EA016410B1 - Pharmaceutical composition based on cyclodextrin nanoparticles comprising rifabutin, method for preparing thereof, method for treating mikobacteriosis and helicobacterial infection (variants) - Google Patents

Abstract

The invention relates to medicine and pharmacy, in particular, to a pharmaceutical composition for the treatment of mikobacteriosis and helicobacterial infection based on dispersed in a pharmaceutically acceptable carrier cyclodextrin nanoparticles from 100 to 1000 nm, polymer free and comprising a therapeutically amount of rifabutin dispersed therein, and also pharmaceutically acceptable auxiliaries. The invention also discloses a method for preparing said pharmaceutical composition and methods for the treatment of mikobacteriosis and helocobacterial infection comprising intravenous and/or intramuscular injection and/or oral administration to a patient in need thereof of said pharmaceutical composition in a therapeutically effective amount. According to the invention the composition can be manufactured both as a lyophilizate applicable (after dispersion in a pharmaceutically acceptable carrier ) for intravenous or intramuscular injection to a patient in need thereof and as a powder applicable for manufacturing solid oral medical dosages and provides effectiveness not less (and in some cases even higher) than administered orally rifabutin substance

Description

Technical field

The present invention relates to a pharmaceutical composition for treating mycobacteriosis and Helicobacter pylori infection based on cyclodextrin nanoparticles dispersed in a pharmaceutically acceptable carrier containing a therapeutically effective amount of rifabutin dispersed therein, as well as a method for producing such a composition and methods for treating mycobacteriosis and Helicobacter pylori infection, including intravenous, and / or intramuscular and / or oral administration of a therapeutically effective amount of such a composition and to the needy patient.

The prior art and objectives of the invention

In recent decades, the problem of increasing the effectiveness of the treatment of tuberculosis and other mycobacterioses has become particularly relevant in connection with an increase in the incidence rate in Russia and around the world. In connection with the spread of HIV infection, a group of patients with an increased risk of developing both tuberculosis and non-tuberculous mycobacterioses, characterized by a severe disseminated course of the disease, was distinguished. Fatal immunodeficiency in HIV-infected patients, as well as secondary immunodeficiency in tuberculosis patients against the background of pathogen polyresistance, cause serious difficulties in the treatment of tuberculosis.

The most promising way to increase the effectiveness of the treatment of tuberculosis and other mycobacterioses (for example, leprosy, which is also a very urgent problem for developing countries) turned out to be the creation of fundamentally new anti-TB drugs by screening new natural molecules or chemical modification of known structures. However, its effectiveness is limited in time due to the continuity of the process of emergence and spread of resistance to new drugs. Nevertheless, the creation in the late 80s of a new semi-synthetic broad-spectrum antibiotic of the rifamycin-rifabutin group was an important achievement in pharmaceutical chemistry [1] (see the List of sources at the end of this description).

By chemical structure, rifabutin is 4-deoxo-3,4- [2-spiro [I-isobutyl-4-piperaidyl] 2,5-dihydro-1H-imidazole]. According to the spectrum and mechanism of action, rifabutin is similar to rifampicin, but significantly exceeds its pharmacodynamic and pharmacokinetic properties [1].

Thus, the better solubility of rifabutin than rifampicin in lipids determines its better penetration into tissues, a larger distribution volume, a smaller difference between the maximum (C _max ) and minimum (C _TT ) concentrations in the blood serum, prolonged half-life from the body [2] .

The antibacterial action of rifabutin, like other antibiotics from the rifamycin group (for example, rifampicin), is based on the suppression of DNA-dependent RNA polymerase of bacteria [1]. It is also assumed that the drug has a direct inhibitory effect on the synthesis of bacterial cell DNA, which determines its activity against rifampicin-resistant mycobacteria [2]. Rifabutin has a wide spectrum of antibacterial action, is active against a wide range of gram-positive and gram-negative bacteria.

The most important property of rifabutin is its activity in relation to Musoacanthis lucensis818 and Musocactylacticus aux-t! Gase11iage sotr1ekh. The values of the minimum inhibitory concentration (MNC) of rifabutin in relation to rifampicin-sensitive strains of Muscoabacillus ulcerida818 are 0.03-0.06 μg / ml, i.e. within the range of concentrations that are maintained in the blood for 24 hours after a single oral administration of 300 mg of the drug . The minimum bactericidal concentration (MBC) is 2-4 times higher than the BMD for the same strains of mycobacteria. This level of antibiotic concentrations in the blood is detected within 6-12 hours after taking the above dose [1], [3], [4].

An important property of rifabutin, which determines the possibility of its use in tuberculosis due to rifampicin-resistant mycobacteria, is the lack of complete cross-resistance of pathogens to these antibiotics. It has been shown that more than 35% of rifampicin-resistant mycobacteria remain susceptible to rifabutin. The effectiveness of rifabutin was confirmed in a model of experimental tuberculosis in mice caused by rifampicin-resistant strains, when the drug used in calculating the average daily doses for humans (300-400 mg) contributed to the rapid elimination of mycobacteria [5].

