WO2012033427A1

WO2012033427A1 - Method for treating mycotic infections and drug for carrying out said method

Info

Publication number: WO2012033427A1
Application number: PCT/RU2010/000492
Authority: WO
Inventors: Михаил Викторович ДАЛИН; Эдуард Георгиевич КРАВЦОВ; Елена Александровна ВАСИЛЬЕВА; Ирина Викторовна АНОХИНА; Надежда Павловна САЧИВКИНА; Наталия Вячеславовна ЯШИНА
Original assignee: Dalin Mikhail Viktorovich; Kravtsov Eduard Georgievich; Vasilieva Elena Aleksandrovna
Priority date: 2010-09-08
Filing date: 2010-09-08
Publication date: 2012-03-15

Abstract

For the treatment of recurring mycotic infections it is proposed to use a lytic exoenzyme of the strain Cellulomonas cellulans VKPM AS-870, which has a molecular mass within the range of 100 to 30 kD and a specific activity of not less than 48.3 active units/mg of a protein, which constitutes lyticase, and local administration means on the basis of the latter. The lytic exoenzyme exhibits a specific action directly on Candida albicans without inhibiting normal microflora on account of acting both on the mannan layer of the cell wall of Candida albicans and on the fibrous layer, with subsequent splitting off of the latter.

Description

METHOD FOR TREATING MYCOTIC INFECTIONS

AND DRUG FOR ITS IMPLEMENTATION

Technical field

The group of inventions relates to medicine, namely to the treatment of mycotic infections and can be used, in particular, in gynecological practice. State of the art

Among mycotic opportunistic infections, candidiasis plays a leading role. About 75% of women in their entire life have at least one episode of vaginal candidiasis (hereinafter VK), the dominant pathogen of which is Candida albicans. Very often, VC occurs as a recurring infection and is difficult to treat. Relapses of candidiasis of the mucous membranes in certain categories of patients are a significant problem (see, for example, the “Practical Guide to Anti-Infectious Chemotherapy,” p / p L.S. Strachunsky, Yu. S. Belousov, S. N. Kozlova. -M ., 2002). The use of antimycotic drugs is not always successful, since chronic infection can be accompanied by the formation of drug resistance in the pathogen. This circumstance determines the relevance of the search for alternative therapies for VK. One of the possible approaches to solving this problem is to study the therapeutic effect of enzymes of microbial origin, affecting morphofunctional structures specific for yeast-like fungus (DPG) cells.

A known method of correcting the microflora of the vagina (RU2131258, Smirnov and others, 06/10/1999). The method consists in the intravaginal administration of Biosporin (in suppositories or tampons). The invention allows to correct the microflora of the vagina in case of its disorders caused by pathogenic and conditionally pathogenic microorganisms of various taxonomic groups. However, the effect of this drug is indirect and, in case of any violation of the microbiota, VC recurrence is possible. A well-known pharmaceutical composition (RU2007105476, Dikovsky et al., 09.09.2008) for the treatment of candidal vulvovaginitis, including as an active component prebiotic lactulose in the amount of 200 mg in the form of vaginal suppositories, and as an active component antimycotics (azoles: intraconazole, terconazole, econazole; nifuratel and natamycin) in an amount of 100 mg or 500 mg. However, it is known that when prescribing antimycotics from the azole group, relapse is observed in 20% of patients within six months.

A method for the complex treatment of VK (RU2254857, Kolomoytseva et al., June 27, 2005) with vaginal suppositories Hexicon is described; Multitabs, Rekitsen-RD drugs, which can significantly improve the condition of patients in the acute period, but are not specific antimycotic therapy, and do not guarantee the absence of relapse within six months after treatment.

A well-known drug is vaginal suppositories (RU2257197, Dulkis et al., May 12, 2004), containing, in particular, benzalkonium chloride, benzoic acid, and vitamin tex with a certain content of ingredients. Benzalkonium chloride has a bactericidal effect on a whole gamut of bacteria, however, it is less active against pathogenic LPG, in particular Candida albicans, and benzoic acid is a pH regulator, it inhibits the growth of pathogenic LPG, but, not related to the means of direct effect on DPG does not lead to the complete elimination of the latter from the vagina.

