A stable antibacterial gynecological composition and gynecological wash thereof

Abstract

The invention discloses a stable antibacterial gynecological composition and a gynecological washing liquid thereof. The antibacterial gynecological composition is composed of bamboo vinegar fermentation liquid, arginine-modified kaempferol nanoparticles and allantoin in a weight ratio of (20-30):(5-10):(1-2). Experiments have shown that the gynecological washing liquid prepared from the antibacterial gynecological composition has a good antibacterial effect on Neisseria gonorrhoeae, Staphylococcus aureus and Candida albicans, and the antibacterial rate reaches more than 98%, can effectively inhibit gynecological pathogens, prevent and treat gynecological infections, and can reduce gynecological inflammation, and has low irritation and good stability.

Description

Technical Field

[0001] The invention belongs to the technical field of preparation of gynecological external medicines, and particularly relates to a stable antibacterial gynecological composition and a gynecological washing liquid thereof. Background Art

[0002] Gynecological inflammation is a common disease among women. Common clinical gynecological inflammations include vaginitis, cervicitis, endometritis, etc. Trichomonas, fungus, and bacterial infections are the main causes of vaginal discharge and vulvar itching in women. Clinically, gynecological inflammation is mainly manifested by vulvar itching, burning and swelling, increased vaginal discharge, frequent urination, urgent urination, and painful urination. It recurs repeatedly and does not heal for a long time, affecting the quality of life.

[0003] At present, the main Western medicine drugs for the treatment of gynecological inflammation are metronidazole and imipenem. The initial effect is obvious, but after long-term use, the antibacterial effect decreases due to the increase of microbial resistance, and patients have to re-purchase. Traditional Chinese medicine lotions are mostly formulated with traditional bactericidal drugs, which are effective in combating microbial resistance. However, the preparation process of traditional Chinese medicine lotions is relatively complicated and involves the interaction of multiple Chinese medicine ingredients. Its stability is poor. Generally, the properties are good after preparation, but the solution gradually becomes turbid after being placed for a few days. This may be due to the precipitation of the co-solvent under light, or the Chinese medicine extraction process may contain some components that are easily discolored and turbid in the light.

[0004] Therefore, it is of great significance to develop a stable, effective antibacterial gynecological composition and gynecological washing liquid. Summary of the invention

[0005] The object of the present invention is to provide an antibacterial gynecological composition and a gynecological washing liquid thereof which have stable properties, significant efficacy, controllable quality and low side effects.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] A stable antibacterial gynecological composition is composed of bamboo vinegar fermentation liquid, arginine-modified kaempferol nanoparticles and allantoin in a weight ratio of (20-30):(5-10):(1-2).

[0008] Furthermore, the bamboo vinegar fermentation liquid is prepared by fermenting bamboo vinegar liquid, inulin and coptis chinensis extract with Lactobacillus acidophilus and Lactobacillus rhamnosus.

[0009] Furthermore, the preparation steps of the bamboo vinegar fermentation liquid are: stirring and mixing the bamboo vinegar liquid and inulin, sterilizing at high temperature, then inoculating Lactobacillus acidophilus and Lactobacillus rhamnosus, fermenting to obtain fermentation liquid, adding Coptis chinensis extract, continuing fermentation for 2 to 4 hours, centrifuging to collect the supernatant, filtering the supernatant, and obtaining the bamboo vinegar fermentation liquid.

[0010] Furthermore, the weight ratio of the bamboo vinegar, inulin and coptis chinensis extract is 1:(0.05-0.1):(0.1-0.2); the inoculation amount of Lactobacillus acidophilus and Lactobacillus rhamnosus is 1% v / v, and the viable count of Lactobacillus acidophilus and Lactobacillus rhamnosus is 1×10 5 cfu / mL.

[0011] Furthermore, the arginine-modified kaempferol nanoparticles are prepared by loading kaempferol onto zein nanoparticles and then modifying the zein nanoparticles loaded with kaempferol with arginine.

[0012] Furthermore, the preparation steps of the arginine-modified kaempferol nanoparticles are as follows:

[0013] (1) dissolving zein in an alkaline solution with a pH of 11 to 13, adding kaempferol, continuing to stir for 1 to 2 hours, adjusting the pH value to neutral, centrifuging to obtain the supernatant, and freeze-drying the supernatant to obtain kaempferol nanoparticles;

[0014] (2) Arginine, EDC and NHS are dissolved in water and activated by stirring for 1 to 2 hours to obtain a mixed solution; the mixed solution is slowly added to the aqueous dispersion of kaempferol nanoparticles and reacted under stirring; the reaction product is dialyzed with distilled water and freeze-dried to obtain arginine-modified kaempferol nanoparticles.

