CN119792358A - LA® gynecological antibacterial gel for improving vaginitis inflammatory response and vaginal environment and its use - Google Patents

Abstract

The present invention relates to the field of biomedical technology, and in particular to a method for improving the inflammatory response of vaginitis and the vaginal environment. Gynecological antibacterial gel and its use. The present invention provides a postbiotic composition, which comprises an inactivated bacterial body of a probiotic composition and its metabolites; the probiotic composition comprises Lactobacillus paracasei ProSci-92 and Lactobacillus rhamnosus ProSci-109; the preservation number of Lactobacillus paracasei ProSci-92 is CGMCC No.26526; the preservation number of Lactobacillus rhamnosus ProSci-109 is CGMCC No.25449; the postbiotic composition can effectively alleviate or treat vaginitis inflammatory response and improve the vaginal environment.

Description

LA ® gel for improving colpitis reaction and vaginal environment and application thereof

Technical Field

The invention relates to the technical field of biological medicine, in particular to a medicine for improving colpitis reaction and vaginal environmentGynecological antibacterial gel and its application are provided.

Background

Mixed vaginitis refers to a local environment where at least two pathogens exist simultaneously and separately or cooperatively to cause abnormal vagina, namely local manifestation of dysbacteriosis of vagina, and few or most of single pathogen infection in clinical practice are mixed infection. Common types of vaginal infections in women of childbearing age mainly include trichomonas vaginitis, bacterial vaginosis, and vulvovaginal candidiasis. In the current clinical work, there is still a phenomenon of inadequate knowledge of the mixed vaginitis and inaccurate diagnosis, which also results in various degrees of over-treatment or under-treatment for mixed vaginitis.

The principle of colpitis treatment is mainly to eliminate inducement, select local or whole body application of antifungal according to the condition of patients, and combine antifungal with antidrip agent and gel disinfectant to avoid feeding irritating food. However, the medicines kill pathogenic bacteria and destroy the vagina environment such as the pH value of the vagina, so that the normal vagina flora with self-cleaning function is killed, and therefore, temporary relief can be only achieved, and new rounds of infection quickly occur due to the loss of the protection of the normal flora, so that lasting is repeated. Therefore, there is a need to find products that are effective in slowing or treating the response to colpitis, improving the vaginal environment, and reducing the number of persistent iterations.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a composition for improving the response of colpitis and the vaginal environmentGynecological antibacterial gel and application thereofThe gynecological antibacterial gel can effectively slow down or treat colpitis reaction, improve the vaginal environment and reduce persistent repetition.

For this purpose, the invention provides the following technical scheme:

embodiments of the present disclosure relate to a metacomposition, the metacomposition comprises inactivated thalli of the probiotic composition and metabolites thereof;

The probiotic composition comprises ProSci-92 of Lactobacillus paracasei (Lacticaseibacillusparacasei) and ProSci-109 of Lactobacillus rhamnosus (Lactobacillus rhamnosus), wherein the preservation number of the ProSci-92 of the Lactobacillus paracasei is CGMCC No.26526, and the preservation number of the ProSci-109 of the Lactobacillus rhamnosus is CGMCC No.25449.

As used herein, a "metabolite" includes any component, compound, substance, or by-product (including but not limited to small molecule secondary metabolites) produced by the probiotic compositions described herein that has any of the beneficial effects described in the present disclosure. The beneficial effects include relieving or treating symptoms such as vaginal itching, vaginal pain, abnormal vaginal secretion and the like, shortening the duration of symptoms, effectively regulating the vaginal microenvironment of a patient, reducing inflammation and the like.

In some embodiments, the total number of cells in the metazoan composition is greater than or equal to 1.0X10 ⁶/g, more preferably, the total number of cells in the metazoan composition is greater than or equal to 2.0X10 ¹¹/g.

In some embodiments, the probiotic composition is inoculated with live bacteria of the lactobacillus paracasei and the lactobacillus rhamnosus in a ratio of 1:1, and fermented to prepare the metacomposition.

The embodiment disclosed by the invention relates to a preparation method of the metacomposite, which comprises the following steps:

Inoculating living bacteria of the probiotic composition into a fermentation medium for culturing to obtain fermentation liquor, and inactivating the fermentation liquor to obtain the metazoan composition.

In some embodiments, the live bacteria of the probiotic composition are inoculated in the fermentation medium in an amount of not less than 1 x 10 ⁶ CFU/mL.

In some embodiments, the probiotic composition, upon inoculation, has a live bacteria ratio of 1:1 of lactobacillus paracasei to lactobacillus rhamnosus.

In some embodiments, the total number of cells in the metazoan composition is not less than 1.0X10 ⁶/g.

In some embodiments, the fermentation broth is subjected to an inactivation treatment, further comprising a step of drying, which is spray drying, freeze drying, or spray freeze drying.

In some embodiments, the fermentation medium may be a common medium, more preferably a fermentation medium widely used for lactobacillus, such as MRS liquid medium, modified BHI liquid medium, etc., and the components of the fermentation medium may also be a medium comprising defatted soybean powder, defatted milk powder, and water.

In some embodiments, the components of the fermentation medium comprise 0.5-2% of defatted soybean flour and 2-10% of defatted milk powder by mass percent, and the balance is water.

