Composition with vaginal antibacterial, anti-inflammatory and repairing effects, gel containing composition and preparation method of gel

Abstract

The invention provides a composition with vaginal antibacterial, anti-inflammatory and repairing effects and an external preparation comprising the composition. The composition comprises carbomer, glycosaminoglycan quaternary ammonium salt and recombinant III-type collagen. The invention also provides a gel with vaginal antibacterial, anti-inflammatory and repairing effects and a preparation method thereof. Carbomer and glycosaminoglycan quaternary ammonium salt are adopted, have synergistic antibacterial and anti-inflammatory effects, are matched with the recombinant III-type collagen, can keep the vagina moisturized and lubricated and keep the vagina moist and compact, have a repairing effect on the vagina, can synergistically enhance the mucous membrane elasticity and repairing capacity and improve the wound healing capacity, and can be used for repairing the vagina. A strong repairing effect is achieved on a genital tract wound surface or an erosion surface; the invention further provides a gel containing the composition with the vaginal antibacterial, anti-inflammatory and repairing effects, and the quality of a gel product is ensured by adopting a double preservative system of sodium methyl p-hydroxybenzoate and sodium propyl p-hydroxybenzoate.

Description

Technical Field

[0001] The present invention belongs to the technical field of female care, and specifically relates to a composition with vaginal antibacterial, anti-inflammatory and repairing effects, an external preparation comprising the composition, and also relates to a gel with vaginal antibacterial, anti-inflammatory and repairing effects and a preparation method thereof. Background Art

[0002] Vaginitis is a common gynecological disease, which is often manifested by vulvar itching, local erosion, increased vaginal discharge, accompanied by symptoms such as frequent urination and dysuria. It is also one of the gynecological diseases with a higher incidence rate. If not treated in time, it can easily cause cervicitis. In severe cases, it can infect the pelvic cavity, upper reproductive tract, abdominal cavity, etc. It is a common and frequently occurring gynecological disease with a high recurrence rate. It is distributed in all age groups and affects the patient's physical and mental health and quality of life.

[0003] Currently, the main routes of administration for vaginitis treatment are oral and topical. Topical vaginal medications are clinically highly regarded due to their high local drug concentration, low dosage, safety, and effectiveness. Topical vaginal preparations for vaginitis treatment primarily include tablets, suppositories, creams, vaginal rings, and hydrogels. Hydrogels are hydrophilic, network-like polymers that swell in water, absorb, and retain large amounts of water. The polymers that form hydrogels primarily include cellulose derivatives, carbomers, polycarbophils, alginates, tragacanth gum, gelatin, and starch, with carbomer being the most widely used in clinical practice.

[0004] Carbomer gel has a smooth surface and good biocompatibility with the vaginal mucosa. However, its antibacterial and antimicrobial effects on vaginal bacteria are limited. It needs to be used in combination with other antibiotics and other drugs to achieve better vaginal antibacterial and antimicrobial effects.

[0005] Chinese patent document CN101049286A discloses a gel formulation containing ciclopirox olamine for vaginal use. Based on a 1000g finished product, the formulation contains 1-50g of ciclopirox olamine, 1-50g of carbomer, 10-500g of ethanol, 20-500g of propylene glycol, 0-50g of triethanolamine, 0.05-2.0g of edetate disodium, and 10-100g of polysorbate 80, with water added to the final volume. In this patent document, ciclopirox olamine, a broad-spectrum antifungal drug, is combined with carbomer and other agents to form a gel formulation. However, due to the use of antibiotics in the gel, long-term use can still lead to drug resistance and damage the beneficial vaginal flora.

[0006] Chinese patent document CN115518029A discloses a carbomer vaginal packing gel. Its raw materials include 50-100 parts carbomer, 20-30 parts plant extract, 5-8 parts polyvinyl alcohol, 7-14 parts glycerin, 3-6 parts benzyl alcohol, and 30-80 parts purified water. The plant extract is made from one or more of the following: Citrus aurantium immaturus, Radix Ophiopogonis, Polygonum cuspidatum, Gentiana scabra, Flos Lonicerae, Salvia miltiorrhiza, and Paeonia lactiflora. In this patent document, the carbomer vaginal packing gel prepared by combining the plant extract with carbomer and other preparations boasts good biocompatibility, long-lasting action, broad-spectrum antibacterial properties, and high safety. However, due to the complex herbal ingredients in the plant extract, its drug release rate and extent are inferior to those of antibiotics, requiring a generally larger dosage and a slower onset of action. Summary of the Invention

[0007] In view of the defects of the prior art, the object of the present invention is to provide a composition with vaginal antibacterial, anti-inflammatory and repairing effects, and an external preparation including the composition.

