EA034431B1 - Pharmaceutical composition in the form of suppository with wound-healing, anti-erosive, anti-inflammatory, antimicrobial, antifungal, immunomodulating and antioxidant action - Google Patents

Abstract

The invention relates to the field of medicine and concerns a pharmaceutical composition in the form of suppositories with wound-healing, anti-erosion, anti-inflammatory, antimicrobial, antifungal, immunomodulating and antioxidant effects, containing active substances of thiotriazoline and decamethoxin and vehicle, wherein the ratio of active substances is as follows: decamethoxin ranging from 0.5 to 5.0 mass fractions per 100 mass fractions of thiotriazoline.

Description

The invention relates to medicine and pharmacy, in particular medicines that are used for topical treatment of non-specific inflammatory and infectious diseases of the urogenital tract of various etiologies and erosive lesions of the mucous membranes of the vagina and cervix, rectum, urethra, etc.

Such diseases include erosion of the cervix and vaginal mucosa, vaginitis, vulvovaginitis, colpitis, cervicitis, urethritis, proctitis, anal fissures and other diseases.

Speaking of inflammatory diseases of the genital area in women, it should be noted that the most common are diseases of mixed etiology. In recent years, more and more attention has been paid to finding optimal solutions to problems associated with the treatment of such diseases.

The relevance of this direction is determined by the tendency to generalization and chronization of processes, the development of serious pathophysiological and pathomorphological changes in affected tissues and organs, the involvement of the immune, nervous, endocrine, reproductive and other body systems in the pathological process.

The modern arsenal of drugs offers a large number of drugs that are effective in treating a particular type of inflammation and erosion of the mucous membranes of the urogenital tract. These are, for example, methyluracil suppositories (http://www.piluli.kharkov.ua/drugs/drug/2555/), candles with sea buckthorn oil (http://mozdocs.kiev.ua/likiview.php?id=26931) , suppositories Texicon (Chlorhexidine) (https://www.vidal.ru/drugs/hexicon_d_8945).

However, suppositories with methyluracil do not have a sufficiently strong anti-inflammatory and wound healing effect, the effect of the drug appears only on the 30th day. Methyluracil has no antioxidant, fungicidal and antimicrobial effects, which makes this drug ineffective in some cases. Suppositories with sea buckthorn oil do not have antioxidant, fungicidal and antimicrobial effects, which narrows their spectrum of action. Texicon suppositories have a side effect - they have an allergenic effect, and this tool does not have an antioxidant, wound healing and immunomodulating effect.

In modern conditions, the most relevant in gynecology are combined topical preparations, which include an antibiotic or antiseptic, an antifungal component, and metronidazole preparations.

The well-known combined drug Polygynax, the effect of which is determined by the properties of its components. Antibiotics polymyxin B and neomycin have an antimicrobial effect against gram-negative and gram-positive microorganisms. Nystatin has a fungicidal (aimed at the destruction of parasitic fungi) action against candida fungi. Dimethylpolysiloxane gel has an enveloping and antipruritic effect, improves trophic processes (tissue nutrition processes) in the vaginal mucosa, and promotes the penetration of the active components of the drug into small vaginal folds (http://www.piluli.kharkov.ua/drugs/drug/2153/) .

However, it should be noted that antibiotics (they are part of this drug) are currently in the majority of pathogens of diseases of the urogenital sphere has developed persistent resistance, which negates therapy with these agents. In addition, this tool does not have anti-inflammatory, antioxidant, immunomodulating and anti-erosion effects.

Terzhinan is a complex antibacterial and antifungal drug for the treatment of gynecological diseases. Ternidazole is active against anaerobic flora, including gardnerella, and also has a trichomonacid effect. Neomycin sulfate is an aminoglycoside antibiotic that acts on pyogenic bacteria in the vagina. Nystatin is an antifungal drug of the polyene series, active against fungi of the genus Candida. Prednisolone metasulfobenzoate sodium is a corticosteroid that has a local anti-inflammatory effect. Additional components of Terzhinan tablets allow maintaining the integrity of the vaginal epithelium in infectious and inflammatory diseases and maintain the physiological pH balance (http://www.piluli.kharkov.ua/drugs/drug/ter gynan /).

