CN111759852A

CN111759852A - Pharmaceutical composition for vagina, pharmaceutical preparation, preparation method and application thereof

Info

Publication number: CN111759852A
Application number: CN202010774323.1A
Authority: CN
Inventors: 郝晓健; 王怡璐; 杨页多
Original assignee: Reindeer Beijing Management Consulting Co ltd
Current assignee: Reindeer Beijing Management Consulting Co ltd
Priority date: 2020-08-04
Filing date: 2020-08-04
Publication date: 2020-10-13

Abstract

The invention relates to a pharmaceutical composition for vagina, which comprises alkyl glycoside or glucoside substances or a mixture thereof as a vagina pathogen inhibitor, prebiotics for promoting the growth and space occupying effect of vagina probiotics and polycarbophil as a vagina mucosa protective film forming substance. Also relates to a vaginal pharmaceutical preparation which comprises the pharmaceutical composition and a pharmaceutically acceptable excipient. The pharmaceutical preparation is gel, ointment, foam, suppository, tablet, hard capsule and soft capsule for vagina. Also relates to the application of the pharmaceutical composition or the pharmaceutical preparation in preparing medicines for restoring the internal environment of the vagina, treating vaginitis and preventing the occurrence and the recurrence of vaginitis.

Description

Pharmaceutical composition for vagina, pharmaceutical preparation, preparation method and application thereof

Technical Field

The invention relates to a vaginal pharmaceutical composition and a pharmaceutical preparation thereof, wherein the vaginal pharmaceutical composition is prepared from alkyl glycoside or glucoside substances as vaginal pathogenic bacteria inhibitors, prebiotics for promoting the growth of vaginal probiotics and polycarbophil as a vaginal mucosa protective film forming substance in a specific proportion, and the vaginal pharmaceutical composition is used for restoring the internal environment of vagina, treating vaginitis and preventing the occurrence and the recurrence of vaginitis.

Background

Female genital tract infection caused by the internal environment disorder of the vagina is a common gynecological disease and affects the physical and mental health of women. According to statistics, the clinic treatment for infectious diseases accounts for more than half of the clinic patients, and the first 3 patients are vaginitis, cervicitis and pelvic inflammation. It follows that vaginitis is the most common gynaecological disease. When various vaginitis (including pure bacterial vaginitis, pure candida infection, trichomonas infection, mixed infection, atrophic/senile vaginitis and the like) occurs, the disappearance of the dominant bacterium lactobacillus and the increase of the pH value are accompanied in the vagina of a patient.

The pH value of the vagina of a female in normal reproductive age is 3.8-4.4, the acidity degree of the vagina is a primary mechanism for controlling the flora composition, and the bacteria planted in the vagina are determined to be acidophilic or acidproof under the general condition. The lactobacillus is beneficial to human body, is the dominant bacterial flora in the vagina of most healthy women and accounts for 95 percent of the bacteria in the vagina. The pH value of 66.2% of the female with lactobacillus as dominant flora is less than or equal to 4.5. Under normal state, a plurality of microorganisms exist in the vagina, wherein the dominant bacteria reduce the pH value of the vagina by producing lactic acid, thereby inhibiting the occurrence of vagina inflammation; and can compete for adhesion to produce H₂O₂And other inhibiting substances such as Lactoferrin, stimulating the immune system, etcThe overgrowth of Escherichia coli, Candida, Gardnerella vaginalis and Campylobacter bacteria is prevented, and the balance of vaginal microecology is maintained. The existence of the lactobacillus can generate biological inhibition, maintain the pH value of the vagina to be acidic, reduce the number of conditional pathogenic bacteria and maintain the normal physiological state of the vagina.

The traditional treatment method of the vaginitis comprises oral or vaginal administration of metronidazole, clindamycin and other antibiotics for treatment, and chitosan medical appliance products with bacteriostatic efficacy are also appeared in recent years, although the antibacterial and bacteriostatic method can effectively treat the vaginitis. However, after treatment, pathogenic bacteria and beneficial bacteria in the vagina are killed at the same time, so that the vagina is not in a healthy and normal state, but the phenomenon of 'empty city' in the vagina occurs, various bacteria can easily enter the vagina in a deficient state, and if the pathogenic bacteria happen to dominate again, vaginitis is relapsed. It is reported that 25% of patients relapse within 4 weeks after antibiotic treatment, while the long-term observed rate of relapse is greater than 70%. Research shows that the lactobacillus becomes dominant flora by adjusting the proportion of the flora in the vagina, the pH value of the vagina is reduced, the cure rate of vaginitis can be effectively improved, the recurrence rate is reduced, and no obvious side effect exists. Meanwhile, the lower pH value is beneficial to the growth of the vaginal dominant flora and the restoration of the internal environment, and the maintenance of the acidic environment in the vagina can improve the treatment effect of the antibiotics. Therefore, the recovery and maintenance of the internal environment of the vagina are better ways to treat and prevent relapse. And for pregnant women, the application of antibiotics for treatment has contraindication, and even the administration by a vaginal route needs to be careful, so the safety of the vaginal environment regulating preparation applied to the pregnant women has obvious advantages.

