CN118126910A - Probiotic agent for improving vaginal infection and preparation method and application thereof - Google Patents

Probiotic agent for improving vaginal infection and preparation method and application thereof Download PDF

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CN118126910A
CN118126910A CN202410575027.7A CN202410575027A CN118126910A CN 118126910 A CN118126910 A CN 118126910A CN 202410575027 A CN202410575027 A CN 202410575027A CN 118126910 A CN118126910 A CN 118126910A
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probiotic
strain
vaginal
lactobacillus casei
lactobacillus acidophilus
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CN118126910B (en
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方曙光
董瑶
唐海峰
顾佳悦
吴智仪
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WeCare Probiotics Co Ltd
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WeCare Probiotics Co Ltd
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Abstract

The invention relates to a probiotic for improving vaginal infection, and a preparation method and application thereof, wherein strains in the probiotic comprise lactobacillus casei Lactobacillus casei LC strain with a preservation number of CGMCC No.24110 and lactobacillus acidophilus Lactobacillus acidophilus LA strain with a preservation number of CCTCC No. M2022572.

Description

Probiotic agent for improving vaginal infection and preparation method and application thereof
Technical Field
The invention belongs to the technical field of microorganisms, and relates to a probiotic agent for improving vaginal infection, and a preparation method and application thereof.
Background
Female vulvovaginal infections are now very common, which are the result of an imbalance in the vaginal ecosystem, caused by different exogenous or endogenous factors that affect the microbiota in the vagina and reduce protective lactobacilli. Lactic acid bacteria are the most abundant dominant microorganisms, 10 7~108 CFU in healthy premenopausal women's vaginal fluid, which have the function of maintaining the healthy environment of the vagina, limiting the growth of pathogenic microorganisms by producing acids, hydrogen peroxide, bacteriocins and modulating local immune responses. The most common vaginal infections are caused by pathogenic bacteria, typically gardnerella vaginalis causing Bacterial Vaginosis (BV), or fungi such as candida causing vulvovaginal candidiasis (VVC). Conventional treatment of BV is oral or intravaginal with metronidazole or clindamycin according to therapeutic guidelines, whereas VVC is typically treated locally and systemically with nystatin. Conventional therapies may produce side effects, the emergence of drug resistant strains, and recurrence of symptoms. In order to minimize these adverse effects, it is important to try to develop different alternative treatments. Therefore, the use of specific probiotic products containing lactic acid bacteria is the first choice for restoring the physiological balance of the vaginal ecosystem.
Probiotics are defined as living microorganisms that, if administered in sufficient amounts, have a beneficial physiological effect on the host. The beneficial effects of lactic acid bacterial strains on BV and VVC have been reported in historical studies, and probiotic formulations have been tested orally and topically so far, however the intravaginal route of administration has so far appeared to be the first choice for probiotic management. Its advantages are: reduces the severity of gastrointestinal enzyme and acid effects, and overcomes pain and tissue damage caused by other parenteral routes. These advantages are mainly manifested in that the patient is given a lower dose, and less frequent administration results in a high absorption rate of the probiotic bacteria, which is not affected by the gastrointestinal tract. Formulation studies that colonize the vaginal environment and maintain relative viability are new challenges.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a probiotic agent for improving vaginal infection, and a preparation method and application thereof.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
In a first aspect, the invention provides a probiotic for improving vaginal infection, wherein strains in the probiotic comprise lactobacillus casei Lactobacillus casei LC strain with a preservation number of CGMCC No.24110 and lactobacillus acidophilus Lactobacillus acidophilus LA strain with a preservation number of CCTCC No. M2022572.
The invention creatively discovers that the related combination of lactobacillus casei LC16 strain and lactobacillus acidophilus LA18 strain can significantly improve the vulvar tract infection, and the effect is larger than that of single strain application. The specific expression is as follows: (1) effective inhibition of pathogenic bacteria growth; (2) a reduction in the inflammatory response of the mouse vulva tract; (3) Reduces the serum inflammatory factors IL-1 beta, TNF-alpha and IL-6 level of BV mice.
Preferably, the ratio of the viable count of the Lactobacillus casei Lactobacillus casei LC strain to the Lactobacillus acidophilus Lactobacillus acidophilus LA strain is (3-9): 1.
