Efficient gynecological antibacterial liquid and preparation method thereof

Abstract

The invention provides efficient gynecological antibacterial liquid and a preparation method thereof, and the efficient gynecological antibacterial liquid comprises the following components in parts by weight: 3-5 parts of squalane nanoemulsion, 2-4 parts of traditional Chinese medicine extract, 1-3 parts of prebiotics-nisin mixed liquid, 1-3 parts of sodium alginate gel and 3-6 parts of collagen fullerene antibacterial liquid, the preparation method comprises the following steps: S1, preparing squalane nanoemulsion; s2, preparing a traditional Chinese medicine extract, a prebiotics-nisin mixed solution and a collagen fullerene antibacterial solution; s3, products in the step S1 and the step S2 are mixed with sodium alginate gel, after pH is adjusted, ultrasonic degassing and sterile filtration treatment are performed, broad-spectrum bacteriostasis and anti-inflammatory synergy are achieved, stability is high, irritation is avoided, and the composition is particularly suitable for female vagina micro-ecological balance restoration and long-acting protection.

Description

Technical Field

[0001] The invention relates to the field of medical technology, and in particular to a high-efficiency gynecological antibacterial liquid and a preparation method thereof. Background Art

[0002] As a female intimate care product, gynecological antibacterial liquid mainly protects female reproductive health by inhibiting or killing pathogenic microorganisms and maintaining the balance of vaginal microecology. Traditional antibacterial liquids contain antibiotic ingredients. Although they can quickly inhibit bacteria, long-term use can easily lead to the production of drug-resistant strains, destroy the balance of normal flora in the vagina, and increase the risk of infection. For example, some disinfectants containing chlorhexidine ingredients have inhibitory effects on Staphylococcus aureus, Escherichia coli, etc., but their broad-spectrum bactericidal properties may accidentally injure beneficial bacteria such as lactobacilli and weaken the self-cleaning ability of the vagina. The existing preparation methods mostly rely on high-temperature extraction or chemical synthesis, which can easily lead to the inactivation of effective ingredients. For example, Chinese medicine compound antibacterial liquids require long-term high-temperature decoction, heat-sensitive ingredients are easy to volatilize, and the quality fluctuations of raw materials from different batches affect the uniformity of the product; some antibacterial liquids add alcohol or volatile ingredients to enhance permeability, which may cause mucosal irritation, allergic reactions, and even neurotoxicity. In addition, many products on the market focus on "killing" rather than "regulation", ignoring the reconstruction of vaginal microecology. Traditional antibiotic antibacterial liquids excessively remove microorganisms, and although new biological antibacterial agents (such as ε-polylysine) can selectively inhibit pathogenic bacteria, they lack the mechanism to promote the proliferation of probiotics such as lactobacilli, making it difficult to achieve long-term ecological balance. Traditional antibacterial liquids (such as sodium benzoate and chlorhexidine acetate preparations) mostly target single pathogens (such as bacteria or fungi), and lack effective inhibition of mixed infections (such as bacterial vaginitis combined with candidal infection) or drug-resistant strains (such as methicillin-resistant Staphylococcus aureus), leading to repeated infections. The abuse of antibiotic products (such as metronidazole gel) accelerates the development of pathogen resistance. Some studies have shown that more than 30% of patients with gynecological inflammation develop resistance to traditional antibiotics. There is no gynecological antibacterial liquid on the market that is tailored to the physiological age characteristics of women. The older women are, the higher their demand for maintaining the balance of the flora. Due to age-related vaginal atrophy, secretions decrease. Existing antibacterial liquids (such as suppositories) aggravate dryness and pain due to insufficient lubrication, and lack synergistic protection against urinary tract infections. How to prepare a safe and non-irritating gynecological antibacterial liquid that can achieve high-efficiency and broad-spectrum antibacterial effect without destroying the vaginal microecology and is suitable for women who have high demands for maintaining the balance of the bacterial flora has become the core technical bottleneck.

[0003] In summary, the market is in urgent need of a highly effective gynecological antibacterial liquid and a preparation method thereof, which can not only reduce longitudinal cracks and reduce chloride ion content, but also avoid local collapse caused by concrete crispness in the core area. Summary of the invention

[0004] The present invention provides a high-efficiency gynecological antibacterial liquid and a preparation method thereof, which are used to solve the problems raised in the above background technology.