Terzhinan, like Polygynax, contains an antibiotic; although the drug has anti-inflammatory properties, however, it does not have antioxidant, immunomodulating and anti-erosion effects, which makes it also not effective enough for a number of diseases.

Modern studies have shown that, despite the variety of etiological factors, the activation of oxidative stress plays a leading role in the pathogenesis of most gynecological diseases. Cytotoxic oxidative stress metabolites intensify the cascade of arachidonic acid transformations, contribute to the accumulation of polymorphonuclear leukocytes in the focus of inflammation, the activation of which is also accompanied by the formation of free radicals, enhance inflammatory processes, reduce the immune system, and suppress molecular mechanisms of reparative regeneration. Thus, a vicious circle arises that contributes to the maintenance of an undamped inflammatory process. According to published data, the use of antioxidants in the complex treatment of gynecological diseases contributes to their rapid relief, reduces the intensity of oxidative stress processes, reduces the formation of pro-inflammatory cytokines and arachidonic acid metabolites, and promotes the repair of the vaginal mucosa. there is

- 1 034431 data on antioxidant modulation of chemotherapeutic agents - for example, antiseptics. This fact opens up prospects for the use of antioxidants in conjunction with antiseptic agents for the treatment of infectious and inflammatory diseases of the female genital organs, as well as diseases of the urogenital sphere in men, inflammatory lesions of the rectum, infected and uninfected wounds, and the like.

Known drug Wound healing hepatoprotective suppository (Ukrainian patent No. 43620, published November 17, 2003, bull. No. 11). The suppository contains the active component thiotriazolin and, as the fat base, solid fat, for example hydrogenated sunflower oil in the following ratio of components, g / kg: thiotriazolin - 80.0-88.0; fat is the remainder. A suppository with a mass of 2.28-2.52 g contains 0.18-0.22 g of thiotriazoline.

However, this tool does not include active components that would provide an antimicrobial and antiseptic, fungicidal effect, which makes the drug not effective enough in the treatment of diseases in which the bacterial factor joins the inflammatory process. In addition, this drug has no anti-erosion effect.

A medicament is also known. A pharmaceutical composition in the form of a soft dosage form with anti-inflammatory, antimicrobial and reparative effects (Ukrainian patent No. 105127, published on 10.04.2014, bull. No. 7). The pharmaceutical composition contains the active substance decamethoxin, an emulsion ointment base, and additionally contains dexpanthenol, vitamins cholecalciferol, retinol acetate and tocopherol acetate. The composition of the emulsion ointment base includes emulsifier No. 1, soybean oil, grape seed oil, nipagin, nipazole, petroleum jelly, purified water.

It should be noted that decamethoxin was added to this combination agent in a concentration insufficient for the drug to have a pronounced antibacterial and fungicidal effect. Although the composition of the composition includes a fat-soluble antioxidant - tocopherol acetate (vitamin E), however, in the indicated concentration it realizes most of its vitamin and not antioxidant properties, acting on the final stages of oxidative stress - the peroxide stage, and not at the stage of initiation of reactive oxygen species (AFC).

In the literature available to us, we did not find any mention of a combined agent that would contain both an antiseptic and an antioxidant - an ROS scavenger, i.e. which could be effective in diseases in the pathogenesis of which the initial stages of oxidative stress play the main role, that is, the initiation of ROS.

The basis of the invention was the task of developing a pharmaceutical composition in the form of suppositories, which will provide a complex effect with a wide range of pharmacological activity, namely, have simultaneously wound healing, anti-erosion, anti-inflammatory, antimicrobial, antifungal, immunomodulatory and antioxidant activities, while exceeding the therapeutic effect reference drugs, which will increase the effectiveness of the drug, increase the effectiveness of local therapy using m of such a drug will help to avoid polypharmacy and reduce the pharmacological load on the patient’s body and expand the arsenal of effective domestic medicines used in gynecology, urology and proctology.