The research shows that although the single effect of inhibiting or killing vaginal pathogenic bacteria is obvious in recent period, the environment in the vagina is not recovered to a normal state, so that the vaginitis is easy to relapse; and the simple supplement of probiotics or prebiotics can promote the probiotics to take the advantage of the position, but needs a long time and the symptom is slowly relieved. In addition, the first-line treatment medicine applied at present is mainly antibiotic, and has application safety risk for people with high vaginitis incidence like pregnant and lying-in women. There is therefore a need for a safe and effective vaginal pharmaceutical composition that can simultaneously meet these needs.

Disclosure of Invention

In view of the above, one aspect of the present invention is to provide a vaginal pharmaceutical composition, which contains alkyl glycoside or glucoside compound or mixture thereof as vaginal pathogenic bacteria inhibitor, prebiotics for promoting the growth of vaginal probiotics, and polycarbophil as vaginal mucosa protective film forming substance.

In the pharmaceutical composition, the natural vaginal pathogenic bacteria inhibiting component and the substance for protecting and promoting the vaginal probiotics are used simultaneously and are in parallel, so that symptoms can be rapidly eliminated in a short time, the normal internal environment of the vagina is promoted to be recovered, and the vaginitis relapse is prevented. In addition, the vagina mucosa is stimulated by pathogenic bacteria to cause congestion and edema during inflammation, and the vagina mucosa protective agent can form a protective film on the surface of the mucosa, physically isolate the stimulus from the mucosa and give the mucosa sufficient repair time. Thus, the vaginal protection device meets the physiological requirements of the vagina in all aspects.

The component for inhibiting pathogenic bacteria in the pharmaceutical composition is alkyl glycoside or alkyl polyglycoside represented by lauric acid monoglyceride. The vaginal antibacterial composition has safe, efficient and broad-spectrum antibacterial action, is not limited by pH, still has better antibacterial effect in an acidic environment (in vagina), can inhibit common vaginal pathogenic bacteria (Gardner bacteria, Candida albicans, staphylococcus aureus and the like) and viruses (reducing susceptibility of viruses related to sexually transmitted diseases), and more importantly, does not influence probiotic flora (lactobacillus) in the vagina. In one embodiment, the alkyl glycoside or glucoside, or mixture thereof, is selected from the group consisting of monoglycerol laurate, glycerol derivatives of capric, caprylic and caproic acids, or mixtures thereof. In a preferred embodiment, it is selected from monoglycerol laurate.

Some special saccharides are substances which can stimulate the growth of endogenous probiotics (lactobacillus in vagina) and regulate the natural ecological microenvironment in the organism. Lactose is the main carbon source available to lactobacilli, and sucrose, fructo-oligosaccharide, gluco-oligosaccharide, etc. can also promote their growth. Studies have shown that the use of the family of glucooligosaccharides enables vaginal endogenous lactobacilli strains to accelerate growth without stimulating the growth of pathogenic flora (e.g. candida, escherichia coli, gardnerella, etc.). However, lactose is more readily available to lactobacilli than monosaccharides typified by glucose when compared to other sugars; compared with the culture medium containing other oligosaccharides (inulin, sucrose, fructose and the like), the culture medium has similar effect of promoting the growth of lactobacillus, and the pH value obtained when lactose is used as a substrate is the lowest and is the best carbon source for forming lactobacillus biofilms. The lactose is prompted to promote the growth of lactobacillus in the vagina, obtain an advantage and be more beneficial to the vagina to maintain a normal acidic environment. And researches show that the proliferation effect of the lactobacillus is improved along with the increase of the lactose concentration, so that the main component for regulating the internal environment of the vagina selected by the product is most preferably high-concentration lactose. In other embodiments, the prebiotic in the pharmaceutical composition that promotes the growth of vaginal probiotics (lactobacillus) may also be selected from one or more of sucrose, fructo-oligosaccharide, and malto-oligosaccharide.

In addition, the polycarbophil is used as a physical protective film forming agent for the vaginal mucosa, has good crosslinking capacity in the acidic environment of the vagina, and is more suitable for being applied to the vagina compared with other gel matrixes (such as carbomer and sodium hyaluronate).