[0008] Another object of the present invention is to provide a gel comprising the above composition having vaginal antibacterial, anti-inflammatory and repairing effects and a preparation method thereof.

[0009] To achieve the above objectives, the solutions adopted by the present invention are as follows:

[0010] In a first aspect, the present invention provides a composition having vaginal antibacterial, anti-inflammatory and repairing effects, comprising carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen.

[0011] Preferably, the mass ratio of the carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen is 1:(0.1-10):(0.0025-1).

[0012] Preferably, the carbomer is selected from at least one of carbomer 934, carbomer 940, carbomer 941, carbomer 934P, carbomer 974P, carbomer 971P and carbomer 1342, preferably selected from at least one of carbomer 934, carbomer 940, carbomer 941 and carbomer 934P; the aminopolysaccharide quaternary ammonium salt is a compound having a chain structure with aminopolysaccharides extracted from natural marine organisms, plants or animals as structural units, and a quaternary ammonium salt group is introduced on the sugar molecule.

[0013] In a second aspect, the present invention further provides a use of the composition having vaginal antibacterial, anti-inflammatory and repairing effects as described above in the preparation of a product for preventing or treating vaginal antibacterial, anti-inflammatory and repairing diseases.

[0014] In a third aspect, the present invention further provides an external preparation having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the composition having vaginal antibacterial, anti-inflammatory and repairing effects as described above.

[0015] Preferably, the external preparation is administered topically into the vagina.

[0016] Preferably, the dosage form of the external preparation includes any one of lotion, suppository, effervescent tablet, ointment, gel and foam.

[0017] Preferably, the external preparation further comprises at least one of an excipient, an antibacterial agent, a preservative, a pH regulator, a moisturizer, an anti-allergic agent, a chelating agent and a flavor.

[0018] In a fourth aspect, the present invention further provides a gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following components in percentage by mass:

[0019] Carbomer 0.5%-2.0%;

[0020] Glycerol 3%-6%;

[0021] Triethanolamine 0.01%-2.0%;

[0022]

[0023] In a fifth aspect, the present invention further provides a method for preparing the above-mentioned gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following steps:

[0024] Step (1), dissolving glycerol, disodium edetate, sodium methyl parahydroxybenzoate, and sodium propyl parahydroxybenzoate in purified water according to a ratio, and stirring to form a uniform solution;

[0025] Step (2), adding carbomer to the solution according to the ratio, and heating at 70-90° C. to promote dissolution;

[0026] Step (3), after cooling to room temperature, add aminopolysaccharide quaternary ammonium salt and recombinant type III collagen according to the ratio and stir evenly;

[0027] Step (4), adding triethanolamine according to the ratio to neutralize, and adjusting the pH value to 4.0-5.0 to obtain.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention provides a composition with vaginal antibacterial, anti-inflammatory and repairing effects, and an external preparation comprising the composition. The composition adopts carbomer and aminopolysaccharide quaternary ammonium salt to have a synergistic antibacterial and anti-inflammatory effect on vaginitis, and is combined with recombinant type III collagen to maintain vaginal moisturizing and lubrication, keep the vagina hydrated and firm, and simultaneously has a repairing effect on the vagina, can synergistically enhance the elasticity and repairing power of the mucosa, improve the healing ability of wounds, and have a strong repairing effect on reproductive tract wounds or erosions. The present invention further provides a gel comprising the composition with vaginal antibacterial, anti-inflammatory and repairing effects, and adopts a dual preservative system of sodium methylparaben and sodium propylparaben to ensure the quality of the gel product. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 These are experimental pictures of the negative control (NC) of the chicken embryo chorioallantoic membrane test in the examples of the present invention (the left is before sample addition, and the right is after sample addition).

[0031] Figure 2 These are experimental pictures of the positive control (PC) of the chicken embryo chorioallantoic membrane test in the examples of the present invention (the left is before sample addition, and the right is after sample addition).

[0032] Figure 3 The experimental picture of the gel sample (T) in the chicken embryo chorioallantoic membrane test in the embodiment of the present invention (the left is before sample addition, and the right is after sample addition);

[0033] Figure 4 This is a test result diagram of an embodiment of the present invention. DETAILED DESCRIPTION

[0034] The invention provides a composition with vaginal antibacterial, anti-inflammatory and repairing effects, comprising carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen.

