RU2751000C1 - Molded carbon sorbent with glycolic acid, a method for its preparation and a method for treating bacterial vaginosis - Google Patents

Abstract

FIELD: pharmacology.SUBSTANCE: group of inventions relates to a molded carbon sorbent, its preparation and use in medicine as an applicator for the treatment of bacterial vaginosis. The modified sorbent is a molded mesoporous carbon sorbent of cylindrical shape and geometric dimensions: diameter 8-10 mm, length 45-60 mm, with one internal channel of circular cross-section, specific adsorption surface no more than 50 m2/g, crushing strength no less than 20 kg/cm2, glycolic acid content in the form of polyglycolide of not less than 5 to 7.4, weight percentage. A method for manufacturing a molded sorbent is described, including impregnation of the initial molded sorbent with a 50% aqueous solution of glycolic acid for 24 hours at room temperature, the ratio of sorbent-modifier 1:2 by weight, followed by drying for an hour at 103-107°C and polycondensation of glycolic acid on a carbon molded sorbent in two stages: at a temperature of 190-200°C for 1 hour, at a temperature of 220-230°C for 5 hours. A method for the treatment of bacterial vaginosis in gynecology, including sanitation with a molded carbon sorbent modified with a glycolic acid polymer, is described.EFFECT: technical result of the invention is the production of a molded carbon sorbent modified with glycolic acid, which has an effective bactericidal effect for the treatment of bacterial vaginosis.4 cl, 3 dwg, 4 tbl, 9 ex

Description

The invention relates to the production of molded carbon sorbents for use in medicine, namely, as an applicator for the treatment of bacterial vaginosis.

Bacterial vaginosis is an infectious non-inflammatory process caused by violations of the qualitative and quantitative ratio of the vaginal biotope, characterized by an excessive increase in the concentration of obligate and facultative anaerobic opportunistic microorganisms and a sharp decrease or complete absence of lactic acid bacteria in the vaginal discharge. There is no single specific causative agent for bacterial vaginosis. The etiological factor is a set of opportunistic microorganisms: G.vaginalis, Atopobium vaginae, Prevotella bivia, P..disiens, P.corporis, Bacteroides urealyticus, Enterococcus, Fusobacterium nuclearum, Mobiluncus, P. In this case, pathogens are capable of forming microbial biofilms. In them, microorganisms acquire other properties than in vitro, which affects the pathogenesis and affects the effectiveness of treatment [Verstraelen H., Swidsinski A. The 1: implications for epidemiology, diagnosis and treatment. // Curr Opin Infect Dis. 2013; 26 (1): 86-9]. In biofilms, microbes are less vulnerable to immunity factors, antibodies and antibiotics - they retain their viability even after exposure to concentrations 500-1000 times higher than therapeutic doses [Antibiotic resistance of bacterial biofilms / Hoiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O . Int J Antimicrob Agents. 2010 Apr; 35 (4): 322-32].

The main method of treating bacterial vaginosis is the appointment of antibacterial drugs [Clinical guidelines for the diagnosis and treatment of diseases accompanied by pathological discharge from the genital tract of women. Russian Society of Obstetricians and Gynecologists / 2nd ed., Revised. and add. M., 2019]. The presence of bacterial films protects the colonies from drug effects, and antibacterial treatment does not always lead to the complete elimination of pathogens. Microorganisms that have retained their viability in the conditions of a bacterial film subsequently cause a relapse of the disease. The reduced number or absence of lactobacilli creates favorable conditions for the reproduction of opportunistic flora, thereby contributing to the occurrence of relapse. Frequent relapses of bacterial vaginosis, repeated courses of antibiotic therapy lead to the emergence of microflora resistance to drugs, a decrease in the effectiveness of the treatment and, as a consequence, to relapses. Thus, the "vicious circle" of the pathogenesis of recurrent bacterial vaginosis is closed.

The main pathogenetic stages of bacterial vaginosis therapy are:

1) overcoming biofilms and antibiotic resistance;

2) restoration of normal biocenosis.

The traditional method of treating bacterial vaginosis in non-pregnant women is the use of antibacterial drugs (metronidazole and clindamycin), which have an anti-anaerobic spectrum of action [Clinical guidelines for the diagnosis and treatment of diseases accompanied by pathological discharge from the genital tract of women. Russian Society of Obstetricians and Gynecologists / 2nd ed., Revised. and add. M., 2019.].

The action of metronidazole is aimed at suppressing the generation of anaerobic microorganisms and gardnerella, that is, the main spectrum of pathogenic microflora characteristic of bacterial vaginosis. Metronidazole penetrates into the microbial cell and blocks the synthesis of nucleic acids. It is known that a single dose of metronidazole per os in a dose of 2 g is as effective as a 5-7-day course of treatment [Barbone FJ "Austin N., Louv WC, et al. Afollow - up study of methods of contraception, sexual activity and rates of trichomoniasis, candidiasis and bacterial vaginosis. Amer J. Obstet Ginec 1990; 163 (2): 510-4] The disadvantage of this method of treatment is that oral administration of metronidazole may cause side effects from the gastrointestinal system (metallic taste in the mouth, diarrhea, dyspeptic disorders), from the nervous system and sensory organs (headache, impaired coordination of movements, syncope, ataxia, confusion), from the genitourinary system (dysuria, polyuria, urinary incontinence), as well as allergic reactions.

Topical administration of metronidazole preparations avoids systemic side effects. However, many studies have proven that Atopobium vaginae is one of the main causative agents of bacterial vaginosis and this microorganism exhibits high resistance to the action of metronidazole [Bradshaw C.S., Tabrizi S.N., Fairley C.K., et al. The association of Atopobium vaginae and Gardnerella vaginalis with bacterial vaginosis and recurrence after oral metronidazole therapy. The Journal of infectious diseases 2006; 194 (6): 828-36; Wathne B., Hoist E., Hovelius B. Erythromycin versus metronidazole in the treatment of bacterial vaginosis. Acta Obstetr. Gynecol. Scand. 1993; 72 (6): 470-474; Ankirskaya A.S., Prilepskaya V.N., Bayramova G.R. et al. Bacterial vaginosis: clinical features, diagnosis and treatment. Russian medical journal. 1998; 6 (5): 276-282].