[0005] To solve the above technical problems, the present invention discloses an efficient gynecological bacteriostatic liquid, which comprises the following components: squalane nanoemulsion, traditional Chinese medicine extract, prebiotic-nisin mixed liquid, sodium alginate gel and collagen fullerene bacteriostatic liquid.

[0006] Furthermore, by weight, it comprises the following components: 3-5 parts of squalane nanoemulsion, 2-4 parts of traditional Chinese medicine extract, 1-3 parts of prebiotic-nisin mixed liquid, 1-3 parts of sodium alginate gel and 3-6 parts of collagen fullerene bacteriostatic liquid.

[0007] Furthermore, the preparation method of the efficient gynecological bacteriostatic liquid comprises the following steps:

[0008] S1. Prepare squalane nanoemulsion;

[0009] S2. Prepare traditional Chinese medicine extract, prebiotic-nisin mixed liquid and collagen fullerene bacteriostatic liquid;

[0010] S3. Mix the products of steps S1 and S2 with sodium alginate gel, adjust the pH, and then perform ultrasonic degassing and sterile filtration treatment.

[0011] Furthermore, the preparation of the squalane nanoemulsion includes: mixing 5 parts of squalane with 2 parts of Tween 80 at 50 °C and stirring to form an oil phase, the aqueous phase contains 3% w / v glycerol, the oil phase and the aqueous phase are pre-emulsified at a volume ratio of 1:9, and then subjected to 6 times of high-pressure homogenization cycling treatment, adding xanthan gum and standing for 24 h to obtain squalane nanoemulsion.

[0012] Furthermore, the traditional Chinese medicine extract is prepared by extracting 35-45 parts of dandelion and 45-55 parts of prunella vulgaris by reduced-pressure reflux extraction for 2.5 h, and the density is 1.2 g / mL.

[0013] Furthermore, in the prebiotic-nisin mixed liquid, the weight ratio of fructooligosaccharide to nisin is 2:1, the dissolution temperature is 40 °C, and the magnetic stirring rate is 600 rpm.

[0014] Furthermore, the collagen fullerene bacteriostatic liquid contains 0.5% w / v recombinant collagen, 1.0% w / v ε-polylysine and 0.01% w / v fullerene-Mg2+ complex, uses propylene glycol as the dispersion medium, and is homogenized at 5000 rpm for 10 min.

[0015] Furthermore, the mixing temperature in step S3 is 35 °C, the stirring rate is 700-900 rpm, and the mixing time is 1-3 h.

[0016] Furthermore, in step S3, the pH is adjusted using 88% lactic acid to adjust the pH to 4.5-4.8.

[0017] Furthermore, the ultrasonic degassing parameters are: frequency 50 kHz, time 10 - 25 min.

[0018] Compared with the prior art, the present invention provides an efficient gynecological bacteriostatic liquid and its preparation method, having the following beneficial effects:

[0019] 1. According to the present invention, due to the decrease of estrogen in elderly women leading to the thinning of the mucosal lipid layer, squalene lipid components are supplemented, recombinant collagen helps with repair and moisturization, ε-polylysine provides antibacterial effects, the fullerene-Mg2+ complex may have antioxidant and anti-inflammatory effects, glycerol is used as a dispersion medium and a moisturizer, and sodium alginate gel (viscosity 800 mPa·s) provides long-term lubrication, thus solving the problem of vaginal dryness in elderly women; the formula is alcohol / hormone-free and has high safety;

[0020] 2. The formula of the present invention has synergistic effects and can inhibit bacteria efficiently and broadly. The collagen fullerene bacteriostatic liquid (ε-polylysine + fullerene-Mg2+) can efficiently kill Candida albicans and drug-resistant bacteria, and the traditional Chinese medicine extract (dandelion + prunella vulgaris) targets and inhibits Neisseria gonorrhoeae and Gardnerella vaginalis. The prebiotic-nisin mixed solution selectively promotes the proliferation of Lactobacillus and maintains the balance of the flora, and the inhibition rate of probiotics is only 9.6%; hormones, antibiotics and heavy metals are abandoned, and natural ingredients are used, which are safe and non-irritating;

[0021] 3. Recombinant collagen and squalane synergistically relieve the inflammatory response, and the reduction rates of IL-1β / IL-6 / IL-8 exceed 78%. Specific Embodiments

[0022] The following describes the preferred embodiments of the present invention. It should be understood that the preferred embodiments described herein are only for the purpose of illustration and explanation of the present invention, and are not used to limit the present invention.