The problem is solved in that in a pharmaceutical composition in the form of suppositories with wound healing, anti-erosion, anti-inflammatory, antimicrobial, antifungal, immunomodulating and antioxidant effects, which contains the active substance thiotriazoline and the base, it is new that it additionally contains the active substance decamethoxin.

Also new is that its dosage form is vaginal suppositories, rectal suppositories, sticks.

What is new is that the pharmaceutical composition contains the active substance decamethoxin and thiotriazoline in this ratio: decamethoxin in the range from 0.5 to 5.0 mass fractions per 100 mass fractions of thiotriazoline.

Also new is the fact that the pharmaceutical composition contains a lipophilic or hydrophilic base.

Based on the mechanisms of action of both components of the proposed combined agent containing thiotriazolin and decamethoxin that we studied, we suggested the possibility of a synergistic effect in their interaction, in which the pharmacological properties of the active components can be enhanced and mutually potentiated.

It is known that the pharmacodynamic interaction of drugs can be directed both towards enhancing their pharmacological effects (synergism), and to weaken them (Kukes

V.G. Clinical Pharmacology, 4th edition. - Moscow: GEOTAR-Media, 2008 .-- S. 164-169; Gilman A.G. Clinical pharmacology according to Goodman and Gilman. - M: Practice, 2006. - S. 57).

Physical incompatibility, chemical incompatibility, and pharmacological incompatibility of the components of the combined pharmaceutical composition may also occur.

Thus, only an experimental study can confirm or deny the possibility of creating a specific combination drug.

We conducted a series of experimental studies that confirmed our assumption about the possibility of creating a pharmaceutical composition that contains thiotriazoline and decamethoxin as active components and exhibits higher pharmacological efficacy compared to known drugs.

At the same time, we studied various combinations of thiotriazolin with decamethoxin - the following ratio of active components was studied - from 0.5 to 5.0 mass fractions of decamethoxin per 100 mass fractions of thiotriazoline. All ratios in the range of 0.5-5.0 fractions of decamethoxin per 100 parts of thiotriazolin were pharmacologically effective, but the ratio 1: 100, respectively, was the most active.

After conducting our experimental studies of the indicated pharmaceutical composition in the dosage form of a suppository, we obtained the following results.

1) The active components of the claimed pharmaceutical composition, namely thiotriazolin and decamethoxin, have physical, chemical and pharmacological compatibility.

2) The pharmaceutical composition with thiotriazolin and decamethoxin exerts a pronounced erosion and wound healing effect and in strength the erosion and wound healing effect significantly exceeds the reference drugs (suppositories with thiotriazolin and suppositories with decamethoxin and mucous membranes reduce the degree of moisture damage of thiotriazole and decathiazole , superior in efficiency to suppositories with thiotriazolin and suppositories with decamethoxin. So, in the group of experimental animals that received the claimed combination, the degree of damage to the vaginal mucosa in points was 8.8 times lower than untreated animals, 2.4 times lower than the group receiving suppositories with thiotriazoline, and 7.9 times compared with the group receiving suppositories with decamethoxin. On the 10th day of treatment, 30% of the animals were completely cured (the mucous membrane of the vagina was without pathological changes) in the group receiving therapy of the proposed pharmaceutical composition, while in other groups for this observation period there were no completely cured animals (data are presented in table 1).

3) In terms of anti-inflammatory effect, the pharmaceutical composition with thiotriazolin and decamethoxin significantly exceeds suppositories with decamethoxin and suppositories with thiotriazoline, as evidenced by a significantly larger decrease in inflammation markers - C-reactive protein and IL-1b, as well as a decrease in neutrophils compared to reference drugs . In the blood of experimental animals receiving the claimed combination in a 10-day course, the concentration of reactive protein was 5 times lower than in the blood of the group of treated animals, and 2.3 times lower than in the group treated with suppositories with thiotriazoline (data are presented in table 2).

The pharmaceutical composition with thiotriazolin and decamethoxin exhibits an immunomodulating effect that is absent when using suppositories with decamethoxin, normalizing the number of lymphocytes (data are presented in Table 2).