Embodiments of the present disclosure relate to the use of the metacomposition comprising:

(1) Use in the preparation of a product for ameliorating or treating vaginitis;

(2) Use in the preparation of a product for improving the vaginal environment;

(3) Use in the preparation of a product for alleviating or treating vaginal itching;

(4) Use in the manufacture of a product for alleviating or treating vaginal pain;

(5) Use in the preparation of a product for alleviating or treating vaginal discharge abnormalities;

(6) The application of the composition in inhibiting colpitis pathogenic bacteria or preparing products for inhibiting colpitis pathogenic bacteria.

In some embodiments, the vaginitis is a mixed vaginitis.

In some embodiments, the product comprises a cosmetic, pharmaceutical, or sanitizing product.

In some embodiments, the sanitary disinfecting article comprises a private care solution, a private care wet wipe, a private care gel, a private care suppository, or a private care spray;

or the medicine comprises an external medicine or an oral medicine, wherein the external medicine comprises cream (cream), ointment, lotion, suppository, tablet, capsule, gel or skin patch, and the oral medicine comprises injection, tablet, capsule, granule or powder;

or, the cosmetic comprises private care liquid, private care wet tissue, private care gel, private care suppository or private care spray.

Embodiments of the present disclosure relate to a method for improving the response to colpitis and the vaginal environmentThe gynecological antibacterial gel comprises the metagen composition.

In some embodiments, the amount of the post-natal composition added in the gynecological antibacterial gel is more than or equal to 0.01% by mass, for example 0.01%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%,45%,50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100% or a range between any two of the above values.

In a preferred embodiment, the added amount of the metacomposition is 5 to 70% by mass, for example, 5%, 7%, 9%, 11%, 13%, 15%, 17%, 19%, 21%, 23%, 25%, 27%, 30%, 34%, 38%, 43%, 48%, 52%, 57%, 60%, 62%, 65%, 67%, 70% or a range between any two of the above values. In a preferred embodiment, the added amount of the metazoan composition is 9-67% by mass.

In some embodiments, theThe gynecological antibacterial gel also comprises acceptable auxiliary materials. The auxiliary materials can be auxiliary materials acceptable in the fields of medicines, cosmetics or sanitary and disinfecting products.

In some embodiments, theThe gynecological antibacterial gel can be cosmetics, medicines or sanitary and disinfecting products. In a preferred embodiment, the gynecological antibacterial gel comprises, by weight, 50-170 parts of matrine, 60-150 parts of fructus cnidii, 0.7-6.3 parts of biological lysozyme, 0.5-4.2 parts of lactoferrin, 57-139 parts of metazoan composition, 3.8-10.1 parts of licorice extract, 2.6-7.4 parts of dandelion extract, 1.9-6.7 parts of honeysuckle extract, 29-73 parts of aloe, 1.1-4.5 parts of borneol, 1.7-5.2 parts of hyaluronic acid, 0.0003-0.001 parts of nano silver and 0.17-3.88 parts of carbomer;

Optionally, the The gynecological antibacterial gel also comprises water which is added to 1000 parts by weight.

The invention also provides a method for improving colpitis reaction and vaginal environmentThe preparation method of the gynecological antibacterial gel comprises the following steps:

weighing according to the formula, and preparing into the final product with or without adjuvants Gynecological antibacterial gel;

Optionally, the method comprises the following steps:

According to the formula, fructus cnidii, licorice extract, dandelion extract and honeysuckle extract are added into water for decoction for 1-4 times, the added water amount is 5-10 times of the total weight of the raw materials, the decoction temperature is 80-100 ℃, the time is 1-5 hours, and the filtrate is combined, filtered and concentrated to obtain concentrated solution;

adding carbomer into water according to a formula, soaking for dissolution, and filtering to obtain a first filtrate;

mixing matrine, borneolum Syntheticum, the metagen composition, biological lysozyme, lactoferrin and hyaluronic acid according to the formula, dissolving in water, and filtering to obtain a second filtrate;

According to the formula, the liquid obtained by crushing and filtering nano silver and aloe, the first filtrate, the second filtrate and the concentrated solution are mixed, fully mixed, added with water to 1000 parts by weight and adjusted to pH 5-7.

The technical scheme of the invention has the following advantages:

1. The invention provides a metagen composition, which comprises inactivated thalli of a probiotic composition and metabolites thereof, wherein the probiotic composition comprises ProSci-92 of Lactobacillus paracasei (Lacticaseibacillus paracasei) and ProSci-109 of Lactobacillus rhamnosus (Lactobacillus rhamnosus), the storage number of the ProSci-92 of the Lactobacillus paracasei is CGMCC No.26526, the storage number of the ProSci-109 of the Lactobacillus rhamnosus is CGMCC No.25449, and the metagen composition can be used for relieving or treating symptoms such as vaginal itching, vaginal pain, abnormal vaginal secretion and the like, shortening the duration of symptoms, effectively regulating the vaginal microenvironment such as pH value of a patient, avoiding affecting normal vaginal flora with self-cleaning function, further reducing the recurrence rate of colpitis, and can effectively slow down or treat colpitis inflammatory reaction and vaginal environment.