Rifabutin is more active than rifampicin in relation to clinical strains of atypical mycobacteria (Musoacobacterium asthma-tigase11iage sotr-ex-MAS) [6]. The values of IPC90 (the minimum concentration that suppresses the growth of 90% of mycobacteria) of rifabutin for most strains of these species ranges from 1.0-2.0 μg / ml. The sensitivity to rifabutin of atypical mycobacteria (Musobaspepit kapzazi, Musobaspompus Goochiyit, Musobaspompus dogbopae, and Musobaspepitum apoptosis) is significantly lower than that of MAS (MPC90, depending on the species, 2.5-8 μg / ml). Muso bacillus siblioe and Muso bacillus 81m bae are moderately sensitive or stable [7].

Most strains of Mycobacterium pylori are also highly sensitive to rifabutin. According to the degree of activity, ίη was eliminated, and with experimental leprosy of white mice, rifabutin was found to be significantly superior to rifampicin. The observed synergistic effect in

- 1 016410 wearing mycobacteria of leprosy with the combined use of rifabutin with fluoroquinolones (sparfloxacin, lomefloxacin, norfloxacin, ofloxacin, perfloxacin, ciprofloxacin, levofloxacin or moxifloxacin) is more pronounced than with the combined use of this group of rifampicin.

One of the most important indications for rifabutin use today is tuberculosis in HIV-infected patients and in patients with AIDS (HIV / AIDS-associated tuberculosis) [9].

In relation to other microbial species, the activity of rifabutin is much less studied than for mycobacteria, however, it is known that rifabutin is similar to rifampicin by the action on many conditionally pathogenic species of gram-positive and gram-negative microorganisms. Rifabutin is active against 81 aryu1ossoss aiteik, coagulase-negative 81 aryuosossik erMetshb, 81 aryuosossik 8artyu11s8, 81aryosossik ayooisyk, 81ter1ossoss5 ruodeik and 81ter1oss015 ml / ml. Sensitive to rifabutin are 81ter1osossi8 rheitoschae and anaerobic cocci; Epilogossossi8 Gaesike strains show only moderate sensitivity to rifabutin (BMD 25 μg / ml) [11].

Rifabutin is active against methicillin-resistant strains of Staphylococcus aureus (MK8A) and coagulase-negative (MKSI8), however, as in the case of rifampicin, staphylococci resistance to риη νίΙΐΌ and ίη νί \ Ό rifabutin develops very quickly, almost after a single contact. Its development can be prevented by the combined use of rifabutin with fusidine, fluorohypolones or erythromycin. Such combinations can be effectively used in the treatment of severe infections caused by methicillin-resistant strains of Staphylococcus aureus (MK8A) and coagulase-negative (ICSI8) [12].

Rifabutin is characterized by the rapid development of resistance in pneumococci, especially in cases of its frequent use in the treatment of tuberculosis, MAS-mycobacteriosis (Musoobacterium austiculitaceae and staphylaxis). Among gram-positive bacteria, rifabutin is highly susceptible to C1O81pb1it crr., Including C1o8BtBGG. Rifabutin is highly active against the neysseries (Jebkepa teshpdShbb, Jebkepa dopotgyoeae), although clinical data on the possibility of its use in these infections are practically absent [11].

Of great interest are the data on the high sensitivity to rifabutin of Neusobacillus elipop [13], [14], [15]; rifabutin MPC90 for a given microorganism is 0.007 μg / ml. Rifabutin is significantly superior to rifampicin in the degree of activity with respect to Neusobacillus rifoputin [13]. Such a property of rifabutin, as stability at wide fluctuations in pH values, gives reason to consider it as an active component in the complex therapy of peptic ulcer.

There is evidence of a high activity of rifabutin (MPC - 0.008 μg / ml), superior to the action of rifampicin, in relation to the United States 1gas1yutO8. Moreover, rifabutin is characterized by a slow formation of chlamydia resistance, while rifampicin ίη ν Нgo resistance is formed in the presence of subinhibitory concentrations for several passages [16].

In the experiment ίη у1уо it was reliably established that rifabutin is a highly effective drug in the treatment of toxocoplasmosis [17]. This feature of the antibiotic is important because antibiotic therapy of toxocoplasmosis causes serious difficulties. A dose of rifabutin 100-200 mg for 10 days protects 100% of mice with experimental toxoplasmosis. This dose is significantly higher than that used in the treatment of tuberculosis or MAS infection (Musoabacepit ausci1Tac11i1age cooper), however, it can be reduced when used in combination with sulfadiazine, pyrimethamine or clindamycin (ibid.). It was also shown in [17] that rifabutin (at a dose of 50 mg / kg) in combination with subtherapeutic doses of pyrimethamine or clindamycin prevented the death of 40 and 90% of animals, respectively.

Rifabutin is also active against many gram-negative bacteria, in particular against Rkeibtoak aetidshoka (Pseudomonas aeruginosa), and its activity against this pathogen has been shown to increase when used together with polymyxin B [18].