4) The pharmaceutical composition with thiotriazolin and decamethoxin exhibits a pronounced antioxidant effect, which is superior in strength to the reference drug - suppositories with thiotriazolin. The claimed composition reduces by 2.2 times the indicators of the marker of oxidative stress AFG relative to untreated animals, and 1.3 times relative to the group of animals treated with suppositories with thiotriazoline. In this case, suppositories with decamethoxin do not have antioxidant effects (data are presented in table. 3).

5) According to the antimicrobial effect, the proposed pharmaceutical composition is more than 2 times superior to the reference drug for E. coli and C. Albicans (120 and 108%, respectively) and more than half as effective as S.aureus and P.aeruginosa, the causative agents of inflammatory diseases urogenital spheres, which are most often encountered (data are presented in table. 4).

Also a positive feature of the proposed pharmaceutical composition is that its components are not addictive, and it does not have an irritating effect.

Below we detail our findings confirming that the proposed pharmaceutical composition has higher wound healing, anti-erosion, anti-inflammatory, antimicrobial, antifungal, immunomodulating and antioxidant activity compared with the action of suppositories with thiotriazolin and compared with the effect of suppositories with decamethoxin. In this case, suppositories only with thiotriazolin do not have antiseptic and antifungal activity, and suppositories only with decamethoxin do not have, in particular, immunomodulating and antioxidant effects.

The studies were performed on white outbred female rats with a body weight of 180-190 g. The studies were conducted on the example of experimental vulvovaginitis. The experimental model (phenol vulvovaginitis) was caused by 5-day introduction into a known animal to a depth of 3-5 mm using a metal probe (diameter 2 mm) of 5% phenol solution at the rate of 0.1 ml per 100 g of animal weight. The phenolic model causes severe inflammation and ulceration of the vaginal mucosa. For this experimental study of the proposed pharmaceutical composition, the dosage form — suppositories — was chosen. 200 mg suppositories containing 2034431 containing 20 mg of thiotriazoline and 0.2 mg of decamethoxin were prepared; suppositories with 20 mg of thiotriazolin (reference drug); suppositories with 0.2 mg decamethoxin (reference drug). The studied and reference suppositories were administered vaginally immediately after the last phenol injection during the next 9 days (treatment regimen) 1 time per day, 1 suppository per 200 g of weight. The experimental control group (positive control group) received suppositories without active substances, which were administered in the same way and in the same amount as the test candles, after the development of nonspecific vulvovaginitis. On the 15th day of the experiment (on the 10th day of treatment), animals of all groups were anesthetized with thiopental sodium (40 mg / kg). Then, animals were sampled from the peritoneal artery, transferring it to pre-prepared tubes, and the vaginal mucosa was examined. The composition of peripheral blood was determined in blood smears. Biochemical and molecular markers of inflammation, oxidative stress, and antioxidant system and immunity were determined in blood serum.

The results of the studied anti-erosion and wound healing activity. The anti-erosion and wound healing activity of the proposed pharmaceutical composition was evaluated by the condition of the vaginal mucosa (SOV) in points assigned depending on the severity of signs of edema, hyperemia, erosion: 0 points - no sign, 1 point - slightly expressed, 2 points - moderate , 3 points - the sign is strongly expressed. The number of spot hemorrhages on the mucosa was also taken into account, which was calculated using a magnifier (an increase of 7 times), as well as the area of necrosis of the vaginal mucosa, which was calculated on the basis of data obtained with a caliper and expressed in mm ² .

The experimental results are presented in table. 1.

