RU2248212C2 - Preparation for treatment of bacterial vaginitis, method for its preparing and method for treatment of bacterial vaginitis - Google Patents

Abstract

FIELD: medicine, gynecology, pharmacology, pharmaceutical industry.

SUBSTANCE: invention proposes a preparation that comprises bacterial mass of live microorganisms as an active component, protecting medium and fat base. The preparation comprises lactobacilli as bacterial mass and one or some eubiotic microorganisms taken among the following group: bifidobacteria, streptococci and lactococci taken in the amount

per a single dose. Also, the preparation comprises additionally an acceptable sorbent and a biologically active supplement. Also, invention relates to a method for preparing this preparation that involves preparing firstly lactobacilli bacterial mass and one or some genus of eubiotic microorganisms taken among the following group: bifidobacteria, streptococci and lactococci. Then the prepared bacterial mass of microorganisms is immobilized on sorbent used in medicine in its ratio to bacterial mass of microorganisms = (9-1):(1-9) followed by addition of the protecting medium and biologically active supplement to formed mass in the necessary amount. Also, invention describes a method for prophylaxis and treatment of bacterial vaginitis that involves intravaginal administration of the preparation described above in the amount 1-3 doses, 1-3 times per 24 h. The treatment course is prescribed individually. Invention provides expanding assortment of agents used for treatment of bacterial vaginitis. Invention can be used in obstetric-gynecological practice.

EFFECT: improved method for vaginitis treatment, valuable medicinal properties of preparation.

13 cl, 4 tbl, 1 ex

Description

The invention relates to medicine, namely to pharmacology, in particular to drugs for the treatment of bacterial vaginosis, and can be used in the pharmaceutical industry and obstetric and gynecological practice.

Currently, diseases caused by genital infections are among the most common gynecological diseases. In recent years, there has been a tendency for an increase in diseases accompanied by a violation of the normal microflora of the vagina, in particular bacterial vaginosis.

In medical practice, antimicrobials, such as antibiotics, are widely used to treat such diseases. However, they are usually toxic, not always effective, in addition, cause the appearance of microorganisms that are resistant to the action of the antibacterial agents used. As a result of treatment with these drugs, additional restoration of the microbiocenosis of the vaginal mucosa is required.

In connection with the foregoing, a very important role is given to the search for new safe and contraindicated medications for the treatment of vaginal diseases accompanied by impaired microbiocenosis.

Closest to the claimed solution is a drug for the treatment of bacterial vaginosis “Lactobacterin in suppositories” (Instructions for the use of lactobacterin in suppositories. Nizhny Novgorod state enterprise for the production of bacterial preparations. Approved by the USSR Ministry of Health, November 29, 1992).

“Lactobacterin in suppositories” is the microbial mass of living lactobacilli, leophilized dried in a culture medium. In addition, it contains a sucrose-gelatinous milk ingredient, which is a protective environment necessary for maintaining the viability of bacteria, and an acceptable fat base.

Living microbial cells (MK) of lactobacilli have antagonistic activity against test strains of pathogenic and opportunistic microbes, including shigella, enteropathogenic Escherichia coli, staphylococcus, protea, which determines the effect of the known drug in cases of normal bacteriocenosis. The prototype preparation has a shelf life of 1 year, provided that it is stored at a temperature of (4 ± 2) ° С.

However, the disadvantage of “Lactobacterin in suppositories” is the insufficiently high pharmacological properties, since they do not provide the proper antagonistic, anti-infectious effects and the necessary level of engraftment of microorganisms on the vaginal mucosa. And, in addition, the drug does not have the necessary bioavailability. These shortcomings are the reason for the relatively low therapeutic effectiveness of the drug.

Another significant drawback of the known drug is the lack of persistence of MK under normal conditions outside the cold chain.

The objective of the present invention is to provide a drug for the treatment of bacterial vaginosis by combining the ingredients included in its composition, resulting in a high viability of live MK and, as a result, increased therapeutic efficacy and increased shelf life of the drug.

The problem is solved in that the preparation for the treatment of bacterial vaginosis, containing as an active principle the microbial mass of live bacteria, a protective environment, a fat base, according to the invention contains lactobacilli and one or more genera of eubiotic bacteria selected from the group: bifidobacteria, lactococci and streptococci in an amount of 10 ⁵ -10 ⁹ MK per dose additionally comprises suitable sorbent in a ratio of its mass to the microbial bacteria (9-1) :( 1-9), the biologically active additive to count honors 0.05-3.0 wt% relative to the weight of the resulting product and as a fat base -. any suitable base containing not more than 5 wt% of water and having a pH of 4.5-7.0..