[0015] Furthermore, the weight ratio of kaempferol to zein is 1:(5-10); the weight ratio of kaempferol nanoparticles, arginine, EDC and NHS is (1-2):5:3:2.

[0016] The present invention also provides the use of the antibacterial gynecological composition in preparing a medicine for treating gynecological inflammation.

[0017] The present invention also provides a gynecological wash comprising the above-mentioned antibacterial gynecological composition.

[0018] Preferably, the gynecological wash further comprises a preservative and / or a buffer.

[0019] Bamboo vinegar is a by-product obtained by pyrolysis of bamboo processing residues. It contains a variety of ingredients with antibacterial effects, including organic acids, phenols, ketones, alcohols and other substances. It has antibacterial activity against a variety of fungi and bacteria. It is a broad-spectrum antibacterial and bactericide, and its properties are safe. Relevant studies have confirmed that bamboo vinegar is safe and reliable for contact with human and animal skin and for oral and nasal use.

[0020] Coptis chinensis extract contains high levels of alkaloids, including berberine, palmatine, coptisine, jatrorrhizine and other ingredients, which have significant inhibitory effects on a variety of bacteria and fungi.

[0021] Inulin is a natural polysaccharide extracted from chicory roots. It has weak direct antibacterial activity, but can indirectly inhibit harmful bacteria by promoting the growth of probiotics.

[0022] The present invention utilizes Lactobacillus acidophilus and Lactobacillus rhamnosus to ferment bamboo vinegar, inulin and coptis chinensis extract to prepare bamboo vinegar fermented liquor. On the one hand, the acidic condition of bamboo vinegar is utilized to promote the effective dissolution of alkaloids in coptis chinensis extract. On the other hand, the prebiotic effect of inulin is utilized to promote the growth of Lactobacillus acidophilus and Lactobacillus rhamnosus. Lactobacillus acidophilus and Lactobacillus rhamnosus are utilized to ferment coptis chinensis extract under the acidic environment of bamboo vinegar to effectively promote the dissolution of active ingredients in coptis chinensis extract. In addition, Lactobacillus acidophilus and Lactobacillus rhamnosus will produce acidic substances during the fermentation process. The increase of these acidic substances can enrich the types of antibacterial components in bamboo vinegar fermented liquor, and the bacteriocins produced during the fermentation process of the two also have an inhibitory effect on multiple pathogens. The presence of Lactobacillus acidophilus and Lactobacillus rhamnosus as probiotics can regulate the ecological microenvironment of the vagina and inhibit the reproduction of harmful microorganisms. In summary, the present invention finally obtains a bamboo vinegar fermented liquor with significant antibacterial activity through the interaction and influence of the above-mentioned components during the fermentation process.

[0023] Kaempferol is a flavonoid compound with antibacterial and antifungal activity. However, it is unstable in nature, has low bioavailability, and poor water solubility, which limits its application in the preparation of gynecological lotion products. Zein is soluble in alkaline solutions and easily aggregates to form nanoparticles under neutral conditions. The present invention uses zein as a nanocarrier material, utilizes the solubility characteristics of kaempferol under specific pH conditions, and encapsulates kaempferol in zein to form nanoparticles, which can significantly improve the stability of kaempferol. However, due to the hydrophobic surface of zein, the formed nanoparticles are difficult to disperse again in water. The inventors have found that further modifying arginine on the surface of zein can improve the redispersibility of nanoparticles in water, and arginine has a positive charge, which enhances affinity with the bacterial surface through electrostatic interaction, thereby improving the transmembrane transport efficiency and antibacterial activity of nanoparticles.

[0024] Allantoin is a natural compound found in the skin and mucous membranes that promotes wound healing and has anti-inflammatory properties.

[0025] Furthermore, after the antibacterial gynecological composition containing the bamboo vinegar fermentation liquid of the present invention is dissolved or diluted with water, the pH value of the prepared gynecological wash is 4.0-5.5, which is close to the pH value of the vaginal private parts, which is beneficial to maintaining the natural defense function of the private parts, avoiding the weak acid environment that destroys the health of the private parts, causing dryness and itching of the private parts, and even causing gynecological inflammation.

