D-lactic acid vaginal micro-ecological repair gel and preparation method and application thereof

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a D-lactic acid vaginal micro-ecological repair gel as well as a preparation method and application thereof. The application provides a D-lactic acid vaginal micro-ecological repair gel, which comprises the following raw materials in percentage by weight: 0.1-3.0% of D-lactic acid, 1-10% of a humectant, 2-4% of a gel base, 0.01-1% of an epithelial repair agent, 0.1-5% of a colonization promoter, a pH regulator and the balance of water; and the pH value of the D-lactic acid vaginal micro-ecological repair gel is 3.8-4.1. The gel of the application can stably supplement D-lactic acid and enhance the competitive colonization ability of lactobacillus; the gel of the application can inhibit the proliferation of pathogenic bacteria, improve the repair ability of the vaginal epithelial barrier and inhibit HPV infection; and the gel is suitable for auxiliary removal of vaginitis and HPV infection.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a D-lactic acid vaginal microecological repair gel, its preparation method, and its application. Background Technology

[0002] Traditional vaginal care products primarily use weakly acidic agents (such as L-lactic acid), but these struggle to restore the dominant D-lactic acid-producing bacteria ecosystem (such as the cristata flora). Bacterial vaginosis (BV) has a high recurrence rate, and persistent HPV infection is associated with microecological imbalance. Therefore, there is a need to develop a safe gel formula that can promote the growth of beneficial lactobacilli and stably maintain pH levels. Summary of the Invention

[0003] In view of this, the purpose of this invention is to provide a D-lactic acid vaginal microecological repair gel and its preparation method. The D-lactic acid vaginal microecological repair gel provided by this invention can increase the supply of D-lactic acid, promote the recovery of lactobacilli, and maintain healthy vaginal acidity, making it suitable for regulating the vaginal microecological system.

[0004] This invention provides a D-lactic acid vaginal microecological repair gel, comprising the following raw materials in weight percentage: D-lactic acid 0.1~3.0%, moisturizer 1~10%, gel matrix 2~4%, epithelial repair agent 0.01~1%, colonization promoter 0.1~5%, pH adjuster and the balance water; The pH value of the D-lactic acid vaginal microecological repair gel is 3.8~4.1.

[0005] Preferably, the weight percentage of D-lactic acid in the raw material is 1.0~3.0%.

[0006] Preferably, the humectant is glycerin; the weight percentage of the humectant in the raw materials is 3-8%.

[0007] Preferably, the gel matrix comprises hydroxypropyl methylcellulose and / or carbomer; the weight percentage of hydroxypropyl methylcellulose in the raw material is 1-2%, and the weight percentage of carbomer in the raw material is 1-4%.

[0008] Preferably, the epithelial repair agent is sodium hyaluronate; the weight percentage of the epithelial repair agent in the raw material is 0.3~0.8%.

[0009] Preferably, the colonization promoter is fructooligosaccharide; the weight percentage of the colonization promoter in the raw material is 1-2%.

[0010] Preferably, the pH adjuster is a sodium hydroxide solution with a concentration of 0.1 mol / L.

[0011] This invention also provides a method for preparing the D-lactic acid vaginal microecological repair gel described in the above technical solution, including method one or method two, wherein method one includes the following steps: The gel matrix and water are mixed to obtain a gel matrix solution; The gel matrix solution, D-lactic acid, moisturizer, epithelial repair agent and colonization promoter are mixed, and the pH of the resulting mixture is adjusted to 3.8~4.1. The mixture is allowed to stand to form a gel, thus obtaining the D-lactic acid vaginal microecological repair gel. The second method includes the following steps: Prepare blank gels from the gel matrix; The blank gel was immersed in the mixed solution for loading, and then the gel surface was rinsed to obtain the D-lactic acid vaginal microecological repair gel; the mixed solution contains D-lactic acid, moisturizer, epithelial repair agent and colonization promoter.

[0012] Preferably, in Method 1, the pH adjustment agent used to adjust the pH value of the resulting mixture to 3.8-4.1 is a sodium hydroxide solution with a concentration of 0.1 mol / L.

[0013] This invention also provides the application of the D-lactic acid vaginal microecological repair gel described in the above technical solution or the D-lactic acid vaginal microecological repair gel obtained by the above preparation method in the preparation of drugs for treating vaginitis or inhibiting HPV infection.