2. The invention provides a method for improving colpitis reaction and vaginal environmentGynecological antibacterial gel, saidThe gynecological antibacterial gel comprises, by weight, 50-170 parts of matrine, 60-150 parts of fructus cnidii, 0.7-6.3 parts of biological lysozyme, 0.5-4.2 parts of lactoferrin, 57-139 parts of metazoan composition, 3.8-10.1 parts of licorice extract, 2.6-7.4 parts of dandelion extract, 1.9-6.7 parts of honeysuckle extract, 29-73 parts of aloe, 1.1-4.5 parts of borneol, 1.7-5.2 parts of hyaluronic acid, 0.0003-0.001 parts of nano silver and 0.17-3.88 parts of carbomer, and the invention verifies that the metazoan composition is prepared by the method of the inventionGynecological antibacterial gel, compared with other metazoan compositionsGynecological antibacterial gel prepared from metazoan composition of the inventionThe gynecological antibacterial gel is more remarkable in treating or relieving symptoms such as vaginal itching, vaginal pain, vaginal secretion abnormality and the like, can remarkably shorten the duration of symptoms, effectively regulate the vaginal microenvironment of a patient, and reduce inflammation, and shows that the inventionThe metazoan composition in the gynecological antibacterial gel has the obvious effect of synergistically improving colpitis reaction and vaginal environment with other components.

Preservation of biological materials

ProSci-92 of Lactobacillus paracasei (Lacticaseibacillus paracasei), designated Lacticaseibacillus paracasei in taxonomy, was deposited in China general microbiological culture Collection center (CGMCC No. 26526) on day 08 of 2023, and was deposited at Seattle No. 3 of Seattle No. 1 of Chaoyang district North Star in Beijing, and mail code 100101.

The ProSci-109 of Lactobacillus rhamnosus (Lactobacillus rhamnosus) has a taxonomy of Lactobacillus rhamnosus and is preserved in China general microbiological culture Collection center (CGMCC No. 25449) on the 08-01-2022 day, and has a preservation address of the No. 3 West Song 1 of the Chao Yangjingku area North Star of Beijing city and a post code of 100101.

Detailed Description

The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.

The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.

Matrine, CAS 519-02-8, commercial product with purity not less than 98%;

fructus Cnidii, chinese medicinal name, is dried mature fruit of fructus Cnidii Cnidium monnieri (L.) Cuss of Umbelliferae, and is commercially available;

Lactoferrin, a commercial product, has a purity of 95% or more;

hyaluronic acid, a commercial product, with purity not less than 96%;

borneol is natural borneol, and is a commercial product, and the purity of the borneol is 96% -98%;

aloe is commercially available (Aloe vera (l.) norm. F.) and is a herb of the africaceae family of liliaceae order.

Biosoluble enzymes were purchased from source leaf biotechnology limited;

the licorice extract was purchased from western Antian Guangdong Biotechnology Co., ltd;

Dandelion extract was purchased from baichuan Kang Ze biotechnology company;

The honeysuckle extract is purchased from Xian Changyue biotechnology Co., ltd;

nano silver was purchased from south kyotona biotechnology limited.

The modified BHI liquid culture medium comprises 10.0g of tryptone, 17.5g of beef heart extract powder, 5.0g of sodium chloride, 2.5g of disodium hydrogen phosphate, 2.0g of glucose and 1000ml of distilled water, and is additionally added with yeast extract with the final concentration of 10% by mass, maltose with the final concentration of 0.1% by mass and glucose with the final concentration of 0.1% by mass, and the pH value is regulated to 7.4. Sterilizing at 121deg.C for 15min. Before use, sterile fetal bovine serum was added to the medium at a final concentration of 10% by mass.

Example 1 acquisition of Lactobacillus paracasei ProSci-92

Taking a natural fermented acid camel milk from Ala as a sample, sucking 0.5mL of the sample, adding the sample into 5mL of MRS liquid culture medium (purchased from Beijing Luqiao technical Co., ltd.) and carrying out anaerobic culture at 37 ℃ for 24 hours for enrichment to obtain an enriched sample; absorbing 0.5mL of enriched sample, adding the sample into 4.5mL of sterile physiological saline to obtain 10 ^-1 diluent, repeating the operation, sequentially carrying out gradient dilution by using 10 ^-1 diluent to obtain 10 ^-2、10^-3、10^-4、10^-5、10^-6 diluent, absorbing 100 mu L of gradient diluent, respectively coating on MRS solid medium, carrying out anaerobic culture for 48 hours at 37 ℃ to obtain colonies, selecting colonies with typical characteristics of Lactobacillus paracasei on the MRS solid medium, picking colonies with an inoculating loop, carrying out streaking on the colonies on the MRS solid medium, carrying out anaerobic culture for 48 hours at 37 ℃ to obtain purified single colonies, picking up the purified single colonies, respectively inoculating the purified single colonies into 5mL of MRS liquid medium, carrying out anaerobic culture for 24 hours at 37 ℃ to obtain bacterial solutions, carrying out gram staining, bacterial identification, physiological biochemical experiment and genome identification analysis after each bacterial strain number corresponds to each bacterial solution, selecting rod-shaped bacteria with typical characteristics of Lactobacillus paracasei to obtain ProSci-92, carrying out identification on ProSci-92 16SrDNA sequences of the strain, displaying the identification results as Lactobacillus paracasei, and carrying out China general preservation of the strain, namely, and carrying out CGMCC No. 26. China general preservation of the strain, and carrying out micro-biological preservation of the strain.