The microorganisms proposed by the authors, which are part of the claimed preparation, are representatives of the normal microflora of the vaginal mucosa and encompass a spectrum of bacteria, in violation of which certain pathologies arise, in particular bacterial vaginosis.

A prerequisite is the presence of lactobacilli in the preparation, since they overwhelmingly determine the microflora of the vaginal mucosa. As for other microorganisms, their presence in the preparation is intended to enhance the biological activity of lactobacilli.

The necessary biological activity and the effectiveness of the treatment is determined by the viability of MK, their adhesive ability and ability to colonize. The authors of the present invention proposed the content of 10 ⁵ -10 ⁹ MK in one dose, which, on the one hand, is associated with the effectiveness of the drug, and on the other - economic feasibility.

In the model of white rats, which were scraped from the vaginal mucosa 24 hours after taking the preparations, 0.1 ml of a suspension of the drug consisting of immobilized lactobacilli and non-immobilized lactobacilli in the same concentrations (from 10 μ / cm ³ to 10 ¹⁰ μ / cm ³ ), it was shown that the adhesive activity of lactobacilli in the composition of the drug reaches a maximum level of 9.1 · 10 ⁷ µ / mm ² with the introduction of the drug containing 10 ⁹ MK. A further increase in the concentration of MK does not lead to an increase in the level of adhesion.

The data in table 1 confirm that the most clinically effective is a drug containing from 10 ⁵ to 10 ⁹ MK in a single dose.

For comparison, it was shown that non-immobilized bacteria are inferior in adhesion to the surface of the mucosa, not exceeding 3.5 · 10 ⁴ µ / mm ² .

Table 1
Adhesive activity of lactobacilli in the claimed preparation and lactobacilli in suspension Lactobacilli The initial concentration of injected MK in 1 cm ³ suspension 10 ² 10 ⁴ 10 ⁵ 10 ⁶ 10 ⁸ 10 ⁹ 10 ¹⁰ Immobilized on a sorbent in the composition of the claimed drug Adhesive activity to epithelium 1,210 2.6 · 10 ² 2.8.0 · 10 ³ 3.8 · 10 ⁴ 6.6 · 10 ⁶ 9.110 ⁷ 9.2 · 10 ⁷ Not immobilized in the composition of the suspension Not 1.7 · 10 ² 1.7 · 10 ² 1.7 · 10 ² 2.8 · 10 ³ 3,510 ⁴ 4.110 ⁴

The protective environment in accordance with this invention should be such that it is able to maintain at the required level the viability of the sorbed bacteria after freeze drying.

The quantitative content of the protective medium, for example, sugar-gelatin medium, is usually determined by the storage conditions of the microorganisms in the preparation and amounts to 0.01-0.02% by weight.

Acceptable sorbents used in the claimed preparation include any sorbents authorized by the Ministry of Health, for example, cellulose derivatives, alumina, in particular sodium smectite, etc.

To achieve the therapeutic efficacy of the claimed drug at the required level, it is necessary to create microcolonies, consisting of a sorbent particle, on which 10-40 MK are adsorbed. This condition, as defined in the present invention, is satisfied when the mass ratio of the sorbent to the microbial mass (9-1) :( 1-9). Under these conditions, the formation of microcolonies of bacteria will be maximum.

The particle size of the sorbent can be any suitable for a preparation for this purpose, however, a size of not more than 100 microns is most acceptable, otherwise mechanical irritation of the mucosa is possible.

As for the dietary supplement, it can have various functions.

The quantitative content of the additive is from 0.05 to 3.0 wt.% Of the microbial mass of bacteria due to the fact that when its content is less than 0.05 wt.%, Its effectiveness sharply decreases, i.e. effects on viability, colonization, and mucosal adhesion of the MK.

At a higher than 3.0 wt.% Content of the biologically active additive, there is a danger of lysis of the introduced bacteria and, in addition, its side effects are possible, for example, irritation of the mucosa from its excess, in particular when using essential oils of plants.

The choice of ingredients of a biologically active additive is very wide and is due to the particular application of the claimed preparation, for example, fungicidal ingredients or, if necessary, ingredients with an antimicrobial effect, etc. can be used. Introduction to the preparation, for example, phyto-ingredients accelerates the process of mucosal colonization and leads to to normalize microbiocenosis by this indicator after 48 hours.

The authors proposed to use as a fat base any base suitable for cream or suppository. However, some requirements are imposed on the fat base - it should not contain more than 5% water and should provide a pH of 4.5-7.0 for the drug, otherwise the bacterial viability drops sharply, which affects the effectiveness of the drug.