The use of clindamycin preparations can significantly reduce the number of multiplying opportunistic microorganisms (Gardnerella vaginalis and Atopobium vaginae), but at the same time, growth is inhibited and the amount of the already reduced content of normal vaginal microflora (lactobacilli) decreases, which contributes to a high percentage of relapses of the disease. In addition, after treatment with antibacterial drugs in 20-30% of cases, candidal vaginitis develops [Prilepskaya V.N., Baramova G.R. Etiopathogenesis, diagnosis and current trends in the treatment of bacterial vaginosis. Russian medical journal. 2002; 10 (18): 795-797]. It should also be noted that with repeated courses of treatment of recurrent bacterial vaginosis with antibacterial drugs, the development of resistance of opportunistic microflora to the drugs used is possible.

There is a known method for the elimination of Atopobium vaginae bacteria in the treatment of bacterial vaginosis (patent No. 2445964), characterized in that the drug "Macmiror Complex" is used, which is administered one candle into the vagina once a day for seven to eight days. There is also a known agent and method for the treatment of bacterial vaginosis (patent No. 2261715), the essence of which consists in the use of Pimafucin (natamycin), which is administered intravaginally in the form of 2% cream 2 times a day in the morning and in the evening for 7-10 days, while in the first 3 days in the evening after the introduction of the cream into the vagina, a vaginal suppository of pimafucin 100 mg is additionally introduced. These methods of treatment make it possible to prevent the development of such a complication as vulvovaginal candidiasis, since the drugs used also have a fungicidal effect, but the possibility of the development of resistance of the main pathogens remains and in order to restore the normal microflora of the vagina it is necessary to carry out an additional second stage of treatment with the use of eubiotics and probiotics, which lengthens the time and increases the cost of treatment.

A known method of treating bacterial vaginosis, which consists in combining traditional antibiotic therapy [Clinical guidelines for the diagnosis and treatment of diseases accompanied by pathological discharge from the genital tract of women. Russian Society of Obstetricians and Gynecologists / 2nd ed., Revised. and add. M., 2019] with additional local use of enterosorbent Enterosgel [Bondarenko K.R., Mazyutov A.R., Ayupova G.V., Fedotova A.A., Gaisina Yu.R. The use of enterosorbent "Enterosgel" in the treatment of bacterial vaginosis // Scientific Bulletin of Belgorod State University. 2012; 4 (123): 26-31]. Disadvantages of this method of treatment: 1) the use of the drug "enterosgel" is carried out in violation of the instructions for its use. The drug belongs to the group of enterosorbents; the instructions do not contain information about the possibility of its use vaginally. 2) "Enterosgel" selectively adsorbs only medium-molecular toxic metabolites, while substances with a low molecular weight (vitamins, mineral salts) and high-molecular weight (proteins, enzymes, immunoglobulins) are not subjected to sorption [Current issues of methodology of preventive medicine, medical prevention and social medicine: Sat. scientific. Art. / Ed. A.E. Agapitova. Irkutsk: RIO IGIUVa, 2011. S. 168].

A known agent for the treatment of bacterial vaginosis (patent No. 2325148), made in the form of a vaginal gel with the main active ingredient - enterosgel - 10.0, preservative - nipagin-nipazol (3: 1) - 0.2, as a basis: carbopol gel 0 , 25%.

A known method of treating bacterial vaginosis, including the introduction into the vagina of tissue carbon sorbent AUT-M (RF patent No. 2150944). First, the vagina is treated with a dry cotton or gauze swab in order to evacuate abundant creamy discharge, then a tissue sorbent AUT-M 5 × 5 cm in size, folded in a "tube" is inserted into the vagina, so that the sorbent over the entire surface encloses the walls of the vagina. Exposure time - 24 hours.

The disadvantage of these methods of treatment using different options for sorbents is that they allow you to eliminate the bulk of the pathogenic microflora, while not contributing to the restoration of normal. This creates favorable conditions for the recurrence of the pathological process.

A known drug for the treatment of bacterial vaginosis, a method for its production and a method for the treatment of bacterial vaginosis (patent No. 2248212). The proposed preparation contains a microbial mass of live bacteria (lactobacilli and one or more genera of eubiotic bacteria), a protective medium, a fatty base, a sorbent acceptable in medicine and a biologically active additive. Derivatives of cellulose, alumina, in particular, sodium smectite, were used as sorbents. However, the disadvantage of this method is the absence of an antimicrobial component in the treatment, an insufficiently high amount of sorbent in its composition, which is used only as a basis for fixing the mass of living bacteria. These disadvantages are the reason for the relatively low therapeutic efficacy of the drug.

The basis of vulnerosorption (application sorption) is the ability of the sorbent to adsorb pathogenic microorganisms on its surface, regardless of their antibiotic resistance, thereby reducing the degree of contamination, in addition, the sorbent, upon contact with a biological surface, absorbs toxins, metabolites, cytokines formed during the inflammatory process and as a result vital activity of pathogens. The use of sorbents helps to reduce the number of pathogens on the biological surface by 100-1000 times in comparison with gauze dressings [Eretskaya EV, Ulchenko V.Yu. Investigation of the antimicrobial properties of application-sorption materials based on activated fibrous carbon // Clinical Surgery. 1985; 3:53].

However, in gynecological practice, as a rule, sorbents are used, which are tissues or powdered materials with granule sizes of less than 0.5 mm. This form of sorption material requires packaging in various types of containers, which significantly reduces the contact of the sorbent with the biological surface, and, accordingly, affects the efficiency of vulnerosorption. In the technical production of sorption materials, certain difficulties arise in terms of giving them the correct shape, which would optimally correspond to the shape and size of the surface or cavity of the human body in order to carry out application sorption.