Rifabutin acts bactericidal on the microbial cell. For Musobacillus aust and Musocabepit tacet11i1age actively proliferating inside the cell, the bactericidal effect of the drug is more correlated with clinical efficacy than the BMD values. When studying the dynamics of the death of Musobactylaaaaaaaaaaaaaaa, the values of MBC (minimum bactericidal concentration causing death of> 99% of bacterial cells) for rifabutin were 8 μg / ml, while BMD (minimal inhibitory concentration) was less than 0.03 μg / ml [6]. To a greater extent, the rifabutin activity characterizes the value of the MBK / MPC ratio, which for Musoacapit aushet ranges from 8 to 128. When determining the rifabutin MBC ίη νίΙΐΌ in a dynamic system that simulates the concentration of antibiotic in the blood when taken orally, by counting the number of CFU (colony forming units) after Exposure of the strain Musobacepit nigasecephalon with 5 μg / ml antibiotic shows a decrease in the number of viable individuals, starting from the 6th day of contact with complete death of the culture by the 8th day. The ratio of MBC to MPC of rifabutin with respect to the reference strain MusoBac1epit 1begi1o818 was 4, with a MBC value of 0.125 μg / ml [19].

The relative activity of ίη of rifabutin compared to rifampicin varies in the range

- 2016410 Lah from 2 to 20%. Plasma antibiotic levels for rifabutin, superior to BMD for rifampicin-sensitive MusoLaSetsti1 and Letsi1O818 strains, are maintained for 24 hours after ingestion of 300 mg; concentrations corresponding to MBC are detected within 6-12 hours after a single dose of rifampicin in the same dose [20].

The values of MBC and IPC of rifabutin and, consequently, the MBC / IPC ratios for other types of bacteria (Lupus erythematosus b., Syshua nigella; Sh8. §1apy1ossoss aiteis) differed by no more than 2 times [1].

Rifabutin is characterized by a prolonged post-antibiotic effect, manifested in the suppression of microbial growth after a short exposure period of the culture with the antibiotic and its subsequent removal from the nutrient medium. The duration of the post-antibiotic effect depended on the type of microorganism, the concentration of the drug, and the duration of exposure. When the culture of MAS with rifabutin was exposed for 1-2 hours at drug concentrations 2-4 higher than MBC, the duration of the post-antibiotic effect ranged from 21.5 to 47.5 hours [6].

The possibility of enhancing the antimicrobial activity of rifabutin when it is combined with other antibacterial drugs has been most thoroughly studied in relation to Muscobacterium arrhythmias and arthritis (MAS), which is due to the difficulties in treating diseases caused by these pathogens. Ιη у1уо rifabutin, used in monotherapy for generalized MAC infection, was characterized by moderate activity, manifested only in a decrease in the seeding of the lungs and spleen of mice, without protecting them from death. With prophylactic administration, the efficacy of rifabutin was comparable to that of clarithromycin or azithromycin [1].

The activity of rifabutin in combination with other anti-tuberculosis drugs was studied in various systems, using when processing the results in some cases, the determination of the index of fractional inhibitory concentration (FIC) (the sum of the MPC of drugs in combination with the MPC of each drug separately), as well as the index of fractional bactericidal concentration (FBK) (the sum of the MBK of each drug in combination, divided by the MBK of each drug separately). The synergistic effect of rifabutin with ethambutol was found for 43-100% of MAS strains (FIC- or FBK-index <0.5) [21].

With a combination of rifabutin with clarithromycin, a significant increase in the sensitivity of MAS strains isolated in patients with HIV infection was observed [22].

Combinations of rifabutin with ethambutol and ciprofloxacin / amikacin (91 and 100% cell death, respectively) were the most active against Musoacobacillus sulacidae spp1ex in the mouse macrophage system. A combination of rifabutin with thiasetasone was also synergistic with respect to MAC strains [23].

Additive or synergistic effects with the combined use of rifabutin and ethambutol have been found in relation to Muso-Shetstat kapsazi [24].

The combinations of rifabutin and ethambutol with clofazimine or cefazolin, as well as rifabutin with cefazolin and streptomycin, showed synergism against strains of Muscobacterium pylori and Steptomycin [25].

Rifabutin is one of the few antibacterial drugs that are highly effective in treating infections caused by intracellular localized pathogens, in particular in the treatment of mycobacteriosis. The basis of such a high activity of rifabutin is its rapid penetration into the cell, in particular into the macrophage, which occurs without a violation of phagocytic function. The ratio between the intracellular and extracellular concentrations of rifabutin in the macrophages of mice is from 9: 1 to 15: 1 compared with the same 5: 1 for rifampicin [26].

Depending on the concentrations used, rifabutin, as shown in the literature, showed inhibitory or bactericidal activity against murine macrophages propagating in the culture of MusoLaSetst ausc-1p1gass11i1age sotrxex and MusoLaSetst hepor1 [27], [28].

When the atypical mycobacteria isolated from AIDS patients with therapeutic concentrations of rifabutin achieved in the blood plasma after its administration as an oral dosage form for 2-5 days were exposed, a bacteriostatic effect was observed with a pronounced decrease in the number of CFU (colony forming units) [29].

With the developed infection of Musobacillus aust-shlagase11iage stp1ex in mice, the effect of rifabutin is manifested in a pronounced decrease in the number of viable mycobacteria in the lungs and spleen; for this indicator, the effect of rifabutin is superior to the similar effect of rifampicin, rifapentin or azithromycin [1].