In addition, the totality of all the ingredients of the claimed drug is aimed at increasing the shelf life and simplifying storage conditions. The results of the relevant studies are shown in table 2.

For the study, the claimed preparations are taken, which differ in the sorbent used and the pH value of the fat base. The studies were carried out after a year of storage at a temperature of 22 ° C.

table 2
The dependence of bacterial viability on the pH of the fat base The inventive drug, characterized by a sorbent The initial number of MK in 1 g of the drug Bacterial viability, MK in 1 g of the drug (one year of storage) Fat base pH 3.0 4,5 6.5 7.0 7.5 Cellulose derivatives 1,0 · 10 ⁹ 1,0 · 10 ⁷ 8.0 · 10 ⁷ 4.010 ⁸ 4.010 ⁸ 5.010 ⁷ Polymethylsiloxane 1,0 · 10 ⁹ 1,0 · 10 ⁶ 4.010 ⁷ 2.010 ⁷ 3.0 · 10 ⁷ 7.0 · 10 ⁷ Smectite Sodium 1,0 · 10 ⁹ 2.010 ⁶ 3.0 · 10 ⁷ 3.010 ⁸ 5.010 ⁸ 1,0 · 10 ⁷

The data in table 2 indicate that the viability of bacteria is highest at pH 4.5-7.0, at values below 4.5 it is insufficient to achieve the necessary results, and at pH values above 7.0, the viability of bacteria decreases markedly.

The dependence of bacterial viability and the pH of the fat base, obtained after a year of storage at a temperature of 22 ° C, indicates an increase in shelf life while simplifying storage conditions.

To prove the clinical effectiveness of the claimed drug, studies were conducted, the results of which are shown in table 3.

As the studied drugs was used the drug (1) corresponding to the claimed. Lactobacilli were taken as bacteria for clarity. The addition of bifidobacteria and / or streptococci and / or lactococci in accordance with the claimed solution to them proportionally enhances the result.

In addition, we studied the drug (2) without phytoadditive and, finally, the drug (3) corresponding to the prototype.

The above drugs were injected into the vagina of women volunteers suffering from bacterial vaginosis, a pronounced decrease in lactoflora.

Table 3
Comparative characteristics of the therapeutic effect of the claimed drug and drug prototype A drug The content of lactobacilli in the secret of the vagina, MK / cm ³ Before treatment After 24 hours After 48 hours 1. A preparation containing lactobacilli immobilized on alumina and herbal supplements 1,2 · 10 ³ 2.510 ⁶ 3.6 · 10 ⁷ 2. A preparation containing lactobacilli immobilized on alumina 1,2 · 10 ³ 3.1 · 10 ⁶ 7.1 · 10 ⁶ 3. Non-immobilized lactobacilli (prototype) 1,2 · 10 ³ 5.0 · 10 ⁶ 4.0 · 10 ⁴

As the results of table 3 show, the best clinical indicators are obtained for the drug (1) corresponding to the claimed.

Another objective of the claimed invention is the creation of a method of obtaining a drug for the treatment of bacterial vaginosis by such a combination of actions and modes of their implementation, which allows to obtain a drug with high therapeutic efficacy and storage resistance under ordinary conditions.

The problem is solved in that in the method for producing a preparation for treating bacterial vaginosis, according to the invention, the microbial mass of lactobacilli and one or more genera of eubiotic bacteria selected from the group of bifidobacteria, streptococci and lactococci is first obtained, and their content in 1 g of microbial mass is 10 ¹⁰ -10 ¹¹ cells. Then, the resulting microbial mass of bacteria is immobilized on a medicine-acceptable sorbent with its ratio to the microbial mass of bacteria (9-1) :( 1-9), after which the protective medium in the required amount and a biologically active additive in the amount of 0.05 are introduced into the resulting mass. -3.0% relative to the mass of the resulting drug. The resulting product is subjected to freeze drying in the mode corresponding to the kind of microorganisms used, then the dried mass is mixed with an acceptable fat base containing less than 5% water and having a pH of 4.5-7.0 and taken in an amount corresponding to the content of 10 ⁵ -10 ⁹ MK in one dose of the drug.

The authors stipulated that 1 g of the microbial mass should contain lactobacilli and bifidobacteria, and / or streptococci, and / or lactococci in an amount of 10 ¹⁰ -10 ¹¹ cells.

This is due to the fact that under these conditions 10-40 MK should be placed on 1 particle of the sorbent, or a conglomerate must be formed, which includes proportional particles of the sorbent and bacteria, which corresponds to the preparation with the most useful properties. Or, in other words, it is possible to obtain a drug with 10 ⁵ -10 ⁹ MK in one dose from the indicated microbial mass, which, as shown above, in combination with other ingredients of the drug corresponds to its greatest clinical effectiveness.