In a preferred embodiment, the vaginal pharmaceutical composition comprises monoglycerol laurate, prebiotics and polycarbophil. In a more preferred embodiment, the vaginal pharmaceutical composition consists of monoglycerol laurate, prebiotics and polycarbophil. In another embodiment, the vaginal pharmaceutical composition comprises 0.1-10% by weight of an alkyl glycoside or a glucoside or mixture thereof, 5-95% by weight of a prebiotic and 1.5-10% by weight of polycarbophil. In a preferred embodiment, the vaginal pharmaceutical composition comprises 0.5-5% by weight of an alkyl glycoside or a glucoside or mixture thereof, 20-90% by weight of a prebiotic and 1.5-5% by weight of polycarbophil. In a more preferred embodiment, the vaginal pharmaceutical composition comprises 1-5% by weight of an alkyl glycoside or a glucoside or mixture thereof, 20-80% by weight of a prebiotic and 1.5-3% by weight of polycarbophil.

In a second aspect, the present invention provides a pharmaceutical vaginal formulation comprising the pharmaceutical vaginal composition of the present invention and a pharmaceutically acceptable excipient, wherein the pharmaceutical formulation has a pH of 3 to 4.5.

In one embodiment, the pharmaceutical formulation is a vaginal gel, vaginal ointment, vaginal foam, pessary, vaginal tablet, vaginal hard capsule, and vaginal soft capsule. In a preferred embodiment, the pharmaceutical formulation is a gel comprising a gel base, a thickener, an emulsifier, a surfactant, and a solvent. In another embodiment, the gel matrix is selected from one or more of carbomer, xanthan gum, alginate, chitosan, the thickener is selected from one or more of methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, cyanoethylcellulose, hydroxypropylcellulose and hydroxypropylmethylcellulose, the emulsifier is selected from one or more of polysorbate, polyglycerol ester, sodium stearoyl lactylate, calcium stearoyl lactylate, diacetyl tartaric acid monoglyceride, the surfactant is selected from one or more of lauryl glucoside, sodium lauryl sulfate, sodium dioctyl sulfosuccinate, sodium dodecyl benzene sulfonate, sodium glycocholate and the solvent is selected from one or more of glycerol, liquid paraffin, water. And adjusting the pH value by using lactic acid, and swelling the gel matrix.

The third aspect of the invention provides a preparation method of the vaginal gel, which comprises the following preparation steps:

1) dissolving alkyl glycoside or glucoside in hot water of 70 deg.C or above, or dissolving in glycerol or liquid paraffin oily solvent;

2) dissolving prebiotics, adding polycarbophil, and adjusting the pH value by lactic acid until full swelling is carried out to form hydrogel;

3) slowly dropping the solution obtained in the step 1) into the hydrogel obtained in the step 2), and fully stirring to form homogeneous emulsion drops of the alkyl glycoside or glucoside type substances in the hydrogel, thereby finally forming emulsion type gel.

The fourth aspect of the invention provides a pharmaceutical composition or a pharmaceutical preparation for vagina, which is used for restoring the internal environment of vagina, treating vaginitis and preventing occurrence and relapse of vaginitis.

In one embodiment, the vaginitis is caused by a vaginal pathogenic bacterium selected from the group consisting of: gardnerella vaginalis, Candida albicans, atorvastatin, Trichomonas vaginalis, Neisseria gonorrhoeae, Staphylococcus aureus, Escherichia coli, said pharmaceutical composition or pharmaceutical preparation being capable of inhibiting or killing said pathogenic bacteria in the vagina.

The pharmaceutical composition or pharmaceutical formulation of the invention is suitable for use in: treating various vaginal infectious diseases (such as bacterial vaginosis, candidal/mycotic vaginitis, trichomonas vaginitis, senile vaginitis, etc.) with antibiotics in combination and sequence; sequential treatment after regular treatment of the antibiotics for the vaginal infectious diseases helps to recover the normal internal environment of the vagina after the antibiotics kill pathogenic bacteria of the vagina, inhibits the pathogenic bacteria from becoming dominant flora again, supports the beneficial flora to obtain an occupying effect, and thus prevents the recurrence of vaginitis; can be used for recovering normal vaginal internal environment and preventing vaginal inflammation when vaginal flora disorder and internal environment disorder are caused by using a large amount of antibiotics in the systemic system; can be used for recovering normal vaginal internal environment and forming vaginal protective film to prevent vaginal inflammation attack when vaginal mucosa atrophy due to hormone level decrease during perimenopause, postmenopausal period or castration operation; can be used for treating pH increase caused by menstrual period and sexual life, and vaginal environment change caused by vaginal flora change, and can help to rapidly recover normal vaginal environment and prevent vaginal inflammation attack; after various intravaginal operations (such as digital vagina diagnosis, vaginal irrigation, transvaginal ultrasound, vaginal midwifery and the like), the vagina can be helped to quickly recover the normal internal environment of the vagina.