[0035] In a composition with vaginal antibacterial, anti-inflammatory and repairing effects provided by the present invention, carbomer (CBM), also known as carbopol or carboxyrinyl polymer, is used. It is a white, loose, acidic, highly hygroscopic powder with a slight odor. It is a type of high-molecular polymer cross-linked with acrylic acid and propylene sucrose. Depending on the degree of polymerization, it includes various models such as carbomer 934, carbomer 940, carbomer 941, carbomer 934P, carbomer 974P, carbomer 971P, and carbomer 1342, forming low-viscosity, medium-viscosity, and high-viscosity products. Among them, carbomer 934, carbomer 940, carbomer 941, and carbomer 934P are suitable for local vaginal gels. Carbomer-based gels are transparent semisolids with a uniform texture and water-soluble properties, making them easy to spread. When applied to mucous membranes such as the skin and vagina, they form a transparent film with strong adhesion, a non-greasy feel, and excellent coupling with the skin and mucous membranes. They are non-irritating to the skin and mucous membranes, can mix with aqueous solutions, and absorb tissue exudates, facilitating the elimination of secretions. They also offer rapid drug release and rapid action, facilitating the localized efficacy of drugs in lesions. When applied to the skin, carbomer forms a barrier that prevents bacteria from reaching the skin, thereby reducing the risk of infection. Furthermore, the anionic nature of carbomer may interact with bacterial cell membranes, altering membrane permeability and, in turn, affecting bacterial metabolism and viability, thus possessing a certain antibacterial effect.

[0036] In a composition with vaginal antibacterial and anti-inflammatory repairing effect provided by the present invention, the aminopolysaccharide quaternary ammonium salt (Quaternary chitosan, QTS) adopted is a class of compounds with aminopolysaccharide as main structural unit, and quaternary ammonium salt groups are introduced on sugar molecules. Aminopolysaccharide is a polysaccharide naturally present in organisms, which can be extracted from sources such as marine organisms, plants and animals. Quaternary ammonium salt groups are quaternary ammonium ions, which are ions formed by four organic groups and a positively charged nitrogen atom. Aminopolysaccharide quaternary ammonium salt has multiple biological activities, including antibacterial, anti-inflammatory, antioxidant, antitumor and other effects, wherein antibacterial effect is one of its most important biological activities. Studies have shown that aminopolysaccharide quaternary ammonium salt has the function of broad-spectrum antibacterial, can significantly inhibit the growth of various bacteria, including common pathogenic microorganisms such as Staphylococcus aureus and Escherichia coli. Moreover, aminopolysaccharide quaternary ammonium salt can basically achieve continuous and same slow-release concentration within 72 hours, ensuring lasting antibacterial. In addition, aminopolysaccharide quaternary ammonium salt also has certain anti-inflammatory effect, can alleviate tissue inflammatory reaction, and promote wound healing. In addition, aminopolysaccharide quaternary ammonium salts have antioxidant and repairing properties, scavenging free radicals to protect cells from oxidative damage, regulating fibroblast proliferation and keratinization, accelerating wound healing, and reducing scarring. They also adsorb iron from platelets, focusing coagulation factors and achieving a hemostatic effect, while also forming a protective film on the wound surface. Consequently, aminopolysaccharide quaternary ammonium salts possess excellent antibacterial, hemostatic, sustained-release, film-forming, and moisturizing properties, leading to their primary medical applications in areas such as antibacterial wound treatment, anticoagulation, sustained-release drug delivery, and skin induction.

