Composition with anti-infectious activity

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and pharmaceutical industry and represents a composition with anti-infectious activity presented in the solid dosage form for oral administration in the form of a powder, representing a balanced complex containing a probiotic agent, an adsorbent and an excipient; the probiotic agent is sterilised cultural fluid containing metabolites of the strain Bacillus subtilis VKPM (Russian National Collection of Industrial Microorganisms) No. B-2335; the adsorbent is zeolite; the excipient is calcium stearate or aerosil; the composition is characterised by the fact that it additionally contains a high-active immunomodulatory agent representing a complex of polysaccharides - products of polymer enzymatic hydrolysis of internal and external layers of brewers' yeast cell coating in the form of β-glucane and mannan with the ingredients of the composition taken in specific proportions, wt %.

EFFECT: invention provides high effectiveness and versatility with respect to dangerous bacterial and viral infections; it is storage safe and stable.

11 ex, 10 tbl, 10 dwg

 

The invention relates to the field of medicine and the pharmaceutical industry, namely the prevention and treatment of infectious pathology of bacterial and viral nature.

Infectious diseases of different nature continues to be one of the urgent problems of modern medicine [1-3]. While the increasing importance of the so-called dangerous infectious diseases (OHI), outbreaks of which is recorded primarily in developing countries with unstable socio-economic conditions [4-7]. Today SIL has not lost its relevance for developed countries, including Russia, as evidenced by the periodic activation throughout the country natural foci of infections (anthrax, hemorrhagic fever with renal syndrome, Crimean-Congo hemorrhagic fever, tick-borne encephalitis), and periodic outbreaks of West Nile, bird flu, swine flu. California encephalitis. Japanese encephalitis, fever Sakhalin, fever Issyk-Kul and other infections [8].

According to experts of the world Health Organization (who) [8] in the near future we should expect epidemics of yellow fever on the Asian continent, a new pandemic of smallpox caused by a transmissible option Monkeypox, the spread of the virus is Ebola, transmitted by airborne. You should not rule out the potential dangers of bioterrorism using as biological agents of infectious agents. According to domestic and foreign experts, the priority among these agents will be given to nontraditional genetic pathogens against which the known means of prevention and treatment may not be effective [9, 10].

Infectious diseases are of great relevance to health, so as to occupy a leading position in terms of the overall incidence and the average prevalence of the population [11], causing significant damage to his disability, and quality of life in General. Analysis of the epidemiological situation in different regions have shown that despite its overall deterioration after the collapse of the USSR there has recently been a stabilization of the situation [12, 13]. Organizational and practical activities of the sanitary-epidemiological service of the country, has created a powerful system of sanitary and epidemiological surveillance, contributing to the improvement of sanitary-epidemiological situation in the regions [12-14].

The above suggests that the problem of creation and development of effective tools and methods for the prevention and treatment of infections, includingincluding hydro SIL remains highly relevant, and at the present time it should be paid attention to.

Intensification of research on pathogenesis OHI, has allowed to establish that in the process of developing this disease is the formation of a functional cell anergy and secondary immune deficiency, the main reasons are the imbalance between the various components of the immune system, especially the imbalance between cyclic nucleotides, inflammatory mediators, cytokines and cell suppressor factors [14].

The induction of cellular immunosuppression occurs, as a rule, when the system hyperproductive these molecules or increase their local concentration higher than the physiological norm. Cellular immunosuppression may also contribute soluble molecules secreted by infected cells. The latter is typical for most infections are viral nature. In particular, the present time is described proteins secreted by cells infected with variola virus, smallpox cows, haemorrhagic fevers Ebola, Las, Marburg and other potent suppressive effect on the production of interferon in the body. With regard to bacterial infections similar suppressive effect region who provide products, secreted by cells that are infected with plague, anthrax, tularemia and other

The main manifestations of imbalance in the immune system are:

- a change in the ratio of cells with phenotypes of CD4+ and CD8+ (change immunoregulatory index) decrease due to reduction under the influence of the pathogen in the number of cells with the phenotype CD4+and increasing the number of cells with a phenotype of CD8+;

the violation ratio of effector and suppressor cells, resulting from failures of the process of their differentiation;

- change the subpopulation of T-lymphocytes-helper cells, leading to modification of the set produced by these cells cytokines and, as a consequence of an imbalance between the cellular and the humoral component of the immune system;

- imbalance between proliferative and apoptotic cellular regulatory programs, manifested in the violation of the relation between the number of cells with phenotypes CD25+, CD30+ and CD95+;

the imbalance caused by the violation of the relationship between the immune, nervous and endocrine systems;

- violation of the principle of clonal activation, manifested by the formation of polyclonal type of immunoreactivity and leading to autocannibalism - aggression immune system against its own cells and tissues [15-17].

So, in the ode infectious process, first of all, the dysfunction of the immune system. This response of the immune system in the organism infection is characterized by the development of systemic Ostrovskogo answer. Pathogens, triggering special mechanisms to counter the formation of anti-infective immunity, have a quite powerful suppressive effect on cellular and humoral components of the immune system, thereby creating the condition for its own reproduction.

In addition to cell violated and humoral mechanisms of the immune system that results in:

- violation of the synthesis of immunoglobulins (Ig) for damage and deformation of the antigen to immunocompetent cells and suppression of production of specific antibodies In-l with the participation of T-l-helper cells, as well as by direct contact of the pathogen with Ig and damage to their molecular structure, leading to the loss of their ability to inactivate the antigen;

- damage to the synthesis and secretion of immune cell mediators, energy and metabolite-protective substances involved in immune processes (prostaglandins, adenylate cyclase, guanilatziklazu, phosphodiesterase, cyclic monophosphate, adenosine triphosphate, and others);

- violation of the absorptive and digestive functions circulating in the blood phagocytes.

The combination of plumage is olenych negative shifts negatively affects the formation of an adequate immune response. Originally violated the processes of recognition of a pathogen of immunocompetent cells, and further intercellular interactions within the immune system. These changes lead to the suppression of specific antitelomerase, production of lymphokines sensitized lymphocytes and the formation of immune deficiency in a compensated or subcompensated form [18].

The depletion of the adaptive capabilities of the immune system is accompanied by severe metabolic disorders and shows the transition of functional-structural immune failure in decompensated form. Further depletion of the reserve capacity of the organism leads to the total immune deficiency, which is characterized by complete disruption of the entire system of immunoreactivity and the simultaneous loss of functionality of the blood-forming organs. This indicates the exhaustion of the last strategic reserves.

Protective response against a number of viral and bacterial pathogens can be enhanced by activation of the immune system [19], when the stimulating signal is nonspecific, not selectively directed against caused the infection of the pathogen, and to increase the immunological resistance.

Currently describes various ways and medium spans the VA correction of immune deficiency. With a certain degree of conditionality means immunopotentiating therapy can be divided into causal (ekstraimmunnaya) and pathogenetic or actually immunotropic implementing its action through the immune system. Action ekstraimmunnaya drugs aimed at improving the General condition of the body and its metabolism, and elimination of the causes of dysfunction of the immune system. These medicines are also called funds mediated immunotherapy. They allow to increase nonspecific immunological resistance to various environmental factors, including infectious agents. Pathogenetic drugs normalize the functioning of the immune system and are intended for the stimulation or suppression of immunoreactivity, correct imbalances in various parts of the immune system of compensation existing violations of immunoreactivity and prevent the development of immune deficiency.

The use of immunotropic drugs in the treatment of SIL or prophylactic seems to be a very relevant and important. Infection occur and intensify immune dysfunction, whereas the possibilities for causal therapy is limited by the emergence of new and transformation known etiopathogenic, the formation of pathogenic viral-tank is arealnykh associations and resistant strains, the change in the ratio between symbiotic and pathogenic flora, increasing the number and severity of resulting complications, duration of treatment, and hence its value [20]. It also increases the frequency of occurrence of immunologically compromised individuals.

Range of immunotropic drugs today is quite extensive and includes as a means of natural or synthetic origin [15, 20, 21]. Among such drugs include probiotics funds that have a positive impact on the homeostasis of the system of the bacterium and its normal microflora, and also has a fairly pronounced immunomodulatory effect, resulting in stimulation of the lymphoid system, synthesis of immunoglobulins, interferons, cytokines (especially IL-1 and IL-2), the processes of synthesis and secretion of humoral factors of nonspecific resistance. In addition, probiotics inhibit colonization of the intestine by pathogenic bacteria and protect macro from potentially harmful factors that fall into the digestive tract from the outside or synthesized in the intestine during a series of processes [22]. Antagonistic effect of probiotics against pathogens based on binding to receptors of the mucous membrane and dense settlement mucinosa layer is of Chechnya, activation synthesis and release of substances with bactericidal or bacteriostatic effect, such as short-chain fatty chains, hydrogen sulfide, hydrogen peroxide, and antibiotics. Bacteriostatic effect of low molecular weight metabolites sacharolytica microflora, primarily short-chain fatty acids (propionic, butyric, acetic, formic, lactic), and lactate.

Given the mechanisms immunotropic action probiotic agents, and immunopathogenesis of SIL are appropriate following strategic directions of use:

for immunoprotective - to prevent the development of immune deficiency. Indication for immunotherapy is the fact the impact of extraordinary factors or situations;

- to immune to compensate for manifestations of immune deficiency that is associated with the cellular component of immunoreactivity and the elimination of regulatory imbalance of the immune system with subsequent restoration of normal algorithms of immunoreactivity, as well as the imbalance of microflora;

for immunorestorative - to recreate elements of immunoreactivity and restore the morphological and functional integrity of the immune system.