The invention has the beneficial effects that:

1) the invention has higher inhibition effect on bacteria, fungi and viruses, and is suitable for female lower genital tract infection; and helps to help repair the normal internal environment of the vagina due to the existence of the prebiotics; the polycarbophil has the functions of absorbing moisture, preserving moisture, protecting and improving vaginal mucosa endothelium and the like. The three components have synergistic effect and accord with the physiological characteristics of treatment and nursing of the female reproductive system;

2) the raw material components of the invention are safe and nontoxic, most of the raw material components are food raw materials, and the medicine is administrated at the vaginal part without the influence of the whole body;

3) the invention belongs to physical antibiosis, has no drug resistance, can be completely degraded, and is a safe and environment-friendly green product;

drawings

FIG. 1: viable count growth curve.

FIG. 2: pH change curve of culture solution.

FIG. 3: gardner zone curve.

Detailed Description

The following description of the preferred embodiments of the present invention is provided for the purpose of illustration and description, and is in no way intended to limit the invention. Also, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or instruments used are conventional products which are not known to manufacturers and are available from normal sources.

A first part: exemplary formulations and methods of making the same

Example 1:

the prescription composition is as follows: (the percentages are the proportion of the final product quality of the preparation form)

The preparation formulation is as follows: gel agent

Example 2:

The preparation formulation is as follows: gel agent

Example 3:

The preparation formulation is as follows: tablet formulation

Example 4:

The preparation formulation is as follows: suppository

Exemplary gel preparation methods:

1) sufficiently dissolving or dissolving lauric acid monoglyceride in hot water of above 70 deg.C in glycerol and liquid paraffin oily solvent;

2) dissolving lactose, adding polycarbophil, and adjusting the pH value by lactic acid until the pH value is fully swelled to form hydrogel;

3) slowly dropping the solution obtained in the step 1) into the hydrogel obtained in the step 2), and fully stirring to form homogeneous emulsion drops of the lauric acid monoglyceride in the hydrogel, thereby finally forming the emulsion type gel. Characteristics of the resulting exemplary gel:

1) pH value: 3.0-4.5;

2) viscosity: 13000 and 13800 centipoises;

3) granularity: 500 nm;

4) the mass percentage concentration of each active component is as follows: 2% of lauric acid monoglyceride, 20% of lactose and 5% of polycarbophil.

A second part: in vitro experiments

1 study design

1.1 target Strain selection

The Lactobacillus vaginalis of healthy female comprises more than 20 sub-species, wherein the species with strong glycogenolytic ability are Lactobacillus acidophilus, Lactobacillus leii and Lactobacillus salivarius, and produce H₂O₂Mainly comprises lactobacillus crispatus, lactobacillus gasseri, lactobacillus jensenii and lactobacillus acidophilus. Therefore, the lactobacillus acidophilus is preferred to synthesize the occupying capacity of each sub-strain, the capability of releasing and inhibiting pathogenic bacteria and the easy accessibility of the strain.

1.2 selection of culture methods

The MRS solid culture medium is used as a basic culture medium, different substances (the product or reference products with the same components on the market) are added into the MRS culture medium, and the inoculation culture of a target strain (lactobacillus acidophilus provided by the microbiological laboratory of university of Dalian medical science) is carried out under the same conditions. The product and the product with the same components on the market are pure dry powder preparations (insoluble auxiliary material components are filtered out), are provided in a powder form, and are directly weighed and dissolved when a system is prepared.

1.3 sample preparation and grouping:

blank group: pure MRS basic culture medium

Example 1 gel (group a): incorporation of example 1 into MRS Medium

Example 3 tablet (group B): incorporation of example 3 into MRS Medium

Ladybalance lactose tablets (group C): is equivalent to the lactose is mixed in the MRS culture medium

Soothing polycarbophil gel (group D): is equivalent to the incorporation of polycarbophil in MRS culture medium

Lauric monoglyceride gel (group E): is equivalent to the addition of lauric acid monoglyceride into MRS culture medium