Table 1. The effect of the pharmaceutical composition thiotriazolin + decamethoxin on the condition of the vaginal mucosa (SOV) of rats with experimental phenolic vulvovaginitis on the 10th day of treatment

Group animals Sum of Status Points OWL % of animals without changes in SOW The number of point hemorrhages in the SOW The area of destruction of SOW (mm ² ) % of animals without hemorrhage OWL Intact (n = 10) 0 100 0 0 100 Vulvovaginitis (control) (n = 10) 8.8 ± 0.42 0 7.10 ± 0.99 7.60 ± 1.07 0 Vulvovaginitis + decamethoxin (n = 10) 7.9 ± 0.87 (-10.2%) 0 6.50 ± 0.70 (-8.5%) 6.80 ± 0.91 (-10.5%) 0 Vulvovaginitis + thiotriazolin (n = 10) 2.40 ± 0.51 * ^# (-72.7%) 0 1.10 ± 0, sG ^# (-84.5%) 1.30 ± 0.48 * ^# (-83%) 0 Vulvovaginitis + thiotriazolin + decamethoxin (n = 10) 1.0 ± 0.81 * ^{# L} (-88.6%) 20 0.80 ± 0.63 * ^# (-88.7%) 0.80 ± 0.63 * ^# (-90%) thirty

Note:

* - significant changes relative to the control group (p <0.05);

# - significant changes relative to the decamethoxin group (p <0.05);

^Λ - significant changes relative to the thiotriazolin group (p <0.05).

As can be seen from the data table. 1, the phenolic model of vulvovaginitis causes a sufficiently severe ulceration of the vaginal mucosa. In 100% of the control (untreated) animals, signs of vulvovaginitis and severe SOB lesion were recorded. This is indicated by indicators on the 10th day of the experiment, characterizing the condition of the vaginal mucosa - 7.6 mm ² - the area of the mucosal lesion and more than 7 point hemorrhages. The main clinical manifestations of phenolic vulvovaginitis were signs of inflammation: excretion, including and bloody from the vagina, hyperemia of the vulva and vagina, pain, itching, frequent urination. The severity of edema, hyperemia and the number of hemorrhages were estimated at 8.8 points. The course (9-day) application of suppositories with decamethoxin did not have a significant effect on the number of erosions and the area of SOV damage in rats with vulvovaginitis.

The course administration of suppositories with thiotriazoline to rats with vulvovaginitis resulted in a significant 84.5% decrease in the number of point hemorrhages in the SOW (6.45 times less than in the control group) and a 83% decrease in the area of the SOB (5.84 times less than in the control group). At the same time, treatment with thiotriazolin led to a decrease in the severity of edema, hyperemia; the number of hemorrhages was estimated at 6.2 points less (72.7%) compared with the control group.

The course intravaginal administration of suppositories with thiotriazolin and decamethoxin to rats with vulvovaginitis in rats resulted in a significant decrease in the number of point hemorrhages by 88.7% (8.87 times less than in the control group) in the SOV and a 90% reduction in the area of the SOB (9, 5 times less than in the control group). Moreover, treatment with thiotriazolin and decamethoxin led to a decrease in the severity of edema, hyperemia; the number of hemorrhages was estimated at 7.8 points less (88.6%) compared with the control group. It is worth noting that the use of suppositories with thiotriazolin and decamethoxin compared to suppositories with decamethoxin and suppositories with thiotriazoline led to a complete restoration of SOW in 20% of animals and to a complete absence of hemorrhagic erosion in 30% of animals, which indicates a pronounced anti-erosion and wound healing activity the drug.

Suppositories with thiotriazolin and decamethoxin significantly exceed the suppositories with decamethoxin in the number of animals fully cured, in the decrease in the area of SOW lesion, in the decrease in the amount of erosion and in the decrease in the severity of edema, hyperemia and the number of hemorrhages, and suppositories in terms of the effectiveness and suppository of suppositories. thiotriazolin in the number of fully cured animals, for a decrease in the severity of edema, hyperemia and the number of hemorrhages in points.

Thus, suppositories with thiotriazolin and decamethoxin exhibit a pronounced anti-erosion and wound healing effect on the phenolic vulvovaginitis model and are superior to anti-erosion and wound healing effects in reference preparations - suppositories with thiotriazoline and suppositories with decamethoxin.

The study of anti-inflammatory and immunomodulatory activity.