[0026] Furthermore, when the antibacterial gynecological composition is made into a gynecological wash, an appropriate amount of a buffer, such as aminomethyl propylene glycol, can be added as needed to adjust the pH value so that the pH of the prepared gynecological wash reaches a level of 4.0 to 5.5.

[0027] The present invention has the following beneficial effects:

[0028] The present invention uses bamboo vinegar fermentation liquid, arginine-modified kaempferol nanoparticles and allantoin as effective ingredients of the antibacterial gynecological composition, which can effectively inhibit gynecological pathogens, prevent and treat gynecological infections, reduce gynecological inflammation, promote the repair of damaged tissues, and restore health, and has obvious antibacterial, anti-inflammatory and repair-promoting effects. DETAILED DESCRIPTION

[0029] The following examples are provided to further illustrate the present invention, but are not intended to limit the present invention. The components of the formulas in the following examples are conventional commercially available products unless otherwise specified. Among them, bamboo vinegar was purchased from Jiangyin Zhongju Biotechnology Co., Ltd., with a pH of about 2.5. Coptis chinensis extract was purchased from Shaanxi Huatai Biological Fine Chemical Co., Ltd.

[0030] The gynecological washes of the following Examples 3-5 were prepared using the bamboo vinegar fermentation liquid prepared in Example 1 and the Arg / Zein / Kae nanoparticles prepared in Example 2 as raw materials.

[0031] Embodiment 1 bamboo vinegar fermentation liquid preparation

[0032] 100 g of bamboo vinegar and 10 g of inulin were mixed and sterilized at 120 °C for 15 min, and then inoculated with 1% by volume of Lactobacillus acidophilus and Lactobacillus rhamnosus, the viable counts of Lactobacillus acidophilus and Lactobacillus rhamnosus were both 1 × 10 5 cfu / mL, fermented at 30°C for 24h to obtain fermentation broth, added 10g of Coptis chinensis extract, continued fermentation at 40°C for 2h, collected the supernatant by centrifugation, and filtered the supernatant to obtain bamboo vinegar fermentation broth.

[0033] Example 2 Preparation and detection of arginine-modified kaempferol nanoparticles

[0034] 1. Preparation of Arginine-modified Kaempferol Nanoparticles

[0035] (1) Disperse 5 g of zein in 500 mL of deionized water, adjust the pH to 12 with NaOH solution, stir to dissolve, then add 1 g of kaempferol (Kae), continue stirring for 2 h, adjust the pH to neutral, centrifuge at 8000 g for 20 min to remove unbound kaempferol, and freeze-dry the supernatant after centrifugation to obtain kaempferol nanoparticles (Zein / Kae nanoparticles).

[0036] (2) dispersing 2 g of kaempferol nanoparticles in 50 mL of deionized water to obtain an aqueous dispersion of kaempferol nanoparticles;

[0037] (3) Dissolve 5 g of arginine (Arg), 3 g of EDC and 2 g of NHS in 50 mL of deionized water, and stir to activate for 1.5 h to obtain a mixed solution; slowly add the mixed solution to the aqueous dispersion of kaempferol nanoparticles in step (2), and react for 12 h under stirring. The reaction product is dialyzed with distilled water and freeze-dried to obtain arginine-modified kaempferol nanoparticles (Arg / Zein / Kae nanoparticles).

[0038] 2. Resolubility test of nanoparticles

[0039] Take 20mg of Zein / Kae nanoparticles and Arg / Zein / Kae nanoparticles and dissolve them in 10mL of deionized water, stir them to disperse them evenly, and observe the dispersion of different nanoparticles after the samples are left to stand for 1 day. The results show that the Zein / Kae nanoparticle dispersion shows obvious stratification after standing, while the Arg / Zein / Kae nanoparticles have good solubility and disperse in water to form a uniform dispersion.

[0040] 3. Stability testing of nanoparticles after reconstitution

[0041] 20 mg of Arg / Zein / Kae nanoparticles were dissolved in 10 mL of deionized water to obtain a 2 mg / mL dispersion. The stability of the Arg / Zein / Kae nanoparticle dispersion was evaluated by observing the changes in the particle size and polydispersity index (PDI) value of the dispersion after fresh re-dissolution and standing for 10 days. The results showed that the particle size of the Arg / Zein / Kae nanoparticles remained almost unchanged after standing for 10 days, while the PDI value decreased, indicating that the particle size distribution of the nanoparticles became more uniform during the standing process. The above results show that the Arg / Zein / Kae nanoparticles have good stability.