As for the drug with the edst, possessing antiviral activity and finds use in the prevention and treatment of dangerous infectious diseases of viral etiology, as you know, currently they can be divided into 3 groups:

1. Chemotherapy drugs commonly used in medical practice for the prevention and treatment of dangerous viral infections: acyclovir (zovirax), foscarnet, ganciclovir, vidarabine with herpes and cytomegalovirus infections; rimantadine and its analogues with influenza; ribamidil, virazole, ribavirin for respiratory-intitially infection; azidothymidine against retroviruses and others [22-24].

2. Interferon inducers (Radostin, Caledon, cycloferon, neovir, and others), which have already found their place in the prevention and therapy of herpes, hepatitis, multiple sclerosis [25-27].

3. Drugs previously used in other areas of medicine and have been diagnosed with biological activity against pathogens of viral infections: Dibazol, nicotinamide [28-30] and others

So far maintained an interest in nucleotide and nucleoside analogues, which are inhibitors of the various stages of the biosynthesis of nucleic acids. Antiviral action of these substances is connected, apparently, with the presence in their structure of hydroxyl groups that can undergo phosphorylation in the body. It is important that phosphoryl the of the to a greater extent proceeds in virusinfection cell than in uninfected. The resulting triphosphates selectively inhibit viruscode DNA polymerase, and this is due to the antiviral effect of these drugs. Typical representatives of this class of compounds are acyclovir, widely used for the treatment of herpetic diseases, and ribavirin, which are becoming more widespread in MS-infections.

One of the ways to increase resistance to viral infections is the stimulation of all parts in the chain of immunity using interferon or inductors. Recent studies conducted in experimental animals and patients, confirmed the fact that interferon inducers are an effective means of prevention and early treatment of viral infections. With this purpose, approved for use in medical practice drugs, Radostin, cycloferon, neovir, anadin and other

At the same time well-known basic circuit emergency prophylaxis and treatment of infectious diseases of bacterial origin are based on the use of broad-spectrum antibiotics [31-34], the application of which is to provide antibacterial protection is associated with a number of difficulties. Thus, there is a marked decline in the number of antibiotics and chemotherapeutic agents, registered in Rossiiu for the last eight years. If in 2002 the total number of permitted use of the main practically significant antibacterial compounds were 65 items, in 2009, only 43 (66%). Especially clearly this negative effect in relation to the item of betalaktamov (28 in 2002, 19 in 2008), aminoglycosides (respectively 11 and 7), tetracycline (5 and 1) and sulfonamides (respectively 8 and 4). The most stable in this respect is the class of fluoroquinolones as the elimination of its part of the same drug (enoxacin) compensated by an appearance on the Russian market in 2008, two new representatives of this class of gemifloxacin and Gatifloxacin [35-38].

The steady decline in the items registered in Russia antibiotics and chemotherapeutic agents are fully correlated with the overall state of production of these drugs in the country. In Russia powerful industry in the production of antibiotics was created in the 50-ies. Production amounted to more than 3000 tons per year, and that was enough to ensure antibiotics all the republics of the former USSR and the countries of the socialist Commonwealth. The production was based on the strains of domestic breeding. In connection with the crisis of the Russian economy by the mid-1990s ggdrivesru domestic substances antibiotics on the basis of microbial synthesis is thawed unprofitable. Currently, a significant part of the fermentation capacity for the production of substances of antibiotics is idle. In 2005, the production capacity of the substances of antibiotics were used by only 17%, for the production of finished dosage forms for injection - 29%. The release of substances of antibiotics in recent years has decreased by almost 4 times (2363 t in 1985, 891 tons in 1995, 624 tonnes in 2004) [10]. The main reasons for the fall production of substances of antibiotics in Russia are not modern technology biosynthesis and chemical transformation of antibiotics, the high cost substances, the lack of government support for producers of substances (loans, tax breaks, etc) and dumping the policy world manufacturers of substances. Due to the sharp reduction in the volume of domestic production of substances antibiotics Russian producers have to buy a significant portion substances abroad, besides their price is 2-3 times lower than the cost of domestic products.

Thus, the disappearance of a significant number of antimicrobial compounds from the market antibacterial chemotherapeutics Russia could not affect the General state of the problem of prevention and treatment of infections, because many of the drugs no longer registered in our country, still on the us in the current regulations, regulating the prevention and treatment of such dangerous diseases.

At the present time to protect against infectious diseases of bacterial origin have been recommended to use a large enough list of antibiotics. Among the basic tools of antimicrobial therapy include doxycycline, rifampicin, defamatory, ampicillin, sulfaton, sizomitsin, gentamicin and other

As a result of recent studies to identify promising drugs, including a number of modern developments, namely:

from the group of penicillins - azlotillin, amoxicillin, piperacillin and others;

from the group of cephalosporins - Cefotaxime, ceftazidime, Ceftriaxone, cefpirome, cefepime, and others;

from the group of carbapenems - imipenem, Meropenem;

from the group of aminoglycosides - sanamycin;

- macrolide - azithromycin, clarithromycin, spiramycin, roxithromycin and others;

- the group of fluoroquinolone - pefloxacin, ciprofloxacin, ofloxacin, norfloxacin and other

However, it is impossible not to note that to achieve the required level of antibacterial effect of these anti-infective drugs, as a rule, you must often take long time and in large doses. Such antibacterial drugs, on the one hand, effectively the turnover is the infectious pathology, but on the other, when a commonly used schemes dosing often cause the development of a number of side effects from various organs and systems of the body, such as a negative impact on the state of microbiocenosis of intestines of humans due to the lack of selectivity of action of antibiotics on microorganisms, allergic reactions, etc. in Addition, basic antibiotics toxic, have nonspecific bactericidal action and cause the emergence of pathogens resistant to antibacterial agents, resulting in antibiotic therapy requires a minimum of subsequent recovery of the intestinal (imbalance in the system macro and normal microflora). We should not forget also about the significant dependence of the pharmaceutical market of Russia from drugs imported goods, which may adversely affect the level of anti-aid.

One of the promising directions to overcome these shortcomings, manifested in the use of antibacterial drugs is not independent of their use, and in combination with the means of correcting intestinal - probiotics. It was found that in the case of a joint appointment with antibiotics priority is learn to give such probiotic means, which contain not holistic living microorganisms (e.g., Bifidumbacterin, Lactobacterin, sporobacterin, laminolact), which is foreign to the digestive tract of a person who will break antibacterial agents, creating additional antigenic strain on the body and the metabolites of probiotic strains of microorganisms [39-41].

Among such tools include probiotic complex Battistin [42]. It is based on (wt.%): sterilized culture fluid (SCI), obtained by cultivation of bacteria of the species Bacillus subtilis and containing metabolites 1,0, hydrolyzed soy flour (POL) - 20,0, natural mineral zeolite with sorption and ion-exchange properties - 78,0 and calcium stearate (CS) or Aerosil to 1.0. Specified probiotic complex composition, mechanism of action on the organism and the achieved effect is closest to the claimed compositions and adopted as a tool prototype.

The Foundation funds the prototype are immobilized on zeolite exogenous and endogenous biologically active substances (BAS), secreted by the bacteria Bacillus subtilis in submerged cultivation, which are largely responsible for therapeutic effect of this probiotic complex. The list of them is quite extensive and includes heterogeneous in chemical composition and biologically the properties of matter: proteolytic and amylolytic enzymes, amino acids, polysaccharides (hexosamine, glucosamine), vitamins (pyridoxine hydrochloride, Riboflavin), nitrogenous bases and their derivatives (adenine, guanine, thymine, uracil, cytosine) [43-45]. Among them are well represented and various natural antibacterial substances (bacteriocins, lysozyme, catalase), which selectively inhibit the growth and reproduction of pathogenic and conditionally pathogenic microorganisms in the gut, while not affecting the symbiotic microflora. All these substance have a positive influence on the organism of man and animals in General.

Zeolite blended with the prototype, has a strong adsorption effect (mainly in relation to compounds with low molecular weight, such as methane, hydrogen sulfide, ammonia and other toxic substances), without direct interaction with vitamins, amino acids, proteins, making these substances remain in the gastrointestinal tract. Ions contained in the body, may be incorporated into the crystalline structure of the mineral, and Vice versa, from mineral body gets those inorganic elements that need. By increasing the height of the intestinal villi, improve digestion and increase the area of absorption of nutrients in the small intestine occurs, so-called, got the active ion exchange [46].

An important part of the funds of the prototype is hydrolyzed soy flour, which is, on the one hand, part of the protective environment of the metabolites, is largely responsible for the strength of their adsorption on the surface of the zeolite and on the other hand, is a source of amino acids, providing the nutritional requirements of normal microflora of the intestine and cells of the microorganism. The main component of hydrolyzed soy flour - soy oligosaccharide (SOE) has bifidogenic properties [47].

Calcium stearate (CS) or Aerosil included in the funds of the prototype as technological additives.