Example 2: metagen composition

The embodiment provides a metagen composition, the components of the metagen composition comprise the inactivated thallus and metabolite of lactobacillus paracasei ProSci-92 and the inactivated thallus and metabolite of lactobacillus rhamnosus ProSci-109, and the preparation method of the metagen composition is as follows:

Seed culture, namely inoculating 0.1mL of lactobacillus paracasei ProSci-92 (with the preservation number of CGMCC No. 26526) into an MRS liquid culture medium (purchased from Beijing Liqiao technology Co., ltd.) and culturing for 30 hours at 30 ℃ to obtain ProSci-92 seed liquid, inoculating 0.1mL of lactobacillus rhamnosus ProSci-109 (with the preservation number of CGMCC No. 25449) into the MRS liquid culture medium, and culturing for 30 hours at 30 ℃ to obtain ProSci-109 seed liquid;

Inoculating the ProSci-92 seed solution and the ProSci-109 seed solution into a fermentation medium, culturing for 30 hours at 30 ℃ after the total viable count of the ProSci-92 and the ProSci-109 bacteria is 2 multiplied by 10 ⁶ CFU/mL to obtain a fermentation broth, wherein the ratio of the viable count of the ProSci-92 bacteria to the ProSci-109 bacteria is 1:1;

performing post-treatment, namely sterilizing and inactivating the fermentation liquor at 85 ℃ for 15min to obtain sterilized and inactivated fermentation liquor, and performing spray drying on the sterilized and inactivated fermentation liquor to obtain a metaplasia composition (the total number of thalli of the lactobacillus paracasei ProSci-92 and the lactobacillus rhamnosus ProSci-109 in the metaplasia composition is more than or equal to 2.0x10 ¹¹/g);

The preparation method of the fermentation medium comprises the following steps:

the fermentation medium comprises 1% of defatted soybean powder, 8% of defatted milk powder and the balance of water by mass percent, wherein the components of the fermentation medium are mixed in proportion, and then are subjected to 58 ℃ material melting (namely stirring at 150 rpm) for 15min to obtain a feed liquid, the feed liquid is homogenized once under 58 ℃ under the primary pressure of 19Mpa and the secondary pressure of 5.0Mpa to obtain a homogenized feed liquid, the homogenized feed liquid is sterilized at 93 ℃ for 30min to obtain a sterilized feed liquid, and the sterilized feed liquid is cooled to 35 ℃ to obtain the fermentation medium.

Example 3: Gynecological antibacterial gel

The embodiment provides gynecological antibacterial gel, which comprises the following raw materials:

110g of matrine, 100g of fructus cnidii, 3.5g of biological lysozyme, 2.9g of lactoferrin, 115g of metazoan composition in example 2, 7g of licorice extract, 7g of dandelion extract, 4.3g of honeysuckle extract, 65g of aloe, 2.8g of borneol, 3.5g of hyaluronic acid, 0.0008g of nano silver and 2g of carbomer.

The preparation method comprises the following steps:

(1) Mixing fructus Cnidii, glycyrrhrizae radix extract, herba Taraxaci extract and flos Lonicerae extract according to the formula, decocting with water for 3 times at 100deg.C for 3 hr each time, mixing filtrates, filtering, concentrating to obtain concentrated solution, and standing.

(2) And placing carbomer in distilled water, soaking and dissolving, and filtering to obtain a first filtrate for later use.

(3) Mixing matrine, borneolum Syntheticum, the metazoan composition, biological lysozyme, lactoferrin, and hyaluronic acid, dissolving in distilled water, and filtering to obtain second filtrate for use.

(4) Mixing the pulverized and filtered solution of nano silver and aloe (leaf), the first filtrate, the second filtrate and the concentrated solution, fully mixing, adding water to 1000g, adjusting pH to 6.0+ -0.2, and canning.

Example 4: Gynecological antibacterial gel

The embodiment provides gynecological antibacterial gel, which comprises the following raw materials:

50g of matrine, 150g of fructus cnidii, 0.7g of biological lysozyme, 4.2g of lactoferrin, 57g of the metazoan composition in example 2, 10.1g of licorice extract, 2.6g of dandelion extract, 6.7g of honeysuckle extract, 29g of aloe, 1.1g of borneol, 5.2g of hyaluronic acid, 0.0003g of nano silver and 3.88g of carbomer.

The preparation method comprises the following steps:

(1) Mixing fructus Cnidii, glycyrrhrizae radix extract, herba Taraxaci extract and flos Lonicerae extract according to the formula, decocting with water for 1 time at 80deg.C for 5 hr, mixing filtrates, filtering, and concentrating to obtain concentrated solution.

(2) And placing carbomer in distilled water, soaking and dissolving, and filtering to obtain a first filtrate for later use.

(3) Mixing matrine, borneolum Syntheticum, the metazoan composition, biological lysozyme, lactoferrin, and hyaluronic acid, dissolving in distilled water, and filtering to obtain second filtrate for use.