A known method of treating VK (NZ539019 (A), HUTMAN, 10.27.2006) using antifungal and antibacterial drugs, such as metro-nidazole and miconazole nitrate, as well as probiotics. However, it is known that for these drugs, the development of resistance of pathogens is possible, and an increase in the dose can cause a toxic effect.

Also known is a method of treating chronic recurrent VC

(RU2309761, Khamadyanov et al., 10.1 1.2007), in which enterosorbent, intranasally thymogen and, in addition, the SolcoTrihovac vaccine, cycloferon liniment intravaginally, euflorin B orally and euflorin L are intravaginally administered in three stages intravaginally and orally. However, with a positive effect of immunomodulating components on nonspecific resistance, there is no specific effect on the causative agents of candidiasis, which does not guarantee complete protection against relapses. A known method of treating mycotic infections using enzymes with a lytic effect on cells (US4062941, Davies, 12/13/1977 - the closest analogue). The complex of lytic exoenzymes was extracted from strains of Corpinus comatus, Licoperdon depressum or Licoperdon pyriforme. Enzymes were identified by laminarinase (chitinase, chitobiasis). However, these enzymes destroy the component of the cell wall of DPG - chitin, but do not affect the mannan layer, which determines the main wall structure of the VK pathogen.

The effect of the commercial drug lithicase (Lyticase, Sigma) on some properties of Candida albicans under isolated in vitro conditions and in model experiments on isolated vaginal epithelial cells of healthy women is described (Sachivkina N.P. et al., Study of the lyticase enzyme as a new antimycotic drug, Vestnik RUDN, Ser. Agronomy and Livestock, 2008, N ° 3, pp. 37-43). The article only suggested a possible therapeutic effect, and the recurring infectious process itself was not modeled.

Disclosure of invention

The invention is directed to the treatment of recurrent mycotic infections, which make it possible to exert a specific effect directly on Candida albicans without inhibiting normal microflora. This ensures the irreversibility of Candida losing its virulent properties and viability, which leads to the absence of relapse.

On the first developed model of recurrent VC, it became possible to evaluate the therapeutic effect of the patented drug. It was found that the lytic exoenzyme obtained from the strain Cellulomonas cellulans VKPM AC-870 has important differences in the mechanism of action, which makes it possible to obtain a drug with an activity more than 2 times higher than that of a commercial drug lyticase. Antimycotic properties are ensured by the destruction of two surface layers - manan and fibrillar, while maintaining the glucan and chitin layers. It is this circumstance that makes it possible to use the lytic exoenzyme - lyticase as an antimycotic etiotropic agent that eliminates the very cause of the disease. The drug does not destroy the cells of the macroorganism and normal microflora, since they do not contain those components that They are targets for the lytic action of the enzyme, which additionally contributes to the selectivity of its action.

A patented method for treating mycotic infections using a lytic exoenzyme in the form of a topical preparation as an active substance is characterized in that they use a lytic exoenzyme of the strain Cellulomonas cellulans VKPM AC-870, having a molecular weight in the range from 100 to 30 kD and specific activity of at least 48.3 U / mg protein.

The method can be characterized by the fact that said exoenzyme is a lyticase, as well as by the fact that with intravaginal and rectal administration, said exoenzyme is used at a dose of 10-50 units once a day for 5-7 days.

The method can also be characterized by the fact that, when applied topically, the mentioned exoenzyme is used in an amount of 2-10 U per 1 g of a biologically acceptable carrier, mainly in the form of an ointment or gel, 1 time per day for 5-7 days.

The method can also be characterized by the fact that a contrical proteinase inhibitor is additionally prescribed, which is administered together with the said exoenzyme in an amount of 500-1000 ATPE per dose of the latter.

