Willidig antibacterial gel and preparation method thereof

Abstract

The invention relates to the technical field of gynecological antibacterial gel, in particular to ulidig antibacterial gel and a preparation method thereof. The invention relates to a hand sanitizer which is prepared from the following components in parts by weight: 1 to 3 parts of chlorhexidine acetate, 4 to 8 parts of parnassia palustris extract, 0.8 to 1.2 parts of carbomer, 0.4 to 0.6 part of triethanolamine, 0.5 to 1.5 parts of penetration enhancer, 0.04 to 0.07 part of modified nano-silver, 5 to 8 parts of glycerol, 0.03 to 0.07 part of hydrogenated polyisobutene, 0.1 to 0.3 part of edetate disodium and the balance of purified water. The chlorhexidine acetate and the ethanol extract of the parnassia palustris are compounded according to the proportion of 1: (2.5-4), flavone and polyphenol substances in the parnassia palustris can destroy the permeability of microbial cell membranes, the chlorhexidine acetate is promoted to enter thalli, and the minimum inhibitory concentration is remarkably reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of gynecological antibacterial gels, and in particular to a Ulidige antibacterial gel and a preparation method thereof. Background Art

[0002] Gynecological inflammation is a common gynecological condition, primarily classified as bacterial vaginosis, candidal vaginitis, nonspecific vaginitis, and trichomonas vaginitis. According to incomplete statistics from the World Health Organization, the incidence of various gynecological diseases among women exceeds 65%, with a trend toward younger women, making them a major health threat to Chinese women. Treatments for gynecological inflammation include physical therapy and medication. Physical therapy can damage epithelial cells, while medications are primarily topical. Because the vagina is rich in capillaries and lymphatic vessels and lacks distinct nerve endings, medication administration is minimally painful, making it an ideal site for drug delivery for specific diseases and medications. Therefore, various dosage forms, including suppositories, tablets, and gels, are available for clinical use, tailored to the specific therapeutic objective. Gels are favored by patients due to their excellent user compliance.

[0003] Traditional medical antibacterial gels, such as those with publication number CN117797195A, mostly rely on a single chemical bactericide, such as chlorhexidine acetate or zinc salts. Long-term use can easily induce microbial resistance. For example, carbomer gynecological gel with publication number CN118576537A directly mixes plant extracts with chemical drugs, resulting in a carbomer gel network encapsulating the active ingredients, with a release rate of ≤60%. At the same time, tannins in the ethanol extract complex with metal ions to produce precipitation, resulting in a significant decrease in light transmittance. In view of this, we propose a Ulidige antibacterial gel and a preparation method thereof. Summary of the Invention

[0004] The purpose of the present invention is to solve the deficiencies mentioned in the above background technology and provide a Ulidige antibacterial gel and a preparation method thereof.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions: A Ulidige antibacterial gel comprises the following components in parts by weight: 1-3 parts of chlorhexidine acetate, 4-8 parts of euphorbia milii extract, 0.8-1.2 parts of carbomer, 0.4-0.6 parts of triethanolamine, 0.5-1.5 parts of a penetration enhancer, 0.04-0.07 parts of modified nanosilver, 5-8 parts of glycerol, 0.03-0.07 parts of hydrogenated polyisobutylene, 0.1-0.3 parts of disodium edetate, and the balance of purified water; Wherein, the mass ratio of the chlorhexidine acetate to the herbaceous elm extract is 1:(2.5-4).

[0006] Preferably, the modified nanosilver is polydopamine-coated nanosilver particles with a particle size of 10-20 nm, which are prepared by in situ reduction of silver nitrate with dopamine under alkaline conditions.

[0007] Preferably, the Herba Eupatorii extract is an ethanol gradient extract, and the extraction process includes: Step 1: Extract with 56-67% ethanol at 50°C for 1.5 hours; Step 2: Extraction at 25°C for 2 hours; Step 3: Extract at 4°C for 12 hours.

[0008] Preferably, the viscosity of the Ulidiger antibacterial gel is 11500-12500 mPa·s.

[0009] Preferably, the penetration enhancer is borneol.

[0010] A method for preparing Ulidige antibacterial gel comprises the following steps: Step 1: Disperse carbomer in 60% purified water and allow to swell for 12 hours to obtain a pre-gel matrix; Step 2: extracting the Herba Lycopodii with 56-67% ethanol in a gradient manner, filtering to obtain a Herba Lycopodii extract, adding chlorhexidine acetate thereto and stirring to activate the extract, thereby obtaining a mixed solution; Step 3: Slowly add the mixture obtained in step 2 to the pre-gel matrix in step 1 and stir at 400 rpm for 15 minutes; Step 4: Then add borneol, ultrasonically disperse for 10 minutes, and then add triethanolamine dropwise to adjust the pH to 5.8-6.2; Step 5: Finally, add glycerol, hydrogenated polyisobutylene, disodium edetate and the remaining purified water, and vacuum degassing at -0.08 MPa for 10 minutes to obtain the Ulidige antibacterial gel.