The remaining conditions for the implementation of the method of obtaining the drug for the treatment of bacterial vaginosis are reduced to a sequence of mixing the components, which ensures high viability of the bacteria, their sufficient colonization on the vaginal mucosa, adhesion and generally high biological activity.

As for the ratio of sorbent to microbial mass, it is due to the cost-effectiveness of obtaining an effective therapeutic dose.

The method of obtaining the drug for the treatment of bacterial vaginosis is confirmed by an example of a specific implementation.

Example 1

The inventive preparation is obtained in several stages. First get the microbial mass of lactobacilli, bifidobacteria, and / or streptococci, and / or lactococci, and their content in 1 g of microbial mass is 10 ¹⁰ -10 ¹¹ cells.

Then, the resulting microbial mass is immobilized on a sorbent, in particular sodium smectite, with a particle size not exceeding 100 microns. This process is carried out by thorough mixing.

The ratio of microbial mass to sodium smectite is 9: 1.

Then, the resulting mass is mixed with sucrose-gelatin medium, taken in an amount of 0.02% relative to the mass of the preparation obtained, and with 2.5% sage essential oil relative to the mass of the preparation obtained.

After that, the resulting product is subjected to freeze drying in pallets.

The dried product is mixed with a fat base, for example, containing 90 wt.% Cocoa butter and 10 wt.% Lanolin, while, for example, citric acid is added to the fat base to pH 6.5. The fat base has a moisture content of less than 5%.

Suppositories weighing from 1 g to 1.5 g are formed from the resulting mass.

The ointment is obtained according to the method described above with the difference that any acceptable ointment base is used as the fat base.

The next objective of the claimed invention is the creation of a method for the treatment and prevention of bacterial vaginosis by using the drug described above, as a result of which high clinical efficacy of treatment is achieved.

The problem is solved in that in the method for the prevention and treatment of bacterial vaginosis according to the invention, a preparation is introduced intravaginally according to claims 1-7, containing lactobacilli and one or more genera of eubiotic bacteria selected from the group: bifidobacteria, streptococci and lactococci in an amount of 10 ⁵ - 10 ⁹ living cells per dose, and the drug is administered 1-3 times a day in the amount of 1-3 doses, and the course of treatment is determined individually depending on the degree of damage to the vaginal mucosa.

In the present method of treating bacterial vaginosis using a preparation containing 10 ⁵ -10 ⁹ MK in 1 dose and the remaining corresponding components, a daily intake of 1-3 doses is determined depending on the severity of the disease. In the case of prophylactic use, the daily intake of the drug can be 1 dose.

The course of treatment is determined in each case individually, depending on objective indicators of the severity of the disease and the results of treatment.

The treatment effect can be enhanced by preliminary irrigation of the vagina, for example, with saline or a decoction of herbs at pH 4.5-5.5. This procedure is advisable, since it is aimed at the mechanical excretion of mucus and conditionally pathogenic microbes. After such cleaning, the access of the claimed drug in the form of a suppository or cream to the affected mucosa is facilitated.

In addition, an additional positive result is observed during pretreatment of the vagina with a suspension of an acceptable sorbent, the role of which is to enhance the cleaning effect and prepare the mucosa for colonizing the introduced bacteria.

In cases where, along with bacterial vaginosis, there are concomitant diseases, it is possible to pre-treat, for example, with antifungicidal or other drugs.

Studies have been conducted aimed at determining the microbiological effectiveness of correcting the microbiocenosis of the vagina of women with symptoms of bacterial vaginosis.

Under observation were 19 women with symptoms of bacterial vaginosis, expressed in a decrease or absence of lactobacilli, in the presence of key cells (more than 10 in the field of view), in the amount of gardnerella in 1 g of secretions more than 10 ⁶ , in itching.

Patients were divided into three randomized groups. Before the introduction of vaginal preparations, they were preliminarily purified with physiological saline, adjusted by adding lemon juice to pH 5.0.

The first group of women was intravaginally administered the claimed drug in the form of an ointment in an amount of 1 g 2 times a day for 5 days.

The second group of women was injected with a suspension of lactobacilli in physiological saline at pH 5.0 for 5 days.

And finally, the third group of women was a control - they were injected with the ointment base of the drug also for 5 days.

The criteria for the positive effect of treatment were:

- bringing lactobacilli to normal concentration (10 ⁷ per 1 g of secretions);

- absence or decrease in the number of key cells;

- a decrease in the population of garnerella to 10 ^2-3 ;

- cessation of itching.

Table 4 presents the results.