In the prevention of mycobacteriosis in infected mice pre-vaccinated with the Calmette-Guerin vaccine, the protective (prophylactic) effect of rifabutin was manifested when this antibiotic was administered at a dose of 10 mg / day for 1 week, while rifampicin had a similar protective effect only after 12 weeks use in the same dose [22].

With respect to the Neisobetanetopian strains resistant to clarithromycin and tinidazole, the three-component therapy, which included the use of a proton pump inhibitor (pantoprazole), amoxicillin, and rifabutin, was most effective [14].

Despite such a wide range of antibacterial activity, the clinical use of reef

- 3 016410 butine is limited to the fact that rifabutin, as already noted above, is sparingly soluble in water, which is why rifabutin preparations suitable for parenteral (intravenous and / or intramuscular) administration to a patient in need do not exist today.

At the same time, traditionally available commercially available oral dosage forms of rifabutin (usually rifabutin is now available in the form of gelatin capsules) are characterized by low bioavailability, and therefore the doses of this antibiotic required for effective antibiotic therapy are quite high, which causes not only significant side effects in relation to the digestive system (diarrhea, heartburn, indigestion, loss of appetite and nausea), but also such side effects various effects such as fever, the appearance of skin itching and skin rashes, thrombocytopenia (according to ΕΌΆ); It has been shown that therapy using traditional oral forms of rifabutin may also be accompanied by the appearance of flu-like symptoms [30] (previously similar symptoms were described in the literature also in patients who received the rifampicin antibiotic).

In this regard, the urgent task is to develop a soluble dosage form of rifabutin suitable for intravenous administration to a patient in need (when dissolved in an appropriate pharmaceutically acceptable carrier), while having a low toxicity and efficacy no less than traditional oral forms of this antibiotic.

One of the methods of increasing the solubility of sparingly soluble drugs that is widely used in the technology of pharmaceutical compositions and dosage forms is the use of cyclodextrins, which form inclusion complexes with said sparingly soluble substances.

Cyclodextrins are a class of cyclic oligosaccharides obtained by enzymatic cleavage of starches and consisting of 6 (α-cyclodextrins, α-ΟΌ), 7 (β-cyclodextrins, βСЭ) or 8 (γ-cyclodextrins, γ-ΟΌ) residues aE (+ ) -glucopyranoses (dextrose) connected into macrocycles by a-O-1,4-glycosidic bonds in such a way that the cyclodextrin molecule has the shape of a truncated cone with a hydrophobic internal cavity. By treating starch with the microbial enzyme, cyclodextrin glucanotransferase, the so-called coarse (1-ga-hdd) cyclodextrins containing nine, ten, eleven or more (up to 30-60) glucose residues in the cycle — so-called δ-cyclodextrins, ε-cyclodextrins, ξ -cyclodextrins, etc.). The structural formulas of the three main types of cyclodextrins (α-ΟΟ, β-ΟΟ, γ-SE) are presented in FIG. one.

The conical shape of the cyclodextrin molecule is stabilized by hydrogen bonds between OH groups, as well as a-O-1,4-glycosidic bonds. All OH groups in cyclodextrins are located on the outer surface of the molecule. Therefore, the internal cavity of cyclodextrins is hydrophobic and is capable of forming inclusion complexes with various molecules of organic and inorganic nature in aqueous solutions. In inclusion complexes, the cyclodextrin ring is a host molecule; the included substance is called a guest.

Inclusion complexes in aqueous solutions dissociate into cyclodextrin and the starting material, exhibiting the basic properties of the latter. When heated above 50-60 ° C, the complexes usually decompose completely and usually recover their structure upon cooling.

The formation of inclusion complexes with various medicinal substances by cyclodextrins makes it possible to give medicinal substances new useful properties, for example, to increase their solubility, physical and chemical stability, and so on. The properties of cyclodextrins, useful for creating soluble dosage forms of sparingly soluble drugs, are described in more detail in sources [31] - [48].

In particular, it was shown that the formation of drug inclusion complexes cyclodextrin is able to increase the physicochemical stability of the drug in solution, as well as protect a number of drugs from biodegradation.

Cyclodextrins are also used in the pharmaceutical industry to mask the bitter / unpleasant taste of a drug, to reduce the volatility of a drug, to produce drugs for oral or parenteral administration with controlled release, etc.

The PCT international publication [49], as well as [50] describe the inclusion complexes of the antibiotic rifampicin with β-cyclodextrin obtained by spray drying and intended for intrapulmonary administration in the treatment of tuberculosis; the publication [49] also showed that rifampicin inclusion complexes with β-cyclodextrin had a higher dissolution rate in a medium with pH 7.4 (corresponding to the physiological pH value) and with pH 5.2 (corresponding to the pH value in alveolar macrophages), Comparison with free rifampicin not included in inclusion complexes. Also in the framework of publication [49], it was shown in the study of ίη νίνο (in rats) that rifampicin as a part of inclusion complexes (with β-cyclodextrin and sucralose) had a higher percutaneous permeability compared to free rifampicin.