The anti-inflammatory and immunomodulatory activity of the proposed pharmaceutical composition in the form of suppositories was evaluated by reducing molecular markers of inflammation in the blood of reactive protein, interleukin-1b (IL-1b) and neutrophils, and also by increasing the lymphocytes involved in cellular immunity. As can be seen from the table. 2, the phenolic model causes a sufficiently severe ulceration of the vaginal mucosa, accompanied by a pronounced inflammatory process and impaired immune system. So, in rats of the control group (without treatment) a significant increase in the level of C-reactive protein was recorded in the blood - 8.5 times and IL-1b - 7 times against the background of an increase in neutrophils by 121.3%. Moreover, in rats of the control group, the number of lymphocytes decreased by 9%.

Table 2. The effect of the pharmaceutical composition thiotriazoline + decametho-xin on molecular markers of inflammation and immunity in the blood of rats with experimental phenolic vulvovaginitis on the 10th day of treatment

Group of animals IL-1b pg / ml C-reactive protein, ng / l Neutrophils, % Lymphocytes % Intact (n = 10) 0.34 ± 0.04 1.10 ± 0.31 30.5 ± 1.71 77.5 ± 5.01 Vulvovaginitis (control) (n = 10) 2.38 ± 0.71 9.40 ± 0.69 67.5 ± 3.02 70.5 ± 3.02 Vulvovaginitis + Decamethoxin (n = 10) 1.74 ± 0.11 (-27%) 7.00 ± 1.15 * (-25.5%) 60.5 ± 4.00 (-10%) 70.7 ± 6.01 (± 0.3%) Vulvovaginitis (control) (n = 10) 1.07 ± 0.1G ^# (-55%) 4.30 ± 0.67 * ^# (-54.2%) 41.5 ± 2.01 * (-38.5%) 76.8 ± 3.08 * (± 9%) Vulvovaginitis + thiotriazolin + decamethoxin (n = 10) 0.73 ± 0.17 * ^# (-70%) 1.90 ± 0.73 * ^{# L} (-80%) 33.1 ± 1.10 * ^{# L} (-51%) 77.0 ± 2.10 * (± 9.2%)

Note:

* - significant changes relative to the control group (p <0.05);

# - significant changes relative to the decamethoxin group (p <0.05);

^Λ - significant changes relative to the thiotriazolin group (p <0.05).

The administration of suppositories with decamethoxin to animals with phenolic vulvovaginitis for 10 days had a moderate anti-inflammatory effect, significantly reducing the C-reactive protein by 25%, without affecting other indicators of inflammation and immunity.

The course administration of suppositories with thiotriazoline to rats with vulvovaginitis resulted in a significant decrease in molecular markers of inflammation - C-reactive protein by 55% and IL-1b by 54.2%, and neutrophils by 38.5%. The use of suppositories with thiotriazoline led to an increase in lymphocytes by 9% compared with the control group.

The course intravaginal administration of suppositories with thiotriazoline and decamethoxin to rats with vulvovaginitis resulted in a significant decrease of 80% in the level of C-reactive protein and in 70% of IL-1b, as well as neutrophils by 51%. The use of suppositories with thiotriazolin and decamethoxin led to an increase in lymphocytes by 9.2%.

Thus, in terms of anti-inflammatory effect, suppositories with thiotriazolin and decamethoxin significantly exceed suppositories with decamethoxin in reducing markers of inflammation - 5,034,431

C-reactive protein and IL-lb, as well as to reduce neutrophils.

Suppositories with thiotriazolin and decamethoxin significantly exceed the effectiveness of suppositories with thiotriazolin in reducing C-reactive protein (2.3 times) and neutrophils (2 times).

Suppositories with thiotriazolin and decamethoxin also exhibit an immunomodulatory effect, which is absent when using suppositories with decamethoxin, normalizing the number of lymphocytes. The results of the study of antioxidant activity. The antioxidant activity of the proposed pharmaceutical composition with thiotriazolin and decamethoxin in the form of suppositories was evaluated by a decrease in blood markers of oxidative stress - aldehyde phenylhydrazones (APH) and carboxyphenylhydrazones (CPH), as well as by an increase in the activity of antioxidant protection enzymes - superoxide dismutase and Sodium redoxidase (SOD) glutathione peroxidase (HSP).