[0014] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a D-lactic acid vaginal microecological repair gel, comprising the following raw materials in weight percentage: D-lactic acid 0.1~3.0%, moisturizer 1~10%, gel matrix 2~4%, epithelial repair agent 0.01~1%, colonization promoter 0.1~5%, pH adjuster and balance water; the pH value of the D-lactic acid vaginal microecological repair gel is 3.8~4.1.

[0015] The D-lactic acid vaginal microecological repair gel provided by this invention can increase the concentration of D-lactic acid in the vagina, reduce the vaginal pH value, inhibit the growth of pathogenic microorganisms, promote the recovery of lactobacilli, and maintain healthy vaginal acidity, making it suitable for regulating the vaginal microecological system. The D-lactic acid vaginal microecological repair gel can be used for vaginal microecological regulation, lactobacillus colonization, prevention of recurrence of vaginitis, and HPV-assisted clearance.

[0016] The D-lactic acid vaginal microecological repair gel provided by this invention has the following functions: (1) increasing the key D-lactic acid in the vaginal natural environment; (2) promoting the colonization of dominant bacteria such as Lactobacillus cristata; (3) reducing the recurrence of BV vaginitis; (4) improving the mucosal repair capacity; (5) increasing the HPV clearance rate; (6) safe, antibiotic-free, and hormone-free; (7) broad-spectrum applicability: suitable for a variety of human microecological sites; (8) multi-dosage form coverage: microspheres, membranes, gels, sprays, and ointments can all use the gel system of this invention; (9) compatible with probiotic preparations: can be used in combination with probiotics to enhance the effect; (10) significantly reducing the recurrence rate: the continuous acidic barrier can be maintained for several hours to several days; (11) strong scalability: can be adapted to different polymer systems. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, 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.

[0018] Figure 1 This is a schematic diagram illustrating the composition of the D-lactic acid vaginal microecological repair gel of the present invention; Figure 2 HE staining image of vaginal tissue after application of gel from Example 1 in Test Example 1; Figure 3 HE staining image of vaginal tissue after gelation of Comparative Example 1 in Test Example 1. Detailed Implementation

[0019] This invention provides a D-lactic acid vaginal microecological repair gel, comprising the following raw materials in weight percentage: D-lactic acid 0.1~3.0%, moisturizer 1~10%, gel matrix 2~4%, epithelial repair agent 0.01~1%, colonization promoter 0.1~5%, pH adjuster and the balance water; The pH value of the D-lactic acid vaginal microecological repair gel is 3.8~4.1.

[0020] Unless otherwise specified, all materials and equipment used in this invention are commercially available products in the field.

[0021] Figure 1 This is a schematic diagram illustrating the composition of the D-lactic acid vaginal microecological repair gel of the present invention.

[0022] In this invention, the weight percentage of D-lactic acid in the raw material is preferably 1.0 to 3.0%. D-lactic acid can supplement the D-lactic acid naturally present in the vaginal environment; help maintain the vaginal pH value below 4.4 (normal state), inhibit the growth of pathogenic microorganisms, and inhibit HPV infection.

[0023] In this invention, the humectant is preferably glycerin; the weight percentage of the humectant in the raw materials is preferably 3-8%, specifically 4% or 5%.

[0024] In this invention, the gel matrix preferably includes hydroxypropyl methylcellulose (HPMC) and / or carbopol; the weight percentage of hydroxypropyl methylcellulose in the raw materials is preferably 1-2%, and the weight percentage of carbopol in the raw materials is preferably 1-4%, specifically 2%.

[0025] In this invention, the epithelial repair agent is preferably sodium hyaluronate; the weight percentage of the epithelial repair agent in the raw materials is preferably 0.3~0.8%, specifically 0.5%.

[0026] In this invention, the colonization promoter is preferably fructooligosaccharide (FOS); the weight percentage of the colonization promoter in the raw materials is preferably 1-2%. The fructooligosaccharide can promote the colonization of lactobacillus.

[0027] In this invention, the pH adjuster is preferably a sodium hydroxide solution, and the concentration of the sodium hydroxide solution is preferably 0.1 mol / L.

[0028] In this invention, the pH value of the D-lactic acid vaginal microecological repair gel is preferably 4.0.