[0023] In addition, in the present invention, descriptions such as "first", "second", etc. are only for descriptive purposes, and do not particularly refer to the meaning of order or sequence, nor are they used to limit the present invention. They are only used to distinguish components or operations described with the same technical terms, and cannot be understood as indicating or implying their relative importance or implicitly indicating the quantity of the indicated technical features. Thus, features defined with "first", "second" may explicitly or implicitly include at least one of such features. In addition, the technical solutions and technical features between various embodiments can be combined with each other, but it must be based on the fact that those skilled in the art can achieve it. When the combination of technical solutions results in contradictions or cannot be achieved, it should be considered that such a combination of technical solutions does not exist and is not within the protection scope required by the present invention.

[0024] Unless otherwise specified, the examples and comparative examples are parallel tests with the same components, component contents, preparation steps, and preparation parameters. The experimental methods in the following examples are all conventional methods unless otherwise specified; the test materials used in the following examples are all analytical reagents (A.R.) and are purchased from commercial channels.

[0025] Sodium alginate gel (viscosity 800 mPa·s).

[0026] Example 1

[0027] S1. Preparation of squalane nanoemulsion: By weight, 5 parts of squalane and 2 parts of Tween 80 are mixed at 50 °C and stirred at 600 rpm for 20 min to obtain the oil phase. Deionized water is preheated to 50 °C, glycerol (3% w / v) is added, and it is stirred at 400 rpm to obtain the water phase. The oil phase is slowly added to the water phase (volume ratio 1:9), pre-emulsified at 12,000 rpm for 3 min, and homogenized under high pressure for 6 cycles at a pressure of 500 bar and a temperature of 25 °C. Xanthan gum (0.1% w / v) is added, and it is stirred at 300 rpm for 15 min and left standing for 24 h to obtain squalane nanoemulsion with a particle size of 50 nm;

[0028] S2. Preparation of active ingredients: 35 parts of dandelion and 45 parts of prunella vulgaris are refluxed and extracted at 55 °C and 15 kPa for 2.5 hours with an ethanol concentration of 70%. The extract is concentrated to a density of 1.2 g / mL to obtain a traditional Chinese medicine extract. Fructooligosaccharide and nisin are dissolved in deionized water at 40 °C in a weight ratio of 2:1 and stirred magnetically at 600 rpm for 15 min to obtain a prebiotic-nisin mixture. Recombinant collagen (0.5% w / v), ε-polylysine (1.0% w / v), and fullerene-Mg2+ complex (0.01% w / v) are dispersed in propylene glycol and treated with a homogenizer at 5000 rpm for 10 min to obtain a collagen fullerene antibacterial solution;

[0029] S3. Preparation of highly efficient gynecological antibacterial solution: By weight, 4 parts of squalane nanoemulsion, 3 parts of traditional Chinese medicine extract, 2 parts of prebiotic-nisin mixture, 2 parts of sodium alginate gel, and 6 parts of collagen fullerene antibacterial solution are added to a reaction kettle at a temperature of 35 °C, a stirring rate of 800 rpm, and a time of 2 hours. 88% lactic acid is added dropwise until the pH reaches 4.6, and it is degassed by ultrasonic wave at 50 kHz for 20 min. It is aseptically filtered through a 0.22 μm membrane at a flow rate of 50 mL / min and a temperature of 25 °C to obtain a highly efficient gynecological antibacterial solution.

[0030] Example 2

[0031] Preparation of highly effective gynecological bacteriostatic liquid: Add 3 parts of squalane nanoemulsion, 2 parts of traditional Chinese medicine extract, 3 parts of prebiotic-nisin mixed solution, 1 part of sodium alginate gel and 3 parts of collagen fullerene bacteriostatic liquid by weight to a reaction kettle. Set the temperature at 35°C, the stirring rate at 700 rpm, and the time at 1 hour. Dropwise add 88% lactic acid until the pH reaches 4.5, and perform ultrasonic degassing at 50 kHz for 10 min. Then, filter aseptically at 25°C through a 0.22-μm membrane with a flow rate of 50 mL / min to obtain the highly effective gynecological bacteriostatic liquid.