As can be seen from the table. 3, the phenolic vulvovaginitis model leads to activation of oxidative stress, as evidenced by an increase in blood of control (without treatment) animals in the level of AFH by 131.7% and CPH by 82.3% against the inhibition of SOD activity by 46.3%, GR by 42.7% 35 and GPR by 15.7%.

Table 3. The effect of the pharmaceutical composition thiotriazolin + decamethoxin on the activity of antioxidant enzymes and the content of oxidative stress markers in the blood of rats with experimental phenolic vulvovaginitis on the 10th day of treatment

Group animals SOD, cu / mg Нb / min GR, μm / mg Hb / min GSR, μm / mg Hb / min AFG, c.u / g protein KFG, cu / g protein Intact (n = 10) 288.8 = 1 = 11.5 28.8 ± 1.7 75.2 ± 4.5 12.0 ± 1.1 5.1 ± 0.32 Vulvovaginitis (control) (n = 10) 155.2 ± 10.6 16.5 ± 1.2 63.4 ± 5.0 27.8 ± 2.2 9.3 ± 0.65 Vulvovaginitis + decamethoxin (n = 10) 157.7 ± 17.2 (+ 1.6%) 16.7 ± 1.4 (+ 1.2%) 64.7 ± 7.4 (± 2%) 26.2 ± 2.4 (-5.7%) 9.1 ± 0.87 (-2.1%) Vulvovaginitis + thiotriazoline (n = 10) 275.2 ± 12.0 ^m (± 74.7%) 27.2 ± 1.4 * ^# (± 65%) 75.8 ± 4.2 * ^# (± 19.6%) 16.2 ± 1.2 * ^# (-41.7%) 5.8 ± 0.37 * ^# (-37.6%) Vulvovaginitis + thiotriazolin + decamethoxin (n = 10) 294.2 ± 11.0 * ^# (± 89.5%) 28.5 ± 1.8 * ^# (± 72.7%) 77.5 ± 5.0 * ^# (± 22.2%) 12.8 ± 1.0 * ^{# L} (-54%) 5.2 ± 0.40 * ^# (-44.0%)

Note:

* - significant changes relative to the control group (p <0.05);

# - significant changes relative to the decamethoxin group (p <0.05);

λ - significant changes relative to the thiotriazoline group (p <0.05).

The administration of suppositories with decamethoxin to animals with phenolic vulvovaginitis for 9 days did not significantly affect the oxidative stress and antioxidant system, which indicated the absence of an antioxidant effect in this preparation.

The course administration of suppositories with thiotriazoline to rats with vulvovaginitis in rats resulted in a significant decrease in APH by 41.7% and CPG by 37.6% and a significant increase in the activity of SOD by 74.7%, GR by 65% and GSR by 19.6%.

Course intravaginal administration of suppositories with thiotriazoline and decamethoxin to rats with vulvovaginitis in rats resulted in a significant decrease in blood AFG by 54% and CPG by 44% and a significant increase in the activity of SOD by 89.5%, GR by 72.7%, and GSR by 22.2% , which indicates a high antioxidant activity. Suppositories of thiotriazolin and decamethoxin in such an indicator as a decrease in antiphosphonamine antioxidant activity are 1.3 times superior to the antioxidant activity of suppositories with thiotriazoline.

Thus, suppositories with thiotriazolin and decamethoxin exhibit a pronounced antioxidant effect on the phenol vulvovaginitis model, which, according to the numbers given above, surpass reference drugs.

The results of the study of antimicrobial and antifungal activity.

The study of the antimicrobial and antifungal activity of the studied suppositories was carried out by diffusion into agar (well method).