However, it is known that rifampicin is readily soluble in an aqueous medium (solubility of rifampicin

- 4 016410 at 25 ° C is 1400 mg / l), while the solubility of rifabutin in water is much lower. The use of cyclodextrins and / or their derivatives to increase the solubility of rifabutin in order to create colloidal forms of this antibiotic suitable for intravenous administration to a patient has not been studied previously.

It should also be noted that among the various types of cyclodextrins, it is β-cyclodextrins used in [49] that have the highest toxicity. Thus, in the studies of ίη νίίτο [51] it was for β-cyclodextrins that the highest hemolytic activity was shown. The hemolytic effect of cyclodextrins, the most pronounced according to this work for β-cyclodextrin and its derivatives - dimethyl-β-cyclodextrin and trimethyl-β-cyclodextrin, to a much lesser extent for hydroxypropyl-β-cyclodextrin, α-cyclodextrin, γ- cyclodextrin; dihydroxypropyl-b-cyclodextrin.

It is believed that the hemolytic activity of cyclodextrins is mainly associated with their ability to solubilize lipids in cell membranes. Indirectly, this assumption is confirmed by the existence of a positive correlation between the hemolytic activity of individual cyclodextrins and their ability to solubilize cholesterol, which is one of the main components of biological membranes [39], [41].

In addition, with respect to β-cyclodextrin and its methyl β-cyclodextrin derivative, it has been shown in the literature that they are able to induce caspase-dependent apoptosis of human keratinocytes by solubilizing cell membrane cholesterol; for α-cyclodextrin, γ-cyclodextrin, and 2hydroxypropyl-β-cyclodextrin, a similar effect was not shown [52].

The above disadvantages are also characteristic of inclusion complexes formed by β-cyclodextrins with the antibiotic rifampicin described in [53].

In recent years, nanoparticles based on biodegradable polymers (polylactides (PBA, PHOA), polyalkyl cyanoacrylates (PACA, PBCA, PPCA, P1CA, etc.)) have become considered promising colloidal drug carriers (in particular, insoluble or practically insoluble in aqueous media funds).

Nanoparticles, in the general case, can be defined as a type of colloidal drug delivery systems, which are solid particles less than 1 micron in size, formed by natural or synthetic polymers.

Depending on the technology of their synthesis among nanoparticles, one can distinguish nanospheres having a polymer matrix in which the therapeutically active and / or diagnostic agent is dispersed, and nanocapsules having a core containing a therapeutically active and / or diagnostic agent in the liquid phase, and the surrounding its shell is a biodegradable polymer. In some cases, the therapeutically active and / or diagnostic agent can also be adsorbed on the surface of these nanoparticles, which can be further modified (in some cases, this modification allows changing not only the physicochemical properties of the nanoparticles, but also their biodistribution in the body).

The literature shows that nanoparticles provide highly efficient transport of adsorbed drugs to macrophages. Once in the circulatory system, the nanoparticles are delivered together with the blood stream to the target organs (which in the case of tuberculosis are primarily the lungs, spleen, liver) and are phagocytosed by resident macrophages and circulating monocytes. In the intracellular environment, the nanoparticles undergo biodegradation under the action of enzymes and secrete the drug substance contained in them, which allows to achieve high intracellular concentrations and, thus, to increase therapeutic efficacy. Nanoparticles have been shown to be able to accumulate in the body in foci of pathology, which are characterized by increased permeability of capillaries and disturbances in lymph flow. These properties of nanoparticles, most fully expressed upon intravenous administration, formed the basis for the idea of a possible increase in the effectiveness of antibacterial drugs using nanoparticles. In particular, it was shown that nanoparticles significantly increase the effectiveness of antibiotics in the treatment of experimental bacterial infections characterized by intracellular localization of the pathogen, including tuberculosis [54], as well as salmonellosis or listeriosis [55].

In the patent of the Russian Federation [56], obtained earlier by the scientific team of the Scientific-Production Complex Nanosystem, LLC, a medicine is disclosed for treating bacterial (intracellular) infections that contains rifabutin adsorbed in a matrix of polymer nanoparticles, potassium cholesteryl phosphate, or sodium glycocholate, or hexadecyl dihydrogen phosphate, or α-tocopheryl succinate, as well as a water-soluble polymer stabilizer (having a molecular weight of not more than 70 kDa and selected from the group consisting of polyvinyl alcohol, polyvinylpyrrolidone, according to isorbat and serum albumin) and fillers. The polymers used to produce nanoparticles with a size of 100-800 nm include a polymer / polymers of lactic acid and / or a copolymer / copolymers of lactic and glycolic acids with a glycolic acid content in said copolymers of up to 50 mol%; the molecular weight of these polymers and copolymers is from 5 to 300 kDa. Specified Tool

- 5 016410 in, disclosed in the patent [56], provides a prolonged action of rifabutin, increasing its bioavailability and, accordingly, the effectiveness of the treatment of bacterial infections.