[0032] Example 3

[0033] Preparation of highly effective gynecological bacteriostatic liquid: Add 5 parts of squalane nanoemulsion, 4 parts of traditional Chinese medicine extract, 1 part of prebiotic-nisin mixed solution, 3 parts of sodium alginate gel and 5 parts of collagen fullerene bacteriostatic liquid by weight to a reaction kettle. Set the temperature at 35°C, the stirring rate at 900 rpm, and the time at 3 hours. Dropwise add 88% lactic acid until the pH reaches 4.8, and perform ultrasonic degassing at 50 kHz for 25 min. Then, filter aseptically at 25°C through a 0.22-μm membrane with a flow rate of 50 mL / min to obtain the highly effective gynecological bacteriostatic liquid.

[0034] Example 4

[0035] When filling the highly effective gynecological bacteriostatic liquid of Examples 1 - 3 into a light-proof PE bottle, nitrogen is filled until the oxygen content in the bottle is <1%.

[0036] Comparative Example 1

[0037] The difference from Example 1 is that it lacks an equal weight portion of squalane nanoemulsion, and the others are the same.

[0038] Comparative Example 2

[0039] The difference from Example 1 is that it lacks an equal weight portion of the prebiotic-nisin mixed solution, and the others are the same.

[0040] Comparative Example 3

[0041] The difference from Example 1 is that it lacks an equal weight portion of collagen fullerene bacteriostatic liquid, and the others are the same.

[0042] Comparative Example 4

[0043] The difference from Example 1 is that it lacks an equal weight portion of sodium alginate gel, and the others are the same.

[0044] Comparative Example 5

[0045] The difference from Example 1 is that it lacks an equal weight portion of traditional Chinese medicine extract, and the others are the same.

[0046] Performance Test

[0047] 1. The stability of the gynecological bacteriostatic solutions prepared in the examples and comparative examples was tested. Using the accelerated experiment method, the gynecological bacteriostatic solutions obtained in the examples and comparative examples were placed in a thermostatic and humidified chamber (40°C / 75% RH, 1 - 3 months), and samples were taken regularly (at 0 and 3 months) to observe the retention rate of the active ingredients. The squalane content was detected by HPLC, and the fullerene-Mg2+ complex was detected by UV-Vis method after dilution 100 times, and the absorbance at 260 nm was measured. Compared with the sample at 0 month, the retention rate (%) = (content after accelerated test / initial content) × 100; Common vaginal probiotics (such as Lactobacillus crispatus, L. gasseri) were selected, and the bacteriostatic solution was co-cultured with the probiotics at a use concentration of 3 mg / mL for 24 hours, and the change in the viable count was detected to obtain the probiotic inhibition rate; The bacteriostatic solution was coated on an artificial skin model (such as ), and the water loss amount within 6 hours was measured at 37°C and a humidity of 50% to obtain the moisturizing rate. The results are shown in Table 1.

[0048] Table 1

[0049]

[0050]

[0051] As can be seen from Table 1, the retention rates of squalane and fullerene-Mg2+ complex in Example 1 were higher than those in the comparative examples during the accelerated test, indicating that the nanoemulsion formulation and the collagen fullerene bacteriostatic solution significantly improved the component stability. The moisturizing rate of Comparative Example 1 (without squalane) was the lowest (62%), proving that squalane nanoemulsion is crucial for long-term moisturization. The content of the fullerene-Mg2+ complex in Comparative Example 3 (without collagen fullerene bacteriostatic solution) was 0, and the probiotic inhibition rate was as high as 45.3%, indicating that the collagen fullerene bacteriostatic solution can selectively inhibit pathogenic bacteria without affecting probiotics.

[0052] 2. The antibacterial activity of the gynecological bacteriostatic solutions prepared in the examples and comparative examples was detected. For the preparation of the bacterial solution, Candida albicans (ATCC 10231), Gardnerella vaginalis (ATCC 49145), Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC29213), and Neisseria gonorrhoeae cultured to the logarithmic growth phase were used to prepare a bacterial solution of 1×10 7 cfu / ml using a nutrient broth medium. 25 ml of agar was added to a sterile petri dish. After it solidified, a sterile cotton swab was used to dip the bacterial solution and evenly coat it on the surface of the agar. Then, 6 holes were punched equidistantly on the culture medium with a puncher with a diameter of 0.6 cm, and 0.2 ml of the gynecological bacteriostatic solution was added to each hole. Three petri dishes were prepared for each medicinal solution and cultured in a 37°C biochemical incubator for 24 h. The diameter of the antibacterial zone was measured in mm, and the average value of the three petri dishes was calculated; The results are shown in Table 2.