[0011] Preferably, the activation condition of step 2 is: stirring at 200 rpm and 35±2° C. for 30 minutes.

[0012] Preferably, the ultrasonic dispersion power in step 4 is 300-400W.

[0013] The invention discloses an application of Ulidige antibacterial gel in the preparation of an external antibacterial preparation for gynecological use, and is used to treat vaginal infections caused by Candida albicans or Staphylococcus aureus.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention uses chlorhexidine acetate and ethanol extract of Herba Eupatorii in a ratio of 1: (2.5-4). The flavonoids and polyphenols in Herba Eupatorii can destroy the permeability of microbial cell membranes, promote the entry of chlorhexidine acetate into the bacteria, and significantly reduce the minimum inhibitory concentration. Therefore, the antibacterial rate of this combination against Staphylococcus aureus is increased by more than 40% compared with the single component, and can delay the development of bacterial resistance. Compared with the existing technology of using only chemical antibacterial agents or common Chinese herbal medicines, this solution achieves broad-spectrum antibacterial effect through a chemical-plant composite antibacterial pathway, especially with specific inhibitory effects on common gynecological pathogens (Candida albicans and Escherichia coli); 2. The present invention extracts the Herba Lycopodii raw material in 56-67% ethanol and adopts a three-stage gradient temperature control (50°C initial extraction → 25°C intermediate extraction → 4°C fine extraction) to maximize the extraction of small molecular polar components with antibacterial activity (such as saponins and flavonoids) and avoid degradation of heat-sensitive components. Compared with the traditional water decoction method, the yield of antibacterial active ingredients is increased by 32%. Chlorhexidine acetate is directly added to the filtered Herba Lycopodii extract, and the ethanol solvent environment is used to depolymerize its molecules into active monomers, avoiding the crystal form change caused by secondary dissolution in the traditional process.

[0015] 3. The present invention adds modified nanosilver to the gel, which is prepared by the dopamine-sodium hydroxide in situ reduction method. The polydopamine layer coated on the surface of the nanosilver can combine with the carboxyl groups in the carbomer network to form a sustained-release unit, thereby extending the release time of chlorhexidine acetate to more than 24 hours. DETAILED DESCRIPTION

[0016] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. In the absence of conflict, the embodiments in this application and the features in the embodiments can be combined with each other. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0017] The present invention describes the above technical solution in detail through the following embodiments: Example 1 A Ulidige antibacterial gel comprises the following components in parts by weight: 1.5 parts of chlorhexidine acetate, 3.75 parts of euphorbia milii extract, 1.0 part of carbomer, 0.5 part of triethanolamine, 1.0 part of borneol, 0.05 part of modified nanosilver, 7 parts of glycerol, 0.05 part of hydrogenated polyisobutylene, 0.2 part of disodium edetate, and the balance of purified water; Wherein, the mass ratio of the chlorhexidine acetate to the herbaceous elm extract is 1:(2.5-4).

[0018] Specifically, the preparation method of modified nanosilver is as follows: 10 mg of dopamine is dissolved in 50 mL of pH 8.5 Tris-HCl buffer, 1 mM silver nitrate solution is added, stirred in the dark at 25°C for 24 h, and centrifuged and washed (12000 rpm×10 min) to obtain polydopamine-coated nanosilver.

[0019] The modified nanosilver is polydopamine-coated nanosilver particles with a particle size of 15 nm, which are prepared by in-situ reduction of silver nitrate with dopamine under alkaline conditions.

[0020] Preferably, the Herba Eupatorii extract is an ethanol gradient extract, and the extraction process includes: Step 1: Extract with 67% ethanol at 50°C for 1.5 hours; Step 2: Extraction at 25°C for 2 hours; Step 3: Extract at 4°C for 12 hours.

[0021] Among them, the viscosity of Ulidige antibacterial gel is 12500 mPa·s.

[0022] A method for preparing Ulidige antibacterial gel comprises the following steps: Step 1: Disperse carbomer in 60% purified water and allow to swell for 12 hours to obtain a pre-gel matrix; Step 2: extracting the Herba Lycopodii with 67% ethanol in a gradient manner, filtering to obtain a Herba Lycopodii extract, adding chlorhexidine acetate to the Herba Lycopodii extract and stirring to activate the extract. The activation conditions are: stirring at 200 rpm and 35±2°C for 30 minutes to obtain a mixed solution. Step 3: Slowly add the mixture obtained in step 2 to the pre-gel matrix in step 1 and stir at 400 rpm for 15 minutes; Step 4: Then add borneol, ultrasonically disperse for 10 minutes, and then add triethanolamine dropwise to adjust the pH to 5.8. The ultrasonic dispersion power is 300W; Step 5: Finally, add glycerol, hydrogenated polyisobutylene, disodium edetate and the remaining purified water, and vacuum degassing at -0.08 MPa for 10 minutes to obtain the Ulidige antibacterial gel.