In the Eurasian patent [57], also obtained earlier by the scientific team of the Scientific-Production Complex Nanosystem, LLC, a pharmaceutical composition is disclosed for the treatment of tuberculosis and diseases mediated by Neurofacial pogo. based on polymer particles with a size of 100-800 nm, containing a therapeutically effective amount of rifabutin antibiotic and excipients suitable for intravenous administration to a patient in need thereof, characterized in that it contains polymer / polymers of lactic acid and / or copolymer / copolymers lactic and glycolic acids with a glycolic acid content in copolymers of up to 50 mol% with or without an additional carboxy group at the end of the molecule or copolymer polymers of lactic acid polymers or copolymers of lactic or glycolic acid with polyethylene glycol; the molecular weight of the polymers and copolymers is from 2 to 200 kDa, and the composition is a lyophilisate, which, when added with water or physiological saline, forms a stable suspension with a particle size of 0.1-0.8 microns.

The specified composition additionally contains a water-soluble natural or synthetic polymer stabilizer with a molecular weight of not more than 70 kDa and, if necessary, a lipid plasticizer and fillers at a certain quantitative ratio of the components.

However, further studies of the polylactide-based nanosomal dosage form of rifabutin showed that this dosage form has a high degree of toxicity, and this toxicity is not due to rifabutin itself, but rather a high concentration of the biodegradable polymer contained in the nanoparticles (PbOL or PbA), which limits the possibility of further development and clinical use of such nanoparticles.

In the Eurasian patent [58], also owned by Nanosystem Research and Production Complex LLC, a pharmaceutical composition is disclosed for treating tuberculosis and diseases mediated by Neusobacillus ruptum, characterized in that it is a lyophilisate of rifabutin, solubilized with human serum albumin, characterized by an average particle size of from 4 to 10 nm, and when a pharmaceutically acceptable diluent or carrier is added, a stable solution suitable for intravenous administration to a patient in need thereof a creature characterized by an average particle size of 4 to 10 nm, as well as a method for producing such a pharmaceutical composition (rifabutin lyophilisate solubilized with human serum albumin), characterized in that the aqueous mixture of rifabutin, albumin and an organic solvent at a temperature of from 0 to + 40 ° C dispersed, subjected to high pressure homogenization to obtain a nanoemulsion, remove the organic solvent from the obtained nanoemulsion, filter, add a cryoprotectant, freeze and lyophilize, comb the specified aqueous mixture of rifabutin and albumin is obtained by dissolving 2-5% (m / v) albumin in demineralized water, then adding to the specified solution of albumin 2-5% (v / v) an organic solvent and rifabutin in an amount of from 0.2 to 20 , 0 wt.%, And a method for the treatment of tuberculosis and diseases mediated by Neusobacillus pylori, comprising the intravenous administration of a therapeutically effective amount of such a pharmaceutical composition to a patient in need thereof.

Although the composition in accordance with [58] does not contain potentially toxic biodegradable polymers, nevertheless, human serum albumin, necessary for obtaining such a composition, being one of the components of human blood plasma, can be infected with viral infections having a parenteral transmission route (HIV, hepatitis B and C, etc.); in addition, in some cases, intravenous administration to a patient of drugs based on albumin of human blood plasma can cause such side effects as urticaria, chills, fever, shortness of breath, tachycardia, lowering blood pressure, pain in the lumbar region.

The use of fully synthetic cyclodextrins for solubilization (ie, increasing solubility) of rifabutin avoids the above disadvantages inherent in the soluble form of rifabutin disclosed in the patent [58].

It should also be noted that in the case of the use of commercially available solutions of albumin for infusion, an additional step is necessary to purify the indicated solutions from the auxiliary components contained in them, which increases the complexity and cost of producing such a composition.

The PCT publication [59], which we offer as the closest analogue of the present invention, describes nanoparticles based on a biodegradable polymer and cyclodextrin (or a cyclodextrin derivative) containing a biologically active substance (in particular a hydrophobic biologically active substance). A copolymer of methyl vinyl ether and maleic anhydride (RUM / MA) is used as a biodegradable polymer in accordance with [59], β-cyclodextrin, or 2-hydroxypropyl-b-cyclodextrin, or 6 mono-deoxy-6-monoamine- is used as a cyclodextrin or its derivative b-cyclodextrin.

- 6 016410

The biologically active substance in accordance with [59] can be a small biologically active molecule, as well as a protein, or peptide, or nucleoside, or nucleotide, or oligonucleotide, or polynucleotide, or nucleic acid, or a drug, or an allergen, or antigen, or a hydrophobic substance, or be a substrate of P-glycoprotein. In particular, the drug according to [59] can be actinomycin Ό, albendazole, amitriptyline, amprenavir, atorvastatin, bunitrolol, camptothecin, carvedilol, celirolol, cyclosporine, clotrimazole, colchicine, cortisone, daunorubicin, debrisoxinazidine, debrisoxinazidine, , docetaxel, domperidone, doxorubicin, epirubicin, erythromycin, estradiol, etoposide, phenytoin, fexofenadine, PK-506, fluorouracil, gentamicin, griseofulvin, imatinib, indinavir, itraconazole, levofloxanzolazolazolazin, lovofloxacinzol, lovofloxacinzol, lovofloxacinzol, lovofloxacinzol, lovofloxacinzol ethylprednisolone, methotrexate, mibefradil, morphine, nelfinavir, ondasetron, paclitaxel, praziquantel, prednisolone, prednisone, quinidine, rapamycin, rifampicin, saquinavir, sirolimus, trimerolinolinolinolinolinoliminolinolinoliminolinolinoliminolinolinoliminolinolinoliminolinolinoliminolinolinoliminolinolinoliminolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinolinopolinolinolinolinolinolinfinolinolinolinolinolinolinolinfinolinolinolinolinolinolinolinolinolinolini vincristine.