[0053] Table 2

[0054]

[0055]

[0056] As can be seen from Table 2, the diameters of the inhibition zones of Example 1 against all pathogens were significantly higher than those of the comparative example, indicating the synergistic effect of the formulation and the broad-spectrum antibacterial effect of the traditional Chinese medicine extract and the collagen fullerene complex.

[0057] 3. The anti-inflammatory effect and safety of the gynecological antibacterial solutions prepared in the examples and comparative examples were tested. A co-culture model of human vaginal epithelial cells (VK2 / E6E7) and Candida albicans was constructed, and the antibacterial solution (10 mg / mL) was added for intervention for 48 hours. The levels of IL-1β, IL-6, and IL-8 in the cell supernatant were detected by ELISA. The experimental subjects were 39 healthy female New Zealand rabbits. 20 μL of the gynecological antibacterial solution was injected into the vaginas of estrogen-deficient female New Zealand rabbits 8 weeks after ovariectomy, and 20 μL of normal saline was injected into the control group. The injection interval was 24 hours, and the injection was continuous for 3 times. After 6 days, the vaginas of the corresponding female New Zealand rabbits were taken out, longitudinally incised, and the physiological conditions of the vaginal sections were observed under a microscope to observe abnormal irritation phenomena such as fluid overflow and erythema edema. The results are shown in Table 3.

[0058] Table 3

[0059]

[0060]

[0061] As can be seen from Table 3, the reduction rates of IL-1β, IL-6, and IL-8 in Example 1 exceeded 80%, proving that it significantly inhibits the release of inflammatory factors through the synergistic effect of the traditional Chinese medicine extract (dandelion, prunella vulgaris) and the fullerene-Mg2+ complex.

[0062] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. If these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention also intends to include these changes and modifications.

Claims

1. A highly effective gynecological antibacterial liquid, characterized in that: The invention comprises the following components: squalane nanoemulsion, Chinese herbal medicine extract, prebiotic-nisin mixed solution, sodium alginate gel and collagen fullerene antibacterial solution.

2. The high-efficiency gynecological antibacterial liquid according to claim 1, characterized in that: The invention comprises the following components by weight: 3-5 parts of squalane nanoemulsion, 2-4 parts of traditional Chinese medicine extract, 1-3 parts of prebiotic-nisin mixed solution, 1-3 parts of sodium alginate gel and 3-6 parts of collagen fullerene antibacterial solution.

3. The high-efficiency gynecological antibacterial liquid according to claim 1, characterized in that: The preparation method of the high-efficiency gynecological antibacterial liquid comprises the following steps: S1. preparing squalane nanoemulsion; S2, preparing Chinese herbal medicine extract, prebiotic-nisin mixed solution and collagen fullerene antibacterial solution; S3, mixing the products of step S1 and S2 with sodium alginate gel, adjusting the pH, and then performing ultrasonic degassing and sterile filtration.

4. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: The preparation of the squalane nanoemulsion comprises: mixing 5 parts of squalane and 2 parts of Tween 80 at 50° C. to form an oil phase, wherein the water phase contains 3% w / v glycerol, the oil phase and the water phase are pre-emulsified at a volume ratio of 1:9, high-pressure homogenization is circulated for 6 times, xanthan gum is added and allowed to stand for 24 hours to obtain the squalane nanoemulsion.

5. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: The Chinese medicine extract is prepared by vacuum reflux extraction of 35-45 parts of dandelion and 45-55 parts of selfheal, the extraction time is 2.5 hours, and the density is 1.2 g / mL.

6. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: The weight ratio of oligofructose to nisin in the prebiotic-nisin mixed solution is 2:1, the dissolution temperature is 40° C., and the magnetic stirring rate is 600 rpm.

7. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: The collagen fullerene antibacterial solution contains 0.5% w / v recombinant collagen, 1.0% w / v ε-polylysine and 0.01% w / v fullerene-Mg2+ complex, uses propylene glycol as a dispersion medium, and is homogenized at 5000 rpm for 10 minutes.

8. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: The mixing temperature of step S3 is 35° C., the stirring speed is 700-900 rpm, and the mixing time is 1-3 h.

9. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: In step S3, the pH is adjusted to 4.5-4.8 using lactic acid with a concentration of 88%.

10. The high-efficiency gynecological antibacterial liquid according to claim 3, characterized in that: The ultrasonic degassing parameters are: frequency 50kHz, time 10-25min.

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