1.4 Observation index

1.4.1 growth kinetics test

Activating lactobacillus acidophilus to prepare seed liquid; dissolving a test sample and a standard sample, performing filtration sterilization, preparing 20% high-concentration mother liquor, and diluting into a corresponding test group with required concentration; inoculating the seed liquid according to 3% inoculation ratio, culturing at 37 deg.C and 120r/min for 24h, and measuring viable count at 0h (i.e. initial viable count), 12-14h, 16-18h and 22-24h respectively. Time point selection description: according to the experience, the growth of the lactobacillus is not obvious before 10h of culture, the lactobacillus enters a platform stage after 15h, and the lactobacillus gradually starts autolysis and dies after 24h, so that 12-14h is selected as an observation point before entering the platform stage to evaluate the growth speed, 16-18h is the comparison between the platform stage and the total amount, and the bacteria amount maintenance condition is compared for 24 h. The specific method comprises the following steps: negating the originally designed OD value method, and because the measured result is not the number of live bacteria, the method also comprises dead bacteria, and particularly after entering the platform period, the difference between the measured result and the real number of the live bacteria is large; therefore, the number of viable bacteria was adjusted to be the number of viable bacteria in streak culture, 3 gradients per group (used for rechecking each other and averaging to finally generate 1 number of viable bacteria data), and 6 groups of 3 number of viable bacteria data (18 data) were obtained in total at 3 time points except for the number of viable bacteria in seed solution (0 h).

1.4.2 acid generation:

culturing at 37 deg.C for 24h, and measuring pH at 12-14h, 16-18h and 22-24h respectively. The pH meter is used for direct measurement, so that the method is more efficient and accurate; similar to viable count, 6 groups of 3 time points, 18 pH data were obtained after final rechecking and averaging.

1.4.3 bacteriostatic ability test:

measuring inhibition zones by an oxford small cup method for 12-14h, 16-18h and 22-24 h; each group and each time point were 3 plates (3 pathogens), and a total of 6 groups and 3 time points and 54 pathogens (including inhibition zone and inhibition zone size) were counted. The target inhibitory strain is the main pathogenic bacteria of vaginitis: gardnerella, Candida albicans and Staphylococcus aureus (Staphylococcus aureus for short), all provided by the microbiological laboratory of university of Dalian medical science.

2 test results and analysis

2.1 count of cultured bacteria and determination of pH

Compared with a blank (MRS conventional culture medium), the bacteria culture, pH value change and bacteriostasis test results of different example groups are basically consistent: the first time point (13h) of the culture is that the viable count is highest, and then the viable count gradually decreases; the pH decreased with increasing incubation time. However, there was a clear difference between groups, and the growth of lactic acid bacteria in group a (example 1), group B (example 2), and group C (lactose) was comparable to that of the blank group at each time point, and was higher than that of group D (polycarbophil) and group E (lauric monoglyceride), and the first three groups were common in that they all contained lactose as a prebiotic for lactic acid bacteria, and lactose replaced the original saccharide compared to the conventional MRS medium, while the absence of prebiotics in group D and group E reduced the amount of saccharide used for lactic acid bacteria by the substituted portion of the original medium, so lactic acid bacteria grew poorly.

Group a (example 1), group B (example 2), and group C (lactose) had lower pH values at each time point than the blank, group D (polycarbophil), and group E (lauric monoglyceride), with the latter three groups having comparable pH values. It is suggested that the addition of prebiotic lactic acid not only provides a metabolic substrate for vaginal probiotics but also facilitates the reduction of vaginal pH.

TABLE 1 viable count of Lactobacillus cultured and pH variation of culture environment

2.2 results of the bacteriostatic test

Bacteriostatic tests on bacteria related to vaginitis show that the groups have obvious difference in bacteriostatic ability. For gardnerella which is the main pathogenic bacterium of bacterial vaginosis, the seed liquid contains the vaginal probiotic lactic acid bacteria, so the gardnerella has a certain inhibiting effect, but the inhibiting effect of the groups A (example 1) and B (example 2) on the gardnerella is obviously stronger than that of other groups, the inhibiting ability of the groups C (lactose) and E (lauric acid monoglyceride) on the gardnerella at each time point is slightly higher than that of a control group, and the inhibiting ability of the group D (polycarbophil) on the gardner is the weakest. For candida albicans, the main pathogen for mycotic vaginitis, inhibitory activity was found only in group a (example 1), group B (example 2) and group E (monoglycerol laurate), whereas the inhibition of candida albicans was not seen in the blank, group C (lactose) and group D (polycarbophil). For staphylococcus aureus, which is a common exogenous pathogenic bacterium in vagina, the inhibiting capability of the group A (example 1), the group B (example 2) and the group C (lactose) is higher than that of the blank group, the group D (polycarbophil) and the group E (lauric acid monoglyceride), and the characteristic is inversely proportional to the pH value. It is possible that the reduction in pH is inhibitory due to the sensitivity of Staphylococcus aureus to acidic conditions.