The main purpose of the funds of the prototype is to restore and maintain optimal microecological parameters in the intestine, the treatment of diseases of the digestive tract by ingestion of substances inhibit the development of pathogenic microflora and stimulate the growth of beneficial microorganisms. The effect of the drug is based on the fact that during its transit through the gastrointestinal tract in a given zone is the destruction of the protective capsule and secretion into the cavity of the intestine immobilized on the particles of the zeolite components of the probiotic. In this case around the particles of the zeolite are formed, the formation of micellar structures in the process of movement through the digestive tract are gradually released from the porous surface of the zeolite. On the one hand, this item is well positioned to support activity in the digestive tract of biological components of the probiotic during the day, that is enough to restore and stimulate the functional activity of the intestinal microflora. On the other hand, the effect of a gradual release from the surface of the zeolite immobilized on its biologically active substances (BAS) leads to the appearance of exposed surfaces of its porous structure that enables the mechanisms of ion exchange and selective sorption of toxic compounds, which helps to cleanse the digestive tract of toxins and decomposition products. This is especially important for overall detoxification of the body.

Thus, due to the complex composition tool prototype contributes to the overall normalization of microecological conditions in the intestine through various mechanisms, such as suppression of conditionally pathogenic flora, sorption and removal of toxins, improving digestion, improving the trophic base for the normal microflora and gastrointestinal tract epithelium.

Clinical trials means the prototype showed its high effectiveness in stopping the phenomena of gastric and intestinal dyspepsia in patients with acid diseases of the upper GI tract. The use of this drug in patients with erosive antral gastritis resulted in positive clinical effect in most patients, manifested in the form of reduction of symptoms of intestinal dyspepsia,developing on the background of eradication therapy. Taking the drug contributed to the improvement of lipid metabolism, increase high density lipoprotein and lower overall blood cholesterol. In addition to improving physical symptoms in patients on background therapy decreased the level of depression and neurotic [48].

Clinical efficacy means the prototype proved and in the appointment of its patients with chronic pancreatitis with exocrine insufficiency of the pancreas sachetana with fermentational therapy (Mezim-Forte, pentitol), manifested in the elimination of violations of the intestine microbiocenosis, correction of secondary exocrine pancreatic insufficiency caused by the excessive activation of the intestinal microflora that develops on the background of primary exocrine insufficiency and significantly aggravate asthma [49].

It is also established whether the destination of the funds of the prototype when implementing a fundamentally new regimens for coronary heart disease (CHD) is a disease that currently has the extremely high clinical and social significance due to the high incidence among people of working age, and the severity of possible complications [50]. Along with the main reasons for the development of this pathology, there are also a number of additional factors, TNA is redoano contributing to its emergence and development. First of all, they are the dysbacteriosis of the intestine, so as to present an opinion has been formed about the relationship of coronary artery disease with quantitative and qualitative disorders of the composition of the microbiota, as well as the key role of the intestinal microbiota with participation mechanisms enterohepatic circulation in the regulation of lipid metabolism. The development of intestinal dysbiosis mediates the formation and progression of adverse changes in lipid metabolism as one of the triggering factors of cholesterol aggression. Lipid-lowering therapy is considered nowadays as one of the main objectives in the treatment of coronary artery disease and involves the use of statins as primary and the most effective cholesterol lowering drugs. However, statins are the latest generation, radically changed the approach to prevention of coronary heart disease and its complications, have side effects, limiting their wide usage: hepatotoxicity, as well as reducing the effectiveness of treatment with long-term use. Additional use of the prototype against the standard baseline therapy, providing for the appointment of nitrates, antiplatelet agents, β-blockers or calcium channel blockers, angiotensin-converting enzyme inhibitors, diuretics, accompanied by lipid-lowering effect greater than the effect in similar conditions simvastatin. This is due to the positive impact of the funds of the prototype due to correction of microbiocenosis of intestines.

The absence of means-prototype living microorganisms can be used in conjunction with antibiotics. Combined their introduction in the digestive tract can improve microecological conditions, which helps to restore the qualitative and quantitative characteristics of symbiotic microorganisms. However, when evaluating the prospects of the combined purpose antibacterial agents (doxycycline) and battistin in cases of emergency prophylaxis and treatment of bacterial SIL nature (anthrax infection) has a low level of effect tools of the prototype [51]. Effect of tool prototype for other bacterial SIL nature were not evaluated.

However, along with the observed effects for the prototype characteristic has the drawback that as the impossibility of the concentration of the culture fluid obtained on soy environment using a fuel because of its high viscosity. In addition, the use in the process of obtaining funds for the prototype as a nutrient medium native soy flour does not allow full use of the nutritional properties of the feedstock. In donnaslut require additional chemical or enzymatic hydrolysis of soy flour, which significantly complicates the technology of obtaining funds for the prototype.

It should also be noted that theoretically expediency and practically proved the possibility to use the prototype mainly only for the correction of violations of state microbiota and treatment of relevant diseases of the gastrointestinal tract by the total normalization microecological conditions in the intestine. About the prospects of the use of probiotics for the prevention and treatment of infectious diseases was reported in the works of Garden N.V. et al. (1998) [52], Ryzhko I.V. et al. (2000) [53], Vorobeychik E.V. et al. (2005) [39]. However, these systematic studies on the immunotropic effects of Battistini (prototype), showing the principal opportunities and prospects of its use in the prevention and treatment of a wide range of infectious diseases of various etiologies, including infectious diseases, both in monotherapy and in combination of course with the appointment of the basic anti-infective means no. At the same time, available information theoretical build on the promising applications of funds on the basis of probiotics for efficient prevention and treatment of such pathology.

The aim of the invention I ilos increase the effectiveness of prevention and treatment of infectious diseases by creating compositions for pathogenetic therapy on the basis of a complex of biologically active substances with probiotic and immunomodulatory activity with high efficiency and flexibility in terms of infections of bacterial and viral nature when orally administered as a monotherapy and when combined with the appointment of causal by means of basic therapy, security and stability during storage.

The achievement of this goal is possible through the creation of compositions for pathogenetic therapy, representing a balanced complex of biologically active substances with probiotic and immunomodulatory activity, qualitative and quantitative composition which will allow the normalization of the functioning of the immune system by rectify dysfunctional changes in the immune system and, above all, overcoming powerful suppressive action of pathogens on her cellular and humoral components that inhibit the formation of anti-infective immunity, indemnification existing violations of immunoreactivity and prevention of the development of immune deficiency.

When forming the qualitative composition of the claimed composition intended for the prevention and treatment of OHI various etiologies, came from the fact that its main components are:

- to exert immunomodulatory effects and to help reduce disfunktion the selected changes in the immune system, developing when exposed to pathogens OHI;

- to be combined with the basic tools etiotropic therapy OHI bacterial and viral etiology;

- to reduce the adverse effects of antibacterial agents on qualitative and quantitative characteristics of the normal microflora of the gastrointestinal tract;

- to increase the body's resistance to viral infections by stimulating all parts in the chain of immunity through induction of the synthesis of endogenous interferon;

- activate the mechanisms of elimination of pathogens from the body;

- to provide anti-toxic effect by binding and excretion of toxic compounds that accumulate in the body as a result of the development of the infectious process and the negative effects of antibiotics in their exchange application.

An important distinguishing feature of the claimed composition is the ability to effectively enhance the nonspecific and specific immune resistance of the organism, which is implemented through complex effects on the immune system of all components of the claimed composition exhibiting together with immunomodulating activity.

For this part of the claimed composition, representing a balanced complex, in addition to the probiotic agent, adsorbent, and auxiliary additives added to them unimodularly agent.

Probiotic agent is a sterilized culture fluid (SCI)containing metabolites resulting from cultivation and sterilization of bacteria of the species Bacillus subtilis. Used strain VKPM No-2335, the metabolites of which are characterized by high antagonistic activity against a broad spectrum of pathogens (Salmonella, Staphylococcus, Shigella, Pseudomonas and others) and opportunistic (Proteus, Klebsiella, Candida and others) microorganisms and can inhibit the growth of pathogenic microorganisms, without exerting inhibitory effect on lactobacilli and bifidobacteria. In addition, metabolites bacilli Bacillus subtilis, increasing the phagocytic activity of macrophages, are able to be together with immunomodulating activity.

Produced by the microorganisms biologically active substances are concentrated in the process of their cultivation it is in the culture fluid. The list of biologically active substances synthesized by Bacillus subtilis in submerged cultivation, sufficiently representative and manifestation of their biological activity is an integral part of therapeutic and preventive properties of the claimed composition. Among them are well represented in various natural antibacterial substances (bacteriocins, lysozyme, catalase), which selectively inhibit the growth and reproduction of pathogenic and conditionally pathogenic microorganisms in to the siechnice, while not affecting the symbiotic microflora. Especially valuable is that when growth and sporoobrazovanie microbes-producing Bacillus subtilis synthesize nitrogenous base (purine and pyrimidine derivatives). These nitrogenous bases and their derivatives, which is a separate group of immunomodulatory substances contained in ski in significant amounts (table 1). This circumstance for the claimed composition is crucial, as nitrogenous bases and their derivatives restore the functional activity of a single macrophage system of the body, the oppressed due to the impact of pathogens, and significantly increased the efficiency of nonspecific and specific protection at different stages of the infectious process.

This pharmacological activity of probiotic agent is expressed by means of mediated interaction produced biologically active substances with immunocompetent organs after overcoming physiological barriers.

The adsorbent in the composition of the claimed composition is suitable primarily for the immobilization of metabolites produced by the bacilli in the process of their cultivation and concentrated in ski, transport and targeted delivery of them throughout the intestine. The gradual release of immobilienbank the x on it BAS allows you to maintain a high level of activity of the claimed composition.