(4) Mixing the pulverized and filtered solution of nano silver and aloe (leaf), the first filtrate, the second filtrate and the concentrated solution, fully mixing, adding water to 1000g, adjusting pH to 7.0+ -0.2, and canning.

Example 5: Gynecological antibacterial gel

The embodiment provides gynecological antibacterial gel, which comprises the following raw materials:

170g of matrine, 60g of fructus cnidii, 6.3g of biological lysozyme, 0.5g of lactoferrin, 139g of metazoan composition in example 2, 3.8g of licorice extract, 7.4g of dandelion extract, 1.9g of honeysuckle extract, 73g of aloe, 4.5g of borneol, 1.7g of hyaluronic acid, 0.001g of nano silver and 0.17g of carbomer.

The preparation method comprises the following steps:

(1) Mixing fructus Cnidii, glycyrrhrizae radix extract, herba Taraxaci extract and flos Lonicerae extract according to the formula, decocting with water for 4 times at 90deg.C for 1 hr each time with water amount of 5 times of the total weight of the above materials, mixing filtrates, filtering, and concentrating to obtain concentrated solution.

(2) And placing carbomer in distilled water, soaking and dissolving, and filtering to obtain a first filtrate for later use.

(3) Mixing matrine, borneolum Syntheticum, the metazoan composition, biological lysozyme, lactoferrin, and hyaluronic acid, dissolving in distilled water, and filtering to obtain second filtrate for use.

(4) Mixing the pulverized and filtered solution of nano silver and aloe (leaf), the first filtrate, the second filtrate and the concentrated solution, fully mixing, adding water to 1000g, regulating pH to 5.0+ -0.2, and canning.

Experimental example 1 determination of the acid producing Capacity of Lactobacillus

The lactobacillus rhamnosus R9639, the lactobacillus rhamnosus LGG and the lactobacillus rhamnosus GR-1 are commercially available, the rest 6 lactobacillus strains are derived from Beijing family Tuotong biotechnology Co., ltd, and specific strain information is shown in Table 1.

The experimental strains are respectively inoculated into MRS liquid culture medium by an inoculating loop, subjected to anaerobic culture at 37 ℃ for 24 hours, and activated for three generations. The culture broth of the third generation strain was centrifuged at 8000g for 10min at 4℃and filtered through a 0.22 μm sterile filter to give a cell-free supernatant. The pH value of the supernatant of lactobacillus cultured for 24 hours was measured by using the pH value measured by the non-inoculated MRS medium as a control, and the acid production capacity of lactobacillus was measured.

TABLE 1 determination of the acid generating ability of Lactobacillus

Lactobacillus strain	PH value of
		Non-inoculated MRS culture medium	6.20±0.20
Lactobacillus rhamnosus R9639	3.91±0.01
		Lactobacillus rhamnosus ProSci-109	3.67±0.02
Lactobacillus rhamnosus LR-82	4.02±0.00
		Lactobacillus paracasei ProSci-92	3.86±0.01
Lactobacillus paracasei PC-101	4.16±0.03
		Lactobacillus paracasei Probiosci-101	3.97±0.00
Lactobacillus paracasei LC-01	4.02±0.01
		Lactobacillus rhamnosus LGG	3.88±0.00
Lactobacillus rhamnosus GR-1	3.90±0.01

The pH of the vaginal environment of healthy women is between 3.8 and 4.5, mainly due to the fact that lactobacillus colonized in the vagina uses glucose in the vaginal epithelium to produce large amounts of organic acids. A large number of in vitro studies prove that the acidic environment of the vagina is unfavorable for the colonization and growth of pathogenic bacteria related to vaginal diseases. The acid production capacity of the above 9 strains was measured in this experiment, and the results are shown in Table 1. The results show that the acid production capacity of each lactobacillus strain is not significantly different, the pH value is between 3.60 and 4.20 and is in the range of the pH of the vaginal environment of healthy women, and the 9 lactobacillus strains used in the experiment have the potential capacity of improving the pH of the vaginal environment of women with bacterial vaginitis.

Experimental example 2 Effect of inactivated Lactobacillus on the proliferation Activity of Gardnerella

The lactobacillus strain in the experimental example 1 is respectively inoculated in MRS liquid culture medium, anaerobic culture is carried out for 24 hours at 37 ℃, the fermentation liquor is sterilized and inactivated for 15 minutes at 85 ℃, the sterilized and inactivated fermentation liquor is obtained, and the number of the thalli is adjusted to be 1.0X10 ⁸ per mL for standby. The gardnerella strain is inoculated in a modified BHI liquid culture medium, anaerobic culture is carried out for 24h at 37 ℃, centrifugation is carried out for 5min at 4 ℃, the supernatant is discarded, the thalli are washed by PBS solution for 3 times, then the thalli are added into the modified BHI liquid culture medium again, and the bacterial concentration is regulated to be 1.0X10 ⁸ CFU/mL for standby.