[0023] It should be noted that carbomer is pre-swelled in 60% of the total water volume for 12 hours to form a uniform hydration layer. Different from the conventional stirring swelling method, to ensure that the gel is particle-free, chlorhexidine acetate and herb extract are first mixed, and then the carbomer matrix is slowly added to avoid the dense network formed when carbomer and triethanolamine are neutralized to wrap the drug and cause delayed release; after adding triethanolamine, degassing is carried out at -0.08 MPa for 10 minutes to eliminate the influence of bubbles on the viscosity of the gel.

[0024] Example 2 The only difference between this embodiment and embodiment 1 is that: in this embodiment, 1.5 parts of chlorhexidine acetate and 4.5 parts of Herba Eupatorii Herba Extract are used, and other conditions are the same.

[0025] Example 3 The only difference between this embodiment and embodiment 1 is that: in this embodiment, 1.5 parts of chlorhexidine acetate and 6 parts of Herba Eupatorii extract are used, and other conditions are the same.

[0026] Example 4 The only difference between this embodiment and embodiment 1 is that: in this embodiment, 1 part of chlorhexidine acetate and 4 parts of Herba Eupatorii extract are used, and other conditions are the same.

[0027] Comparative Example 1 The only difference between this comparative example and Example 1 is that modified nanosilver is not added in this comparative example, and other conditions are the same.

[0028] Comparative Example 2 The only difference between this comparative example and Example 1 is that borneol is not added in this comparative example, and other conditions are the same.

[0029] Comparative Example 3 The only difference between this comparative example and Example 1 is that in this comparative example, the Herba Eupatorii extract was subjected to a single extraction at 50° C. instead of a gradient extraction, and other conditions were the same.

[0030] Comparative Example 4 The only difference between this comparative example and Example 1 is that in this comparative example, the carbomer is omitted from being pre-swelled in 60% of the total water for 12 hours, and other conditions are the same.

[0031] Test items: 1. Minimum Inhibitory Concentration (MIC) Test Standard: GB / T 20944.3-2008 Evaluation of antibacterial properties, oscillation method Procedure: 1. Bacterial activation: Candida albicans (ATCC 10231) and Staphylococcus aureus (ATCC 6538) were inoculated into SDA / TSB medium and cultured at 37°C for 24 h.

[0032] Two-fold dilution method: dilute the gel extract with MH broth to a gradient concentration of 0.39-50 μg / mL.

[0033] Add bacteria culture: add 1×10 6 CFU / mL bacterial solution, and culture at 37℃ with shaking for 24 h.

[0034] Judgment: The lowest concentration without turbidity observed with the naked eye is the MIC.

[0035] 2. Biofilm Removal Rate Test Standard: Modified CLSI M27-A3 protocol (quantification by laser confocal microscopy) Process: 1. Biofilm construction: Candida albicans was inoculated into a 96-well plate (RPMI 1640 containing 10% FBS) and cultured at 37°C for 48 h to form a mature biofilm.

[0036] Gel treatment: Add 0.2 mg / mL gel extraction solution to each well and incubate for 24 h.

[0037] Fluorescent staining: SYTO9 / PI double staining (live bacteria - green, dead bacteria - red).

[0038] CLSM analysis: Calculate the percentage of red pixels (mortality rate) = clearance rate.

[0039] 3. Cumulative release rate test Standard: Chinese Pharmacopoeia 2020 Edition 0931 Release Determination Method Procedure: 1. Dialysis bag method: Take 1.0 g of gel and place it into a dialysis bag with a molecular weight cut-off of 8-14 kDa.

[0040] Receiving medium: 500 mL pH 4.5 acetate buffer (simulating vaginal environment), 37°C ± 0.5°C, rotation speed 50 rpm.

[0041] Sampling points: 5 mL of receiving solution was collected at 2, 4, 6, 12, and 24 h (with simultaneous rehydration).

[0042] HPLC detection: C18 column (4.6×250 mm), mobile phase acetonitrile-0.1% phosphoric acid (35:65), flow rate 1.0 mL / min, detection wavelength 254 nm.

[0043] 4. Rabbit mucosal irritation test Standard: "Disinfection Technical Specifications 2002 Edition" Procedure: 1. Animal grouping: New Zealand white rabbits (female, 2.5-3.0 kg), 6 rabbits per group.

[0044] Dosage: 0.2 g of gel is injected vaginally daily for 7 consecutive days.

[0045] Scoring: 24 hours after the last administration, the mucosa was dissected and observed: 0 point: no redness or swelling, 1 point: mild congestion, 2 points: moderate congestion / edema, 3 points: severe congestion / ulceration, 4 points: necrosis.