[0037] In a composition with vaginal antibacterial, anti-inflammatory and repairing effects provided by the present invention, carbomer and aminopolysaccharide quaternary ammonium salt are combined to have a synergistic effect on the antibacterial and antibacterial effects (Note: This is reflected in Example 1 and Comparative Example 1 in Table 2). It was found through the examples of the present invention that the composite gel containing carbomer and aminopolysaccharide quaternary ammonium salt has stronger antibacterial and antibacterial abilities than the gel containing only carbomer, and the antibacterial abilities against Escherichia coli, Staphylococcus aureus and Candida albicans are increased by 44.9%, 51.9% and 49.3%, respectively. This synergistic antibacterial effect may be that the addition of carbomer makes the composite gel easily adsorbed on the bacterial cell membrane and adsorbed on it for a long time, thereby increasing the permeability of the cell membrane and facilitating the penetration of the antibacterial components in the composite gel into the cell, further enhancing the antibacterial effect. Moreover, carbomer and aminopolysaccharide quaternary ammonium salt undergo a cross-linking reaction, causing the molecular chain state of the aminopolysaccharide quaternary ammonium salt to diffuse and stretch into a greatly expanded state. The thickened gel improves the shortcomings of the aminopolysaccharide quaternary ammonium salt gel with poor performance. The synergistic effect of the two also improves the shortcomings of the aminopolysaccharide quaternary ammonium salt such as low mechanical strength and poor spreading performance, thereby preparing a gel with lower cost, more stable properties and easier application.

[0038] In a composition with vaginal antibacterial, anti-inflammatory and repairing effects provided by the present invention, the recombinant type III collagen (Recombinant human collagen III, Rhc-III) used is a type of protein that is identical to the corresponding part of the natural type III collagen amino acid sequence obtained by optimizing and modifying the recombinant expression using genetic engineering technology and taking the human type III collagen gene sequence as a template. It has high biological activity and biocompatibility and good water solubility. Studies have shown that recombinant type III collagen can promote full-thickness wound healing and angiogenesis, promote the orderly arrangement of collagen fibers in the new dermal tissue, and has the function of repairing skin trauma and improving the quality of wound healing. In the present invention, the addition of recombinant type III collagen can maintain the moisturizing and lubrication of the vagina, while enhancing the elasticity and repair power of the mucosa, keeping the vagina moist and firm, improving the healing ability of the wound, and having a repairing effect on the wound or erosion surface of the reproductive tract.

[0039] In the composition with vaginal antibacterial, anti-inflammatory and repairing effects provided by the present invention, the mass ratio of the carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen is 1:(0.1-10):(0.0025-1).

[0040] The present invention further provides the use of the above-mentioned composition having vaginal antibacterial, anti-inflammatory and repairing effects in the preparation of a product for preventing or treating vaginal antibacterial, anti-inflammatory and repairing effects.

[0041] The present invention also provides an external preparation with vaginal antibacterial, anti-inflammatory and repairing effects, comprising the composition with vaginal antibacterial, anti-inflammatory and repairing effects as described above. The external preparation is locally administered into the vagina, and its dosage form includes any one of a lotion, suppository, effervescent tablet, ointment, gel and foam.

[0042] In the present invention, the "composition" and "product for preventing or treating vaginal antibacterial and anti-inflammatory repair" are generally compositions comprising a composition having active ingredient components (referred to as active ingredient) and pharmaceutically acceptable excipients.

[0043] In the present invention, the active ingredients include carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen.

[0044] In the present invention, the "pharmaceutically acceptable excipients" include inert diluents, dispersants and / or granulating agents, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or lubricating oils.

[0045] The present invention provides a product for preventing or treating vaginal bacteriostasis, anti-inflammatory and restorative effects comprising the above-mentioned composition, wherein the female subject being treated for vaginal bacteriostasis exhibits symptoms of dysbiosis of the microbiome in the urogenital tract, such as bacterial vaginosis, candidiasis, human papillomavirus infection, urinary tract infection, sexually transmitted infection, etc. The product for preventing or treating vaginal bacteriostasis, comprising the above-mentioned composition, provided by the present invention, is formulated for vaginal application in the form of a cream, spray, douche, or suppository. This can be achieved by adding at least one of an excipient, a bacteriostatic agent, a preservative, a pH adjuster, a moisturizer, an anti-allergic agent, a chelating agent, and a fragrance to the composition to prepare a vaginal external preparation, such as a douche. During application, the douche is applied to the vaginal cavity via a tool such as an applicator.

[0046] The present invention further provides a gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following components in percentage by mass:

[0047]

[0048] The present invention also provides a method for preparing the above-mentioned gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following steps:

[0049] Step (1), dissolving glycerol, disodium edetate, sodium methyl parahydroxybenzoate, and sodium propyl parahydroxybenzoate in purified water, and stirring to form a uniform solution;

[0050] Step (2), adding carbomer to the solution and heating at 70-90° C. to promote dissolution;

[0051] Step (3), after cooling to room temperature, add aminopolysaccharide quaternary ammonium salt and recombinant type III collagen and stir evenly;

[0052] Step (4), adding triethanolamine for neutralization and adjusting the pH value to 4.0-5.0 to obtain the product.