[0029] In this invention, the D-lactic acid vaginal microecological repair gel preferably further includes auxiliary regulatory factors, which preferably include one or more of buffers, probiotic metabolites, organic acids, oligosaccharides, microbial growth promoting factors, barrier repair factors, and antioxidants. This invention does not have any special requirements on the amount of auxiliary regulatory factors added.

[0030] This invention also provides a method for preparing the D-lactic acid vaginal microecological repair gel described in the above technical solution, including method one or method two, wherein method one includes the following steps: The gel matrix and water are mixed to obtain a gel matrix solution; The gel matrix solution, D-lactic acid, moisturizer, epithelial repair agent and colonization promoter are mixed, and the pH of the resulting mixture is adjusted to 3.8~4.1. The mixture is allowed to stand to form a gel, thus obtaining the D-lactic acid vaginal microecological repair gel. The second method includes the following steps: Prepare blank gels from the gel matrix; The blank gel was immersed in the mixed solution for loading, and then the gel surface was rinsed to obtain the D-lactic acid vaginal microecological repair gel; the mixed solution contains D-lactic acid, moisturizer, epithelial repair agent and colonization promoter.

[0031] When the gel matrix is ​​carbomer, method one is preferred. The following is a detailed explanation of method one (pH-triggered gelation method): In this invention, after mixing the gel matrix and water, an alkali is preferably added, preferably a sodium hydroxide solution, with a concentration of 0.1 mol / L. Adding the alkali promotes the dissolution of the gel matrix, and the sodium hydroxide solution is slowly added dropwise until the pH value reaches 4.0.

[0032] In this invention, the preferred method for mixing the gel matrix solution, D-lactic acid, moisturizer, epithelial repair agent, and colonization promoter is to first mix the gel matrix solution, moisturizer, epithelial repair agent, and colonization promoter, and then add the D-lactic acid solution; the preferred mixing temperature for the gel matrix solution, moisturizer, epithelial repair agent, and colonization promoter is ≤40℃ to avoid the system being destroyed and deactivated.

[0033] In this invention, the preferred temperature for gelation is 2-6°C, specifically 4°C, and the preferred time is 6 hours. This invention employs a pH-triggered gelation method, where a decrease in pH triggers the self-assembly of the gel matrix to form a three-dimensional network. The conditions are mild, requiring no heating, making it suitable for loading various active ingredients. pH changes are controllable, preventing localized precipitation.

[0034] In this invention, the step of allowing the gel to stand and solidify preferably includes homogenization to make the gel finer.

[0035] When the gel matrix is ​​hydroxypropyl methylcellulose, method two is preferred. The following is a detailed explanation of method two (preparing a gel network first and then loading the active ingredient): The present invention does not have any special requirements for the method of preparing the gel matrix into a blank gel.

[0036] In this invention, the soaking temperature is preferably 2~6℃, specifically 4℃, and the soaking time is preferably 24~48 hours; this invention allows the components to diffuse into the gel network through soaking.

[0037] In this invention, deionized water is preferably used to rinse the gel surface in order to remove unadsorbed components.

[0038] This invention also provides the application of the D-lactic acid vaginal microecological repair gel described in the above technical solution or the D-lactic acid vaginal microecological repair gel obtained by the above preparation method in the preparation of drugs for treating vaginitis or inhibiting HPV infection.

[0039] In this invention, the drug is formulated as a hydrogel, and the hydrophilic polymers form a three-dimensional network that is insoluble in water through chemical or physical cross-linking.

[0040] The D-lactic acid vaginal microecological repair gel of the present invention can regulate the pH value and bacterial composition of the female vaginal microecological system. By lowering the local pH value through D-lactic acid, it selectively inhibits the reproduction of conditionally pathogenic bacteria and promotes the dominant growth of lactic acid bacteria, thereby realizing an environment-feedback type microecological repair mechanism.

[0041] To further illustrate the present invention, the D-lactic acid vaginal microecological repair gel, its preparation method, and its application provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0042] Example 1 D-Lactic Acid Vaginal Microecological Repair Gel, with a total weight of 100g, includes the following ingredients: 3.0g D-lactic acid, 2.0g carbomer, 5.0g glycerin, 0.5g epithelial repair agent (sodium hyaluronate), 1.0g colonization promoter (fructooligosaccharide), and the remainder water.

[0043] The preparation method (pH-triggered gelation method) includes the following steps: 1. Prepare the gel matrix solution (carbomer swelling). Disperse the gel matrix (carbomer 2.0g) in 70g of water, slowly sprinkle it in while stirring, and let it stand to swell for 4~12 h until there are no obvious particles.