Table 4
The results of treatment of patients with bacterial vaginosis Treatment method The number of women in the group Recovery Criteria Itching Number of key cells Number of Gardnerell Number of Lactobacilli The inventive drug 4 Not Not 10 ² > 10 ⁷ Lactobacilli in suspension 4 + 6-9 > 10 ³ <10 ⁷ Ointment base 4 ++ 10-12 > 10 ⁷ <10 ³ Before treatment 12 +++ > 10 > 10 ⁷ <10 ³

The results obtained (table 4) indicate that when applying the treatment using the inventive preparation after 5 days, vaginal microbiocenosis normalizes, which is confirmed by microbiological indicators and one of the elements of the clinical characteristic.

For comparison, during treatment with a suspension of lactobacilli, a slight improvement was noted according to the studied criteria, however, normalization of microbiocenosis did not occur.

The ointment base, in comparison with the initial state of patients, did not give an improvement.

Thus, the proposed drug for the treatment of bacterial vaginosis showed high clinical characteristics, the method of its production is aimed at ensuring high treatment efficiency. And, in addition, the composition of the drug and the features of its preparation provide simplified conditions for its storage with a simultaneous increase in shelf life.

A method of treating bacterial vaginosis has proven high clinical results of the drug, which is confirmed by objective criteria for the recovery of patients.

Claims (13)

1. A drug for the treatment of bacterial vaginosis, containing as an active principle the microbial mass of live bacteria, a protective environment, a fat base, characterized in that the microbial mass contains lactobacilli and one or more genera of eubiotic bacteria selected from the group: bifidobacteria, streptococci and lactococci in an amount of 10 ⁵ -10 ⁹ MK per dose, additionally contains an acceptable sorbent with its mass ratio to the microbial mass of bacteria (9-1) :( 1-9), a biologically active additive in an amount of 0.05-3.0 wt. .% relative but the masses of the resulting product and as a fat base -. any suitable base containing not more than 5 wt% of water and having a pH of 4.5-7.0. 2. The drug according to claim 1, characterized in that the acceptable sorbent has a particle size of not more than 100 μm and has immobilizing activity for MK from 10 to 100% of its surface. 3. The drug according to claim 1 or 2, characterized in that the biological additive contains one or more ingredients having a fungicidal and / or antimicrobial effect, and / or an action that increases the activity of microorganisms introduced into the preparation. 4. The drug according to claim 1, characterized in that it is a suppository or ointment. 5. A method of obtaining a drug for the treatment of bacterial vaginosis, characterized in that the microbial mass of lactobacilli and one or more genera of eubiotic bacteria selected from the group of bifidobacteria, streptococci and lactococci is first obtained, and their content in 1 g of microbial mass is 10 ¹⁰ -10 ¹¹ cells, then the resulting microbial mass of bacteria is immobilized on a medicine-acceptable sorbent with its ratio to the microbial mass of bacteria (9-1) :( 1-9), after which a protective medium is introduced into the resulting mass in the required amount As a biologically active additive in an amount of 0.05-3.0% relative to the mass of the obtained preparation, the resulting product is subjected to freeze drying in the mode corresponding to the type of microorganisms used, then the dried mass is mixed with an acceptable fat base containing less than 5% water and having a pH 4.5-7.0 and taken in an amount corresponding to the content of 10 ⁵ -10 ⁹ MK in one dose of the drug. 6. The method according to claim 5, characterized in that they use a sorbent with a particle size of not more than 100 microns. 7. The method according to claim 5 or 6, characterized in that they use a biologically active additive containing one or more ingredients having a fungicidal and / or antimicrobial effect, and / or an action that increases the activity of microorganisms introduced into the preparation. 8. The method according to claim 5, characterized in that they use a fat base suitable for obtaining a cream. 9. The method according to claim 5, characterized in that they use a fat base suitable for producing a suppository. 10. A method for the prevention and treatment of bacterial vaginosis, characterized in that the drug is administered intravaginally according to claims 1 to 4, containing lactobacilli and one or more genera of eubiotic bacteria selected from the group of bifidobacteria, streptococci and lactococci in the amount of 10 ⁵ -10 ⁹ live cells per dose, and the drug is administered 1-3 times a day in the amount of 1-3 doses, and the course of treatment is determined individually depending on the degree of damage to the vaginal mucosa. 11. The method according to claim 10, characterized in that the vagina is previously irrigated with physiological saline or a decoction of herbs. 12. The method according to claim 10 or 11, characterized in that a suspension of an acceptable sorbent is previously introduced into the vagina. 13. The method according to claim 10, or 11, or 12, characterized in that antifungicidal drugs are preliminarily injected into the vagina.