A preparation for treating mycotic infections using a lytic exoenzyme as an active substance in the form of a topical application is characterized in that they use a lytic exoenzyme of the strain Cellulomonas cellulans VKPM AC-870, having a molecular weight in the range from 100 to 30 kD and specific activity of not less than 48.3 U / mg protein, as well as the fact that said exoenzyme is a lyticase.

A preparation for local, intravaginal and rectal administration contains 10-50 units of the mentioned exoenzyme per unit pharmaceutical form, mainly a suppository, tampon or sanitary pad.

The preparation may be characterized in that it contains the said exoenzyme in an amount of 2.0-10.0 UNITS per 1.0 g of a biologically acceptable carrier, and is in the form of an ointment or gel, and also contains a proteinase inhibitor in an amount of 500-1000 ATPE per dose of said lytic exo-enzyme. EFFECT: reduced recurrence rate in the treatment of mycotic infections, increased antimycotic activity without inhibition of normal microflora. This is ensured, as indicated above, due to the action of the mentioned exoenzyme both on the mannan layer of the cell wall of Candida albicans and on the fibrillar layer with its subsequent cleavage. An additional technical result is a synergistic effect when using the aforementioned lytic exoenzyme and contrikal together: a developed surface is formed in the Candida genetically modified form, however, it does not exhibit pathogenic properties.

Brief Description of the Drawings

The invention is illustrated in the graphs, where in FIG. 1 - presents the growth dynamics of the producer culture and changes in the proteolytic activity of the culture fluid (QL); in FIG. 2 - change in the activity of lyticase in the dynamics of cultivation of Cellulomonas cellulans; in FIG. 3, 4 - vaginal clearance in relation to DPG under the action of lithicase at a dose of 2 U / ml and 10 U / ml, respectively.

Embodiments of the invention

As an active substance, a lytic exoenzyme is used - lithase of the strain Cellulomonas cellulans VKPM AC-870. Before inoculation on mattresses, C. cellulans culture is stabilized by creating a cold shock (inoculum exposure for 20 minutes at a temperature of + 4 ° С). Next, the strain is cultivated with constant juggling for 2.5-3 days at a temperature of 37 ° C on a modified cardiac cerebrum broth (SMB) with the addition of killed baking yeast in the amount of 9.5-10.5 g per 1 liter of medium as an inducer of lithicase. Growth dynamics is controlled by optical density (OD). It was found that the activity of lithicase increased during the period of exponential growth of the culture and reached a maximum in the stationary phase, after which it decreased. The optimal period of effective cultivation is 65-72 hours, since during this period there is the greatest accumulation of the desired product (figure 1, 2).

At the end of cultivation, QOL in a volume of 1.5 liters is centrifuged at 3000 rpm for 15 minutes. and passed through a sterilizing filter with a pore diameter of 0.02 μm. Further, QOL is subjected to diafiltration on AMICON cells using XM-300, XM-100 and RM-30 membranes. Each of fractions are concentrated to a volume of 20 ml, after which the activity of lithium is determined. It was found that the enzyme activity was absent in fractions containing biopolymers with a molecular mass of more than 100 kD and 300 kD, but was fully manifested in the range from 100 to 30 kD. The obtained parameters are consistent with the characteristics described in the literature. After washing and concentrating this fraction, the specific activity of the final product is not less than 48.3 U / mg protein, which is more than 2 times the specific activity of the commercial drug Lyticase from Sigma (20 U / mg protein), which we adopted as a standard.

Acute toxicity of the preparation was evaluated in animals, for which 0.4 mice of a solution containing 117 mg of protein were injected into 10 mice in the dorsal tail vein. The animals remained healthy for 10 days, from which it was concluded that the drug is not toxic at doses that are hundreds of times higher than the therapeutic ones.