TABLE 2 results of the bacteriostatic test

Remarking: the zone of inhibition is measured as the distance from the edge of the oxford cup to the outer edge of the zone of inhibition, and the diameter of the oxford cup is 7.80 mm.

If the diameter of the whole inhibition zone is required, the diameter is 7.80+ 2A. A is the measured data in the table. Jingu grape restrainer

The bacteria colony basically has no difference, and the bacteria colony is not measured one by one

The result of the bacteriostatic test shows that the formula has bacteriostatic zones on Gardner bacteria, Candida albicans and Staphylococcus aureus, and the bacteriostatic effect is stronger than that of a blank control group, pure lactose, polycarbophil or lauric monoglyceride, so that the culture product is prompted to have inhibitory effect on bacteria, and the treatment effect on various vaginitis is supported.

And a third part: animal experiments

1. Making of experimental animal model

The experimental animal is purchased from animal institute of Chinese medical science institute, and the establishment of rabbit animal model is completed with the assistance of animal institute of Chinese medical science institute.

1.1 bacterial vaginosis animal models

Detecting pH of rabbit vagina to 7.2 + -0.2 before making animal model, subculturing pure Staphylococcus aureus and Escherichia coli to obtain high-concentration pathogen suspension with concentration of 1 × 10⁹0.2mL of CFU/mL staphylococcus aureus and escherichia coli suspension is injected into the vagina of a rabbit through a catheter, 1 time per day, after 3 days of continuous molding, the red swelling and secretion condition of the vulva and the vagina of the animal are clinically checked, the vagina is taken to be clinically checked whether the vagina contains infectious pathogenic bacteria or not, the vulva and the vagina generate obvious edema, the vagina secretion contains the infectious pathogenic bacteria through clinical microbial inspection, and the vaginitis is judged to be manufacturedAnd (4) success.

1.2 animal model of mycotic vaginitis

Culturing Candida albicans with medium to obtain high-concentration bacteria suspension with concentration of 1 × 10⁹0.2mL of the suspension of the CFU/mL candida albicans is injected into the vagina of the rabbit by a catheter, 1 time per day, and after continuous molding for 3 days, the edema, the swelling and the secretion of the vulva and the vagina of the animal are clinically checked. The red swelling and secretion condition of the vulva and the vagina of the animal are clinically checked, and the vaginal and cervical secretions are taken to be tested by clinical microorganisms to determine whether the vaginal and cervical secretions contain pathogenic bacteria. The vagina secretion contains infectious pathogenic bacteria through clinical microbiological examination to judge the success of vaginitis.

2. Group administration and Observation

In the vaginitis test, 10 successfully molded rabbits, 5 bacterial vaginosis and 5 mycotic vaginitis are treated by randomly dividing each vaginitis into 5 preparations of group A (example 1), group B (example 2), group C (lactose), group D (polycarbophil) and group E (lauric monoglyceride), and 2 female rabbits without molding are taken as a blank control group, the corresponding medicine is given to each rabbit once a day for 5 days continuously, the administration dose is controlled by filling the vagina and coating the periphery of the vulva, and the blank control group is given with sterile physiological saline. Observing general behaviors and vulva conditions of animals every day, clinically checking red and swollen vulva and vagina and secretion condition of animals after 6 hours of non-administration, and taking vagina and cervix secretion to test whether containing infectious pathogenic bacteria by clinical microorganisms. The vulva and vagina red swelling is judged to be positive (+), and divided into light, medium and severe degrees according to the red swelling degree, which are respectively indicated by +++, ++, and plus, otherwise, judged to be (one), the secretion is judged to be positive (+) by clinical microbiological examination containing positive bacteria, divided into light, medium and severe degrees according to the quantity of the positive bacteria, which are respectively indicated by +++, ++, and plus, otherwise judged to be (one).

3. Vaginal irritation mucosae test in rabbits

The method selects 12 healthy and adult female white New Zealand rabbits with the weight of 2.0 kg-2.5 k g, wherein the female vaginal orifice of the animals is checked to have secretion, congestion, edema and other injury conditions before experiments. Animals were randomized into: negative control group (saline), group a (example 1), group B (example 2), group C (lactose), group D (polycarbophil) and group E (lauric monoglyceride), 2 per group. The administration is finished once a day for 6 days continuously, 24 hours after the last administration of the experimental animal, the animal is killed by an air embolism method, the complete vagina is taken out after the abdominal incision, the longitudinal incision is carried out, whether congestion, edema and other symptoms exist or not is observed by naked eyes, the reference is provided for pathological material taking, then the vagina is put into 10% formalin solution for fixation for more than 24 hours, tissues at two ends and 3 parts in the center of the vagina are selected for flaking, and histopathological examination is carried out after HE staining. And (4) pathological anatomy examination, namely scoring according to a vaginal mucosa stimulation response scoring standard, and determining the stimulation intensity according to a vaginal mucosa stimulation intensity grading standard.