In modern medicine, the most famous materials with a pronounced compatibility with biological systems are carbon sorbents, which have a high chemical purity, a developed porous texture, providing increased sorption activity. These sorbents are produced at the Center for New Chemical Technologies of the Federal State Budgetary Institution of Science "Federal Research Center" Institute of Catalysis named after G.K. Boreskov of the Siberian Branch of the Russian Academy of Sciences ”(Omsk Branch) (TsNKhT IC SB RAS). This institution has developed technologies that make it possible to obtain carbon sorbents of the optimal shape (in the form of cylinders) for use in obstetrics and gynecology for application sorption therapy. They are administered intrauterinely, intravaginally, painlessly.

Closest to the proposed method of treatment is a molded carbon sorbent VNIITU-1 (RF patent No. 2516878), developed for the prevention of septic complications in obstetrics. This medical product has the form of a cylinder with a diameter of 8-13 mm, a length of 50-80 mm, with one internal channel of circular cross-section and is a nanodispersed mesoporous carbon material containing at least 99.5% of the mass. carbon, not more than 0.15% of the mass. ash, not more than 0.30% of the mass. sulfur, with a total pore volume of at least 0.4 cm ³ / g, a specific surface for nitrogen adsorption of 200-300 m ² / g and a crushing strength of at least 50 kg / cm ² . Sorbent VNIITU-1 is placed in the uterine cavity in the postpartum period in women of the infectious risk group in order to carry out application sorption for the prevention of postpartum endometritis. Shaped sorbent VNIITU-1: TU 9398-043-71069834-2013 with rev. No. 1 (TU 21.20.10.171-043-36704515-2013). Registration certificate for a medical device dated September 16, 2019 No. RZN 2015/2967, permitted for production and sale on the territory of the Russian Federation. Toxicological conclusion No. 1998.013Р dated 08/14/2013: non-toxic, sterile, pyrogen-free. According to the assessment of the results of technical tests (protocol No. 12.404-2.013 of 08/26/2013): meets the requirements of regulatory documents. However, this medical device does not meet the basic requirements for the treatment of bacterial vaginosis. In particular, with a good detoxification ability and the ability to reduce the number of pathogenic and opportunistic microorganisms on biological surfaces, the VNIITU-1 sorbent does not contribute to the restoration of normal microflora.

Known molded sorbent with antibacterial properties, proposed for the treatment of endometritis (patent No. 2545743). Carbon sorbent with antibacterial properties is a nanodispersed mesoporous carbon material in the form of cylinders with a diameter of 2-4 mm, a length of 15-25 mm with one internal circular channel and is characterized by a polyvinylpyrrolidone content of at least 5.0%, a specific adsorption surface of not more than 50 m ² / g, crushing strength not less than 20 kg / cm ² . Shaped sorbent VNIITU-1PVP: TU 9398-044-71069834-2013 with rev. No. 1 (TU 21.20.10.171-044-36704515-2013). Registration certificate for a medical device dated September 17, 2019 No. RZN 2015/2969, permitted for production and sale on the territory of the Russian Federation. Toxicological conclusion No. 2099.013Р dated 08/14/2013: non-toxic, sterile, pyrogen-free. According to the evaluation of the results of technical tests (protocol No. 12.405-2.013 of 08/26/2013: complies with the requirements of regulatory documents. Modification with polyvinylpyrrolidone allows an additional effect of the sorbent on the pathogenic microflora, contributing to the inhibition of its growth. bacterial vaginosis.

The closest technical solution is a carbon sorbent with antibacterial and antimycotic properties (RF patent No. 2541103, prototype). The carbon sorbent with antibacterial properties is round-shaped granules, contains a polymer of glycolic acid (polyglycolide) in an amount of at least 5%, is characterized by a specific adsorption surface of less than 250 m ² / g and a total pore volume of less than 0.50 cm ³ / g. The method of obtaining a carbon sorbent with antibacterial properties includes impregnation of granules of carbon hemosorbent with 10-50% aqueous solution of glycolic acid for 7-9 hours at room temperature, the ratio of hemosorbent: modifier solution 1: 1, followed by drying for an hour at 100-110 ° C ... Polycondensation of glycolic acid on a carbon hemosorbent takes place in 2 stages: at a temperature of 185-205 ° C for 1 hour, at a temperature of 215-235 ° C - at least 5 hours.

Glycolic acid (hydroxyacetic acid, hydroxyethanic acid) - organic compound C ₂ H ₄ O ₃ , 99% (wt.) Produced by "Merk Schuchardt OHG" (Germany). Well soluble in water, alcohols, ethers. Meets the requirements of medicine: non-toxic; has oxygen-containing groups that are capable of entering into a polycondensation reaction with the formation of a biologically active polymer (oligomer). Glycolic acid and its derivatives obtained by polycondensation (oligo-, polymers) exhibit anti-inflammatory, antibacterial properties. It is widely used in cosmetology and medicine (masks, creams, ointments, medicines, etc.).

The technical result of the claimed invention is to obtain a molded carbon sorbent modified with a polymer of glycolic acid in the form of a polyglycolide, which has an effective bactericidal effect for the treatment of bacterial vaginosis.

The proposed modified sorbent is a formed mesoporous carbon sorbent of cylindrical shape and geometric dimensions: diameter 8-10 mm, length 45-60 mm, with one internal channel of circular cross-section, specific adsorption surface no more than 50 m ² / g, crushing strength not less 20 kg / cm ² , ash content not more than 0.30%; with a glycolic acid content in the form of a polyglycolide of at least 5%. These dimensions of the sorbent provide ease of intravaginal use. The presence of the channel ensures the accessibility of the outer and inner surfaces for the sorption of bacterial cells.

The problem is solved by using a molded sorbent as a starting material with dimensions: diameter 8-10 mm, length 45-60 mm, with one internal channel of circular cross-section; mass fraction of ash not more than 0.30%, specific adsorption surface 220-300 m ² / g, crushing strength not less than 20 kg / cm ² .