Mixing 0.5mL of lactobacillus inactivated bacterial liquid with the bacterial number of 1.0X10 ⁸/mL and 0.5mL of gardnerella bacterial liquid, setting a control group (without adding lactobacillus bacterial liquid), carrying out anaerobic culture at 37 ℃ for 4 hours, taking 0.2mL of mixed bacterial liquid, adjusting the concentration to 0.5 McMaillard turbidity standard, carrying out gradient dilution (the dilution multiple is 10 ^-9), inoculating the diluted bacterial liquid to an improved BHI agar plate, carrying out anaerobic culture at 37 ℃ for 24 hours, and calculating the viable count according to the gardnerella bacterial colony number.

TABLE 2 Effect of inactivated Lactobacillus on Gardnerella proliferation Activity

The influence of 9 inactivated lactobacillus strains on the proliferation activity of gardnerella vaginalis is detected, the activity of gardnerella vaginalis is inhibited to different degrees, wherein lactobacillus rhamnosus ProSci-109 and lactobacillus paracasei ProSci-92 with better inhibition effect further show that the inactivated lactobacillus strains have better antagonistic effect on gardnerella vaginalis and can be used as potential probiotics for treating colpitis.

Experimental example 3 Effect of inactivated Lactobacillus combination on the proliferation Activity of Gardnerella vaginalis

Experiments on the influence of different inactivated lactobacillus combinations on the proliferation activity of gardnerella vaginalis are prepared according to the method of experiment example 2, wherein the difference is that in the lactobacillus combinations, the inoculation proportion of each lactobacillus strain is that the viable bacteria number is inoculated in a proportion of 1:1, and finally the fermentation liquor after sterilization and inactivation is obtained, and the total number of the bacteria is adjusted to be 1.0x10 ⁸/mL for standby. The results are shown in Table 3.

TABLE 3 Effect of inactivated Lactobacillus on Gardnerella proliferation Activity

As can be seen from the above table, the effect of the inactivated lactobacillus composition of group 31 comprising lactobacillus rhamnosus + lactobacillus rhamnosus, lactobacillus rhamnosus + lactobacillus paracasei and lactobacillus paracasei + lactobacillus paracasei on the proliferation activity of gardnerella vaginalis was examined altogether, the activity of gardnerella vaginalis being inhibited to a different extent, wherein the most significant inhibition is the inactivated lactobacillus composition of the combination of lactobacillus rhamnosus ProSci-109+ lactobacillus paracasei ProSci-92, further illustrating that the inactivated thallus of the combination of lactobacillus rhamnosus ProSci-109+ lactobacillus paracasei ProSci-92 of the present invention and its metabolites have a better antagonistic effect against gardnerella vaginalis.

Experimental example 4 adding the metacompositionInfluence of gynecological antibacterial gel on colpitis reaction and vaginal environment

1 Experimental method

120 Cases of vaginitis patients collected and treated by auxiliary hospitals of university of Mongolian medical science in 2 months of 2023 to 5 months of 2023 are selected, and the ages of the patients are 25-40, and are randomly divided into three groups:

40 metagroups of the metagroup A, The combination of the post-natal composition of Lactobacillus rhamnosus ProSci-109+Lactobacillus paracasei PC-101 in Experimental example 3 was prepared as in example 2; the preparation method of the gynecological antibacterial gel is the same as that of example 3;

40 metagroups of the metagroup B, The post-element composition is the post-element composition of rhamnose cheese bacillus ProSci-109+paracasei cheese bacillus ProSci-92 prepared in example 2; the preparation method of the gynecological antibacterial gel is the same as that of example 3;

control group 40 cases, without added metazoan composition Gynecological antibacterial gel is prepared by the same method as in example 3, except that no metazoan composition is added.

Each group of the aboveGynecological antibacterial gel with specification of 3g×6 pieces/box.

Among the cases of the above groups, the metazoan group A had an average age (31.90.+ -. 1.07), the metazoan group B had an average age (30.70.+ -. 1.16), the control group had an average age (31.05.+ -. 2.15), and the case age difference between the three groups had no statistical significance (P > 0.05). Approved by the Independent Ethics Committee (IEC). The subject can clean hands and pudendum with warm water before using during menstrual periodOne gynecological antibacterial gel product is prepared by slightly lifting buttocks in supine position, slowly inserting gel catheter carrying gel liquid into deep vagina, pushing into pudendum with push rod, keeping posture for about 15 minutes, and continuously using for 28 days, one for each night. During exhort family members must not perform sexual life and other vaginal invasive procedures and must not use other antibiotics, probiotics, etc. during treatment. The difference is that between the three groupsWhether the post-metazoan composition or the post-metazoan composition is added into the gynecological antibacterial gel is different.

Three groups of patients were compared with general data, and differences were statistically significant (P > 0.05). The exclusion criteria were ① for infection-induced vaginitis, ② for 6 months before treatment with the relevant hormone drugs, ③ for allergy to the treatment.

