation product, 1 portion of Coenzyme Q10, and 1 portion of Centella Asiatica extract; all other components are the same.

[0087] The preparation method of an anti-aging, moisturizing and antibacterial vaginal gel is the same as in Example 2, except that lactobacillus fermentation product, coenzyme Q10 and centella asiatica extract are added in step 5.

[0088] Comparative Example 1

[0089] The difference between Comparative Example 1 and Example 2 is that Comparative Example 1 does not contain β-nicotinamide mononucleotide, but all other aspects are the same.

[0090] Comparative Example 2

[0091] The difference between Comparative Example 2 and Example 2 is that Comparative Example 2 does not contain... PE 9010, everything else is the same.

[0092] Comparative Example 3

[0093] The difference between Comparative Example 3 and Example 2 is that Comparative Example 3 does not contain β-nicotinamide mononucleotide and PE9010, everything else is the same.

[0094] Comparative Example 4

[0095] The difference between Comparative Example 4 and Example 2 is that Comparative Example 4 does not contain β-nicotinamide mononucleotide. PE9010 and xanthan gum are the same, everything else is the same.

[0096] Test Example 1

[0097] Testing the antibacterial effects of different vaginal gels

[0098] Test samples: Examples 1, 2, 3, 4, 5, and 6; Comparative Example 2 (none) PE 9010), Comparative Example 3 (None) PE 9010 and NMN)

[0099] Test method: WS / T 650-2019 "Evaluation Methods for Antibacterial and Bacteriostatic Efficacy" 5.1.2 Carrier Immersion Quantitative Antibacterial Test. Contact time: 30 min.

[0100] Antibacterial rate: The antibacterial rate against Candida albicans and Staphylococcus aureus was tested according to ISO 20743 standard for 24 hours.

[0101] Selective antibacterial activity: Detection of the survival rate of vaginal lactobacillus crispatus.

[0102] Table 2. Results of antibacterial rate tests for different gels.

[0103]

[0104] The testing standards stipulate that an antibacterial rate of ≥50%-90% is considered to have an antibacterial effect; an antibacterial rate of ≥90% is considered to have a strong antibacterial effect.

[0105] As shown in Table 2, compared with Comparative Examples 2 and 3, Examples 1-6 exhibited higher antibacterial rates, and the antibacterial rate increased with the addition of β-nicotinamide mononucleotide and... The increased concentration of PE 9010 leads to a higher antibacterial rate, indicating that the vaginal gel of this invention has good antibacterial properties.

[0106] Test Example 2

[0107] Testing the anti-aging function of different vaginal gels

[0108] Test samples: Examples 2, 5, and 6; Comparative Example 1 (without β-nicotinamide mononucleotide), Comparative Example 4 (without β-nicotinamide mononucleotide + xanthan gum)

[0109] I. Elastase

[0110] Elastase has the ability to degrade various proteins such as collagen and elastin. Changes in elastin structure during skin aging are a major cause of wrinkles and sagging skin. The tightening effect of vaginal gels was analyzed using elastase inhibition rates. A higher elastase inhibition rate indicates a stronger tightening effect.

[0111] Test method:

[0112] Sample pretreatment: Vaginal gels were prepared to concentrations of 5%, 10%, 15%, 20%, and 25%, respectively. Then, lichen red elastin and borate buffer solution were added, and the mixture was kept at a constant temperature and shaken in a 37°C water bath for 10 min. Then, 1 mL of elastase solution was added, and the absorbance was measured at 495 nm to calculate the elastase inhibition rate.

[0113] Test method: MC-ZY-162 elastase inhibition rate method (source: Zhongke Testing Technology Service (Guangzhou) Co., Ltd.)

[0114] Depend on Figure 1As shown, the vaginal gels all have a significant inhibitory effect on elastase, and the inhibition rate of elastase increases linearly with the increase of sample concentration. In particular, the vaginal gels of Examples 5 and 6, which contain coenzyme Q10 and Centella asiatica extract, have the highest inhibition rate of elastase, indicating that the vaginal gel of the present invention has a significant tightening effect.