The proposed method for manufacturing a sorbent includes:

- modification of the surface of the formed sorbent with an aqueous solution of glycolic acid, followed by polycondensation during heat treatment. The modification process is carried out in several stages:

- impregnation of the starting material with a 50% aqueous solution of glycolic acid under static conditions for 24 hours at room temperature, the ratio of sorbent: modifier 1: 2 (by weight);

- drying the sorbent impregnated with an aqueous solution of glycolic acid for an hour at 103-107 ° C;

- polycondensation of glycolic acid on a molded carbon sorbent in 2 stages: at a temperature of 190-200 ° C for 1 hour, at a temperature of 220-230 ° C for 5 hours.

The determination of the physicochemical characteristics of carbon sorbents was carried out by standard methods used in the study of carbon materials: the specific adsorption surface, total pore volume, and pore size distribution were determined from nitrogen adsorption-desorption isotherms obtained on a Gemini 2380 device (Micromeritics, USA). The calculation of the adsorption specific surface area was carried out according to the BET equation. The amount of glycolic acid polymer applied to the molded sorbent was determined by differential thermal analysis based on weight loss, which was recorded from thermograms in the temperature range of 300-360 ° C. The thermal analysis method makes it possible to directly control the process of modification of the carbon sorbent, as well as to register the products of glycolic acid polycondensation on its surface, and to quantify their mass. Thermal analysis (TG-DTG-DTA) of the studied sorbents was carried out on a DTG-60H device (SHIMADZU, Japan). The temperature was determined with an accuracy of 1 ° C, and the weight change was 0.1%. The surveys were carried out in an air atmosphere, in the temperature range from room temperature to 1000 ° C, the sample weight was 10 mg, the temperature rise rate was 10 ° / min. Using pH-metric methods, a decrease in the pH of a physiological solution upon its contact with a modified sorbent from 6.4 to 1.8 was established due to the cleavage of polymer chains by hydrolysis of ester bonds to monomer units.

The results illustrating the effect of the modification process parameters (concentration of the modifier solution, impregnation time, sorbent: modifier ratio, polycondensation temperature, and sorbent size) on its physicochemical properties (specific surface area, total pore volume, glycolic acid content in the form of polyglycolide) are given in table 1.

Example 1 (prototype)

A sample of granular carbon hemosorbent weighing 1 g is impregnated with a 50% aqueous solution of glycolic acid at a hemosorbent: modifier solution 1: 1 ratio, dried at a temperature of 105 ° C for 1 hour, polycondensation at a temperature of 195 ° C for 1 hour, a temperature of 225 ° From within 5 hours. The main characteristics of this sample are given in table. one.

Example 2

The initial molded sorbent is impregnated with a 50% aqueous solution of glycolic acid at a sorbent: modifier ratio of 1: 2 (by weight) for 24 hours at room temperature, dried at 105 ° C for 1 hour, polycondensation at 195 ° C for 1 hour, temperature 225 ° C for 5 hours. The main characteristics of this sample are given in table. 1. Under the selected modification conditions, the content of glycolic acid reaches a predetermined value (not less than 5%) with partial preservation of the original porous structure, which determines the dispersive nature of the sorption of toxic compounds. These conditions are selected to obtain a modified sorbent.

Example 3

Similar to example 2, but the concentration of an aqueous solution of glycolic acid is 40%. Reducing the concentration of the glycolic acid solution and the impregnation time reduces the content of the modifier on the sorbent surface (less than 5%).

Example 4

Similar to example 2, but the time of impregnation of the sorbent with a solution of glycolic acid is 18 hours. The content of glycolic acid on the surface of less than 5% indicates that the time of impregnation of the sorbent with an acid solution is not enough to create the required amount of polymer during heat treatment.

Example 5

Similar to example 2, but the ratio of the impregnation of the sorbent: the solution of glycolic acid is 1: 1. A change in the ratio of components leads to a decrease in the content of the modifier on the surface.

Example 6

Similar to example 2, but the concentration of the modifier for impregnation is 40%, while maintaining the sorbent: solution ratio of 1: 2, the impregnation time is 24 hours. These conditions (decrease in the concentration of the modifier) do not allow the modifier to be fixed on the surface in the required amount.

Example 7

Similar to example 2, but the concentration of the modifier solution and the polycondensation temperature of the first stage were 30% and 180 ° C, respectively. A decrease in the concentration of the modifier solution and the process temperature do not allow reaching the specified value of the modifier on the surface.

Example 8

Similar to example 2, but the length of the molded sorbent is 65 mm. On the surface of a sorbent of this size, even with an increase in the duration of the modification process (impregnation stage), the concentration of the modifier does not reach the specified value of 5%.

Example 9

Similar to example 2, but the length of the molded sorbent is 35 mm. The concentration of the modifier on the surface of the sorbent of this size leads to the closure of its internal channel, which reduces the accessibility of the surface for the adsorption of pathogenic microorganisms.

Based on the results of the tests carried out, the main dimensions of the carbon sorbent and the parameters of the process of its modification with glycolic acid were determined: the length of the sorbent is 45-60 mm, the diameter is 8-10 mm; the concentration of the modifier in the solution for impregnation is 50%, the ratio of sorbent: aqueous solution of glycolic acid is 1: 2; impregnation time 24 hours. An increase in the length of the sorbent does not allow reaching 5% glycolic acid in the form of polyglycolide. Diameter 8-10 mm, which provides close contact of the sorbent with the surface of the vaginal wall and creates ideal conditions for the elimination of pathogenic microorganisms. A decrease in the concentration of the modifier in the impregnation solution reduces its amount on the surface of the sorbent in the form of polyglycolide (less than 5%). The same effect is observed with a decrease in the impregnation time.