[0053] In the above-mentioned gel with vaginal antibacterial, anti-inflammatory and repairing effects and its preparation method provided by the present invention, in addition to the composition with vaginal antibacterial, anti-inflammatory and repairing effects provided by the present invention, two preservatives, sodium methylparaben and sodium propylparaben, are also included to ensure the quality and long-term effectiveness of the gel product. Triethanolamine, as a neutralizer of the carbomer acidic polymer gel, can adjust the pH value of the gel to 4.0-5.0 to maintain the microecological balance of the private parts.

[0054] The present invention provides a composition with vaginal antibacterial, anti-inflammatory and repairing effects and a gel comprising the composition. The composition is a repair gel that can be used for the auxiliary treatment of itching, pain, increased leucorrhea and odor symptoms caused by female cervicitis and vaginitis. While providing long-lasting antibacterial and anti-inflammatory effects, it can regulate fibroblast proliferation and keratinization formation, accelerate wound healing, and improve gynecological leucorrhea abnormalities, vulvar harassment and other problems. The prepared gel has good antibacterial and anti-inflammatory effects, is non-irritating to the mucosal system, and has good safety.

[0055] The present invention is further illustrated below with reference to the examples, the purpose of which is to better understand the content of the present invention and to embody the essential features of the present invention. Therefore, the examples given should not be construed as limiting the scope of protection of the present invention. It is also particularly noted that the specific experimental methods and equipment involved in the examples are conventional methods or carried out under the conditions recommended by the manufacturer's instructions unless otherwise specified, and the reagents involved are commercially available unless otherwise specified.

[0056] Example 1:

[0057] This embodiment provides a gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following components in percentage by mass:

[0058]

[0059] This embodiment further provides a method for preparing the above-mentioned gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following steps:

[0060] Step (1), dissolving glycerol, disodium edetate, sodium methyl parahydroxybenzoate, and sodium propyl parahydroxybenzoate in purified water, and stirring to form a uniform solution;

[0061] Step (2), adding carbomer to the solution and heating at 70-90° C. to promote dissolution;

[0062] Step (3), after cooling to room temperature, add aminopolysaccharide quaternary ammonium salt and recombinant type III collagen and stir evenly;

[0063] Step (4), adding triethanolamine for neutralization and adjusting the pH value to 4.0-5.0 to obtain the product.

[0064] Example 2:

[0065] This embodiment provides a gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following components in percentage by mass:

[0066]

[0067] This embodiment further provides a method for preparing the above-mentioned gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following steps:

[0068] Step (1), dissolving glycerol, disodium edetate, sodium methyl parahydroxybenzoate, and sodium propyl parahydroxybenzoate in purified water, and stirring to form a uniform solution;

[0069] Step (2), adding carbomer to the solution and heating at 70-90° C. to promote dissolution;

[0070] Step (3), after cooling to room temperature, add aminopolysaccharide quaternary ammonium salt and recombinant type III collagen and stir evenly;

[0071] Step (4), adding triethanolamine for neutralization and adjusting the pH value to 4.0-5.0 to obtain the product.

[0072] Example 3:

[0073] This embodiment provides a gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following components in percentage by mass:

[0074]

[0075] This embodiment further provides a method for preparing the above-mentioned gel having vaginal antibacterial, anti-inflammatory and repairing effects, comprising the following steps:

[0076] Step (1), dissolving glycerol, disodium edetate, sodium methyl parahydroxybenzoate, and sodium propyl parahydroxybenzoate in purified water, and stirring to form a uniform solution;

[0077] Step (2), adding carbomer to the solution and heating at 70-90° C. to promote dissolution;

[0078] Step (3), after cooling to room temperature, add aminopolysaccharide quaternary ammonium salt and recombinant type III collagen and stir evenly;

[0079] Step (4), adding triethanolamine for neutralization and adjusting the pH value to 4.0-5.0 to obtain the product.

[0080] Comparative Example 1:

[0081] Based on the formula of the gel in Example 1, no aminopolysaccharide quaternary ammonium salt is added, and the rest is the same as in Example 1.

[0082] Efficacy trials:

[0083] 1. pH value determination:

[0084] Basis: "Technical Specifications for Disinfection" (2002 edition) 2.2.1.4 Determination of pH value.

[0085] Testing instrument: FE28 pH meter.