[0115] II. Collagen and Mitochondrial ATP

[0116] Test method:

[0117] Collagen synthesis: Type I collagen content was detected by ELISA in VK2 / E6E7 cells (after 48 h of treatment);

[0118] Mitochondrial activity: ATP content determination ( Law).

[0119] Table 3. Results of Anti-aging Function Test

[0120] Group Collagen content increase rate Increased mitochondrial ATP production rate Example 2 38.5％ 42.1％ Example 5 45.2％ 48.7％ Example 6 47.8％ 51.2％ Comparative Example 1 5.3％ 6.8％ Comparative Example 4 4.1％ 5.2％

[0121] As shown in Table 3, the collagen enhancement rate and mitochondrial ATP production enhancement rate of Examples 2 and Examples 5-6 were significantly higher than those of Comparative Examples 1 and 4 which lacked β-nicotinamide mononucleotide or xanthan gum. Furthermore, Examples 5 and 6, which added coenzyme Q10 and Centella asiatica extract, showed the highest enhancement rates, indicating that the addition of coenzyme Q10 and Centella asiatica extract can effectively improve the anti-aging effect.

[0122] Test Example 3

[0123] Testing the moisturizing properties and rheological characteristics of different vaginal gels

[0124] Test samples: Examples 1, 2, 3, 4, 5, and 6; Comparative Example 4 (without xanthan gum); Commercially available gel (metronidazole vaginal gel).

[0125] Test method:

[0126] Long-lasting moisturizing effect: Moisture content of isolated porcine vaginal mucosa 12 hours after application ( );

[0127] Adhesion: Storage modulus (G') and loss modulus (G”) were determined by rotational rheometer.

[0128] Table 4. Results of Moisturizing Performance and Rheological Properties Tests

[0129] Group G'(Pa, 37℃) G (Pa, 37℃) 12-hour moisture retention rate Example 1 1250 320 78.3％ Example 2 1450 290 82.4％ Example 3 1680 260 84.1％ Example 4 1420 310 81.9％ Example 5 1500 285 83.2％ Example 6 1550 275 84.5％ Comparative Example 4 480 180 54.6％ Commercially available gels 620 210 48.9％

[0130] As shown in Table 4, compared with Comparative Example 4 and commercially available gels, the vaginal gel of the present invention has a water retention rate of over 75%, indicating that the vaginal gel of the present invention has better moisturizing properties. Furthermore, the storage modulus and loss modulus test results show that the storage modulus and loss modulus of Examples 1-6 are much greater than those of Comparative Example 4 and commercially available gels, indicating that the vaginal gel of the present invention has high stability, can maintain a certain shape within the vagina, and will not be easily squeezed out by slight body movements. It also has good fluidity, allowing for better contact with the vaginal wall during use and even distribution within the vagina, thus better exerting its medicinal or nursing effects.

[0131] Test Example 4

[0132] Microecological regulation and stability testing

[0133] Test subjects: Examples 4, 5, and 6; Example 2 (no fermentation products)

[0134] Test method:

[0135] Probiotic proliferation: Co-culture with vaginal lactobacilli for 24 hours, and detection of viable bacteria count and pH;

[0136] Fermentation product stability: The retention rate of active small molecules was detected after accelerated testing (40℃ / 75%RH, 3 months).

[0137] Table 5 Results of Microecological Regulation and Stability Test

[0138]

[0139] As shown in Table 5, Examples 4-6 significantly improved the proliferation of probiotics, and the stability of the active ingredients was >93%. In particular, Example 6 contained Centella asiatica extract with anti-inflammatory effects, which further promoted the balance of the gut microbiota.

[0140] Test Example 5

[0141] Safety and irritation assessment

[0142] Test subjects: Examples 1-6; Comparative Examples 1-4; Commercially available benzalkonium chloride-containing gels.