Table 1 - Characteristics of the studied samples of molded carbon sorbent

Sample Modification modes Sorbent dimensions, mm Specific surface S _BET m ² / g Total pore volume, cm ³ / g Polyglycolide content,% Concentration of HA modifier for impregnation,% Sorbent / HA aqueous solution ratio Impregnation time, h Temperature ° С and polycondensation time, h Example 1 (prototype) fifty 1: 1 eight 195 ± 5 ° C 1 h
225 ± 5 ° C 5 h diameter 0.5-1.0 8.8 0.418 8.8 Example 2 fifty 1: 2 24 195 ± 5 ° C 1 h
225 ± 5 ° C 5 h diameter 8 length 60 7.4 0.19 7.4 Example 3 40 1: 2 12 195 ± 5 ° C 1 h
225 ± 5 ° C 5 h diameter 8 length 60 3.7 0.12 3.7 Example 4 fifty 1: 2 eighteen 195 ± 5 ° C 1 h
225 ± 5 ° C 5 h diameter 8 length 60 4.8 0.25 4.8 Example 5 fifty 1: 1 24 195 ± 5 ° C 1 h
225 ± 5 ° C 5 h diameter 8 length 60 3.2 0.20 3.2 Example 6 40 1: 2 24 195 ± 5 ° C 1 h
225 ± 5 ° C 5 h diameter 8 length 60 4.5 0.14 4.5

During the manufacturing process, the sorbent is placed in a mesh nylon container with a pore size of no more than 1 mm, which does not create obstacles for the adsorbate to pass through it. A thread is attached to the nylon container, which is necessary for the subsequent removal of the sorbent. This medical device is produced in a sterile form in a single-use package (Fig. 1A, 1B, 1C).

A molded carbon sorbent modified with glycolic acid is shown in the images: FIG. 1A - sorbent rod with one internal circular section; FIG. 1B - sorbent in a mesh nylon container; FIG. 1B - sorbent in disposable sterile packaging.

The proposed method for the treatment of bacterial vaginosis is to use a molded carbon sorbent modified with glycolic acid intravaginally for 5 days. The exposure time of one sorbent is 24 hours, after which it must be removed by pulling on the thread attached to it and a new sorbent must be introduced.

The effectiveness of this medical device lies in the ability of carbon sorbents to adsorb microbial cells, toxins secreted by microorganisms, waste products, thereby reducing the percentage of contamination of the vaginal mucosa with opportunistic flora, regardless of the resistance of pathogens to antibacterial drugs. These properties of the sorbent make it possible to solve the problem of massive seeding of the vaginal wall by opportunistic microorganisms.

The problem of treating bacterial vaginosis associated with the development of bacterial biofilms is also successfully solved using a carbon sorbent, since its physical properties (size, mesoporous structure) allow the adsorption of substances with a high molecular weight. And the bacterial biofilm is a slimy extracellular matrix consisting of proteins, polysaccharides and nucleic acids, which protects microorganisms from external influences. The adsorption of the main components of the biofilm starts the process of degradation of this extracellular polymer matrix, microbial cells begin to detach from the community and undergo sorption in the same way.

I. Microbiome. 2017 Sep 13; 5(1):119. Thermoplastic polyurethane-based intravaginal rings for prophylaxis and treatment of (recurrent) bacterial vaginosis. Verstraete G, Vandenbussche L, Kasmi S¹, Nuhn L, Brouckaert D, Van Renterghem J,

W, Vanhoorne V, Coenye T, De Geest BG, De Beer T, Remon JP, Vervaet C. Int J Pharm. 2017 Aug 30; 529(1-2):218-226. The role of lactobacilli and probiotics in maintaining vaginal health. Borges S, Silva J, Teixeira P. Arch Gynecol Obstet. 2014 Mar; 289(3):479-89. The role of lactic acid production by probiotic Lactobacillus species in vaginal health. Tachedjian G, Aldunate M, Bradshaw CS, Cone RA. Res Microbiol. 2017 Nov-Dec; 168(9-10):782-792]. Полноценное восстановление нормального влагалищного биотопа после проведенной терапии способствует снижению частоты рецидивов заболевания, улучшению качества жизни пациентки.The molded carbon sorbent proposed for the treatment of bacterial vaginosis is modified with glycolic acid, which is capable of acidifying the vaginal environment, which leads to additional damage to the bacterial biofilm and creates favorable conditions for the restoration of normal lactoflora. Thus, there is a double pathogenetic effect on the treatment of bacterial vaginosis [Treatment of biofilms in bacterial vaginosis by anamphoteric tenside pessary-clinical study and microbiota analysis. Gottschick C, Deng ZL, Vital M, Masur C, Abels C, Pieper DH, Rohde M, Mendling W,

I. Microbiome. 2017 Sep 13; 5 (1): 119. Thermoplastic polyurethane-based intravaginal rings for prophylaxis and treatment of (recurrent) bacterial vaginosis. Verstraete G, Vandenbussche L, Kasmi S ¹ , Nuhn L, Brouckaert D, Van Renterghem J,

W, Vanhoorne V, Coenye T, De Geest BG, De Beer T, Remon JP, Vervaet C. Int J Pharm. 2017 Aug 30; 529 (1-2): 218-226. The role of lactobacilli and probiotics in maintaining vaginal health. Borges S, Silva J, Teixeira P. Arch Gynecol Obstet. 2014 Mar; 289 (3): 479-89. The role of lactic acid production by probiotic Lactobacillus species in vaginal health. Tachedjian G, Aldunate M, Bradshaw CS, Cone RA. Res Microbiol. 2017 Nov-Dec; 168 (9-10): 782-792]. Full restoration of the normal vaginal biotope after the therapy helps to reduce the frequency of disease relapses and improve the patient's quality of life.

A clinical trial of the proposed method for treating bacterial vaginosis was carried out.