[0044] 2. Add active ingredients Add 5.0g of moisturizer (glycerin), 0.5g of epidermal repair agent, and 1.0g of colonization promoter to the above gel matrix solution in sequence, and stir to dissolve. Note that the temperature should be controlled ≤40℃ to avoid inactivation of the ingredients.

[0045] 3. Add D-lactic acid Dilute 3.0g of D-lactic acid with 10g of purified water, then slowly add it to the gel matrix solution and stir at low speed until homogeneous.

[0046] 4. Adjust the pH value to trigger gelation. Prepare a pH adjuster solution (0.1M NaOH solution or D-lactic acid stock solution). While stirring, slowly add the pH adjuster dropwise until the pH of the system reaches the gelation point (pH 4.0±0.1). Add purified water to bring the total volume to the target 100g. Continue stirring until the system is homogeneous and free of bubbles. Stop stirring and let stand. The system will gradually form a gel.

[0047] 5. Homogenization and Filling Low-speed homogenization makes the gel fine, and it is then filled after standing to remove bubbles.

[0048] Comparative Example 1 The only difference from Example 1 is that the D-lactic acid in Example 1 is replaced with an equal mass of L-lactic acid, while the composition, ratio and preparation process of the other raw materials remain the same.

[0049] Comparative Example 2 The only difference from Example 1 is that D-lactic acid was not added, while the composition, ratio and preparation process of the other raw materials remained the same, and a blank gel was prepared.

[0050] Comparative Example 3 The only difference from Example 1 is that a gel sustained-release system is not constructed. Instead, the D-lactic acid from Example 1 is directly dissolved in purified water to prepare a D-lactic acid solution, and the D-lactic acid concentration is kept consistent with that of Example 1.

[0051] Test Example 1 1. Detection of viral load in a persistent HPV infection model Establishment of an animal model of persistent HPV infection: Because HPV has strict racial and tissue tropism, it cannot complete a full infection cycle in the vagina or cervix of mice under natural conditions. Therefore, the most commonly used HPV16 / 18 pseudovirus carrying luciferase was used for vaginal inoculation to observe viral entry and short-term infection. 0.1 ml of gel from Example 1, Comparative Example 1, and Comparative Example 2 were administered vaginally every other day for 4 weeks. The HPV load in cervical tissue was then measured, and the results are shown in Table 1.

[0052] Table 1 HPV load test results

[0053] The results showed that the D-lactic acid vaginal microecological repair gel of Example 1 could significantly reduce HPV load, and its clearance effect was significantly better than that of the comparative group.

[0054] 2. Vaginal mucosal tissue repair experiment Vaginal tissue samples were collected after drug administration and subjected to HE staining for observation. The results are as follows: Figure 2 (Example 1) and Figure 3 As shown in (Comparative Example 1).

[0055] The results showed that the epithelial structure in the model group was incomplete, with local inflammatory cell infiltration and tissue damage. In the control group (Group 1), the inflammatory response was significant, with partial epithelial shedding. In the example group (Group 1), the vaginal epithelial structure was intact, cells were regularly arranged, inflammatory cells were significantly reduced, and the tissue morphology was close to normal.

[0056] Test Example 2 1. D-lactic acid in vitro release experiment 1.0 g of each of the samples from Example 1 and Comparative Example 3 were placed in dialysis bags. 50 mL of simulated vaginal fluid (pH 4.0) was used as the release medium. An in vitro release experiment was conducted under constant temperature and shaking conditions at 37 °C. The concentration of D-lactic acid was measured at different time points, and the cumulative release rate was calculated. The results are shown in Table 2.

[0057] Table 2. Cumulative D-lactic acid release rate (%)

[0058] The results showed that the D-lactic acid in Example 1 could be released stably for more than 72 hours, while the D-lactic acid in Comparative Example 3 was basically released completely within 24 hours, indicating that the D-lactic acid vaginal microecological repair gel of the present invention has good sustained-release performance.

[0059] 2. Vaginal acidity maintenance experiment The gel samples of Example 1, Comparative Example 1 and Comparative Example 2 were added to simulated vaginal fluid (pH 4.0) and cultured at 37°C for 72 hours. The pH changes of the system were measured at different time points, and the results are shown in Table 3.