[0143] Test method:

[0144] Rabbit vaginal stimulation test: scored according to OECD 405 standards (0-4 grades);

[0145] Cytotoxicity: VK2 / E6E7 cell viability (MTT assay, 48h treatment).

[0146] Table 6 Safety and Irritation Assessment Table

[0147] Group Stimulation Index (Rabbit) Cell viability (%) Examples 1-6 0.2-0.4 (non-irritating) 98.5-101.2％ Comparative Examples 1-4 0.3-0.5 (non-irritating) 97.8-99.1％ Commercially available gels 2.1 (Moderate Stimulation) 82.3％

[0148] As shown in Table 6, the vaginal gels of Examples 1-6 and Comparative Examples 1-4 all meet the safety requirements and have significantly lower irritation levels than commercially available products.

[0149] Test Example 6

[0150] Evaluation of the synergistic antifungal effects of clotrimazole in combination therapy

[0151] Test subject:

[0152] Experimental group: In Example 6, the gel was used in combination with clotrimazole (50 mg, 75 mg, 100 mg) at mass ratios of 1:20, 1:50, and 1:100, respectively; clotrimazole was added to the gel in preparation step (5).

[0153] Negative control: Example 6 gel (without clotrimazole).

[0154] Commercially available combination control group: commercially available ordinary gel (same matrix) and clotrimazole (100 mg, mass ratio 1:50).

[0155] Positive control: Commercially available clotrimazole suppositories (containing 100 mg of clotrimazole).

[0156] Test method:

[0157] 1. In vitro antifungal activity: The minimum inhibitory concentration (MIC) and 24-hour inhibition rate against Candida albicans were determined according to CLSIM27-A3 standard;

[0158] 2. Synergistic effect index (FICI): The combined antibacterial effect of clotrimazole and the gel of the present invention was calculated (FICI≤0.5 indicates synergy).

[0159] 3. Transdermal absorption and maintenance of drug efficacy:

[0160] The cumulative penetration of clotrimazole into isolated porcine vaginal mucosa was determined using a Franz diffusion cell (24 hours).

[0161] The release rate of clotrimazole was measured in a simulated vaginal environment (pH 4.8, 37℃) (0-12 hours).

[0162] 4. Stimulation test:

[0163] Rabbit vaginal mucosal irritation test (OECD 405): After 7 consecutive days of administration, the reactions such as erythema and edema were evaluated.

[0164] 1. Antifungal activity and synergistic effect, results are shown in Table 7.

[0165] Table 7 Results of Antifungal Activity and Synergistic Effect Tests

[0166]

[0167] Note: Example 6 itself achieved a 99.7% inhibition rate against Candida albicans (due to the presence of...). PE 9010 and carboxymethyl chitosan).

[0168] FICI determination: The combination groups in Example 6 all showed significant synergistic effects (FICI<0.5), while the commercially available combination groups showed no synergistic effect (FICI>0.5).

[0169] 2. Transdermal absorption and release performance; results are shown in Table 8.

[0170] Table 8. Results of Transdermal Absorption and Release Performance Tests

[0171]

[0172] 3. Stimulation test; test results are shown in Table 9.

[0173] Table 9. Results of Stimulation Tests

[0174]

[0175] Results analysis:

[0176] Table 7 shows that when clotrimazole (100 mg, 1:100) was used in combination in Example 6, the MIC was as low as 1.2 μg / mL, the antibacterial rate was 99.8%, and the FICI was 0.28 (strong synergistic effect). Table 8 shows that the gel in Example 6, containing PEG-40 hydrogenated castor oil (a transdermal penetration enhancer), increased clotrimazole penetration by 100% compared to commercially available gels, indicating that the vaginal gel of this invention has high permeability and a 12-hour release rate >89%, significantly better than suppositories (72.8%). Table 9 shows that the irritation index of clotrimazole used in combination in Example 6 was only 0.4, with no mucosal damage, indicating that the gel of this invention not only has good antibacterial properties, transdermal permeability, and safety, but also does not affect efficacy, and can significantly reduce drug dosage (50 mg is equivalent to 100 mg of commercially available suppositories).