The effect of the obtained exoenzyme - lyticase is manifested in the form of lysis of DPG cells, is expressed in a decrease in the number of viable cells and depends on the dose of the enzyme used in the experiment. When processing a suspension of C. albicans with lithase, a decrease in the optical density of the suspension to 53% relative to the control within 1 hour is observed. A dose-dependent effect is observed when evaluating the results of seeding of the remaining viable cells. Both processes are described by two regression equations that reflect the dependence of the lytic effect on the dose of the enzyme:

y (x) = -10.54978561η (χ) +67.1981 181, with a standard deviation of

5.128;

where: y is the magnitude of the decrease in the optical density of DPG suspension under the influence of lithase, compared with the control; x is the dose of lithicase;

Y (X) ⁼ -1 1.641 17331η (χ) +83.5368459, with a standard deviation of

5,945; where: Y is the percentage of viable cells relative to the control; x is the dose of lithase.

The equations make it possible to calculate the 50% effect of the action of lyticase: 5.0 IU - according to the indicator of lysis of the culture, 7.8 IU - according to the cytostatic effect.

Studies have shown that viable DPG remaining after exposure to lithicase loses its virulence. This is due to a decrease in their adhesion to vaginal epithelial cells, due to a decrease in their colonizing effect. abilities. The effect of lithicase on the surface structures of the cell wall of DPG by the destruction of the mannan and fibrillar layers enhances their capture by phagocytes by about 1, 5 times and promotes the activation of intracellular digestion. Thus, as a result of studies, it was found that after contact of native DPG with macrophages, 41.0 ± 0.4% of the cells were digested. As a result of lithification treatment, this indicator increased to 48.0 ± 1.0%. The data obtained indicate that the digesting phase of phagocytosis proceeds more efficiently in the case of the action of the lyticase lyticase exoenzyme that damages the cell wall. Experiments with isolated macrophages revealed that the efficiency of the phagocytic process is manifested in the development of complete phagocytosis. A decrease in the virulence of DPG under the action of lyticase due to cleavage of the fibrillar layer and destruction of the manan layer of their cell walls leads to a loss of adhesion of the latter and induces an increase in the efficiency of DPG phagocytosis by macrophages. In addition, it was found that the antimycotic effect of lithicase can be supplemented by the introduction of the drug "Contrical". The drug “Contrical” (Aprotinin / APROTININ), an inhibitor of proteolytic enzymes and plasma proteases, is usually used to treat pancreatitis (manufactured by AWD pharma GmbH & Co., Germany). The applicant for the first time established the previously unknown property of contracal to inhibit the production of aspartyl proteinase aggression enzymes in DPG. Inhibition of aspartyl proteinases by contrical prevents the realization of the invasive properties of the pathogen, blocks its transition to a pathogenic mycelial form and promotes the elimination of the pathogen. The counter-counter is administered in an amount of 500-1000 ATPE together with lithase, per dose of the latter. The patented preparation contains lithicase and counter-cal in a single composition. The synergistic antimycotic effect of the two drugs is proved by the mechanism found in an in vitro experiment. Litikase provides a lysing effect, and contracal blocks the aggression enzymes of aspartyl proteinase in DPG, which have remained viable.

The study of the therapeutic effect of lithicase was carried out on a VK model using a clinical strain of Candida albicans N ° 40, which was isolated from the vagina of a pregnant woman with a diagnosis of VK. The specified strain was introduced

about

in a volume of 20 µl intravaginally at a dose of 10 CFU / ml, against a background of dysbiosis caused by antibiotics and artificially induced estrus through the drug Messaline containing estrodiol. After elimination of the pathogen from the vagina, a relapse of the infection was provoked by repeated treatment of animals with antibiotics and a hormone without introducing the culture of the pathogen. The infection course was monitored by two indicators of seeding: by the number of CFU in 1 μl of the vaginal discharge and clearance calculated by the formula: K = (Co - Q) / Co · 100%; where: K - clearance (purification), Co - the number of DPG in 1 μl of vaginal discharge of mice at the initial time after infection; C _t - the number of DPG in 1 μl of vaginal discharge after a known period of time.

At the height of relapse, the animals of the experimental group with an interval of 2 days were intravaginally injected with lyticase. Remediation control in the experimental and control groups was carried out by seeding DPG. Two series of experiments were carried out using different doses of the enzyme: 2 and 10 IU / ml (in Fig. 3.4). The provocation is marked with the letters A (antibiotic) and M (hormone mesaline).