Wherein, the specific point values in (3-9) can be selected from 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, etc., and other specific point values in the above numerical range can be selected, and will not be described in detail herein.
Preferably, the viable count of the lactobacillus casei Lactobacillus casei LC strain and the lactobacillus acidophilus Lactobacillus acidophilus LA strain is independently not lower than 1×10 9 CFU/mL, such as 1×109CFU/mL、2×109 CFU/mL、3×109 CFU/mL、4×109 CFU/mL、5×109 CFU/mL、6×109 CFU/mL、7×109 CFU/mL、8×109 CFU/mL、9×109 CFU/mL、1×1010 CFU/mL、2×1010 CFU/mL、5×1010CFU/mL、8×1010 CFU/mL、1×1011 CFU/mL、1×1012 CFU/mL、1×1013 CFU/mL, and other specific values in the above numerical range are selected, and will not be described in detail herein.
Preferably, the dosage form of the probiotic comprises a tablet.
Preferably, the probiotic agent further comprises a protective agent, a quick release component and a slow release component.
Preferably, the protective agent comprises any one or a combination of at least two of acetic acid, bovine serum albumin, propylene glycol, lactic acid or alginate.
Preferably, the probiotics comprises 0.1-1% of acetic acid, 0.1-0.5% of bovine serum albumin, 0.1-0.4% of propylene glycol and 0.1-1% of lactic acid by mass percent.
The mass percentage of the acetic acid may be selected from 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, etc., the mass percentage of the bovine serum albumin may be selected from 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, etc., the mass percentage of the propylene glycol may be selected from 0.1%, 0.2%, 0.3%, 0.4%, etc., and the mass percentage of the lactic acid may be selected from 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, etc., and other specific values in the above numerical ranges may be selected, and will not be repeated here.
Preferably, the retarded release component comprises any one or a combination of at least two of stearic acid, sodium carboxymethyl cellulose, ascorbic acid, lactose, talc, magnesium stearate, chitosan or carbomer.
Preferably, the quick release component comprises any one or a combination of at least two of lactose, adipic acid, sodium bicarbonate or corn starch.
In a second aspect, the present invention provides a method of preparing a probiotic for ameliorating vaginal infections according to the first aspect, the method comprising:
(1) Mixing the activated and fermented strain with a protective agent, and freeze-drying to obtain composite probiotic particles;
(2) Mixing the composite probiotic granule with the slow release component, granulating, mixing with the quick release component, and tabletting.
In a third aspect, the present invention provides the use of a probiotic for the amelioration of a vaginal infection according to the first aspect in the manufacture of a product for the amelioration of a vaginal infection.
Compared with the prior art, the invention has the following beneficial effects:
The invention creatively discovers that the related combination of lactobacillus casei LC16 strain and lactobacillus acidophilus LA18 strain can significantly improve the vulvar tract infection, and the effect is larger than that of single strain application. The specific expression is as follows: (1) effective inhibition of pathogenic bacteria growth; (2) a reduction in the inflammatory response of the mouse vulva tract; (3) Reduces the serum inflammatory factors IL-1 beta, TNF-alpha and IL-6 level of BV mice.
Drawings
Figure 1 is a graph of the probiotic tablet swelling index over 24 hours.
FIG. 2 shows the results of bacterial viability tests in different environments.
Detailed Description
The technical scheme of the invention is further described by the following specific embodiments. It will be apparent to those skilled in the art that the examples are merely to aid in understanding the invention and are not to be construed as a specific limitation thereof.
The Lactobacillus casei Lactobacillus casei LC strain related in the following is classified and named as Lactobacillus casei Lactobacillus casei, the preservation number is CGMCC No.24110, the preservation date is 2021, 12 and 15, the preservation unit is China general microbiological culture Collection center, and the preservation address is North Star Xila No.1 and 3 in the Korean region of Beijing city.
Lactobacillus acidophilus Lactobacillus acidophilus LA strain related in the following is classified and named Lactobacillus acidophilus LA, the preservation number is CCTCC NO: M2022572, the preservation date is 2022, 05 and 09, the preservation unit is China center for type culture Collection, and the preservation address is China, university of Wuhan.