The possibility and feasibility of using zeolite as a component of the claimed composition (adsorbent) is defined accurate understanding of the nature and the mechanism of its influence on the body. So, due to the unique chemical structure of molecules zeolite provides selective binding and excretion of low molecular weight toxins: methane, hydrogen sulfide, ammonia, heavy metals, radionuclides, etc. there is no direct interaction with useful nutrients: vitamins, amino acids, proteins and other Zeolite contains minerals and has ion-exchange properties, as the main skeleton of its crystal lattice consists primarily of a tetrahedron and has a cavity in which are ions (sodium, potassium, calcium), easy to communicate among themselves and with the surrounding substrate. Zeolite is not absorbed in the digestive tract and passing in transit, is involved in the selective ion exchange and selective sorption, as an additional source of a wide spectrum of essential micronutrients. It is also important that the specified natural mineral improves digestive processes in General, due to the activation of biochemical reactions in the gut, sorption of low molecular weight metabolites and normalization of intestinal microflora. Particles used zeolite have dimensions of not more than 500 is miles and an oval shape crystal that, in turn, eliminates the formation of micro traumas in the intestine and is completely safe [54-56].

The essential difference between the claimed composition from the funds of the prototype is that in its composition additionally introduced highly active immunomodulatory agent, which is a complex of polysaccharides - products of enzymatic hydrolysis of polymers of inner and outer layers of the cell membrane of yeast, namely β-glucan and mannan. These easily digestible in the body substance is complex and consists primarily in the fact that β-glucan and mannan through immunomodulatory effects on immunocompetent cells effectively inhibit the growth and reproduction of germs and viruses trapped in the body.

Immunomodulatory effects used β-glucan yeast due to the fact that, he is transported through the walls of the cells of the intestinal tract in the lymph, which interacts with the key cells of the immune system - macrophages - by attaching to specific receptors on their cell membranes and activates macrophages. Acting on macrophages, β-glucan enhances their phagocytic activity, mobility, and ability to find and eliminate the alien, including pathogenic microorganisms. In addition, β-glucan beer drug what she stimulates these cells as neutrophils NK (natural killer cells) and LAK (lymphokineactivated killer cells), significantly increasing antibacterial activity of the claimed composition. It also allows you to intensify production in cells of the macrophage system such cytokines as interferon, thereby increasing the antiviral activity of the claimed composition.

At the same time, the polysaccharide mannan, being able antigenic stimulation, increases the production in lymphocytes of different classes of immunoglobulins, which are responsible for the specific protection of the body from exposure to pathogens.

To improve the technological process of obtaining the claimed composition in solid dosage form for oral administration in the form of powder in its composition an additive. As an auxiliary additive to prevent adhesion of the inventive compositions to be used in the process equipment used calcium stearate or Aerosil.

As can be seen, the qualitative composition is selected in such a way that the claimed composition is a balanced complex with pronounced immunomodulatory activity in the following ratio of ingredients, wt.%:

Sterilized culture fluid
containing metabolites of the strain VKPM No-233
bacteria Bacillus subtilis1,6-2,3
Zeolite43,0-46,0
Complex polysaccharides (β-glucan and mannan)51,0-54,0
Calcium stearate or Aerosil0.5 to 1.5

The possibility of achieving the goal of the invention is confirmed by the research results presented in the following examples.

Example 1. Obtaining the claimed composition in the form of solid dosage forms for oral administration (powder).

To obtain the claimed composition in powder form perform the following algorithm. Grow the microorganisms Bacillus subtilis by the method of deep cultivation in the biological reactor (fermenter). After completion of cultivation, the culture fluid (QOL) with microorganisms initially centrifuged for separation and subsequent removal of living cells of microorganisms, and then sterilized. Before sterilization QOL is mixed with the adsorbent is a zeolite, the pre-crushed to a particle size of not more than 500 μm. Then in the resulting sterilized culture fluid (SCI)containing metabolites producer, enter the comp is CEN polysaccharides, as well as calcium stearate or Aerosil. The resulting mixture is subjected to freeze-drying, in which the immobilization of biologically active metabolites producer on the zeolite particles. The thus prepared paste is put into the tank setup for granulating the purpose of mixing and dried.

Used complex polysaccharides (LLC "BITRA, Moscow) includes biologically active products of enzymatic lysis by polymers inner and outer layers of the cell membrane of yeast: β-glucan (30,7%) and mannan (10,3%).

As the adsorbent using natural zeolite - natural microporous silicate mineral Golynskogo field (Buryatia) (CJSC SPC "NOV", Novosibirsk) [57, 58].

As an auxiliary additive use calcium stearate [59] or Aerosil [60].

The inventive composition can also be made in solid dosage form for oral administration in the form of capsules or tablets that not only greatly simplify its use, but will provide reliable protection of its constituent components from exposure to factors causing their degradation.

As you can see, the process of obtaining the claimed composition in the form of solid dosage forms for oral administration (powder) is very simple and the ingredients are inexpensive, the issue of what are the domestic industry and available.

Example 2. Safety assessment of the claimed composition.

The security of the claimed composition was evaluated by carrying out sanitary-chemical analysis, sanitary-microbiological research, as well as determining important Toxicological characteristics acute toxicity.

Sanitary-chemical analysis.

The sample preparation was carried out according to GOST 26929-94 Raw materials and food products. Preparation of samples. Mineralization for the determination of toxic elements. International standard. Lead and cadmium was determined according to GOST 30178-96 Raw materials and food products. Atomic absorption method for the determination of toxic elements. International standard. Arsenic was determined according to GOST 26930-86 Raw materials and food products. Method for the determination of arsenic". Mercury was determined according to the "Methodological guidelines for the detection and determination of total mercury content in food products by the method of flameless atomic absorption (No. 5178-90).

The content of organochlorine pesticides (toxicants) was determined according to the "Methodological guidelines on the determination of residues of organochlorine pesticides (No. 1766-77) and official methods of analysis of AOAC ("Official methods of analysis of AOAC", 1984, 14th ed., Chapter 29, pp.537-538).

Studies were performed using laboratory scales VL-210, complex voltammetric HUNDRED, chromatograph "Cree is taluks 4000" PID detectors, ASD, atomic absorption spectrometer "quantum-2A", fotoelektrokalorimetry CPK-2, the spectrophotometer SF-26, radiomir spectrometer gamma and beta radiation ICBG-01 "RADEK".

The results of the measurements are presented in tables 2 and 3 and showed that in the inventive composition, the content of toxicants (organochlorine pesticides GHCT and DDT), hazardous substances (lead, cadmium, mercury, arsenic, radionuclides (90Sr137Cs) does not exceed the allowable levels set by "the Uniform sanitary and epidemiological and hygienic requirements for goods subject to sanitary-epidemiological supervision (control)" of the EurAsEC Customs Union (Chapter II, section I, index 10.6) (standard EM (Chapter II, section I, index 10.6)). The same pesticides like Aldrin and heptachlor in the claimed composition was not found.

Sanitary-microbiological research.

Microbiological studies were performed in accordance with the requirements of TORMENT 4.2.577-96 Methods of microbiological control of products for children, therapeutic foods and their components", GOST R 52814-2007 "food Products. Method of identifying bacteria of the genus Salmonella, GOST R 52815-2007 "food Products. Methods for detection and quantification coagulasepositive staphylococci and Staphylococcus aurous", GOST R 52816-2007 "food Products. A method for the detection and determination of amounts of the bacteria of the coli group (coliforms)", GOST 10444.8-88 "food Products. Method for the determination of Bacillus cereus", GOST 10444.12-88 "food Products. Method for determination of yeasts and moulds, GOST 30726-2001 "food Products. Methods for detection and quantification of bacteria of the species Escherichia coli.

The results of studies on the sanitary-microbiological safety indicators are presented in table 4. As you can see, on sanitary-microbiological safety performance of the inventive composition corresponds to the Uniform sanitary and epidemiological and hygienic requirements for goods subject to sanitary and epidemiological supervision (control)" of the EurAsEC Customs Union (Chapter II, section I, index 10.6,10.10) (standard EM (Chapter II, section I, index 10.6,10.10)).

Study of acute toxicity.

To measure the acute toxicity of the inventive composition was administered to healthy outbred white mice-males weighing 22-24 g and healthy outbred rats-females weighing 130-150 g orally in the form of an aqueous suspension, stabilized by tween-80, in the amount of not more than 0.5 ml per animal mice and not more than 5.0 ml - rats. Studies were performed using probit analysis according to Litchfield-Wilcoxon signed modification Srota [61]. Of samples of each species, 50 species were formed by 2 groups of 25 animals each: control (injected with similar volumes of di is fillerbunny water) and experienced (has introduced the claimed composition twice daily at a dose of 0.33 mg for mice and 2.0 mg for rats).

Observation of test animals within 8 days has allowed to establish that no animal was killed, visible changes in their behavior was not. Duration of follow-up was 1 month since the introduction of the claimed composition. The results of the study of the dynamics of body weight of experimental animals within 21 days of observation showed that in none of the experimental groups was not statistically significant in t-criterion of student differences from the corresponding control group (table 5).

As the maximum usable dose of the claimed composition of the toxic effect was not detected, in accordance with the requirements of farmkomiteta Ministry of health-social development of Russia [62] introduced the dose was increased 10 times. The effect of the claimed composition was evaluated in young adult white mice-males weighing 24-27 g using the test for acute toxicity. Experimental animals were kept in groups of 10 animals in a cage under standard conditions: room temperature 20±2°C, feed PC-121-2 (000 "INFORMKOM"), drink at libitum. An hour before the start of the experiment, animals were no longer fed with free access to water.