[0177] In summary, this invention provides an anti-aging, moisturizing, and antibacterial vaginal gel comprising glycerin, sodium hyaluronate, xanthan gum, hydroxyethyl cellulose, carboxymethyl chitosan, β-nicotinamide mononucleotide, an antibacterial agent, and PEG-40 hydrogenated castor oil. The carboxymethyl chitosan and antibacterial agent exhibit a 24-hour inhibition rate of ≥99.5% against pathogenic bacteria such as Candida albicans and Staphylococcus aureus, while selectively preserving the activity of vaginal lactobacilli, thus avoiding the risk of microecological imbalance. Furthermore, the vaginal gel also includes coenzyme Q10, Centella asiatica extract, or lactobacillus fermentation products. The β-nicotinamide mononucleotide, combined with coenzyme Q10, effectively improves vaginal tissue laxity and decreased elasticity by activating mitochondrial energy metabolism (activation increased by ≥40%) and promoting collagen synthesis (content increased by ≥35%), filling a gap in the market for anti-aging functions. Simultaneously, the Lactobacillus plantarum fermentation products (<10kDa active small molecules) promote the proliferation of beneficial vaginal bacteria, and the trehalose / mannitol freeze-drying protection technology ensures the stability of the active ingredients, thereby maintaining bacterial diversity. The vaginal gel of this invention achieves four effects in one: antibacterial, anti-aging, moisturizing, and microecological regulation. It can also be adapted to different needs through optimized component ratios (such as NMN 0.5-3% gradient). At the same time, when used in combination with clotrimazole, it can reduce the dosage of drugs by 50% and improve the efficacy (MIC reduced by 60%), showing broad application prospects.

[0178] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. An anti-aging, moisturizing, and antibacterial vaginal gel, characterized in that, Includes solutions A, B, and C; by weight: Solution A: 15-25 parts glycerin, 10-15 parts sodium hyaluronate, 1-5 parts xanthan gum, 50-100 parts water; Solution B: 0.5-2 parts hydroxyethyl cellulose, 1-2 parts carboxymethyl chitosan, 10-20 parts water; Solution C: 0.5-3 parts β-nicotinamide mononucleotide, 0.5-1.5 parts antibacterial agent, 0.1-0.8 parts PEG-40 hydrogenated castor oil, 0.1-0.3 parts fragrance, 30-60 parts water.

2. The anti-aging, moisturizing, and antibacterial vaginal gel according to claim 1, characterized in that, The anti-aging, moisturizing, and antibacterial vaginal gel also contains at least one ingredient selected from the following: 0.5-2 parts Coenzyme Q10; 0.5-1.5 parts Centella Asiatica extract; 1-5 portions of Lactobacillus fermentation products.

3. The anti-aging, moisturizing, and antibacterial vaginal gel according to claim 2, characterized in that, The preparation of the lactobacillus fermentation product includes the following steps: S1. Inoculate Lactobacillus plantarum into MRS medium containing 1-2 wt% inulin, 1-3 wt% soybean peptide and 0.1-0.3 wt% glutamine, and culture anaerobically at 35-40℃ until OD600 = 3.0-3.5; S2. Fermentation medium was loaded into the bioreactor, and the glucose flow rate was controlled by dissolved oxygen feedback to maintain the DO value at 10%-15%. The cells were collected after 48 hours of fermentation. S3. After the bacterial suspension is homogenized under high pressure, it is sequentially passed through a 0.22μm ceramic membrane for ultrafiltration and a 10kDa tangential flow ultrafiltration membrane for fractionation, and the <10kDa active small molecules are collected. S4. Add 3-8% trehalose and 0.5-1.5% mannitol to the filtrate, and freeze-dry under vacuum to obtain a powdered fermentation product.