Alginate, corn starch, ascorbic acid, stearic acid, sodium carboxymethylcellulose, ascorbic acid, lactose, talc, magnesium stearate, chitosan, adipic acid, sodium bicarbonate were all purchased from the Ming Xin chemical plant.
Acetic acid, and bovine serum albumin, propylene glycol, lactic acid were all purchased from Sigma.
Example 1
The embodiment provides a composite probiotic fermentation liquor, wherein the ratio of the viable count of lactobacillus casei LC16 strain to lactobacillus acidophilus LA18 strain in the fermentation liquor is 4:1, and the total viable count is 1X 10 9 CFU/mL.
The preparation method comprises the following steps:
inoculating Lactobacillus casei LC16 strain and Lactobacillus acidophilus LA18 strain into MRS culture medium, culturing at 37deg.C for 24h, adjusting viable count with sterile physiological saline, and mixing.
Example 2
The embodiment provides a composite probiotic fermentation liquor, wherein the ratio of the viable count of lactobacillus casei LC16 strain to lactobacillus acidophilus LA18 strain in the fermentation liquor is 1:1, and the total viable count is 1X 10 9 CFU/mL.
The preparation method is described in example 1.
Example 3
The embodiment provides a composite probiotic fermentation liquor, wherein the ratio of the viable count of lactobacillus casei LC16 strain to lactobacillus acidophilus LA18 strain in the fermentation liquor is 10:1, and the total viable count is 1X 10 9 CFU/mL.
The preparation method is described in example 1.
Comparative example 1
This comparative example provides a complex probiotic fermented liquor differing from example 1 only in that the lactobacillus casei LC16 strain was replaced with lactobacillus casei ATCC393 strain, the ratio of viable count of lactobacillus casei to lactobacillus acidophilus and total viable count were kept unchanged.
The preparation method is described in example 1.
Comparative example 2
This comparative example provides a complex probiotic fermented liquid differing from example 1 only in that lactobacillus acidophilus LA18 strain was replaced with lactobacillus acidophilus ATCC4356 strain, and the ratio of the viable count of lactobacillus casei to lactobacillus acidophilus and the total viable count were kept unchanged.
The preparation method is described in example 1.
Comparative example 3
The comparative preparation example provides a probiotic fermentation broth, wherein the viable count of the Lactobacillus casei LC16 strain in the fermentation broth is 1X 10 9 CFU/mL.
The preparation method is described in example 1.
Comparative example 4
The comparative preparation example provides a probiotic fermentation broth, and the viable count of Lactobacillus acidophilus LA18 strain in the fermentation broth is 1×10 9 CFU/mL.
The preparation method is described in example 1.
Test example 1
Influence of in vitro probiotic bacterial liquid on Escherichia coli
The E.coli standard strain (ATCC 25922) was inoculated into LB liquid medium, cultured at 37℃for 18 hours, centrifuged (3000 rpm,5 min) to collect the bacteria, and diluted 10-fold with PBS to prepare a desired bacterial suspension. Coli density was 10 4 CFU/mL.
1ML of 2 XPTY liquid medium was added to each well of a 24-well microplate, and 500. Mu.L of the E.coli suspension prepared as described above was inoculated. Then 500. Mu.L of the fermentation broths described in examples 1-3 and comparative examples 1-4, respectively, were added as experimental groups, and 500. Mu.L of physiological saline was used as a growth control. As a blank, 1mL of 2 XPTY liquid medium and 1mL of sterile physiological saline were additionally used. All experiments were performed in triplicate to ensure reproducibility. After anaerobic culture at 37℃for 48 hours, the number of viable E.coli cells was measured by plate counting, and the results are shown in Table 1.
TABLE 1
The in vitro experiment result shows that lactobacillus casei LC16 and lactobacillus acidophilus LA18 fermentation liquid have different degrees of inhibition effects on the growth of escherichia coli, and the LC16 and the LA18 can be matched with each other in the effect, and the combined use effect of the two is superior to that of the single use of the two strains.