In the experiment, the claimed composition is administered orally in the form of an aqueous suspension by means of a metal probe. Immediately before each introduction of the sample was mixed the ü to homogeneous turbidity. Control group animals were injected with saline - 0.9% NaCl (placebo).

The volume of injected sample was calculated for each animal based on its mass according to the formula:

where V - volume of sample, ál; m is the mass of the animal,

In the experiment, the claimed composition was injected at 6.6 mg per animal, which corresponds to the maximum value allowed for testing [63]. Experimental and control group consisted of 20 animals. The criterion of selection was the survival of animals experimental and control groups, which was evaluated after 1 day, 2 days, 4 days, 8 days, 16 days from the moment of introduction of the claimed composition. The number of animals within a specified time of the observation are shown in table 6. In the course of the experiment in the control and experimental groups of deaths of animals were observed. Thus, it was experimentally established that the claimed composition does not differ acute toxicity, because it does not cause any visible changes in the overall health of experimental animals and does not cause their death.

Example 3. Evaluation of the stability of the inventive compositions during storage.

Investigated the possibility of saving activity and safety developed a solid dosage form of the inventive compositions in powder form and has established a guaranteed shelf life if stored under different the temperatures and without using special equipment.

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (1.6 wt.%), zeolite (43,4 wt.%), complex polysaccharides (53,6 wt.%), calcium stearate (1.4 wt.%).

In the experiment, the powder was put in 80 consumer packaged in double plastic bags and kept them for 2 years at temperatures of 5±2°C, 10±3°C 25±5°C, 35±6°C, 20 packs at each temperature storage mode.

Every 6 months storage were selected on 5 packages from the appropriate temperature and determine the characteristics of the powder: moisture, amylolytic and proteolytic activity, antagonistic activity and microbiological purity. To estimate the changes in these characteristics during storage was determined by their values at the beginning of the research.

The data presented in table 7, indicate that at the selected test temperature storage (5 to 40°C) evaluated the performance of the claimed composition are virtually unchanged in over 2 years. The average values of the indicators were: moisture - 6,8±2,5%, the amylolytic activity of 4.8±3,5% AA/g, proteolytic activity of 0.66±3,0% PA/, Microbiological purity of the claimed composition, evaluated by the absence of pathogenic microor the mechanisms (Pseudomonas aeruginosa, Enterobacteriaceae, Staphylococcus aureus and Bacillus cereus), was maintained for 2 years, that is, the entire period of research. Antagonistic activity of the inventive compositions against Staphylococcus aureus, Candida albicans, Shigella sonnei and Salmonella typhimurium were also preserved during the whole period of storage.

Thus, the guaranteed shelf life of the claimed composition, made in solid dosage form (powder), is not less than 2 years at a temperature not exceeding 40°C, subject to preservation packaging. Special temperature storage conditions or the use of special equipment are required.

Example 4. The study immunotropic effects tools for the prototype and the claimed composition.

The aim of the research was to assess the ability of the funds of the prototype and the claimed composition to activate the mechanisms of anti-infective immunity and above all influence factors of nonspecific resistance of the organism. Defining the role of active components (β-glucan and mannan), optionally included in the claimed composition, since it is known that the metabolic products of bacilli Bacillus subtilis are themselves powerful enough stimulus for immune cells, leading to their activation. The study was conducted in comparison with the antibiotic doxycycline is m - antibacterial drug, widely used in clinical practice.

The investigations were carried out on outbred laboratory mice (kennel "Rapolano RAMS weighing 18-23 g were used 200 animals were distributed into four groups of 50 animals each: control (received isotonic sodium chloride solution) and three Guinea, in which animals were injected with the antibiotic doxycycline, the tool is a prototype or of the inventive composition.

Doxycycline was administered intragastrically once a day, 1 at a dose of 0.05 mg/mouse.

The tool prototype was administered orally 1 time a day at a dose of 150 μg/mouse.

The claimed composition is administered orally 1 time a day at a dose of 150 μg/mouse.

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (2.3 wt%), zeolite (45,3 wt.%), complex polysaccharides (51,7 wt.%), calcium stearate (0.7 wt.%).

The duration of antibiotic, means the prototype and the claimed composition was 7 days. The impact assessment of the means was performed after 1, 5 and 7 days. To do this, before the introduction of the drugs (control)and after 1, 3 and 7 d after injection in animals of each group were collected blood for research. At each time material was collected from 10 animals from a group, individually the t of each mouse by the method of decapetala.

Assessment of the dynamics of changes of factors of nonspecific resistance was carried out by determination of the functional state of cells phagocytic system (digestive activity of granular leukocytes), and concentrations of serum lysozyme and myeloperoxidase, which has not only a direct bactericidal effect, but which are mediators of intercellular interaction in the development of the immune response to any antigen exposure.

Digestive activity of granular leukocytes in the peripheral blood of experimental animals was evaluated by the method of Telenovas Mrs x and Nikiforov, V.A. (1972). Serum was obtained by a standard method, separating the blood clot from the liquid phase. To assess the functional status faguoqitirute cells blood was taken in tubes pre-treated with silicone (Serva, Germany), containing a heparin solution at the rate of 20 UNITS per 1 ml of blood.

Phagocytic index (PI), representing the ratio of the intensity of the absorptive and digestive functions faguoqitirute cells was determined after 15 min and 30 min of incubation with the test object phagocytosis - test microbe (M lysodeicticus), which were grown on mesopatamia agar in thermostat at 37°C for 24 h as assessed indicator served as the index of the digestive activity of gr is olyarnik leukocytes (PI), represents the ratio of the number faguoqitirute cells in a smear after 30 min incubation of blood with the test object phagocytosis to the number faguoqitirute cells in a smear after 15 min incubation of blood with the test object phagocytosis.

Values of FE was determined by the formula:

where a is the number faguoqitirute cells 100, calculated in a granular leukocyte that has consumed the test microbe within 15 min of incubation;

In the number faguoqitirute cells 100, calculated in a granular leukocyte that has consumed the test microbe within 30 min of incubation.

Upon the satisfactory functioning of the granular leukocytes of peripheral blood values PI is always greater than 1. The value of PI expressed in arbitrary units (figure 1).

The concentration of lysozyme in the serum was determined by the method of Bukharin O.V. et al. (1974). The blood of the experimental animal was collected in a centrifuge tube and received the serum. Then 0.1 ml of serum was transferred into a bacteriological test tube and added to 0.9 ml of saline. The resulting mixture was stirred and added to 1.5 ml of the suspension of the test microbe, which was previously standardized on fotoelektrokalorimetry FEC-56M to extinction 0,66.

The prepared suspension containing the test microbe and analyzed serum, mixed and is emali in a thermostat at 30 min at 37°C. After a time of incubation, the tubes were removed from thermostat and determined the extinction of material on FEC-56M. The obtained values of the extinction using special tables translated into values that characterize the concentration of lysozyme in the serum, and expressed in µg/ml.

The level of myeloperoxidase (MPO) in the serum was determined by the method developed on the basis of generally accepted approaches to the identification of the substrate in the serum and immunocompetent cells (P.G.H. Gell and others (1968); Page R.C. and others (1978)). To determine the concentration of MPO in serum in 96-well plates to immunological reactions with a flat bottom, starting from the second wells of row a, was poured into 0.1 ml of serum taken from experimental animals. Then they were added to 0.1 ml of the reagent, which is a mixture of benzidine and 3% hydrogen peroxide. In the first hole series And panel (control) instead of whey poured into 0.1 ml of saline. After adding reagent panel was shaken, placed in a thermostat and kept at 37°C for 60 min after the incubation time the panels were removed from thermostat and viewed on a vertical spectrophotometer "Dynatech" (Germany) at a wavelength of 495 nm. The concentration of the enzyme in serum were received in units of extinction and expressed in arbitrary units.

As witness Tvout presented in figure 2-3 data immunotropic effects tools for the prototype and the claimed composition are manifested at the level of phagocytic cells of the blood system and processes associated with the synthesis and secretion into the blood of the enzymes lysozyme and myeloperoxidase. Dynamics of changes in the values of the factors of nonspecific resistance in experimental animals experimental groups, which were introduced by means of a prototype or the claimed composition are almost identical, as evidenced by the absence of statistically significant differences (p>0,05). Identified effects, consisting in a reliable increase of the functional activity of factors of nonspecific immunity, was detected already after 1 day after administration of these funds, and they lasted at least 7 days from the moment of introduction. It has a stimulating effect as a means of the prototype, and the claimed composition on the immunological mechanisms of resistance took place already after a single introduction of them.

The values of the factors of nonspecific resistance of animals treated with doxycycline for 1, 3 and 7 days, remain virtually unchanged relative to the control, indicating that the lack of this basic antimicrobials ability to stimulate the functional activity of immunocompetent cells.

Introduction tools the prototype for 1, 3 a day leads to a significant increase in the activity of factors of nonspecific resistance in comparison with the control. This is manifested in the increase in the index digestive activity of granular leukocytes, as well as the concentration of lysozyme and myeloperoxidase. Quite a significant increase in the functional activity of these factors is observed after a single injection of funds-prototype already during the first day. On the one hand, this indicates the activation of factors of nonspecific immunity and the possibility of the rapid nonspecific defense against pathogens. On the other hand, this means that the primary mechanisms of action tools of the prototype are related not only to its direct influence on the microflora of the gastrointestinal tract, but also with a direct impact on the components of local immunity, which, including that related to the regulation of the balance of endogenous cytokines that enhance protective response.