From these experiments shown in FIG. 3 (1-5 days - antibiotic; 5 days - infection with C. albicans; 36-40 days - antibiotic; 43-72 days - lithicase), it follows that after the first administration of lithicase there is a significant decrease in the contamination of the vagina with candida compared with control.

When using lithicase at a dose of 10 PIECES / ml (Fig. 4), rehabilitation is completed 8 days earlier than at a dose of 2 PIECES / ml. In Fig. 4, experimental data: 1-5 days - antibiotic and hormone; 5 day - infection with C. albicans; 38-42 days - antibiotic and hormone (mesaline); 47-73 days - lithicase; 76-80 days - antibiotic and hormone. When VK was provoked by antibiotics and hormones, repeated relapses did not develop, which is evidence of complete sanitation of the vagina under the influence of lithicase.

When determining the dose, we proceeded from the experimental VK model. A calculation has been performed allowing extrapolation of experimental data obtained in vitro and in vivo in animals to therapeutic doses of treatment for humans. We proceeded from the following. The volume of fluid injected into the mouse vagina was 25 μl (25.10 ^"6 L). The therapeutic therapeutic dose introduced in the experiment was 2–10 U / ml (page 6 of the description) for 7 days. The volume of fluid injected into the vagina women - 0.05 L. Accordingly, extrapolating the dose to the woman’s vaginal volume, we obtain the indicated range of values of 10-50 U (p. 3 of the formula), and 2-10 U per gram of carrier. It must be recalled that the strain C. albicans N ° 40 is clinical, isolated from a pregnant woman with VK. Therefore, experiments with lithase and C. albicans strain N ° 40 in vitro and in vivo on the VK model can be considered as experimentally substantiated evidence of the drug's applicability for the treatment of mycotic infections.

Investigations by electron microscopy made it possible to record precisely what changes the cell cell of the C. albicans strain undergoes the action of lyticase. In terms of chemical composition, the cell wall is represented by chitin, glucan and mannan, and the fibrillar layer is clearly marked. It was found out that even if the destruction of the manan layer does not occur, its noticeable thinning is observed, as well as the destruction of the surface of the fibrillar layer and its cleavage. Around the DPG there is a clear zone, in some places the cell wall ruptures, while the glucan and chitin layers remain unchanged.

The result of this destruction is a change in the adhesive properties of Candida genus DPG: their adhesion on the surface of target cells (vaginal epithelium) decreases, but the adhesive activity against representatives of the normal microflora of the vagina (lactobacilli) does not change. The recommended dose of lithicase is much lower than necessary for cell lysis. This allows to reduce the drug load while maintaining the therapeutic effect.

The proposed preparation uses lithicase, the initial activity of which is at least 48.3 U / mg protein. The specified lytic exoenzyme can be introduced in various amounts into the composition of different pharmaceutical forms of the drug to achieve an activity of 10-50 units per dose.

The drug lithicase can be used for local, vaginal or rectal use in the form of an aqueous solution obtained by stationary or industrial technology. As a solvent, buffered saline, pH 7.2 can be used.

The drug lyticase may also optionally contain a filler, usually used in the manufacture of the corresponding prescription forms. In the manufacture of suppositories or suppositories, for example, confectionery fat, lanolin, cocoa butter, aluminum hydroxide gel, paraffin, and the like are used as filler. Upon receipt of an ointment or gel - petrolatum, lanolin, vitepsol, alginate sodium, methyl cellulose, metocel, gispagel, HPMC, etc. The drug can also be lyophilized.

The invention is illustrated by the following examples.

Example 1. Obtaining a liquid preparation of lithicase. In industrial technology, the cultivation of the Cellulomonas cellulans strain VKPM AC-870 is carried out in a reactor / fermenter with culture medium at a temperature of 35-37 ° C for 10-24 hours until the culture reaches the stationary growth phase. At the end of the incubation, QOLs are separated and, by fractionation on cartridges, an extracellular biopolymer with a molecular weight in the range from 100 to 30 kD is isolated and concentrated. The activity of lyticase is determined by the lysis of yeasts of the genus Sacharomyces, and the resulting preparation with a specific activity of not less than 48.3 U / mg protein is considered suitable for subsequent use and poured into vials with the introduction of known stabilizing additives.