[59] also discloses a pharmaceutical composition comprising said nanoparticles containing a biodegradable polymer (such as RUM / MA), cyclodextrin or its derivative and biologically active substance (such as actinomycin Ό, albendazole, amitiriptilin, amprenavir, atorvastatin, bunitrolol, camptothecin, camptothecin , celirolol, cyclosporine, clotrimazole, colchicine, cortisone, daunorubicin, debrisoquin, dexamethasone, digitoxin, digoxin, diltiazem, docetaxel, domperidone, doxorubicin, epirubicin, erythromycin, estradiol fen, etopide Toin, fexofenadine, RC-506, fluorouracil, gentamicin, griseofulvin, imatinib, indinavir, itraconazole, levofloxacin, losartan, lovastatin, mebendazole, methylprednisolone, methotrexate, mibefradil, morphine, nelfinavir, ondansetron, paclitaxel, praziquantel, prednisolone, prednisone, quinidine, rapamycin, rifampicin, saquinavir, sirolimus, sulfamethizole, ritonavir, tacrolimus, talinolol, teniposide, terfenadine, topotecan, triamcinolone, verapamil, vinblastine or vincristine), as well as a pharmaceutically acceptable excipient, carrier or adjuvant the teachings of these nanoparticles, including co-incubation in an organic solvent of a biodegradable polymer (such as RUM / MA) and an inclusion complex (cyclodextrin or its derivative: biologically active substance), followed by desolvation of the indicated biodegradable polymer in an aqueous-alcoholic solution, or (as an alternative ) comprising the incubation of nanoparticles of the specified biodegradable polymer in an aqueous solution containing an inclusion complex (cyclodextrin or its derivative: biologically active eschestvo).

The most preferred cyclodextrins according to [59] are β-cyclodextrin, or 2-hydroxypropyl-β-cyclodextrin, or b-monodeoxy-β-monoamine-β-cyclodextrin, or γ-cyclodextrin, or 2-hydroxypropyl-γ-cyclodextrin, or mixtures thereof.

Although the use of a copolymer of methyl vinyl ether and maleic anhydride (RUM / MA) in the composition of nanoparticles for drug delivery is described not only in the international PCT publication [59], but also in earlier publications of the authors' data [60], it is well known that, although maleic anhydride copolymers have also been studied clinically (in particular, it was shown, in particular, that such polymers themselves also have therapeutic activity, namely, antitumor, anti-inflammatory, immunopotentiating and interferonind tsiruyuschey activity [61]), but their use in medical applications, including for the manufacture of pharmaceutical compositions it was impossible due to their high toxicity [62], [63].

So, when using a copolymer of methyl vinyl ether and maleic anhydride (RUM / MA), toxic effects typical of many synthetic anionic polymers were observed - pyrogenicity, thrombocytopenia, inhibition of microsomal enzymes, sensitization to endotoxins, toxic liver damage, organomegaly, and inhibition of the reticuloendothelial system [reticuloendothelium 64].

Moreover, it is known from the literature that even safer and significantly better studied toxicologically than RUM / MA, polymers (for example, (co) polymers of lactic and glycolic acids POBA and / or PBA) when used in the form of nanoparticles can toxicity.

So, in a toxicological study [65] previously synthesized by a team of authors of the Scientific and Production Complex Nanosystem of nanosomal forms of rifabutin obtained on the basis of polylactides (РОАА or РАА) and not containing cyclodextrins (dosage forms of rifabutin described in Eurasian patents [56] - [58 ]), it was shown, on the one hand, the weakening of the toxic properties inherent in the substance of rifabutin, and on the other, the emergence of new types of toxicity, due precisely to the fact that a small amount of rifabutin was incorporated It was incorporated into nanoparticles from the slowly biodegradable polymer PbOA or PbA.

In addition, the hybrid nanoparticles described in the PCT international publication [58] consist of a biodegradable polymer (RUM / MA) rather than cyclodextrins and are characterized by a low drug loading (less than 20%); it should also be noted that the materials [58] do not contain any indications of the possibility of using such nanoparticles for the treatment of intracellular

- 7 016410 infections, in particular for the treatment of mycobacterioses (including tuberculosis).

All of the above limits the possibility of using the nanoparticles disclosed in [58] to create colloidal (nanosomal) forms of rifabutin suitable for intravenous administration to a patient.

Objectives of the invention

Given the above, it would be preferable to create such a dosage form of rifabutin that would combine the advantages of inclusion complexes with cyclodextrins on the one hand, and colloidal carriers, such as nanoparticles, on the other, without containing biodegradable polymers, which can cause additional species toxicity, would be toxicologically safe and would be no less effective than the currently known dosage forms of this antibiotic.