The observation index is ① curative effect observation, and the effect before treatment (day 0) and after treatment (day 28) are compared. The clinical treatment effects are analyzed by combining the symptom manifestation of a patient, the condition of vaginal secretion and the like, and the clinical treatment effects are divided into three cases of obvious effect (uncomfortable symptoms such as pruritus and the like completely disappear, the mucous membrane of the outer wall of the vagina is light pink, the cleanliness of secretion is I-II degrees), the effect (the uncomfortable symptoms such as pruritus and the like are obviously relieved, the congestion problem of the mucous membrane of the outer wall of the vagina is improved, the cleanliness of secretion is II-III degrees), and the effect (the uncomfortable symptoms are not obviously changed or even aggravated), wherein the total effective rate is equal to the effective rate+the effective rate, the effective rate is equal to the effective rate/the total rate multiplied by 100 percent. ② Symptoms and signs duration symptoms and signs observations include vaginal itching, vaginal pain, abnormal secretions. And observing the disease symptom change conditions of three groups of patients before and after treatment (day 0 and day 28), and setting scores of 0-10, wherein 0 is asymptomatic, 10 is symptomatic, and the scores are positively correlated with symptoms. ③ And (5) detecting the pH value. The changes in vaginal pH before and after treatment (day 0 and day 28) were measured in three groups of patients using a test paper (merck SuPelcopH paper), and the measurement method was according to the instructions of the test paper. ④ And detecting inflammatory factors. Three groups of patients were drawn 3mL of fasting venous blood before and after treatment (day 0 and day 28), and the inflammatory factor levels were detected by ELISA, the indicators including tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), and hypersensitive C-reactive protein (hs-CRP). ⑤ Recurrence rate for 6 months, recording vaginal inflammation recurrence (at least one of clinical symptoms, pH value, and inflammatory factor is recovered before treatment), recurrence rate = number of recurrence/total number of recurrence x 100%.

The data processing method adopts SPSS25.0 for statistical analysis.

2 Experimental results

After treatment, the results are shown in Table 4 below, the total effective rate of the metatuple is above 95%, which is significantly higher than that of the control group, indicating the use of the added metatuple compositionThe gynecological antibacterial gel can effectively strengthen the overall treatment effect and accelerate the recovery of physical signs. Compared with the metagen A group, the metagen B group has higher total effective rate and higher effective rate, which indicates that the treatment effect of the metagen composition B group is obviously higher than that of the metagen A group.

Table 4 effects of patients versus [ examples (%) ]

The three groups of patients after treatment all had lower scores for each symptom than before treatment, and the metatuple was lower than the control group, while shortening the duration of the symptoms, see table 5. Further description of the use of added metacompositionsThe gynecological antibacterial gel can treat or relieve vaginal itching, vaginal pain, abnormal vaginal secretion and other symptoms, and shorten the duration of symptoms. Compared with the metagroup A, the metagroup B has lower score and shorter duration of each symptom after treatment and has different statistical significance, which indicates that the metagroup B has obviously better effect of treating the symptoms such as vaginal itching, vaginal pain, abnormal vaginal secretion and the like and shortening the duration of the symptoms than the metagroup A.

Table 5 patient symptom evaluation comparison

Note that differences in comparison to the control groups represent p <0.05, p <0.01, p <0.001, respectively, are statistically significant. In each index, p <0.05, p <0.01, p <0.001 are shown in #, #, and # # respectively, and the difference is statistically significant compared with the pre-treatment index.

After treatment, the vaginal pH was lower for all three groups of patients than before treatment, and the metazoan group was lower than for the control group, see table 6. The therapeutic effect is probably due to the fact that the metazoan composition contains bioactive substances such as organic acid, antibacterial peptide, short-chain fatty acid and the like, which inhibit the growth and proliferation of harmful flora in the vagina, promote the decomposition and conversion of glycogen to lactic acid in the vagina, provide favorable conditions for the growth of lactic acid bacteria, keep the acidic environment in the vagina stable and enhance the antibacterial capability.

Table 6 patient vaginal pH comparison

Note that differences in comparison to the control groups represent p <0.05, p <0.01, p <0.001, respectively, are statistically significant. In each index, p <0.05, p <0.01, p <0.001 are shown in #, #, and # # respectively, and the difference is statistically significant compared with the pre-treatment index.

After treatment, TNF- α, IL-6 and hs-CRP were lower in the three groups of patients than before treatment, and the metazoan index was lower in the control group, as shown in Table 7. As a further explanation of the present invention,The gynecological antibacterial gel has the effects of sterilizing and diminishing inflammation and improving colpitis inflammation, and the added metaplasia composition has the synergistic effect. Compared with the metagen A group, the metagen B group has lower TNF-alpha level after treatment and has differential statistical significance, which proves that the effects of sterilizing and diminishing inflammation and improving colpitis inflammation of the metagen B group are obviously better than the metagen A group.

TABLE 7 comparison of patient inflammation index

Note that differences in comparison to the control groups represent p <0.05, p <0.01, p <0.001, respectively, are statistically significant. Of the indices, ^#、^##、^### represents p <0.05, p <0.01, and p <0.001, respectively, and the differences are statistically significant compared with those before the treatment.

In the recurrence rate comparison, the recurrence rate of the metagroup subjects at 6 months of follow-up was less than 10%, while the recurrence rate of the control group was greater than 50%, and the metagroup recurrence rate was overall lower than that of the control group, further illustrating the addition of the metagroup compositionThe gynecological antibacterial gel has the effects of improving colpitis reaction and vagina environment effectively and permanently.