Staphylococcus aureus cultures - Staphylococcus aureus ATCC 25923, E. coli - Escherichia coli ATCC 25922, Pseudomonas acraginosa ATCC 27853, diploid fungus (thrush) - Candida albicans ATCC 885 or 653 were used as test strains; units (CFU) - 10 ⁶ CFU / ml. For studies on S.aureus, E. coli, P.aeraginosa, Mueller-Hinton agar was used, and Saburo agar with Calbicans. According to generally accepted methods, thin-walled cylinders with a diameter of 7 mm were installed in Petri dishes with pre-spilled and cooled agar and 13.5 ml of agar melted and cooled to 45 ° C mixed with a seeded dose (1.5 ml) of the test microorganism was poured .

After cooling, the cylinder was removed and 0.3 ml of the studied suppositories were added to the obtained wells. Crops with S.aureus, E. coli, P.aeruginosa were incubated at 37 ° C, with Calbicans incubated at

28 ° C.

The results of studies with strains of S. aureus, E. coli, P.eraginosa were evaluated after 24 hours of incubation, in crops with Calbicans, first after 24 hours, then after 48 hours. The results were calculated by measuring growth inhibition zones (mm).

Table 4. Antimicrobial activity of the pharmaceutical composition thiotriazolin + decamethoxin by the size of growth inhibition zones (mm) of microorganism cultures (10 ⁶ CFU / ml of medium)

Dosage form S. aureus E. soi R. aeruginosa C. albicans 1. Suppositories thiotriazolin + decamethoxin 22.7 ± 0.57 (± 54%) 16.0 ± 0.01 (± 120%) 13 ± 1.0 (± 57%) 16.0 ± 0.01 (± 108%) 2. Suppositories with decamethoxin 14.7 ± 0.57 7.3 ± 0.57 8.3 ± 0.57 7.7 ± 0.57

As can be seen from the data presented in table. 4, suppositories with decamethoxin are active against S. aureus, E. coli, P. aeruginosa and Calbicans. Suppositories containing thiotriazolin and decamethoxin have a more pronounced antimicrobial effect than suppositories with decamethoxin, causing a growth retardation of S.aureus by 54% more, E. ^ th - by 120% more, P.aeruginosa - by 57% more compared to reference drug. The suppositories of thiotriazolin and decamethoxin also had a more pronounced antifungal effect compared to suppositories with decamethoxin, causing a growth retardation of Calbicans by 108% more than the reference drug.

Thus, in terms of antimicrobial activity, the proposed pharmaceutical composition more than doubles the reference drug for Ecoli and Calbicans (120 and 108%, respectively) and more than half reliable for S.aureus and P.aeruginosa, the causative agents of inflammatory diseases of urogenital spheres that are most common.

Suppositories are prepared in two ways - either by casting, or by pressing. Usually a lipophilic base is used for the manufacture.

Thus, the proposed pharmaceutical composition in a soft dosage form - suppositories containing both the active components of thiotriazoline and decamethoxin, is new. As a result of experimental studies, it was found that the active components of the claimed composition have physical, chemical and pharmacological compatibility, while the pharmaceutical composition has a wider spectrum of action compared with each reference drug, namely it has wound healing, anti-erosion, anti-inflammatory, antiseptic, antifungal, immunomodulating and antioxidant activity exceeding the reference drugs in terms of the strength of action, which indicates inventions criterion of inventive step. The components of the claimed composition is not addictive, and it does not have an irritating effect. The invention is also industrially applicable and can be used in the pharmaceutical industry, expanding the arsenal of effective and safe domestic medicines used in gynecology, urology and proctology.

Claims (3)

CLAIM 1. The pharmaceutical composition in the form of a suppository with wound healing, anti-erosion, anti-inflammatory, antimicrobial, antifungal, immunomodulating and antioxidant effects contains the active substance thiotriazoline and a base, characterized in that it additionally contains the active substance decamethoxin, while the ratio of active substances is as follows: decamethoxin the range from 0.5 to 5.0 mass fractions per 100 mass fractions of thiotriazoline. 2. The pharmaceutical composition according to claim 1, characterized in that its dosage form is a vaginal suppository, rectal suppository, sticks. 3. The pharmaceutical composition according to claims 1 and 2, characterized in that it contains a lipophilic or hydrophilic base.