The study included 22 women with bacterial vaginosis. The diagnosis of bacterial vaginosis was made using the criteria of R. Amsel et al. [Amsel R, Totten PA, Spiegel CA, et al. Nonspecific vaginitis: diagnostic criteria and microbial and epidemiologic associations. Amer J Med. 1983; 74 (1): 14-22. Charonis G, Larsson PG. Use of pH / whiff test or QuickVue Advanced pH and Amines test for the diagnosis of bacterial vaginosis and prevention of postabortion pelvic inflammatory disease. Acta Obstet Gynecol Scand. 2006; 85: 837-843. Guidelines for laboratory diagnosis of urogenital tract infections. Ed. Domeyki M., Savicheva A., Sokolovsky E., Ballard R., Unemo M. 2012. St. Petersburg: Publishing house N-L, 2012; 287. Clinical guidelines for the diagnosis and treatment of diseases accompanied by pathological discharge from the genital tract of women. Russian Society of Obstetricians and Gynecologists / 2nd ed., Revised. and add. M., 2019]. When 3 out of 4 Amsel criteria were combined, the diagnosis of bacterial vaginosis was made.

The patients were divided into 2 groups: the main group (n = 11) and the comparison group (n = 11). The randomization was carried out at random. All patients underwent preliminary examination of the following: microscopy of the vaginal discharge; Real-time PCR (INBIOFLOR - Comprehensive Study of Microflora Composition of Urogenital Tract (UGT)); microbiological examination of the vaginal discharge. The growth of microflora was assessed as poor, when the level of contamination was up to 10 CFU on a Petri dish, moderate - from 10 to 100 CFU, massive - more than 100 CFU.

Therapy for bacterial vaginosis was carried out in two stages:

Stage 1: all patients included in the study (n = 22), at the first stage, received the same antibiotic therapy [Clinical guidelines for the diagnosis and treatment of diseases accompanied by pathological discharge from the genital tract of women. Russian Society of Obstetricians and Gynecologists / 2nd ed., Revised. and add. M., 2019]:

a) nitroimidazole systemically, Metronidazole tablets 500 mg orally 2 times a day for 7 days were used;

b) Clindamycin 2% vaginal cream one full applicator (5 g) once at night for 7 days.

Stage 2: the patients of the main group (n = 11) after antibiotic therapy were intravaginally injected with a molded carbon sorbent modified with glycolic acid for 5 days. Patients in the comparison group (n = 11), in accordance with clinical guidelines, received lactic acid therapy 100 mg per vaginam, 1 suppository 1 time per day for 10 days.

After the treatment, in 1 week the following control was carried out: microscopy of the vaginal discharge; Real-time PCR (INBIOFLOR - Comprehensive Study of Microflora Composition of Urogenital Tract (UGT)); microbiological examination of the vaginal discharge with the determination of the growth of colonies of microorganisms.

Data processing was carried out using the STATISTIKA-6.1 application package and standard Microsoft Excel mathematical tables. Comparison of two dependent variables (before and after treatment) was carried out using the McNemar test; comparison of two independent groups was performed using χ ² .

Tables 2, 3 show the results of treatment of patients included in the study. After the treatment, the number of complaints in the patients of both groups significantly decreased, and the microscopy indices of the vaginal discharge improved. In particular, the number of key cells decreased and the number of lactobacilli increased (Tables 2, 3). When evaluating the effectiveness of the treatment in terms of the microflora state in both groups, the results were comparable (Table 4). A decrease in the growth of gardnerella vaginalis and atopobium vaginae, streptococci, staphylococci, enterobacteria was achieved, however, the persistence of Prevotella (p = 0.023) and Escherichia coli (p = 0.028) remained in the comparison group. While in the main group, a complete restoration of normal microflora was achieved.

During further observation of the patients included in the study, three months after the therapy for bacterial vaginosis, relapses were diagnosed in two (18.2%) women in the comparison group. No relapses were observed in the main group three months after treatment.

Six months after treatment, three more patients (27.3%) came from the comparison group with recurrent complaints, who were diagnosed with recurrence of bacterial vaginosis during follow-up examination. While from the main group, after 6 months, only one woman (9.1%) applied with relapses.

Both groups received the same course of antibacterial drugs, and at the second stage, the patients of the main group used a molded carbon sorbent modified with glycolic acid, in contrast to the comparison group, where lactic acid was used. The preservation of the growth of Prevotella and Escherichia coli in the vaginal discharge in women of the comparison group indicates the resistance of these microorganisms to the antibacterial agents used. At the same time, the carbon sorbent, which was injected into the vagina in the women of the main group at the second stage of treatment, made it possible to achieve the elimination of these pathogens. Glycolic acid on the surface of the sorbent and the drug containing lactic acid used in the comparison group, by changing the pH of the medium, contributed to the restoration of lactobacilli colonies. Within six months, 45.5% of relapses were recorded in the comparison group, which could be provoked by opportunistic pathogenic microorganisms that retained their vital activity.

Thus, based on the results of the treatment and follow-up for 6 months, the following conclusions could be drawn: the use of a molded carbon sorbent modified with glycolic acid as the second stage in the treatment of bacterial vaginosis makes it possible to completely eliminate opportunistic pathogens, including strains resistant to antibacterial drugs, and to achieve a full restoration of the vaginal microflora, to reduce the frequency of recurrence of the disease within 6 months by 3 times.

Clinical examples:

Patient K., 24 years old. She complained of vaginal discharge with an unpleasant odor.

Medical history: these complaints appeared for the first time two weeks ago.

Gynecological history: Menstruation from 11 years old, regular, come in 28-30 days, 4-5 days each, moderate, painless. The last menstruation was on time, without any peculiarities. As a contraception, he has been taking JES for 1.5 years. There were no pregnancies. Not married, the beginning of sexual activity at the age of 18, currently there is no permanent sexual partner, the total number of sexual partners in the last six months is 3.

Life history: chronic gastritis, without exacerbation. Denies trauma, surgery, hepatitis, tuberculosis, blood transfusion. He denies taking antibacterial drugs for the last month.