[0086] Standard buffer for calibration: potassium hydrogen phthalate pH (25°C) = 4.00; mixed phosphate pH (25°C) = 6.86; sodium tetraborate pH (25°C) = 9.18.

[0087] Test conditions: 25°C, relative humidity 50%. The test was repeated twice.

[0088] Procedure: Allow the PXS-270 ion meter to stabilize for 20 minutes. Rinse the electrode with deionized water and calibrate with a pH standard solution of 4.00. Then, rinse the electrode with deionized water and calibrate with a pH standard solution of 9.18. After calibration, rinse the electrode with deionized water and test the standard solution for pH 6.86. Calibration is complete if the pH reading is within ±0.02. Measure the sample pH (25°C). Repeat twice for each batch of samples and take the average.

[0089] The average pH (25°C) values ​​of the highest concentration solutions of the sample stock solutions are shown in Table 1.

[0090] Table 1 pH measurement results of gel samples of the present invention

[0091]

[0092] 2. Antibacterial performance test:

[0093] Basis: "Hygiene Standard for Disposable Sanitary Products" GB15979-2002 (Appendix C) C3.

[0094] Test bacteria: Escherichia coli (8099) fourth generation (provided by Guangdong Microbial Culture Collection Center), Staphylococcus aureus (ATCC6538) fourth generation (provided by Guangdong Huankai Microbial Technology Co., Ltd.), Candida albicans (ATCC10231) fourth generation (provided by Guangdong Huankai Microbial Technology Co., Ltd.).

[0095] Testing conditions: ambient temperature is 18℃~26℃, relative humidity is 45%~65% PH.

[0096] Instruments and equipment: SPX-490 biochemical incubator.

[0097] Antibacterial experiment: 0.1 mL of the test bacterial suspension was dropped into 5.0 mL of the control sample solution (PBS), and the number of recovered bacteria was 1x10 4 --9x10 4cfu / mL. Dilute the test sample with sterile standard hard water. Pipette 5.0 mL of the original test sample solution or its dilution into a sterile test tube and incubate at 20°C for 5 minutes. Pipette 0.1 mL of the test bacterial solution into the test tube containing 5.0 mL of the sample, mix rapidly, and immediately start the timer. After the set time has elapsed, take 0.5 mL of the test bacteria and sample mixture and add it to a test tube containing 4.5 mL of sterile PBS. Mix thoroughly. After incubation for 10 minutes, pipette 1 mL of the sample solution into a sterile plate. Inoculate two sterile plates for each sample solution or dilution. Pour 15 mL of nutrient agar (bacteria) or Sabouraud agar (Candida albicans) cooled to 40°C-45°C, rotate the plate to allow for even distribution, and flip it over after the agar solidifies. Incubate at (35±2)°C for 48 hours (bacteria) or 72 hours (Candida albicans), and then count the viable colonies. Substitute PBS for the test sample and follow the above steps as a control sample. Repeat the experiment three times and calculate the average value.

[0098] The test results are shown in Table 2.

[0099] Table 2 Antibacterial performance test results of the gel samples of the present invention

[0100]

[0101] 3. Microbiological testing:

[0102] Basis: Appendix B of "Hygiene Standard for Disposable Sanitary Products" GB 15979-2002.

[0103] Testing conditions: ambient temperature 18℃~26℃, relative humidity 45%~65%RH.

[0104] Table 3 Microbiological index test results of the gel sample of Example 1 of the present invention

[0105] Detection indicators Technical requirements Test results Total bacterial colony count (CFU / mL) ≤200 <20 Total fungal colony count (CFU / mL) ≤100 <20 coliform group Not to be detected Not detected Staphylococcus aureus Not to be detected Not detected Pseudomonas aeruginosa Not to be detected Not detected Hemolytic Streptococci Not to be detected Not detected Fungal qualitative analysis Not to be detected Not detected

[0106] 4. Vaginal mucosal irritation test:

[0107] Basis: "Procedures and methods for toxicological evaluation of safety of disinfectants" (GB / T 38496-2020).

[0108] Environment: Temperature is 18℃~26℃, relative humidity is 40%~70%.