Test example 2
BV model mouse model establishment and animal grouping and treatment
88 Female ICR mice were weighed and randomly divided into 11 groups (n=8), namely normal, estrogenic, model, antibiotic control (kanamycin 7 mg/kg daily for 6 days), probiotic (vaginal infusion 1mL daily for 6 days of fermentation broth treatment as described in examples 1-3 and comparative examples 1-4). Except for the normal group, the mice were subcutaneously injected with estrogen (estradiol benzoate) (0.1 mg/mouse) in the lower abdomen 3 days before the bacteria were inoculated in the vagina, and then 30 μl (10 8 CFU/mL) of an escherichia coli suspension was intravaginally injected into the mice of the model group, the antibiotic control group and the probiotic group each day, and BV models were established in succession of 5D.
The 3 clinical indexes of vaginal congestion, edema and hemorrhage of the mice are respectively scored: no congestion, edema, hemorrhage, infiltration of 0 minutes; a score of 0.1 with a slight effect; an effect of 0.2 points; severe 0.3 minutes; extremely severe 0.4 minutes. The severity of the inflammation and the therapeutic effect were then quantitatively analyzed.
TABLE 2
The results of relieving clinical symptoms of BV are shown in Table 1, and each clinical index of the mice in the normal group is in the normal range. The clinical symptoms of BV such as vaginal inflammation, mucosal congestion, secretion turbidity and the like of the mice in the model group are more obvious than those of the mice in other groups, and the clinical index score is increased than that of the mice in other groups. Chronic mild redness occurred in the vaginal opening of the estradiol group mice. After treatment of 3d, the vaginal redness and swelling and the secretion turbidity of the antibiotic group and the probiotic group are obviously improved. The lactobacillus casei LC16 and lactobacillus acidophilus LA18 fermentation liquid has a certain improvement effect on vaginal inflammation and secretion turbidity, and the two strains can be matched with each other in the effect.
Test example 3
The mice described in test example 2 were sacrificed after the end of the treatment and blood samples were collected. Serum is collected after centrifugation and detected by a double-antibody sandwich ABC-ELISA method according to ELISA kit instructions. The therapeutic effect of the antibiotic and probiotic bacterial suspensions was evaluated by detecting the expression levels of the inflammatory factors IL-1 beta, TNF-alpha and IL-6, and the results are shown in Table 3 without comparison in the B-group model set-up control group.
TABLE 3 Table 3
The colpitis caused by colibacillus has inhibiting effect on the expression of serum inflammatory factors IL-1 beta, TNF-alpha and IL-6 of BV mice, and the treatment effect of the group of the example 1 is superior to that of the antibiotics group and other groups. The combined use of lactobacillus casei LC16 and lactobacillus acidophilus LA18 fermentation liquid proves that the inflammatory reaction of BV mice can be reduced, and the two strains can be matched with each other in the effect.
Test example 4
Freeze-dried probiotic particles: mixing the fermentation broth obtained in example 1 with 0.5% acetic acid, 0.3% bovine serum albumin, 0.2% propylene glycol and 0.5% lactic acid to form granules, mixing with 1% alginate, emulsifying, and freeze drying.
A particulate substrate: 2% corn starch was prepared as a 10% paste for wetting the pellet mixture, and 2% sodium bicarbonate, 10% adipic acid, 10% corn starch and the balance lactose were added. The pellets were dried through a 40 mesh screen in an oven at 37 c until the weight was constant and after drying the pellets the agglomerates were removed through a 20 mesh screen.
The lyophilized probiotic particles and the granular matrix were then formulated into tablets in the proportions shown in table 4.
TABLE 4 Table 4
The preparation method comprises the following steps:
mixing the freeze-dried probiotic particles with ascorbic acid, lactose, sodium carboxymethyl cellulose, carbomer, chitosan, talcum powder, magnesium stearate and sodium bicarbonate, and granulating;
mixing the product obtained in the step (2) with a granular substrate, drying, and tabletting.
1. Tablet swelling index study
The preparation method of the simulated vaginal fluid comprises the following steps: 3.5 g/L NaCl, 1.4 g/L KOH, 0.2 g/L Ca (OH) 2, 2.0 g/L lactic acid, 1.0 g/L acetic acid, 0.2 g/L glycerol, 0.4 g/L urea, 5.0 g/L glucose, 0.02 g/L bovine serum albumin (all available from Shanghai, biotechnology Co., ltd.) the mixture was adjusted to pH 5.5.