[0060] Table 3 Results of pH value changes

[0061] The results showed that the D-lactic acid vaginal microecological repair gel of Example 1 could maintain the vaginal environment in the normal weakly acidic range for a long time, while the ability of Comparative Examples 2 and 3 to maintain pH stability was significantly reduced.

[0062] Test Example 3: Lactobacillus Proliferation Experiment The gel samples from Example 1, Comparative Example 1, and Comparative Example 2 were added to the Lactobacillus culture system and anaerobically cultured at 37°C for 24 hours. The OD600 value and colony forming units (CFU) were then measured. The results are shown in Table 4.

[0063] Table 4 Results of Lactobacillus proliferation

[0064] The results showed that the D-lactic acid vaginal microecological repair gel of Example 1 could significantly promote the proliferation of lactobacilli, and its colony count was significantly higher than that of the control group.

[0065] In summary, the pH-responsive D-lactic acid sustained-release vaginal microecological repair gel provided by this invention has excellent sustained-release performance, which can continuously release D-lactic acid and maintain the weakly acidic environment of the vagina, promote the proliferation of lactobacilli, inhibit HPV infection, reduce HPV load, and promote the repair and reconstruction of the vaginal mucosal barrier, thereby achieving the effects of promoting vaginal microecological repair and assisting HPV clearance.

[0066] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, not all embodiments. People can obtain other embodiments based on the present invention without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A D-lactic acid vaginal microecological repair gel, characterized in that, The ingredients include the following ingredients by weight percentage: D-lactic acid 0.1-3.0%, moisturizer 1-10%, gel matrix 2-4%, epithelial repair agent 0.01-1%, colonization promoter 0.1-5%, pH adjuster, and the balance water; The pH value of the D-lactic acid vaginal microecological repair gel is 3.8~4.

1.

2. The D-lactic acid vaginal microecological repair gel according to claim 1, characterized in that, The weight percentage of D-lactic acid in the raw material is 1.0~3.0%.

3. The D-lactic acid vaginal microecological repair gel according to claim 1, characterized in that, The humectant is glycerin; the weight percentage of the humectant in the raw materials is 3-8%.

4. The D-lactic acid vaginal microecological repair gel according to claim 1, characterized in that, The gel matrix comprises hydroxypropyl methylcellulose and / or carbomer; the weight percentage of hydroxypropyl methylcellulose in the raw material is 1-2%, and the weight percentage of carbomer in the raw material is 1-4%.

5. The D-lactic acid vaginal microecological repair gel according to claim 1, characterized in that, The epithelial repair agent is sodium hyaluronate; the weight percentage of the epithelial repair agent in the raw materials is 0.3~0.8%.

6. The D-lactic acid vaginal microecological repair gel according to claim 1, characterized in that, The planting promoter is fructooligosaccharide; the weight percentage of the planting promoter in the raw materials is 1-2%.

7. The D-lactic acid vaginal microecological repair gel according to claim 1, characterized in that, The pH adjuster is a sodium hydroxide solution with a concentration of 0.1 mol / L.

8. A method for preparing the D-lactic acid vaginal microecological repair gel according to any one of claims 1 to 7, comprising method one or method two, wherein method one comprises the following steps: The gel matrix and water are mixed to obtain a gel matrix solution; The gel matrix solution, D-lactic acid, moisturizer, epithelial repair agent and colonization promoter are mixed, and the pH of the resulting mixture is adjusted to 3.8~4.

1. The mixture is allowed to stand to form a gel, thus obtaining the D-lactic acid vaginal microecological repair gel. The second method includes the following steps: Prepare blank gels from the gel matrix; The blank gel was immersed in the mixed solution for loading, and then the gel surface was rinsed to obtain the D-lactic acid vaginal microecological repair gel; the mixed solution contains D-lactic acid, moisturizer, epithelial repair agent and colonization promoter.

9. The preparation method according to claim 8, characterized in that, In Method 1, the pH value of the resulting mixture is adjusted to 3.8-4.1 using a sodium hydroxide solution with a concentration of 0.1 mol / L.

10. The use of the D-lactic acid vaginal microecological repair gel according to any one of claims 1 to 7 or the D-lactic acid vaginal microecological repair gel obtained by the preparation method according to claim 8 or 9 in the preparation of drugs for treating vaginitis or inhibiting HPV infection.

Images