In case of use of the claimed composition, which also includes the complex polysaccharides in the form of biologically active products of hydrolysis of the polymers of the cell wall yeast (ß-glucans and mannan), was traced its most effective in the established effects in comparison with the tool prototype. Thus, the inventive composition had a significantly more pronounced effect on the phagocytic cells of the blood system and oxygendependent bactericidal mechanism (increased to ncentrate myeloperoxidase serum). However, it is known that increasing the concentration of serum myeloperoxidase shows not only the activation of nonspecific immunological resistance, but also increase the probability of its activating effect on immunocompetent cells, since it is the membrane has a corresponding specific receptor.

The claimed composition on activity surpassed the tool is a prototype and in relation to the actual bactericidal mechanisms of action of blood serum, which include the activity of serum lysozyme. Observed more pronounced effects of the inventive compositions for the specified component of immune protection against the studied infectious diseases of bacterial origin allow to give her preference as a means of anti-infective protection.

Thus, studies have shown that the inventive composition is substantially activates the mechanisms of bacterial protection. In further studies conducted to evaluate the feasibility and prospects of application of the inventive composition as a means of prevention and treatment of dangerous infections of bacterial etiology, including generalized Salmonella and anthrax infections.

Example 5. The study interferonogenna the aphrodisiac properties of the prototype and declare HDMI is tion.

Because nonspecific immunological resistance is designed to provide protection not only in bacterial, and viral infections, we investigated the extent to which the tool is a prototype and the claimed composition can influence interferonogenez.

The investigations were carried out on outbred laboratory mice (kennel "Rapolano RAMS weighing 18-23, Experimental groups of animals were formed as described in example 4.

In the private case, the composition of the claimed composition: CCM containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (2.1 wt.%), zeolite (44,6 wt.%), complex polysaccharides (52.4 wt.%), calcium stearate (0.9 wt.%).

To determine interferonogenna the aphrodisiac properties of the prototype and the claimed composition in dynamics were blood collection in laboratory animals from retroorbital sine, combining in one sample material from three animals. Obtained after centrifugation at 1000 rpm./min for 10 min, the serum was aspirated and stored at minus 20°C. the Internal organs of mice triturated with the addition of saline solution, the homogenate was centrifuged (4000 rpm./min) for 20 min and supernatant was prepared 10% suspension in saline. Thus obtained samples were stored at minus 20°C. Titration of interferon (IFN) in p is obtained samples were micromethods in cell culture. Culture cells L-929, diluted growth medium to the desired concentrations (3-4·105cells/ml)were made in the wells of 96-well plastic panels, and incubated in an incubator for 24 hours throughout the titration process incubation of cell cultures were held under identical conditions: temperature 37°C in an atmosphere with 5% CO2and 98% humidity. After the formation at the bottom of the wells of the cell monolayer growth environment of the panel was removed and made supportive environment in which produced a serial twofold dilution of test samples (paired holes). At the end of the daily incubation of the test sample was removed and made the test virus encephalomyocarditis (EMC) working dilution average PD50(cytopathogenic dose at which there is a loss of 50% of the cells). The next day was counting results using an inverted microscope. The titer of IFN - reciprocal of the last serum dilution or sample homogenate body, where there is a 50% protection against the test virus.

It is established that the tool prototype practically does not possess interferonogenna properties (figure 4). In laboratory animals, which has introduced a tool prototype, the level of serum interferon practically did not change compared to control values, although somewhat increased (up to 20 U/ml), one is about these differences with the control level (5 U/ml) were not statistically significant. At the same time, a single injection of the inventive composition in the same dose as a tool prototype (150 μg/mouse) had quite pronounced interferonogenna action. After 1 day after administration of the inventive composition, the level of serum interferon was increased to 160 IU/ml (the difference being statistically significant (p<0.05) as compared to control and the level of serum interferon in animals treated with the vehicle-prototype). In further observed decrease in the level of serum interferon, however, and beyond the time frame of the study assessed value significantly exceeded the control group and the group of animals treated with the vehicle-prototype (p<0,05).

Thus, studies have shown that the claimed composition in addition to the activation of mechanisms of bacterial protection significantly activated and interferonogenez is a key mechanism of antiviral defense. Fairly quick production of endogenous interferon allows early on to protect the body from infectious diseases of viral etiology.

In further studies conducted to evaluate the feasibility and prospects of application of the inventive composition as a means of prevention and treatment of dangerous viral infections, including tick-borne encephalitis and fever of the Western Nile is, having osseetilaisia is, agents which are among the potential biological damaging agents and agents of bioterrorism.

Example 6. Study of specific anti-infective activity funds of the prototype and the claimed composition on the generalized model of experimental Salmonella infection.

The experiments were carried out on outbred laboratory mice (kennel "Rapolano RAMS at the age of 2-3 MESI weighing 18-23, Before the experiment, control animals were obliged to endure quarantine for 1 week.

The most adequate experimental model for studying the effectiveness of means-prototype Isabaev composition is a model of salmonellosis outbred mice, vyzvannogo oral introduction of the pathogen. For modeling generalized experimental Salmonella infection used collection strain of microorganism of the family Enterobacteriaceae (Salmonella typhimurium breslau, strain No. 20). By determining the dose of the pathogen that causes the death of 50% of infected animals (LD50/ml), it was found that LD50/ml S.typhimurium for outbred mice is 107M.K./ml In further studies to infect animals used, the dose of the causative agent, equal to 5 LD50/ml.

In the experiment the formation of the Ali 3 groups of animals (control and 2 experimental) of 10 animals each.

Animals of the first group no expense has been appointed, was administered isotonic solution of sodium chloride (placebo) and were used as control.

Animals of the second group was administered vehicle prototype in the form of mist powder (oral dose of 150 μg/mouse).

Animals of the third group was administered the inventive composition in the form of suspended powder (oral dose of 150 μg/mouse).

For oral administration of the suspension means of the prototype and the claimed composition used syringes with a special probe.

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (1.8 wt.%), zeolite (46,0 wt.%), complex polysaccharides (51,4%by weight), calcium stearate (0.8 wt.%).

Schemes means the prototype and the claimed composition relative to infection by the pathogen: once for 24 h before infection (prophylactic); once within 24 hours after infection (treatment); three times: 24 hours before and 24 h after infection (combined).

The criterion for anti-infective activity of the means was the survival of animals in the experimental groups. The percentage of surviving animals was determined using the tables of Genes V.S. Watched infected animals within 21 days from the moment of infection, daily record the number of living and pawsitivity in the experimental groups.

As presented on figa data, a single prophylactic administration tool prototype to infect experimental animals pathogen Salmonella infection dose LD50had some protective effect against this type of infection, as manifested in ensuring the survival rate of 20% of the infected animals compared with 100% mortality in the control. At the same time, prophylactic efficacy in similar conditions the claimed composition was higher against the background 100% mortality in the control of survival of animals was 40%, that is 2.0 times exceeded the prophylactic efficacy means the prototype.

During therapeutic use of the prototype and the claimed composition (figb) found that the effectiveness of the use of both drugs was 10% less pronounced than when a single prophylactic use. However, in this case the effectiveness of the claimed composition in comparison with the tool prototype was 3.0 times higher.

The most pronounced protective effect as a means of the prototype, and the claimed composition was recorded in their application for a combined scheme (pigv). In the case of application of the prototype survived 60%, and the claimed composition - 100% of infected animals on a background of 100% mortality in the rear the Le. The protective properties of the claimed composition not only were significantly higher than control values (p<0,05), and 1.7 times higher than the specified rate means the prototype.

Thus, the protective properties of the claimed composition in relation to generalized experimental Salmonella infection is more pronounced than the means of the prototype. Single prophylactic use excess was 2.0 times, a single treatment and 3.0 times, and when combined application - 1.7 times.

Example 7. Study of specific anti-infective activity funds of the prototype and the claimed composition in experimental anthrax infection.

The investigations were carried out on outbred laboratory mice (kennel "Rapolano RAMS aged 2-3 months and weighing 18-23 gperez experiment experimental animals were obliged to endure quarantine for 1 week.

To simulate experimental anthrax infection used collectible strain of Bacillus anthracis 71/12 (second vaccine of Tsenkovsky). The spore form of the pathogen was prepared as follows: mesopatamia broth (pH 7.2 to 7.4) at 37°C for 24 h grown uterine material. Daily broth culture was evenly distributed across the surface of wheat agar, aslauga in mattresses, and cultivated in an incubator at a temperature of 32±0.5°C within 72 hours after culturing in an incubator mattresses were wrapped in dark paper and kept in a dark place at a temperature of 10°C for 5-7 days. The grown culture was washed from the surface of the agar with sterile distilled water. The resulting spore suspension was transferred into a sterile flask with a glass stopper, which was dissolved in sutel machine within 60 minutes after the time shaking the contents of the flask were filtered through sterile cotton-gauze filter was placed in a sterile vial with beads and kept at a temperature of 4±0,5°C.

By determining the dose of the pathogen that causes the death of 50% of infected animals (LD50/ml) found that the magnitude of LD50/ml for Bacillus anthracis outbred mice was 20 spores/ml In further studies to infect animals used, the dose of the causative agent, equal to 10 LD50/ml.

Experimental groups of animals were formed as described in example 6. The tool is a prototype and the claimed composition was injected at a dose of 150 μg/mouse of each.

Studies were conducted to assess the protective properties means the prototype and the claimed composition in a single of them at different times relative to infection causative agent of anthrax infection. Criterion against which infektsionnoi the effectiveness of the means used was the survival of animals in the experimental groups.