The tablets were weighed (G1) and placed in a pre-weighed 200 mesh pore size stainless steel basket. The tablet-containing mesh was immersed in 25 mL simulated vaginal fluid in a glass beaker at 37 ℃ to expand the tablet. The stainless steel basket was removed from the beaker, and after removing excess water with filter paper, it was re-weighed (G2) at a predetermined time. The experiment was performed in three times and the final data of the swelling index (%) was calculated as follows:
As a result, as shown in fig. 1, it is necessary to conduct hydration studies to understand the swelling capacity of the tablet, which is directly related to the adhesion mechanism, because of the presence of the expandable bioadhesive polymer in the formulation, the stronger the swelling capacity, the better the tablet adhesion. The swelling index study proves that two different release curves, one is rapid bacterial release of the effervescent layer and the other is prolonged release of the slow-release matrix layer until the end of 24 hours, indicate that in the probiotic tablet for improving vaginal infection, the slow-release layer tablet swells and decreases due to small carbomer and chitosan addition amount in the formula tablet A after the proper blending proportion of sodium carboxymethyl cellulose is changed, and the slow-release layer tablet swells and increases when the carbomer and chitosan addition amount in the formula tablet B is relatively more.
2. Tablet adhesion study
New Zealand female white rabbits with dead weights greater than 2.5kg were euthanized, the vaginal tissue exposed at 2cm×2cm was dissected, washed with phosphate buffer (PBS, pH 6.8), and the mucosa integrity was maintained. The formulation B of the above experiment was selected and 2 drops of cyanoacrylate glue were added dropwise to the tablets, which were attached to a dynamometer support. The vaginal tissue was kept moist by washing with PBS solution, and then the free side of the tablet was moistened with simulated vaginal fluid, with the fingertip forced to bring its matrix layer into contact with the vaginal tissue mucosal surface for 20s. After 2 minutes the tablet forms an adhesion with the tissue. The adhesion strength of the tablets (the force required to detach the tablets from the rabbit vaginal mucosa) was measured using a dynamometer. The bioadhesive strength of the vaginal tablet was measured by a load cell in grams (g).
Adhesion (N) =bioadhesive strength (g) ×0.0098
The 4 formulation tablet adhesion test was 3 times, 3 tablets were randomly drawn each time for study.
The adhesion and adhesion time results are shown in Table 5.
TABLE 5
The adhesion of the tablet is very important, and the main function is to enable probiotics to be attached to the administration place for a long time and prevent the probiotics from being discharged without reaching the action effect. As the results in table 5 show, the results of the adhesion strength and adhesion are expressed as the average of three tablet measurements per completion of the test (x represents p < 0.05). To check whether the adhesion achieved guarantees a sustained tablet in the vagina until the probiotic load is completely released, it can be seen from table 5 that the adhesion test results for each tablet studied are similar. Formulation C had the lowest tablet adhesion, which may be related to the vaginal mucosa condition used, since in this case its surface was more even and smooth, formulation B had the highest tablet adhesion of 0.0983N. Formulation B tablets were selected for subsequent study.
The test also investigated the adhesion time, and the results of the analysis of the tablets of 4 different formulations were expressed as the time average of the measurements of the number of adhesion hours tested for each formulation. The adhesion time average was 12.75.+ -. 0.42 h. The results indicate that probiotic tablets to ameliorate vaginal infections can be administered 2 times daily.
3. Tablet probiotic storage and vitality test study
Group B probiotic formula tablets, 10 tablets each, were stored in a room temperature environment and in a closed desiccator for 60 days, respectively. The tablets were moisture protected in a closed desiccator using silica gel as the desiccant. During this time, the microorganisms in the tablets were cultured every 30 days to evaluate the viability of the bacteria. Specifically, the tablets were dissolved in 9mL of sterile purified water, 1mL was aspirated onto MRS solid medium at 37℃and incubated for 48h for plate counting.
The results are shown in FIG. 2, and the bacterial activity was measured after 60 days of storage at room temperature in a dry box. n=10, all points are expressed as mean±sd, stability studies indicate that formulation B tablet bacterial viability remains unchanged overall for 60 days at desiccator and room temperature.