[0042] Example 3 Preparation of antibacterial gynecological wash

[0043] An antibacterial gynecological washing liquid comprises the following components in percentage by weight: 40% bamboo vinegar fermentation liquid, 10% Arg / Zein / Kae nanoparticles, 2% allantoin, 0.1% sodium benzoate, an appropriate amount of aminomethyl propylene glycol, and the balance is deionized water.

[0044] The preparation steps are as follows: dissolving Arg / Zein / Kae nanoparticles, allantoin and sodium benzoate in deionized water, adding bamboo vinegar fermentation liquid, stirring and mixing, and adjusting the pH to about 5 with aminomethyl propylene glycol to obtain a gynecological wash.

[0045] Example 4 Preparation of antibacterial gynecological wash

[0046] An antibacterial gynecological washing liquid comprises the following components in percentage by weight: 45% of bamboo vinegar fermentation liquid, 15% of Arg / Zein / Kae nanoparticles, 1.5% of allantoin, 0.1% of sodium benzoate, an appropriate amount of aminomethyl propylene glycol, and the balance of deionized water.

[0047] The preparation steps are as follows: dissolving Arg / Zein / Kae nanoparticles, allantoin and sodium benzoate in deionized water, adding bamboo vinegar fermentation liquid, stirring and mixing, and adjusting the pH to about 5 with aminomethyl propylene glycol to obtain a gynecological wash.

[0048] Example 5 Preparation of antibacterial gynecological wash

[0049] An antibacterial gynecological washing liquid comprises the following components in percentage by weight: 50% of bamboo vinegar fermentation liquid, 10% of Arg / Zein / Kae nanoparticles, 4% of allantoin, 0.1% of sodium benzoate, an appropriate amount of aminomethyl propylene glycol, and the balance of deionized water.

[0050] The preparation steps are as follows: dissolving Arg / Zein / Kae nanoparticles, allantoin and sodium benzoate in deionized water, adding bamboo vinegar fermentation liquid, stirring and mixing, and adjusting the pH to about 5 with aminomethyl propylene glycol to obtain a gynecological wash.

[0051] Comparative Example 1 Preparation of antibacterial gynecological wash

[0052] The difference with Example 5 is that bamboo vinegar fermented liquid of the present invention is replaced with commercially available bamboo vinegar liquid.

[0053] Comparative Example 2 Preparation of Antibacterial Gynecological Wash

[0054] The difference from Example 5 is that bamboo vinegar fermentation liquid is not contained and an equal amount of deionized water is used to supplement it.

[0055] Comparative Example 3 Preparation of Antibacterial Gynecological Wash

[0056] The difference from Example 5 is that no Arg / Zein / Kae nanoparticles are contained and an equal amount of deionized water is used to supplement the mixture.

[0057] Comparative Example 4 Preparation of Antibacterial Gynecological Wash

[0058] An antibacterial gynecological washing liquid comprises the following components in percentage by weight: 45% bamboo vinegar, 5% coptis root extract, 10% Arg / Zein / Kae nanoparticles, 4% allantoin, 0.1% sodium benzoate, 2% Tween 80, an appropriate amount of aminomethyl propylene glycol, and the balance is deionized water.

[0059] The preparation steps are as follows: dissolving Arg / Zein / Kae nanoparticles, allantoin, sodium benzoate and Tween 80 in deionized water, adding bamboo vinegar and coptis root extract, stirring and dissolving, and adjusting the pH to about 5 with aminomethyl propylene glycol to obtain a gynecological wash.

[0060] The difference between this comparative example and Example 5 is that the bamboo vinegar fermentation liquid of the present invention is replaced by bamboo vinegar liquid and coptis root extract.

[0061] Example 6 Safety Testing

[0062] Take the gynecological washing liquid prepared in Example 3-5, set up a test group (Example 3-5), a negative control group and a blank control group. Six adult healthy female SD rats were included in each group. The test group took 10mL of the gynecological washing liquid prepared in Example 3-5, soaked the corresponding gynecological washing liquid with sterilized cotton wool of appropriate size and placed it in the rat vagina, and the negative control group soaked the same size cotton strip in sterilized saline and placed it in the rat vagina, all of which were in contact with the animal vaginal mucosa for 24 hours. The blank control group was not given any treatment. After 24 hours, the animals were killed, the entire vagina was taken out, the congestion and edema reaction phenomena of the local vaginal tissue were observed, and the stimulation reaction was evaluated. It was found that no abnormal reactions such as congestion and edema were found in the local vaginal tissue of the rats in the test group of Example 3-5 and the control group, and no other symptoms were found. It shows that the gynecological washing liquid of the present invention has low stimulation to the vaginal mucosa and is safe in nature.