Table 8 patient recurrence rate comparison [ number of cases (%) ]

In conclusion, the metacomposition is addedThe gynecological antibacterial gel has definite effect of improving colpitis, can treat or relieve symptoms such as vaginal itching, vaginal pain, abnormal vaginal secretion and the like, shortens the duration of symptoms, effectively adjusts the vaginal microenvironment of a patient, reduces inflammation, and has remarkable effect.

It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.

Claims (10)

1. A metacomposition, characterized in that the metacomposition comprises inactivated cells of a probiotic composition and metabolites thereof;

The probiotic composition comprises ProSci-92 of Lactobacillus paracasei (Lacticaseibacillus paracasei) and ProSci-109 of Lactobacillus rhamnosus (Lactobacillus rhamnosus), wherein the preservation number of the ProSci-92 of the Lactobacillus paracasei is CGMCC No.26526, and the preservation number of the ProSci-109 of the Lactobacillus rhamnosus is CGMCC No.25449.

2. The metacomposition of claim 1, wherein the metaplastic comprises,

The total number of thalli in the metazoan composition is more than or equal to 1.0x10 ⁶/g;

optionally, the total number of thalli in the metazoan composition is not less than 2.0X10 ¹¹/g.

3. The metacomposition according to claim 1 or 2, wherein,

The probiotic composition has a ratio of viable count of Lactobacillus paracasei to Lactobacillus rhamnosus of 1:1 at the time of inoculation.

4. A process for the preparation of a metacomposition as claimed in any one of claims 1 to 3,

Inoculating living bacteria of the probiotic composition into a fermentation medium for culturing to obtain fermentation liquor, and inactivating the fermentation liquor to obtain the metazoan composition.

5. A metacomposition according to any one of claims 1-3 comprising the use of:

(1) Use in the preparation of a product for ameliorating or treating vaginitis;

(2) Use in the preparation of a product for improving the vaginal environment;

(3) Use in the preparation of a product for alleviating or treating vaginal itching;

(4) Use in the manufacture of a product for alleviating or treating vaginal pain;

(5) Use in the preparation of a product for alleviating or treating vaginal discharge abnormalities;

(6) The application of the composition in inhibiting colpitis pathogenic bacteria or preparing products for inhibiting colpitis pathogenic bacteria.

6. The use according to claim 5, wherein the vaginitis is mixed vaginitis;

and/or the product comprises a cosmetic, pharmaceutical or sanitary disinfecting product.

7. The use of claim 6, wherein the sanitary disinfecting article comprises a private care solution, a private care wet wipe, a private care gel, a private care suppository, a private care sanitary towel, or a private care spray;

Or the medicine comprises an external medicine or an oral medicine, wherein the external medicine comprises cream (cream), ointment, lotion, suppository, gel or skin patch, and the oral medicine comprises injection, tablet, capsule, granule or powder.

8. A composition for improving colpitis reaction and vaginal environmentGynaecological bacteriostatic gel, characterized in that it comprises a metaplastic composition according to any one of claims 1-3.

9. The method according to claim 8The gynecological antibacterial gel is characterized in that the added amount of the metazoan composition in the product is more than or equal to 0.01 percent by mass;

optionally, the added amount of the metagen composition is 5-70% by mass;

Optionally, the The gynecological antibacterial gel also comprises acceptable auxiliary materials, wherein the auxiliary materials can be acceptable auxiliary materials in the fields of medicines, cosmetics or sanitary disinfection products;

Preferably, the said The gynecological antibacterial gel comprises, by weight, 50-170 parts of matrine, 60-150 parts of fructus cnidii, 0.7-6.3 parts of biological lysozyme, 0.5-4.2 parts of lactoferrin, 57-139 parts of a metaplastic composition, 3.8-10.1 parts of licorice extract, 2.6-7.4 parts of dandelion extract, 1.9-6.7 parts of honeysuckle extract, 29-73 parts of aloe, 1.1-4.5 parts of borneol, 1.7-5.2 parts of hyaluronic acid, 0.0003-0.001 parts of nano silver and 0.17-3.88 parts of carbomer;

Optionally, the The gynecological antibacterial gel also comprises water which is added to 1000 parts by weight.

10. A method for improving colpitis response and vaginal environment as claimed in any one of claims 8 to 9The preparation method of the gynecological antibacterial gel is characterized by comprising the following steps:

weighing according to the formula, and preparing into the final product with or without adjuvants Gynecological antibacterial gel;

Optionally, the method comprises the following steps:

According to the formula, fructus cnidii, licorice extract, dandelion extract and honeysuckle extract are added into water for decoction for 1-4 times, the added water amount is 5-10 times of the total weight of the raw materials, the decoction temperature is 80-100 ℃, the time is 1-5 hours, and the filtrate is combined, filtered and concentrated to obtain concentrated solution;

adding carbomer into water according to a formula, soaking for dissolution, and filtering to obtain a first filtrate;

mixing matrine, borneolum Syntheticum, the metagen composition, biological lysozyme, lactoferrin and hyaluronic acid according to the formula, dissolving in water, and filtering to obtain a second filtrate;

According to the formula, the liquid obtained by crushing and filtering nano silver and aloe, the first filtrate, the second filtrate and the concentrated solution are mixed, fully mixed, added with water to 1000 parts by weight and adjusted to pH 5-7.