The applicant states that the present invention is illustrated by the above examples as a probiotic agent for the amelioration of vaginal infections, and its preparation and use, but the invention is not limited to, i.e. it is not meant that the invention must be practiced in dependence upon the above examples. It should be apparent to those skilled in the art that any modification of the present invention, equivalent substitution of raw materials for the product of the present invention, addition of auxiliary components, selection of specific modes, etc., falls within the scope of the present invention and the scope of disclosure.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.

Claims (10)

1. The probiotics for improving vaginal infection is characterized in that strains in the probiotics comprise lactobacillus casei Lactobacillus casei LC strain with a preservation number of CGMCC No.24110 and lactobacillus acidophilus Lactobacillus acidophilus LA strain with a preservation number of CCTCC No. M2022572.
2. The probiotic for improving vaginal infection according to claim 1, wherein the ratio of viable count of lactobacillus casei Lactobacillus casei LC strain and lactobacillus acidophilus Lactobacillus acidophilus LA strain is (3-9): 1.
3. The probiotic for improving vaginal infection according to claim 1, wherein the viable count of lactobacillus casei Lactobacillus casei LC strain and lactobacillus acidophilus Lactobacillus acidophilus LA strain is not lower than 1 x 10 9 CFU/mL independently.
4. The probiotic for ameliorating vaginal infections of claim 1, wherein the dosage form of the probiotic comprises a tablet.
5. A probiotic for ameliorating a vaginal infection according to claim 1, wherein said probiotic further comprises a protective agent, a quick release component and a slow release component.
6. The probiotic agent for improving vaginal infections according to claim 5, characterized in that said protective agent comprises any one or a combination of at least two of acetic acid, bovine serum albumin, propylene glycol, lactic acid or alginate.
7. A probiotic for ameliorating a vaginal infection according to claim 5, wherein said retarded release ingredients comprise any one or a combination of at least two of stearic acid, sodium carboxymethylcellulose, ascorbic acid, lactose, talc, magnesium stearate, chitosan or carbomer.
8. The probiotic for ameliorating vaginal infections according to claim 5, wherein said quick-release ingredients comprise any one or a combination of at least two of lactose, adipic acid, sodium bicarbonate or corn starch.
9. A method of preparing a probiotic for ameliorating vaginal infections according to any of claims 1-8, characterized in that the method of preparation comprises:
(1) Mixing the activated and fermented strain with a protective agent, and freeze-drying to obtain composite probiotic particles;
(2) Mixing the composite probiotic granule with the slow release component, granulating, mixing with the quick release component, and tabletting.
10. Use of a probiotic according to any one of claims 1-8 for the preparation of a product for ameliorating vaginal infections.
CN202410575027.7A 2024-05-10 2024-05-10 Probiotic agent for improving vaginal infection and preparation method and application thereof Active CN118126910B (en)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140234259A1 (en) * 2012-12-11 2014-08-21 Suzanah Juras Natural Intra-Vaginal Inserts to Control Imbalanced pH
CN114634901A (en) * 2022-05-18 2022-06-17 微康益生菌(苏州)股份有限公司 Lactobacillus casei LC16 for promoting bone health and culture method and application thereof
CN114990030A (en) * 2022-07-18 2022-09-02 微康益生菌(苏州)股份有限公司 Lactobacillus acidophilus LA18 and application thereof in preparing product for regulating intestinal flora or immunoregulation
CN115768452A (en) * 2020-04-28 2023-03-07 伊比利亚护理健康有限公司 Coriolus versicolor extract for treating vaginal or cervical disorders caused by infectious pathogens

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140234259A1 (en) * 2012-12-11 2014-08-21 Suzanah Juras Natural Intra-Vaginal Inserts to Control Imbalanced pH
CN115768452A (en) * 2020-04-28 2023-03-07 伊比利亚护理健康有限公司 Coriolus versicolor extract for treating vaginal or cervical disorders caused by infectious pathogens
CN114634901A (en) * 2022-05-18 2022-06-17 微康益生菌(苏州)股份有限公司 Lactobacillus casei LC16 for promoting bone health and culture method and application thereof
CN114990030A (en) * 2022-07-18 2022-09-02 微康益生菌(苏州)股份有限公司 Lactobacillus acidophilus LA18 and application thereof in preparing product for regulating intestinal flora or immunoregulation

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