TABLE 3 vaginal mucosal irritation response Scoring criteria

TABLE 4 vaginal mucosal irritation Strength grading

4. Results of the experiment

4.1 treatment of vaginitis

After the bacterial vaginosis rabbit is treated by the groups A \ B \ C \ D \ E, the red and swollen condition of the vaginal mucosa is obviously improved, the positive bacteria can still be detected except the group D, and the treated bacteria of other groups are turned into negative. The mycotic vaginosis rabbits of the groups A (example 1), B (example 2) and E (lauric monoglyceride) basically regressed the vaginal mucosa redness after being treated by each group, while the vaginal mucosa redness of the rabbits of the groups C (lactose) and D (polycarbophil) is improved by the age of the year and still shows moderate redness. And the detection of bacteria in vaginal secretion of group A (example 1), group B (example 2) and group E (lauric monoglyceride) turned negative, but the vagina of rabbit of group C (lactose) and group D (polycarbophil) is still positive, and the positive bacteria amount of group D is equivalent to that after modeling.

TABLE 5 treatment and improvement of vaginitis in rabbits

4.2 vaginal mucosa irritancy in rabbits

The vaginal mucosa irritation response test was performed in 12 rabbits and found that none of the negative control group (saline), group a (example 1), group B (example 2), group C (lactose), group D (polycarbophil), and group E (lauric monoglyceride) had significant irritation to the vaginal mucosa. However, mild irritation was still seen in the vaginal mucosa of rabbits in the negative control group (saline), group C (lactose), and group E (monolaurate), suggesting that repeated vaginal irrigation was not a good choice for protecting vaginal mucosa, whereas continuous use of lactose and monolaurate, although having therapeutic effect on vaginitis, still produced some irritation in normal mucosal state. The group A (example 1), the group B (example 2) and the group D (polycarbophil) play a good role in protecting vaginal mucosal epithelium due to the existence of the polycarbophil component, have no irritation even if continuously used, and can isolate other components existing in the product, such as lactose and lauric monoglyceride, through a physical isolation effect, so that the vaginitis is treated while the mucosa is not irritated.

Table 6 vaginal mucosal irritation response scores (n-12, two per group, numbered as group and group')

5. Conclusion of animal experiments

Groups a (example 1), B (example 2), C (lactose), D (polycarbophil) and E (monolaurin) all had some therapeutic effect on vaginitis, with minimal irritation of the vaginal mucosa. In contrast, group a (example 1), group B (example 2), group C (lactose) and group E (monolaurin) treatments were predominantly effective for bacterial vaginosis; whereas for mycotic vaginitis, group a (example 1), group B (example 2) and group E (lauric acid monoglyceride). Groups a (example 1), B (example 2) and D (polycarbophil) showed prominence in vaginal mucosal protection.

By comprehensive analysis, the product (group A-example 1-gel type, group B-example 2-tablet) in any dosage form has better curative effect on the most common vaginal infection diseases (bacterial vaginosis and mycotic vaginitis), while lactose alone is only effective on the bacterial vaginosis and cannot inhibit the mold, and lauric monoglyceride alone has more remarkable curative effect on the mycotic vaginitis, while polycarbophil cannot effectively treat the vaginal infection diseases because of only having the function of protecting vaginal mucosa and having no bacteriostatic ability.

Clinically, bacterial vaginosis and mycotic vaginitis exist in a mixed mode, so that the problem that the product containing lactose or lauric acid monoglyceride is difficult to completely solve is difficult to solve for the part of patients. And mucosal damage, which may otherwise occur due to the presence of inflammation, may be more irritating when therapeutic drugs are applied. Therefore, the product combines the vaginal prebiotics lactose, the lauric acid monoglyceride and the polycarbophil, can solve the problems of bacterial vaginosis and mycotic vaginitis simultaneously, can protect injured mucosa, reduce further stimulation and accelerate the environmental recovery in the vagina and the repair of the vaginal mucosa.

It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims

1. A vaginal pharmaceutical composition, which is characterized in that the pharmaceutical composition contains alkyl glycoside or glucoside substances or a mixture thereof as vaginal pathogenic bacteria inhibitors, prebiotics for promoting the growth of vaginal probiotics and polycarbophil as a vaginal mucosa protective film forming substance.