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (1.7 wt%), zeolite (45.7 wt.%), complex polysaccharides (51,1 wt.%), calcium stearate (1.5 wt%).

As follows from the presented data (figa), a single prophylactic insertion of the prototype or the claimed composition is most effective, then if the infection occurred after 1 d after injection of funds. In this case, the background 100% mortality in the control groups of experimental animals survived 40% (p<0,05). Efficiency means the prototype and the claimed composition in these conditions was the same. With increasing time interval between a single introduction of each tool and the subsequent infection of animals causative agent of anthrax infection their protective efficacy was falling. Moreover, the dynamics of reducing the protective properties of both funds was the same: in the introduction as a means of the prototype, and the claimed composition for 2 days before infection survived 10% of the animals used in the future with increasing time these funds anti-infective efficacy have not been shown.

Somewhat different results were obtained in a single therapeutic use of the prototype or the claimed composition (figb). So, the tool prototype showed practically no the efficiency is, providing only 20% protection of infected animals on a background of 100% mortality in the control with the introduction of it in 1 day after infection. In similar conditions the claimed composition was provided by a more pronounced level of protection. At its introduction in 1 day after infection, the survival rate of infected animals was 50%, after 2 days - 30%, after 3 days - 10%. The claimed composition was not practically effective for its introduction through 4 days after infection.

The most effective was the use of two tools in terms of a single injection simultaneously with infection (pigv), which was manifested in the increased survival of infected mice by 40% (medium-prototype) and 60% (of the inventive composition on the background 100% mortality in the control. As can be seen, the effectiveness of the claimed composition in 1,5 times in comparison with the tool prototype.

The combination of the presented results shows that a single use of the claimed composition for protection against experimental anthrax infection is more effective than a single application of the prototype.

Example 8. The study of protective properties of the claimed composition on the model of experimental tick-borne encephalitis.

The experiments were performed on white weinbrenner mice-males weighing 16-18, Before experimentality animals obligatory kept quarantined for 1 week.

Experimental tick-borne encephalitis was modeled using tick-borne encephalitis virus (TBE) (pathogenic strain Absettarov). Accumulation vaccinated material to infect laboratory animals were performed on mice-suckers. Originally prepared by five serial tenfold dilutions vaccinated material, which served as centrifugal 10% suspension of the brain previously infected mice or registrationentry of the lyophilic state vaccinated material. 0.02 ml of each dilution was injected into the mice brain-suckers, which have established observation in 24-48 h, after which animals were sacrificed ether, extracted the brain and deposited him on three samples in penicillin vials, which were stored in a freezer at a temperature of minus 20±0,5°C. In a further 10% suspension of the brain was used as vaccinated material. The initial titer of the virus was 102-103LD50/ml.

In the experiment formed 3 groups of animals (control and 2 experimental) 14 animals each. Experimental animals of all groups were infected with virus at a dose of LD50.

Animals of the control group no expense has been appointed and used as control.

Animals of the first and second experimental groups were administered the inventive composition of peror the flax in a volume of 0.5 ml and a dose of 150 μg/mouse. When this animal first experimental group claimed composition was injected for 5 day, 4 day, 3 day, 2 day, 1 day before infection (prophylactic scheme), and the animals of the second experimental group - together with the infection and then 1 time per day during 5 days in a row (medical scheme).

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (2.2 wt%), zeolite (43,8 wt.%), complex polysaccharides (53,6 wt.%), calcium stearate (1.4 wt.%).

The criterion for anti-infective efficacy of the claimed composition was survival of animals in the experimental groups. In the course of the experiment set (table 8)that with the introduction of the claimed composition for prophylactic or therapeutic purposes, the stability of the experimental animals to infection with tick-borne encephalitis virus was, respectively, 64,28±10,52% or 57,14±12,34% at 100% mortality in the controls (p<0,05). This fact suggests that the claimed composition exhibits antiviral activity due to its interferonogenna properties.

Example 9. Study of specific anti-infective activity of the claimed composition on experimental models of West Nile fever.

The investigations were carried out on outbred mice-males weighing 18-23 g, obtained from the kennel "Rapolano the RAMS. Before the experiment, control animals were obliged to endure quarantine for 1 week.

Experimental West Nile fever was modeled using West Nile virus (West Nile virus) (pathogenic strain NR). Accumulation vaccinated material to infect experimental animals were performed on mice-suckers. Originally prepared by five serial tenfold dilutions vaccinated material, which served as centrifugal 10% suspension of the brain previously infected mice or registrationentry of the lyophilic state vaccinated material. 0.02 ml of each dilution was injected into the mice brain-suckers and set watching them within 24-48 h, after which animals were sacrificed ether, extracted the brain and deposited him on three samples in penicillin vials, which were stored in a freezer at a temperature of minus 20±0,5°C. In a further 10% suspension of the brain was used as vaccinated material. The initial titer of the virus was 103-104LD50/ml.

In the experiment formed 3 groups of animals (control and 2 experimental) of 10 animals each. Virulent strain of the pathogen was used in infecting dose of 10 LD50and was injected subcutaneously in a volume of 0.3 ml

Animals of the control group no means is not appointed.

Animals of the first and second experimental groups were administered the inventive composition is administered orally in a volume of 0.5 ml and a dose of 150 μg/mouse. When this animal first experimental group claimed composition was injected for 5 day, 4 day, 3 day, 2 day, 1 day before infection (prophylactic scheme), and the animals of the second experimental group - together with the infection and then 1 time per day during 5 days in a row (medical scheme).

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (1.9 wt.%), zeolite (43,2 wt.%), complex polysaccharides with 53.4 wt.%), calcium stearate (1.5 wt%).

The criterion for anti-infective efficacy of the claimed composition was survival of animals in the experimental groups. Observation of infected animals was carried out for 21 days, every day given the number of live and dead animals.

Found that all experimental animals treated with the inventive composition oral repeatedly, in terms of their infection by West Nile virus in a dose LD50survived in 100% mortality in the control group (table 9). Moreover, in the control group the deaths of two animals (20%) was observed after 5 days after infection, after 7 days - 8 animals (80%), and in 10 days - all 10 animals (100%) (table 10).

Thus, once again proved that use the output of the claimed composition as a means for prevention and therapy of viral infections, including dangerous virus infection West Nile fever.

Example 10. Study of the efficiency of the combined purpose of the claimed composition and the antibiotic doxycycline in experimental anthrax infection.

The investigations were carried out on outbred laboratory mice (kennel "Rapolano RAMS weighing 18-23, Before the experiment, control animals were obliged to endure quarantine for 1 week.

To simulate experimental anthrax infection used collectible strain of Bacillus anthracis 71/12 (second vaccine of Tsenkovsky). The spore form of the pathogen was prepared as described in example 7. To pathogen infection in animals injected doses of 10, 100 and 1000 LD50/ml.

In the experiment we formed 4 groups of animals 10 animals in each.

Animals of the first group no expense has been appointed, was administered isotonic solution of sodium chloride (placebo) and were used as control.

Animals of the second group was administered the antibiotic doxycycline capsules 0.1 g production Ferane", the contents of which were dissolved in 0.9% sodium chloride solution and administered orally 1 time a day. Dose antibacterial agent was equivalent to a human dose was 1 mg/mouse.

Animals of the third and fourth groups were societegenerale antibiotic doxycycline and the claimed composition and introduced them orally. Doxycycline was administered according to the standard scheme, that is, only after infection. Introduced doxycycline orally 1 time a day and at a dose of 1 mg/mouse. The claimed composition was administered in a dose of 150 μg/mouse in a volume of 0.5 ml. Animals of the third group of the claimed composition was administered for preventive scheme (before infection), and the animals of the fourth group - medical scheme (after infection).

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (2.3 wt%), zeolite (45,4 wt.%), complex polysaccharides (51,2 wt.%), calcium stearate (1.1 wt.%).

As shown in Fig.7, the average life expectancy of animals from the control group infected with the causative agent of anthrax infection at doses of 10, 100 and 1000 LD50, respectively 4,1 day, 3,5 days and 2.5 days.

It was established that in contrast to monotherapy with combined antibiotic therapeutic administration of antibiotics and the claimed composition contributes significant (p<0,05) increase the average life span of experimental animals when infecting doses of the pathogen 10 and 100 LD50. So, oral co-administration of antibiotics and the claimed composition in 1 day or 2 days after infection increases the average life span of experimental animals, respectively, at 56.6 per cent is whether to 35.6% at the dose of the pathogen 10 LD 50and by 26.6% or 25,0% at the dose of the pathogen 100 LD50. When a high dose of the pathogen, component 1000 LD50the increase in the average life expectancy of animals was only 14.3% or 13.6% and had no statistical significance (p>0,05).

Thus, it is found experimentally that the medical appointment of a combination of doxycycline and the claimed composition suitable after 1 day or 2 days after infection with the pathogen at low doses. For a reliable increase in the average life span of experimental animals infected with high doses of the pathogen (1000 LD50), the combined introduction of the antibiotic and the claimed composition should begin no later than 0,5 days after infection (Fig). This will increase the average lifespan of experimental animals by 25.0%.

In addition, it was found that the combined purpose of doxycycline and the claimed composition regardless of the scheme introduction the claimed composition was more effective in terms of the protection against infection by the causative agent of anthrax infection than using only the claimed composition or doxycycline. Thus, the appointment of the claimed composition sachetana with doxycycline before infection with a high level of stability of the experimental animals to the infection was provided not only with Sarai is the fact that the dose of the pathogen 10 LD 50but 100 LD50i.e. when monophospate the claimed composition was ineffective, and doxycycline has provided protection at the level of 70% (figa).