[0046] The specific data are as follows Table 1: Table 1 *Note: Comparative Example 4 release stagnation after 6 hours (gel particles blocked the dialysis membrane) From the data in the above table, it can be seen that according to the data of Example 1 and Example 4, when the ratio of chlorhexidine acetate to herba sylvestris extract is 1:2.5, the MIC is the lowest (0.78 μg / mL) and the biofilm clearance rate is the highest, reaching 91.2%. When the ratio of chlorhexidine acetate to herba sylvestris extract is increased to 1:4, the MIC increases by 100% and the clearance rate decreases by 5.6%.

[0047] From the data of Comparative Examples 1 and 2, it can be seen that when modified nanosilver is not added, the release rate plummets by 31.1% (92.3%→61.2%), proving the key role of sustained release of nanosilver. The biofilm clearance rate decreased by 21% due to the lack of synergistic penetration of silver ions, and when borneol is not added, the biofilm clearance rate decreased by 25.4% due to the loss of the penetration-enhancing function.

[0048] The data from Comparative Examples 3 and 4 show that when a single extraction is used, the MIC soars to 3.13 μg / mL, and gradient temperature control ensures the activity of the thermosensitive component. When the carbomer is not pre-swelled in 60% of the total water volume for 12 hours, the release rate decreases by 23% due to clogging of the gel particles, and the transmittance decreases by 7.7%, resulting in turbidity due to precipitation.

[0049] In summary, Example 1 (chlorhexidine acetate: herba sylvestris = 1:2.5) reached peak values in antibacterial efficacy (MIC 0.78 μg / mL), biofilm clearance (91.2%), and release rate (92.3%).

[0050] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

Claims

1. A Ulidige antibacterial gel, characterized in that: The composition comprises the following components in parts by weight: 1-3 parts of chlorhexidine acetate, 4-8 parts of euphorbia pulegium extract, 0.8-1.2 parts of carbomer, 0.4-0.6 parts of triethanolamine, 0.5-1.5 parts of penetration enhancer, 0.04-0.07 parts of modified nanosilver, 5-8 parts of glycerol, 0.03-0.07 parts of hydrogenated polyisobutylene, 0.1-0.3 parts of disodium edetate and the balance of purified water; Wherein, the mass ratio of the chlorhexidine acetate to the herbaceous elm extract is 1:(2.5-4).

2. The Ulidige antibacterial gel according to claim 1, wherein: The modified nanosilver is polydopamine-coated nanosilver particles with a particle size of 10-20 nm, which are prepared by in-situ reduction of silver nitrate with dopamine under alkaline conditions.

3. The Ulidige antibacterial gel according to claim 1, wherein: The herb extract is an ethanol gradient extract, and the extraction process includes: Step 1: Extract with 56-67% ethanol at 50°C for 1.5 hours; Step 2: Extraction at 25°C for 2 hours; Step 3: Extract at 4°C for 12 hours.

4. The Ulidige antibacterial gel according to claim 1, wherein: The viscosity of the Ulidige antibacterial gel is 11500-12500 mPa·s.

5. The Ulidige antibacterial gel according to claim 1, characterized in that: The penetration enhancer is borneol.

6. A method for preparing Ulidiger antibacterial gel, applicable to the Ulidiger antibacterial gel according to any one of claims 1 to 5, characterized in that: The following steps are involved: Step 1: Disperse carbomer in 60% purified water and allow to swell for 12 hours to obtain a pre-gel matrix; Step 2: extracting the Herba Lycopodii with 56-67% ethanol in a gradient manner, filtering to obtain a Herba Lycopodii extract, adding chlorhexidine acetate thereto and stirring to activate the extract, thereby obtaining a mixed solution; Step 3: Slowly add the mixture obtained in step 2 to the pre-gel matrix in step 1 and stir at 400 rpm for 15 minutes; Step 4: Then add borneol, ultrasonically disperse for 10 minutes, and then add triethanolamine dropwise to adjust the pH to 5.8-6.2; Step 5: Finally, add glycerol, hydrogenated polyisobutylene, disodium edetate and the remaining purified water, and degas under -0.08 MPa vacuum for 10 minutes to obtain the Ulidige antibacterial gel.

7. The method for preparing the Ulidige antibacterial gel according to claim 6, wherein: The activation conditions of step 2 are: stirring at 200 rpm and 35±2°C for 30 minutes.

8. The method for preparing the Ulidige antibacterial gel according to claim 6, wherein: The ultrasonic dispersion power in step 4 is 300-400W.

9. Use of the Ulidige antibacterial gel according to claim 1 in the preparation of an antibacterial preparation for external use in gynecology, characterized in that: For the treatment of vaginal infections caused by Candida albicans or Staphylococcus aureus.

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