4. The anti-aging, moisturizing, and antibacterial vaginal gel according to claim 3, characterized in that, The fermentation medium contains 20-30 g / L trehalose, 15-20 g / L whey protein hydrolysate, 0.05-0.1 g / L MnSO4·H2O, 2 g / L potassium dihydrogen phosphate, 0.2 g / L magnesium sulfate, 0.01 g / L vitamin B1, and 10 g / L initial glucose. The inoculum size of *Lactobacillus plantarum* is 5%-10% (v / v); the homogenization pressure is 800-1000 bar.

5. The preparation method of the anti-aging, moisturizing, and antibacterial vaginal gel as described in claim 1, characterized in that, Includes the following steps: (1) Add glycerin, sodium hyaluronate and xanthan gum to water for injection at 60±2℃ in sequence, stir at 500-600rpm for 15-20min, cool to 30℃ and let stand to remove bubbles to obtain solution A; (2) Disperse hydroxyethyl cellulose and carboxymethyl chitosan in water for injection at 40±2℃, heat to 37±1℃ and stir at 200-300 rpm for 10-20 min, cool to 20-25℃ to form a pregel, and obtain solution B; (3) Slowly add solution A to solution B, stir at 100-150 rpm for 10-15 min, and adjust the pH to 4.8-5.2 with lactic acid to obtain solution AB; (4) Dissolve the antibacterial agent, PEG-40 hydrogenated castor oil, β-nicotinamide mononucleotide and cucumber flavoring in water for injection at 30±2℃ and disperse by ultrasonication for 5-10 min to obtain solution C; (5) Add solution C dropwise to the AB mixed gel, homogenize under vacuum for 10-15 minutes, and then fill the gel.

6. The method for preparing the anti-aging, moisturizing, and antibacterial vaginal gel according to any one of claims 2-4, characterized in that, Includes the following steps: (1) Add glycerin, sodium hyaluronate and xanthan gum to water for injection at 60±2℃ in sequence, stir at 500-600rpm for 15-20min, cool to 30℃ and let stand to remove bubbles to obtain solution A; (2) Disperse hydroxyethyl cellulose and carboxymethyl chitosan in water for injection at 40±2℃, heat to 37±1℃ and stir at 200-300rpm for 10-20min, cool to 20-25℃ to form a pregel, i.e. solution B; (3) Slowly add solution A to solution B, stir at 100-150 rpm for 10-15 min, and adjust the pH to 4.8-5.2 with lactic acid to obtain solution AB; (4) Dissolve the antibacterial agent, PEG-40 hydrogenated castor oil, β-nicotinamide mononucleotide and cucumber flavoring in water for injection at 30±2℃ and disperse by ultrasonication for 5-10 min to obtain solution C; (5) Final product mixing: Add solution C dropwise to the AB mixed gel, and simultaneously add coenzyme Q10, Centella asiatica extract or lactobacillus fermentation product. After vacuum homogenization for 10-15 minutes, fill the gel.

7. The anti-aging, moisturizing, and antibacterial vaginal gel according to any one of claims 1-4, characterized in that, The vaginal gel exhibits a 24-hour inhibition rate of ≥99.5% against Candida albicans or Staphylococcus aureus, and has no significant inhibitory effect on vaginal lactobacilli.

8. The anti-aging, moisturizing, and antibacterial vaginal gel according to any one of claims 1-4, characterized in that, After treating VK2 / E6E7 cells with the vaginal gel for 48 hours, the collagen content in the cells increased by ≥35%, and the mitochondrial activity increased by ≥40%.

9. The use of the anti-aging, moisturizing, and antibacterial vaginal gel as described in any one of claims 1-4 in the preparation of care products / medications for preventing vaginal infections, delaying vaginal tissue aging, or maintaining vaginal microecological balance.

10. The use of the vaginal gel according to any one of claims 1-4 in the preparation of clotrimazole vaginal gel in combination with clotrimazole, characterized in that, The clotrimazole dosage is 50-100 mg, and the mass ratio of clotrimazole to vaginal gel is 1:20 to 1:100.

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