A similar pattern was observed with the introduction of the claimed composition sachetana with doxycycline after infection (figb). However, in this case a high protective effect from the combined purpose of these funds is noted not only at low infecting dose of the causative agent of anthrax infection (10 LD50and 100 LD50), but 1000 LDO. In the latter case, monotherapy using doxycycline provided protection at the level of 40%, and the claimed composition was not effective. The effect on co-administered drugs in a similar situation reached 90 percent, while 100% mortality in the controls (p<0,001).

Thus, the totality of the experimental data shows the viability of combined antibiotic and probiotic agents in cases of emergency prevention and etiotropic treatment of infectious diseases of bacterial origin, in particular of anthrax infection. Using in such a situation, the claimed composition has a high efficiency, which is associated with the development between the claimed composition and the applied antimicrobial agent specific vzaimoposhi tiraumea effect. This effect is caused by the presence of the claimed composition of polysaccharides in the form of a complex of biologically active products of enzymatic hydrolysis of polymers of the cell wall yeast (ß-glucans and mannan), which have a more significant activating effect on immunocompetent cells, and cause the observed potentiating effect.

Example 11. Study of the efficiency of the combined purpose of the claimed composition and the antibiotic doxycycline to model generalized Salmonella infection.

The investigations were carried out on outbred laboratory mice (kennel "Rapolano RAMS weighing 18-23 gperez experiment experimental animals were obliged to endure quarantine for 1 week.

Used collectible strain of microorganism of the family of Enterobac-teriaceae (S.typhimurium breslau, strain No. 20). Infecting dose of the pathogen was 5 LD50/ml. Formed 4 groups of animals, as described in example 10.

In the private case, the composition of the claimed composition: ski containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis (1.6 wt.%), zeolite (43,7 wt.%), complex polysaccharides (53,2 wt.%), calcium stearate (1.5 wt%).

During the experiment found that to increase the effectiveness of protection in case of infection with Salmonella, infecti the th appropriate to assign the claimed composition sachetana with doxycycline, and to put it before infection (figa). In this case, the protective effect is provided even in terms of infection by the pathogen at a dose of 1000 LD50that is , when monotherapy with the use of the claimed composition is not effective, and the effect of antibacterial drug is reduced by 60%. The preventive purpose of the claimed composition is provided as a direct effect on pathogens its probiotic component, and significant activation of immunocompetent cells. As a result of such exposure to pathogenic organisms lose their stability and antibacterial exhibits a more pronounced anti-infective action.

Both funds after infection by therapeutic scheme is also more effective than monotherapy with antibiotica (figb).

Confirmed the results obtained in terms of infection anthrax infection, which showed the feasibility of the combined purpose of the claimed composition and the antibiotic also generalized infection Salmonella infection.

Thus, a set of experimental data testifies to the achievement of the purpose of the invention is a composition for efficient pathogenesis prevention and treatment of infectious diseases, significantly act is virusa mechanisms of antibacterial protection and interferonogenez (the key mechanism of antiviral defense) and universal against infections of bacterial and viral etiology when administered orally as when monotherapy and in combination with the appointment of the basic causal means. The claimed composition is safe, because the content of toxicants (organochlorine pesticides GHCT and DTST)and hazardous substances (lead, cadmium, mercury, arsenic and radionuclides (90Sr and137Cs) does not exceed permissible levels, and it has no acute toxicity. Guaranteed shelf life of the claimed composition is not less than 2 years at a temperature not exceeding 40°C and the storage does not require the use of special equipment.

The claimed invention satisfies the criterion of "novelty"as a first for the prevention and treatment of infectious diseases is proposed to use the song, providing the combined effect of the metabolites of probiotic microorganisms and the products of enzymatic hydrolysis of polymers of inner and outer layers of the cell membrane of yeast with immunomodulatory activity, characterized by high specific efficiency and flexibility in terms of infections of bacterial and viral nature.

The claimed invention satisfies the criterion of "inventive step"as available sources of information is not obvious expediency integrated use of the metabolites probiotics the microorganisms and their products of enzymatic lysis by cell membrane yeast (ß-glucans and mannan) in the composition of the claimed composition, as well as their optimal content to ensure effective anti-infective protection.

The compliance of the claimed invention, the criterion of "fitness for use" is confirmed by examples, showing relatively simple technology for the production of solid dosage forms of the claimed composition in powder form, the ingredients are inexpensive, are produced by the domestic industry and available, and the results of numerous studies that have confirmed its safety and stability when stored for 2 years without use of special equipment, as well as the presence of immunotropic effects interferonogenna properties and high anti-infective efficacy, indicating that the inventive composition will be widely used in clinical practice for the prevention and treatment of dangerous infections both bacterial and viral etiology in the case of the introduction of under the scheme monotherapy or in combination appointment with traditionally used basic drugs.

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Composition with anti-infective activity, performed in a solid dosage form for oral administration in the form of powder, representing a balanced complex comprising probiotic agent, adsorbent, and an auxiliary additive, to the m as a probiotic agent use sterilized culture fluid containing metabolites of the strain VKPM No-2335 bacteria Bacillus subtilis, as adsorbent zeolite as a support Supplement, calcium stearate or Aerosil, characterized in that it further includes a highly active immunomodulatory agent, which is a complex of polysaccharides - products of enzymatic hydrolysis of polymers of inner and outer layers of the cell membrane of yeast - in the form of β-glucan and mannan, in the following ratio of ingredients, wt.%:

Sterilized culture fluid containing
metabolites of strain VKPM No-2335 bacteria Bacillus
subtilis1,6-2,3
Zeolite43,0-46,0
Complex polysaccharides: β-glucan and mannan51,0-54,0
Calcium stearate or Aerosil0.5 to 1.5



 

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19 cl, 8 ex, 6 tbl, 19 dwg

FIELD: medicine.

SUBSTANCE: what is described is a new pharmaceutically solid preparation of a vasopressin antagonist which contains: (a) 7-chlor-5-hydroxy-1-[2-methyl-4-(2-methylbenzoylamino)benzoyl]-2,3,4,5-tetrahydro- 1H-benzoazepine and/or its salt; (b) hydroxypropylcellulose containing a hydroxypropoxyl group in the amount of 50% or more; and (c) at least one of the ingredients specified in a group consisting of carmellose, sodium carboxymethyl starch, crospovidone and hydroxypropylcellulose with a lower degree of substitution with an average diameter of particles 30 to 70 mcm and 90% of the cumulative diameter of 100 to 200 mcm.

EFFECT: pharmaceutical solid preparation shows excellent desintegration properties and solubility leading to adequate gastrointestinal absorbability of the active ingredients.

17 cl, 4 tbl, 21 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to solid pharmaceutical preparation of matrix type, which contains: (a) enterosoluble polymer based on methacrylic acid; and (b) sugar and/or sugar-alcohol, in which 1 g of sugar and/or sugar-alcohol can be dissolved in not more than 4 g of water at water temperature from 20 to 25°C. Pharmaceutical preparation does not contain softener. Pharmaceutical medicine is produced by extrusion method. Application of sugar and/or sugar-alcohol with specified properties ensures acceptable plasticity of mixture with high content of enterosoluble polymer based on methacrylic acid without addition of softener.

EFFECT: preparation by invention has satisfactory controllability of medicine release, suppressing release in upper part of gastrointestinal tract and insuring immediate release in lower part of gastrointestinal tract.

7 cl, 4 dwg, 31 tbl, 26 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to method of obtaining highly dispersed meloxicame by fast cooling of meloxicame solution in 1,4-dioxane to the temperature of liquid nitrogen with further removal of solvent from formed mixture of solid phases in vacuumed thermostated vessel at pressure <5·10-2 mm Hg and temperature lower than temperature of 1,4-dioxane melting. Fast cooling of solution is performed by dispersion of solution into vessel with liquid nitrogen or spraying solution on copper plate at liquid nitrogen temperature. Solvent removal is performed mainly at temperature 3°C, drying being carried out in flow of dry gas (nitrogen of argon) at pressure 100 Pa.

EFFECT: obtaining of highly dispersed meloxicame.

5 cl, 3 dwg, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and deals with hard pharmaceutical composition, which includes escitalopram or its pharmaceutically acceptable salt and/or solvate.

EFFECT: invention ensures absence of disadvantages as to fluidity and technical characteristics.

12 cl, 8 ex, 1 dwg, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: dosage form contains oxycodone hydrochloride as a physiologically active substance (A), optionally one or more physiologically combined excipients (B), a synthetic or natural polymer (C) and optionally natural, semisynthetic or synthetic wax (D).

EFFECT: dosage form of oxycodone hydrochloride possess degradation resistance of at least 400 N to less than 500 N, and releases max 99% of oxycodone hydrochloride in the physiological conditions after 5 h.

14 cl, 7 dwg, 17 ex

FIELD: biotechnologies.

SUBSTANCE: application describes compositions and methods of nerve cells visualisation. A composition contains a virus component bound with a fluorescent dye to produce a complex, which is capable of penetration into nerve cells, where the virus component is selected from a neurotropic replication-defect virus, a virus protein of the neurotropic virus and capsid of the neurotropic virus.

EFFECT: reduced risk of iatrogenic damage of nerves in medicine.

19 cl, 23 dwg, 1 tbl, 5 ex

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