[0063] Example 7 Antibacterial Effect Detection

[0064] According to the antibacterial effect test method (suspension quantitative method) of antibacterial daily chemical products in QB / T 2738-2012 "Evaluation method for antibacterial and antibacterial effects of daily chemical products", the antibacterial rates of the gynecological washes prepared in Examples 3-5 and Comparative Examples 1-4 against Neisseria gonorrhoeae (ATCC 49226), Staphylococcus aureus (ATCC 29213), and Candida albicans (ATCC 10231) were tested. The experiment was repeated 3 times and the average value was taken. The results are shown in Table 1.

[0065] Table 1 Antibacterial effect test results

[0066]

[0067]

[0068] The results show that the gynecological wash prepared in Examples 3-5 has good antibacterial effects on Neisseria gonorrhoeae, Staphylococcus aureus, and Candida albicans, with an inhibition rate of more than 98%. It can effectively inhibit gynecological pathogens and prevent and treat gynecological infections, and the effect is better than the gynecological wash prepared in Comparative Examples 1-4.

[0069] Example 8 Stability Test

[0070] The gynecological wash prepared in Examples 3-5 was filled into a transparent container and sealed, and placed in a dark place at room temperature of 30±2°C and relative humidity of 60±5% for 24 months, and the changes in the appearance of the gynecological wash were observed. The antibacterial activity of the gynecological wash against Neisseria gonorrhoeae, Staphylococcus aureus, and Candida albicans was tested again with reference to the method of Example 7. The results showed that no stratification was observed after the gynecological wash was placed for 24 months, and the antibacterial activity against Neisseria gonorrhoeae, Staphylococcus aureus, and Candida albicans was still maintained at more than 95%. The results showed that the gynecological wash provided by the present invention had good stability.

[0071] The above are only preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be regarded as limiting the present invention, and the protection scope of the present invention should be based on the scope defined by the claims. For ordinary technicians in this technical field, several improvements and modifications can be made without departing from the spirit and scope of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A stable antibacterial gynecological composition, characterized in that: The composition is composed of bamboo vinegar fermentation liquid, arginine-modified kaempferol nanoparticles and allantoin in a weight ratio of (20-30): (5-10): (1-2); The preparation steps of the bamboo vinegar fermentation liquid are as follows: mixing the bamboo vinegar liquid and inulin, sterilizing at high temperature, inoculating Lactobacillus acidophilus and Lactobacillus rhamnosus, fermenting to obtain fermentation liquid, adding Coptis chinensis extract, continuing fermentation for 2-4 hours, collecting supernatant by centrifugation, filtering the supernatant, and obtaining the bamboo vinegar fermentation liquid; The weight ratio of the bamboo vinegar, inulin and coptis extract is 1: (0.05-0.1): (0.1-0.2); the inoculation amount of Lactobacillus acidophilus and Lactobacillus rhamnosus is 1% v / v, and the number of viable bacteria of Lactobacillus acidophilus and Lactobacillus rhamnosus is 1×10 5 cfu / mL; The preparation steps of the arginine-modified kaempferol nanoparticles are as follows: (1) Dissolve zein in an alkaline solution with a pH of 11 to 13, add kaempferol, continue stirring for 1 to 2 hours, adjust the pH value to neutral, centrifuge to obtain the supernatant, and freeze-dry the supernatant to obtain kaempferol nanoparticles; (2) Dissolving arginine, EDC and NHS in water, stirring and activating for 1 to 2 h to obtain a mixed solution; slowly adding the mixed solution into the aqueous dispersion of kaempferol nanoparticles, reacting under stirring, and dialyzing the reaction product with distilled water and freeze-drying to obtain arginine-modified kaempferol nanoparticles; The weight ratio of kaempferol to zein is 1:(5-10); the weight ratio of kaempferol nanoparticles, arginine, EDC and NHS is (1-2):5:3:

2.

2. Use of the antibacterial gynecological composition according to claim 1 in the preparation of a medicament for treating gynecological inflammation.

3. A gynecological wash, characterized in that: The antibacterial gynecological composition according to claim 1 is used as the only active ingredient.

4. The gynecological wash according to claim 3, characterized in that: Preservatives and / or buffers may also be included.

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