During treatment, a liquid preparation in an amount containing a single dose (for example, 5 ml of buffered saline (PBS), pH 7.2, containing 50 IU of lyticase) is applied to a tampon or pad. Apply topically, rectally or intravaginally.

Example 2. Obtaining the drug in the form of a suppository. The liquid preparation obtained in Example 1 is dehydrated on a spray or vacuum-drying unit, combined with fillers (confectionery fat, lanolin, paraffin, vitepsol, etc.) in an amount providing at least 10-50 UNITS of activity lithicases per suppository. So, for the preparation of 100 suppositories weighing 5 g. each, it is necessary to introduce 103.5 mg of lyticase per 500 g of excipient (with initial activity of 48.3 U / mg of protein). If suppositories contain a concical, then it is administered simultaneously with lithase according to the calculation: for one suppository - 500-1000 ATre. Further, suppositories are formed on a special installation.

The stability of 8 experimental production series of the drug was studied. As the results show, immediately after the formation of suppositories, the activity of lithicase is at least 40 PIECES, and after nii 12 months storage - activity of less than 10 PIECES per suppository was not recorded.

Example 3. Obtaining the drug in the form of an ointment or gel. The liquid preparation obtained in Example 1 is dehydrated on a spray or vacuum dryer, combined with an ointment (petrolatum, lanolin, etc.) or gel (methyl cellulose, sodium alginate, etc.) base in an amount that provides not less than 2-10 PIECES of lithicase activity per gram of base. So, for the preparation of an ointment (gel), it is necessary to introduce 20.7 mg of lyticase per 100 g of base (with an initial activity of at least 48.3 U / mg of protein). If the ointment (gel) contains contrycal, then it is administered simultaneously with lithase in accordance with the calculation: for one gram of ointment (gel) 100 - 200 ATPE. Next, the ointment (gel) is Packed in tubes or bottles.

The stability of 6 experimental-production series of the drug was studied. As the results show, immediately after preparation, the activity of lyticase is not less than 10 PIECES per 1 gram of base, and after 12 months. storage - there were no series of the drug with activity less than 2 PIECES per 1 gram of base.

The preparations obtained by the above examples are characterized by harmlessness. To determine harmlessness, mice are administered intravaginally. The drug is in a volume of 20 μl. For each experiment, at least 10 mice weighing 15-16 g are used. The drug is considered harmless if all the mice remain alive for 5 days of observation and no disease is detected.

The drug lithicase is characterized by fungicidal activity against the test strain C. albicans N ° 40, which is an internal standard for determining the antimycotic effect of the claimed drug.

The fungicidal effect of lithicase was determined by the method of serial dilutions in a liquid nutrient medium (Labinskaya). A suspension containing 10 ⁵ microbial bodies in 1 ml of broth was prepared from a culture of C. albicans using a bacterial optical standard. Then a series of dilutions of lithicase (containing from 10 to 5120 PIECES) in 2 ml meat-peptone broth was carried out, 0.2 ml of C. albicans culture was added to all tubes. Two controls were carried out: control of the viability of C. albicans culture without lithicase administration and control of the microbiological purity of the enzyme containing 5120 units of lyticase without culture. After 48 hours of incubation, the results of the presence or absence of the thermostat were taken into account. growth. Of all the tubes, 0.1 ml of lawn was sown in a dense Saburo medium. After 24 hours of incubation, the number of CFU per plates was counted in a thermostat. It was found that the minimum concentration of lithicase, which completely inhibits the growth and reproduction of C. albicans, was 1280 units, which was confirmed by seeding on a solid medium. With activity over 640 PIECES, in no case of the experiment was observed growth of C. albicans N ° 40.