Survey results:

Status genitalis: the external genital organs are formed, correctly, female-pattern hairiness, the skin and visible mucous membranes of the usual color, clean. Examination in the mirrors: the cervix is without visible pathology, the pharynx is rounded, there is no discharge from the pharynx; the vaginal mucosa is pale pink, the discharge is grayish, sticky, abundant, thick, with a fishy odor. When carrying out the Kolpo-Test with test strips (manufacturer: Biosensoran), pH - 7.0. The vaginal discharge was applied with a spatula on a glass slide, with the addition of 10% KOH, the fishy odor intensified. Bimanual: The vagina is narrow, the cervix is conical, up to 3 cm long, the uterus is centered in anteversio - flexio, not enlarged, the contour is even, of tight elastic consistency, mobile, no painful, the appendages are not enlarged, painless, deep vaults, parametria free.

Microscopy of the discharge: key cells were found, the content of leukocytes was 2-3 in the field of view, lactobacilli were absent, the microflora was represented mainly by coccal microorganisms.

PCR result: DNA of Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum (quantitative), Ureaplasma parvum (quantitative), Neisseria gonorrhoeae, Trichomonas vaginalis - not detected.

PCR Real-time PCR: Gardnerella vaginalis> 10%; Atopobium vaginae> 10%; Prevotella <10%; Leptotrichia amnionii> 10%; Mobiluncus curtisi <10%; Mobiluncus mulieris <10%; Lactobacillus <20%.

Bacteriological examination (sowing of vaginal discharge): massive growth of Enterococcus faecalis and Escherichia coli (more than 100 CFU); moderate growth of Enterobacterium cloaca and Gardnerella vaginalis (from 10 to 100 CFU).

Treatment: Metronidazole tablets 500 mg orally 2 times a day for 7 days; Clindamycin 2% vaginal cream one full applicator (5 g) once at bedtime for 7 days. Then the second stage of treatment was carried out: Suppositories Femilex (Femilex) containing lactic acid 100 mg 1 suppository 1 time per day per vaginam for 10 days.

After the treatment, the patient did not show any complaints.

Control studies after therapy:

Microscopy of the vaginal discharge: key cells were not found, the leukocyte count was normal, and lactobacilli were absent.

PCR Real-time PCR: Gardnerella vaginalis <10%; Atopobium vaginae <10%; Prevotella <10%; Leptotrichia amnionii> 10%; Mobiluncus curtisi <10%; Mobiluncus mulieris <10%; Lactobacillus <20%.

Bacteriological examination (sowing of vaginal discharge): the poor growth of Enterococcus faecalis and Staphylococcus epidermidis (less than 10 CFU) is determined.

After 4 months, the patient returned with recurrent complaints of pathological discharge. Upon further examination, a diagnosis of recurrence of bacterial vaginosis was made.

Patient V. 29 years old. Complaints of itching, discomfort in the vagina.

Anamnesis of the disease: complaints bother for a month.

Gynecological history: Menstruation from 13 years old, regular, come in 25-26 days, 5-6 days each, moderate, painful on the first day. The last menstruation was on time, without any peculiarities.

Sexual life from the age of 20. Married. Currently, there is 1 permanent sexual partner, the number of sexual partners in the last 6 months is 1.

Does not use contraception for 6 months. Pregnancy alone, ended in urgent childbirth two years ago, without complications.

Anamnesis of life: denies somatic diseases. Denies trauma, surgery, hepatitis, tuberculosis, blood transfusion. He denies taking antibacterial drugs for the last month.

Survey results:

Status genitalis: the external genital organs are formed, correctly, female-pattern hairiness, the skin and visible mucous membranes of the usual color, clean. Examination in mirrors: cervix without visible pathology, slit pharynx, no discharge from the pharynx; the vaginal mucosa is pale pink, the discharge is abundant, sticky, whitish, thick, with a fishy odor. When carrying out the Kolpo-Test with test strips (manufacturer: Biosensoran), pH - 6.5. The vaginal discharge was applied with a spatula on a glass slide, with the addition of 10% KOH, the fishy odor intensified. Bimanual: The vagina is capacious, the cervix is cylindrical, up to 3 cm long, the uterus is centered in anteversio-flexio, not enlarged, the contour is smooth, tight-elastic consistency, mobile, painless, the appendages are not enlarged, painless, the fornices are deep, the parametria are free.

Microscopy of the discharge: key cells are present, the content of leukocytes is 25, lactobacilli are absent.

PCR result: DNA of Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum (quantitative), Ureaplasma parvum (quantitative), Neisseria gonorrhoeae, Trichomonas vaginalis - not detected.

PCR Real-time PCR: Gardnerella vaginalis> 10%; Atopobium vaginae> 10%; Prevotella <10%; Leptotrichia amnionii <10%; Mobiluncus curtisi <10%; Mobiluncus mulieris <10%; Lactobacillus <20%.

Bacteriological examination (sowing of vaginal discharge): massive growth of Escherichia coli (more than 100 CFU); moderate growth of Gardnerella vaginalis (from 10 to 100 CFU); moderate growth of Enterococcus faecalis (10 to 100 CFU); moderate growth of Streptococcus agalactia (10 to 100 CFU); poor growth of Proteus mirabilis (less than 10 CFU).

Treatment: Metronidazole tablets 500 mg orally 2 times a day for 7 days; Clindamycin 2% vaginal cream one full applicator (5 g) once at bedtime for 7 days. Then the second stage of treatment was carried out: after the performed antibacterial therapy, a molded carbon sorbent modified with glycolic acid was intravaginally injected for 5 days.

After the treatment, the patient had no complaints.

Control studies:

Microscopy of the vaginal discharge: key cells were not found, the content of leukocytes 2-3 in the field of view, lactobacilli were found.

PCR Real-time PCR: Gardnerella vaginalis <10%; Atopobium vaginae <10%; Prevotella <10%; Leptotrichia amnionii <10%; Mobiluncus curtisi <10%; Mobiluncus mulieris <10%; Lactobacillus> 20%.

Bacteriological examination (sowing of the vaginal discharge): the growth of pathogenic microflora was not detected.

After 6 months, no relapses of the disease were registered.