[0109] Animals and feeding: 24 New Zealand rabbits, female virgin rabbits, weighing 2.18 kg to 2.31 kg, were purchased from Huadong Xinhua Experimental Animal Farm, Huadu District, Guangzhou, with an experimental animal production license number of SCXK (Guangdong) 2019-0023 and an animal qualification certificate number of 44007600010181. They were randomly divided into a control group, Examples 1-3, and Comparative Examples 1-4, with 3 rabbits in each group. Feed was purchased from Beijing Huafukang Biotechnology Co., Ltd. with a qualification certificate number of 1103222200092765 and an experimental animal use license number of SYXK (Guangdong) 2022-0198.

[0110] Detection method: The test group animals were fixed on their backs, with the vagina and vaginal opening exposed. The catheter was moistened with the test solution and gently inserted into the vagina (4cm to 5cm). 2ml of the test solution was slowly injected with a syringe, and the catheter was withdrawn to complete the exposure. The exposure was repeated every 24 hours for 5 consecutive days. The control group animals were treated with normal saline in the same way. 24 hours after the last exposure, the animals were killed by gas embolism, and the intact vagina was removed by laparotomy. It was cut open longitudinally and observed with the naked eye for signs of congestion, edema, etc. for reference when taking pathological samples. The vagina was then placed in a 10% formalin solution and fixed for more than 24 hours. Tissues from the two ends and three central parts of the vagina were selected for preparation. After HE staining, histopathological examination was performed.

[0111] Histopathological examination results were scored according to the vaginal mucosal irritation response scoring standard. The irritation response scores for the three sites of the three animals in the experimental group (control group) and the control group were summed and divided by the total number of animals observed (number of animals × 3) to obtain the average score of the vaginal mucosal irritation response for the experimental group (control group). The irritation index was calculated by subtracting the average score of the control group from the average score of the experimental group. The irritation intensity was then graded according to the vaginal mucosal irritation intensity grading table. The results are shown in Table 4.

[0112] Table 4 Results of New Zealand rabbit vaginal mucosal irritation test on the gel sample of Example 1 of the present invention

[0113]

[0114] 5. Safety testing:

[0115] According to the chicken embryo chorioallantoic membrane test (SN / T2329-2009) for cosmetic eye irritation / corrosion, the eye irritation and corrosiveness of the samples were tested. The experimental process is as follows:

[0116] 5.1 Preparation:

[0117] Preparation of the chorioallantoic membrane (CAM): Purchase chick embryos of appropriate age. At 9 days of incubation, inspect and discard any defective embryos. Mark the location of the air cells on the surface of the eggshell of a healthy chick. Peel back the shell to expose the white membrane. Carefully remove the inner membrane with forceps, ensuring that the vascular membrane is not damaged.

[0118] Negative control (NC): 0.9% NaCl.

[0119] Positive control (PC): 0.1 mol / L NaOH.

[0120] 5.2 Test:

[0121] Stimulation scoring method: 0.3 mL of the sample was directly dripped onto the CAM surface in each group of 6 chicken embryos. The CAM reaction was observed and the time of bleeding, coagulation and vascular melting within 5 minutes was recorded, as well as the degree of reaction.

[0122] Positive control, negative control and samples were all operated using the stimulation scoring method.

[0123] 5.3 Data Analysis

[0124] Stimulation score (IS): Record the time and degree of each endpoint and calculate the stimulation score (IS) using the following formula, with the result rounded to two decimal places:

[0125] IS=(301-H)+7(301-L)+9(301-C) / 300

[0126] sec H, sec L, and sec C represent the average time (in seconds) to the onset of bleeding, vascular lysis, and coagulation, respectively, observed on the CAM membrane.

[0127] The evaluation criteria for the stimulation scoring method are shown in Table 5.

[0128] Table 5 Evaluation results of stimulation scoring method

[0129] Stimulus score Irritation classification IS<1 Non-irritating 1≤IS<5 Mild irritation 1≤IS<5 Moderate irritation IS≥10 Strong irritant / corrosive

[0130] 5.4 Experimental Results

[0131] 5.4.1. The scoring results of the control group are shown in Table 6. Figure 1 and Figure 2 .

[0132] Table 6

[0133]

[0134] 5.4.2. The scoring results of the gel samples of Example 1 of the present invention are shown in Tables 7 and Figure 3 .

[0135] Table 7

[0136] Group Stimulus score Stimulus classification sample 0 Non-irritating

[0137] According to the results of SNIT 2329-2009, under the conditions of this experiment, the IS score of the sample is 0, which is non-irritating.

[0138] 6. Anti-inflammatory test

[0139] The anti-inflammatory efficacy of the samples was tested by laboratory methods using the ability to inhibit lipoxygenase activity as the evaluation index.