2. The vaginal pharmaceutical composition according to claim 1, characterized in that it comprises 0.1-10% by weight of alkyl glycoside or glucoside or mixture thereof, 5-95% by weight of prebiotic and 1.5-10% by weight of polycarbophil, preferably wherein it comprises 0.5-5% by weight of alkyl glycoside or glucoside or mixture thereof, 20-90% by weight of prebiotic and 1.5-5% by weight of polycarbophil, more preferably wherein it comprises 1-5% by weight of alkyl glycoside or glucoside or mixture thereof, 20-80% by weight of prebiotic and 1.5-3% by weight of polycarbophil.

3. The vaginal pharmaceutical composition according to claim 1 or 2, characterised in that the alkyl glycoside or glucoside is selected from the group consisting of monoglycerol laurate, glycerol ester derivatives of capric acid, caprylic acid and caproic acid, or mixtures thereof, and the vaginal probiotic is lactobacillus.

4. The vaginal pharmaceutical composition according to any one of claims 1 to 3, characterized in that it comprises monoglycerol laurate, prebiotics and polycarbophil.

5. The vaginal pharmaceutical composition according to any one of claims 1-4, wherein the prebiotics comprise one or more of lactose, sucrose, fructo-oligosaccharides, malto-oligosaccharides.

6. A pharmaceutical formulation for vaginal use, characterized in that it comprises a pharmaceutical composition according to any one of claims 1 to 5 and a pharmaceutically acceptable excipient, said pharmaceutical formulation having a pH of 3 to 4.5, preferably said pharmaceutical formulation is a vaginal gel, vaginal ointment, vaginal foam, pessary, vaginal tablet, vaginal hard capsule or vaginal soft capsule.

7. The pharmaceutical preparation of claim 6, wherein the pharmaceutical preparation is a gel containing a gel matrix selected from one or more of carbomer, xanthan gum, alginate, chitosan and hyaluronic acid, a thickener selected from one or more of methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, cyanoethylcellulose, hydroxypropylcellulose and hydroxypropylmethylcellulose, an emulsifier selected from one or more of polysorbate, polyglycerol ester, sodium stearoyl lactylate, calcium stearoyl lactylate and diacetyl tartaric acid monoglyceride, a surfactant selected from one or more of lauryl glucoside, sodium lauryl sulfate, sodium dioctyl sulfosuccinate sodium dodecylbenzene sulfonate and sodium glycocholate, and a solvent selected from glycerol, One or more of liquid paraffin and water, and lactic acid is used for adjusting pH and swelling the gel matrix.

8. A method of preparing the vaginal gel of claim 6 or 7, comprising the steps of:

9. Use of the vaginal pharmaceutical composition as claimed in any one of claims 1 to 5 or the vaginal pharmaceutical preparation as claimed in claim 6 or 7 for the preparation of a medicament for restoring the vaginal environment, treating vaginitis, preferably caused by vaginal pathogenic bacteria selected from the group consisting of: gardnerella vaginalis, Candida albicans, atorvastatin, Trichomonas vaginalis, Neisseria gonorrhoeae, Staphylococcus aureus, Escherichia coli, said pharmaceutical composition or pharmaceutical preparation being capable of inhibiting or killing said pathogenic bacteria in the vagina.

10. The use according to claim 9, wherein the pharmaceutical composition or the pharmaceutical preparation is used for sequential treatment after antibiotic formal treatment of vaginal infectious diseases, helps to restore the normal vaginal internal environment after antibiotic kills vaginal pathogenic bacteria, inhibits pathogenic bacteria from becoming dominant bacterial flora again, supports beneficial bacterial flora to take place space effect, and prevents the onset of vaginitis, or the pharmaceutical composition or the pharmaceutical preparation is used for restoring the normal vaginal internal environment when the vaginal flora is disturbed and the internal environment is disordered after the antibiotic is used in a whole body system in a large amount, and prevents the onset of vaginitis, or the pharmaceutical composition or the pharmaceutical preparation is used for restoring the normal vaginal internal environment and forming the vaginal protective membrane when the vaginal mucosa is atrophied due to the reduction of the hormone level during the perimenopause, the postmenopause or the castration operation, thereby preventing the onset of vaginal inflammation, or the pharmaceutical composition or the pharmaceutical preparation is used for the increase of pH value caused after the menstrual period and after sexual life, and the change of vaginal internal environment caused by the change of vaginal flora, and helps to rapidly recover the normal vaginal internal environment, thereby preventing the onset of vaginal inflammation, or the pharmaceutical composition or the pharmaceutical preparation is used for the operation in various vaginas, helps to rapidly recover the normal vaginal internal environment, thereby preventing the onset of vaginal inflammation.