Table 2 - Dynamics of indicators during treatment in the main group (n = 11)

Indicator Before treatment After treatment McNemar criterion p abs % abs % 1.The presence of complaints 10 91 0 0 10 0.002 2.pH> 4.5 eleven 100 one nine 10 0.002 3.microscopic examination of the vaginal discharge the presence of key cells eleven 100 one nine 10 0.002 leukocyte count> 20 6 54 0 0 6 0.015 number of lactobacilli <5 eight 72 one nine 7 0.009 4. PCR test "Inbioflor" Gardnerella vaginalis eleven 100 0 0 eleven 0.001 Atopobium vaginae 7 64 0 0 7 0.009 Prevotella eight 72 one nine 7 0.009 Leptotrichia amnionii 7 64 2 eighteen five 0.026 Mobiluncus curtisi four 36 0 0 four 0.046 Mobiluncus mulieris 3 27 0 0 3 0.084 Lactobacillus <20% 7 64 0 0 7 0.009 Lack of lactobacilli DNA 3 27 0 0 3 0.084 5.bacteriological examination of the vaginal discharge Escherichia coli 6 54 0 0 6 0.015 Streptococcus agalactiae five 45 0 0 five 0.026 Staphylococcus epidermidis four 36 0 0 four 0.046 Enterococcus faecalis five 36 one nine four 0.046 Enterobacterium cloacae 2 eighteen 0 0 2 0.158 Proteus mirabilis 2 nine 0 0 2 0.158 Gardnerella vaginalis 3 27 0 0 3 0.084

Table 3 - Dynamics of indicators during treatment in the comparison group (n = 11)

Indicator
n = 11 Before treatment After treatment McNemar criterion p abs % abs % 1.The presence of complaints 10 91% four 36% 6 0.015 2.pH> 4.5 eleven 100% 2 eighteen% nine 0.003 3.microscopic examination of the vaginal discharge the presence of key cells eleven 100% 2 eighteen% nine 0.003 leukocyte count> 20 five 45% four 36% one 0.318 number of lactobacilli <5 eight 72% four 36% four 0.046 4. PCR test "Inbioflor" Gardnerella vaginalis eleven 100% 2 eighteen% nine 0.003 Atopobium vaginae 7 64% 3 27% four 0.046 Prevotella eight 72% 6 54% 2 0.158 Leptotrichiaamnionii 6 54% four 36% 2 0.158 Mobiluncus curtisi four 36% 3 27% one 0.318 Mobiluncus mulieris 3 27% 2 eighteen% one 0.318 Lactobacillus <20% 7 64% 3 27% four 0.005 Lack of lactobacilli DNA 3 27% 3 27% - - 5.bacteriological examination of the vaginal discharge Escherichia coli 6 54% four 36% 2 0.158 Streptococcus agalactiae five 45% 2 eighteen% 3 0.084 Staphylococcus epidermidis four 36% 0 0 four 0.046 Enterococcus faecalis four 36% 3 27% one 0.318 Enterobacterium cloacae 2 eighteen% one nine% one 0.318 Proteus mirabilis one nine% one nine% - - Gardnerella vaginalis 3 27% one nine% 2 0.158

Table 4 - Comparison of the results of the treatment carried out in both groups

Indicator Main group (n = 11) Comparison group
(n = 11) Criterion χ ² p abs % abs % 1.The presence of complaints 0 0 four 36 4.889 0.028 2.pH> 4.5 one nine 2 eighteen 2,220 0.139 3.microscopic examination of the vaginal discharge the presence of key cells one nine 2 eighteen 3.841 0.139 leukocyte count> 20 0 0 four 36 4.889 0.028 number of lactobacilli <5 one nine four 36 2,329 0.127 4. PCR test "Inbioflor" Gardnerella vaginalis 0 0 2 eighteen 2.200 0.139 Atopobium vaginae 0 0 3 27 3.474 0.063 Prevotella one nine 6 54 5.238 0.023 Leptotrichiaamnionii 2 eighteen four 36 0.917 0.339 Mobiluncus curtisi 0 0 3 27 3.474 0.063 Mobiluncus mulieris 0 0 2 eighteen 2.2 0.139 Lactobacillus <20% 0 0 3 27 3.474 0.063 Lack of lactobacilli DNA 0 0 3 27 3.474 0.063 5.bacteriological examination of the vaginal discharge Escherichia coli 0 0 four 36 4.889 0.028 Streptococcus agalactiae 0 0 2 eighteen 2,432 0.119 Staphylococcus epidermidis 0 0 0 0 - - Enterococcus faecalis one nine 3 27 1,222 0.269 Enterobacterium cloacae 0 0 one nine 1,048 0.307 Proteus mirabilis 0 0 one nine 1,048 0.307 Gardnerella vaginalis 0 0 one nine 1,048 0.307

Claims (3)

1. Shaped carbon sorbent with antibacterial properties for the treatment of bacterial vaginosis, which is a nanodispersed mesoporous carbon material containing a polymer of glycolic acid, with a specific adsorption surface of not more than 50 m ² / g, characterized in that it is made in the form of cylinders with a diameter of 8-10 mm , 45-60 mm long, with one internal circular channel and is characterized by a glycolic acid polymer content in the form of a polyglycolide of at least 5 wt.% to 7.4 wt.%; crushing strength of at least 20 kg / cm ² . 2. A method of manufacturing a molded carbon sorbent described in claim 1, which comprises impregnating the original molded carbon sorbent with a 50% aqueous solution of glycolic acid for 24 hours at room temperature, the ratio of sorbent: glycolic acid solution 1: 2 by weight followed by drying for an hour at 103-107 ° C and polycondensation of glycolic acid on a molded carbon sorbent in two stages: at a temperature of 190-200 ° C for 1 hour, at a temperature of 220-230 ° C for 5 hours. 3. A method of treating bacterial vaginosis in gynecology, including sanitation using a sorption material, characterized in that the molded carbon sorbent described in claim 1 or prepared by the method according to claim 2, modified with polymer glycolic acid, in a mesh nylon container with an attached thread for subsequent extraction in a sterile form intravaginally injected for 5 days for 24 hours.

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