The fungistatic and fungicidal effect of the enzyme in relation to C. albicans was also determined in an experiment on a dense AGV medium. 10 samples of 1 ml of a lithicase solution in PBS with an initial specific activity of 10, 20, 40, 80, 160, 320, 640, 1280, 2560, 5120 IU were layered on AGV medium and kept for 5 hours at a temperature of 37 ° С to absorb liquid. PBS was added to the control sample. Thereafter, 10 μl of a suspension of C. albicans was seeded, providing growth of about 50 CFU in the control. The results were taken into account after 24 hours of incubation in a thermostat according to the percentage of viable C. albicans after treatment with the enzyme. The experiment was repeated ten times. Complete suppression of candida growth was observed with lithicase in excess of 640 units in relation to candida suspension. The fungistatic action of lithicase began to manifest itself with values corresponding to 10 PIECES of activity. At 40 IU, growth was suppressed in more than 50% of C. albicans cells, at 640 IU - more than in 98%, and at 1280 IU in 100% of cases the complete bactericidal effect of lyticase was recorded.

Thus, the conducted studies allow us to conclude that the patented method of treating mycotic infections using lithicase, a lytic exoenzyme of the strain Cellulomonas cellulans VKPM AC-870, with a specific activity of at least 48.3 U / mg of protein contained in extracellular fractions of a biopolymer with a molecular mass in the range from 100 to 30 kDa and selectively destroying the main structure of the cell wall of DPG, contribute to the complete rehabilitation of the infectious focus. Moreover, the elimination of the pathogen from the vagina is so effective that provoking a relapse of the infection becomes impossible.

Industrial applicability

The drug is based on the strain Cellulomonas cellulans VKPM AC-870, deposited in the All-Russian collection of industrial microorganisms FSUE State Research Institute of Genetics, can be obtained through traditional technologies for microbiological and pharmaceutical industry, equipment and equipment.

Claims

SUMMARY OF THE INVENTION 1. A method for treating mycotic infections using a lytic exoenzyme as an active substance in the form of a topical preparation,

characterized in that they use the lytic exoenzyme of the strain Cellulomonas cellulans VKPM AC-870, having a molecular weight in the range from 100 to 30 kDa and a specific activity of at least 48.3 U / mg protein.

2. A method according to claim 1, characterized in that said lytic exoenzyme of the Cellulomonas cellulans strain is a lyticase.

3. The method according to p. 1 or 2, characterized in that with intravaginal and rectal use, the aforementioned lytic exoenzyme is used at a dose of 10-50 ED once a day for 5-7 days.

4. The method according to p. 1 or 2, characterized in that when applied locally, the mentioned lytic exoenzyme is used in an amount of 2-10 ED per 1 g of a biologically acceptable carrier, mainly in the form of an ointment or gel, 1 time per day for 5-7 days.

5. The method according to p. 1 or 2, characterized in that they additionally prescribe a contrical proteinase inhibitor, which is administered together with the aforementioned lytic exoenzyme in an amount of 500-1000 ATPE per dose of the latter.

6. A preparation for the treatment of mycotic infections in the form of a topical preparation containing as an active substance a lytic exoenzyme, characterized in that it contains a lytic exoenzyme of the strain Cellulomonas cellulans VKPM AC-870, having a molecular weight in the range of 100 up to 30 kDa and specific activity of at least 48.3 U / mg protein.

7. The preparation according to claim 6, characterized in that said lytic exo-enzyme of the Cellulomonas cellulans strain is a lyticase.

8. The drug in p.p. 6 or 7, characterized in that for intravaginal and rectal administration it contains 10-50 units of the aforementioned lytic exoenzyme of the strain Cellulomonas cellulans per unit pharmaceutical form, preferably a suppository, tampon or sanitary towel.

9. The drug in p.p. 6 or 7, characterized in that it contains the mentioned lytic exoenzyme of the strain Cellulomonas cellulans in an amount of 2.0-10.0 UNITS per

1.0 g of a biologically acceptable carrier, and is in the form of an ointment or gel.

10. The drug according to any one of paragraphs 6-9, characterized in that it further comprises a contrical proteinase inhibitor in an amount of 500-1000 ATPE per dose of said lytic exoenzyme.