[0140] 6.1 Treatment of test samples

[0141] Test sample: Dilute with anhydrous ethanol to the reference concentration (10 mg / mL, 1%).

[0142] 6.2 Test operation process

[0143] Prepare 1.5 mL centrifuge tubes and set up a sample group (A), a positive control group (B), a negative control group (C), and a blank control group (D). Set up three parallel tubes for each sample group (A) and positive control group (B). For the zileuton control group, follow the same protocol as for sample group (A) and pair it with the corresponding group C.

[0144] Table 2 Experimental operation flow chart (unit / μL)

[0145]

[0146] 6.3 Calculation formula

[0147]

[0148] D A : The absorbance value of the sample tube, that is, the absorbance value of the solution after the sample reacts with lipoxygenase;

[0149] D B : The average of three absorbance values ​​in the enzyme reaction tube, i.e., the absorbance value of the reaction between lipoxygenase and substrate linoleic acid when no sample is added;

[0150] D C : Sample blank absorbance

[0151] D D : Solvent blank absorbance value.

[0152] 6.4 Experimental Results

[0153] The experimental results are detailed in Table 3 and Figure 4The test results showed that the positive control (zileuton) had a lipoxygenase inhibition rate of >50%, indicating that the reaction system was effective. The lipoxygenase inhibition rate of carbomer gynecological gel was 72.305±0.357%, indicating that it has certain anti-inflammatory effects.

[0154] Table 3 Test results

[0155]

[0156] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A composition with vaginal antibacterial, anti-inflammatory and repairing effects, characterized in that: Includes carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen.

2. The composition having vaginal antibacterial, anti-inflammatory and repairing effects according to claim 1, characterized in that: The mass ratio of the carbomer, aminopolysaccharide quaternary ammonium salt and recombinant type III collagen is 1: (0.1-10): (0.0025-1).

3. The composition having vaginal antibacterial, anti-inflammatory and repairing effects according to claim 1, characterized in that: The carbomer is selected from at least one of carbomer 934, carbomer 940, carbomer 941, carbomer 934P, carbomer 974P, carbomer 971P and carbomer 1342, preferably at least one of carbomer 934, carbomer 940, carbomer 941 and carbomer 934P; the aminopolysaccharide quaternary ammonium salt is a compound having a chain structure with aminopolysaccharides extracted from natural marine organisms, plants or animals as structural units, and a quaternary ammonium salt group is introduced on the sugar molecule.

4. Use of the composition having vaginal antibacterial, anti-inflammatory and repairing effects as claimed in any one of claims 1 to 3 in the preparation of a product for preventing or treating vaginal antibacterial, anti-inflammatory and repairing effects.

5. An external-use preparation with vaginal antibacterial, anti-inflammatory and repairing effects, characterized in that: The invention comprises a composition having vaginal antibacterial, anti-inflammatory and repairing effects as described in any one of claims 1 to 3.

6. The external preparation having vaginal antibacterial, anti-inflammatory and repairing effects according to claim 5, characterized in that: The external preparation is administered topically into the vagina.

7. The external preparation having vaginal antibacterial, anti-inflammatory and repairing effects according to claim 5, characterized in that: The dosage form of the external preparation includes any one of lotion, suppository, effervescent tablet, ointment, gel and foam.

8. The external preparation having vaginal antibacterial, anti-inflammatory and repairing effects according to claim 5, characterized in that: The external preparation further comprises at least one of an excipient, an antibacterial agent, a preservative, a pH regulator, a moisturizer, an anti-allergic agent, a chelating agent and a fragrance.

9. A gel with vaginal antibacterial, anti-inflammatory and repairing effects, characterized in that: The following components are included in mass percentage:

10. A method for preparing the gel having vaginal antibacterial, anti-inflammatory and repairing effects as claimed in claim 9, characterized in that: The steps include: Step (1), dissolving glycerol, disodium edetate, sodium methyl parahydroxybenzoate, and sodium propyl parahydroxybenzoate in purified water according to a ratio, and stirring to form a uniform solution; Step (2), adding carbomer to the solution according to the ratio, and heating at 70-90° C. to promote dissolution; Step (3), after cooling to room temperature, add aminopolysaccharide quaternary ammonium salt and recombinant type III collagen according to the ratio and stir evenly; Step (4), adding triethanolamine according to the ratio to neutralize, and adjusting the pH value to 4.0-5.0 to obtain.

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