With respect to rifabutin, such safe (toxicologically) and highly effective dosage forms of rifabutin, in particular dosage forms suitable for intravenous administration, have not been previously investigated.

SUMMARY OF THE INVENTION

In the framework of the present invention, the team of authors of the Nanosystem Scientific-Production Complex LLC synthesized a pharmaceutical composition for the treatment of mycobacteriosis and helicobacter infection based on cyclodextrin nanoparticles dispersed in a pharmaceutically acceptable carrier from 100 to 1000 nm in size, containing a therapeutically effective amount of polymer-free rifabutin dispersed in them ; this composition, as was shown in the framework of this work, is suitable both for intravenous administration to a patient (with the addition of a pharmaceutically acceptable carrier and / or diluent), and for the manufacture of solid oral dosage forms.

Thus, within the framework of the present invention, there is provided a pharmaceutical composition for treating mycobacterioses and Helicobacter pylori infection based on dispersed in a pharmaceutically acceptable carrier, cyclodextrin nanoparticles ranging in size from 100 to 1000 nm, containing a therapeutically effective amount of rifabutin dispersed in them, as well as pharmaceutically acceptable excipients, characterized in that these nanoparticles do not contain a polymer.

Said pharmaceutical composition can be either a lyophilizate when dispersed in a pharmaceutically acceptable carrier, forming a suspension suitable for intravenous and / or intramuscular administration in a therapeutically effective amount to a patient in need thereof, and a powder suitable for the manufacture of solid oral dosage forms (such as tablets capsules or dragees).

A pharmaceutically acceptable carrier suitable for use in accordance with the claimed invention can be either a 0.9% aqueous solution of sodium chloride, or a 0.010.1% aqueous solution of polysorbate-80, or a 0.05-5% aqueous solution of poloxamer-188, or 0.05-5% aqueous solution of poloxamer-407; Additionally, the pharmaceutically acceptable carrier may contain a small amount of ascorbic acid.

In the framework of this work, no less (and in some cases higher) efficacy of oral and intravenous dosage forms of rifabutin based on cyclodextrin nanoparticles obtained in the framework of the present invention was shown in the treatment of acute mycobacterial infection (experimental mouse model of acute tuberculosis caused by intravenous administration Mycobacteria Musobac! etst 1bbci1O818, strain Η37Βν), as well as the safety of this form in toxicological terms in comparison with the substance of rifabutin.

Mycobacteriosis in accordance with the claimed invention is a mycobacteriosis caused by one or more pathogens selected from Musoacobacterium tuberculosis or Musocobacterium tuberosus 35. or MusoBas! stst kaikaki, or MusoBas1stst 5sgoGi1aseit. or MusoBas! ergst a \ 's-sh1gase11i1age sotr1ekh, or Musobas! ergst 1ergay, or Musobas! ergst raga! ergs1o818.

Helicobacter pylori infection in accordance with the claimed invention is an infection caused by Hepatobacter pylori infection

If necessary, the surface of the nanoparticles forming the inventive pharmaceutical composition may be further modified by one or more surface-active substances (surfactants); such a modification, as shown in the literature [66] - [68], allows one to change such parameters of nanoparticles as the duration of circulation of nanoparticles in the body, the biodistribution of the drug substance, the duration and severity of the therapeutic effect, and ensure targeted delivery of nanoparticles to target organs (target organs) etc.

Suitable surfactants for this purpose include, but are not limited to, Poloxamer-101, Poloxamer-105, Poloxamer-108, Poloxamer-122, Poloxamer-123, Poloxamer-124, Poloxamer-181, Poloxamer-182 Poloxamer-183, Poloxamer-184, Poloxamer-185, Poloxamer-188, Poloxamer-212, Poloxamer-215, Poloxamer-217, Poloxamer-231. poloxamer-238, poloxamer-282, poloxamer-288, poloxamer-331, poloxamer-401. poloxamer-407, sorbitan monostearate

- 8 016410 (δΡΑΝ 60), sorbitan tristearate (δΡΑΝ 65), sorbitan monolaurate (δΡΆΝ 20), sorbitan monooleate (δΡΆΝ 80), sorbitan monopalmitate (δΡΑΝ 40), sorbitan trioleate (δΡΑΝ 85) or mixtures thereof.

The most preferred surfactants for use in the framework of the claimed invention are polysorbate-80 (trademark T \ usp® 80), poloxamer-188 (trademark ΡΙιίΓοπίο® R-68) and poloxamer-407.

Poloxamers (see the general structural formula of poloxamers in Fig. 2) are copolymers of polyoxyethylene and polyoxypropylene, exist in the form of liquids, pastes or in solid form, and have been used in pharmaceuticals since the end of 1950 [68]; each poloxamer is characterized by a certain ratio of the number of polyoxyethylene and polyoxypropylene units.

It has been shown previously in the literature that surface modification (coating) of nanoparticles based on a biodegradable polymer (a copolymer of lactic and glycolic acids and / or a polymer of lactic acid) containing a pharmacologically active substance, poloxamer-188 allows you to prolong the circulation time of such nanoparticles in the blood, so and enhance the pharmacological effect of this substance, for example, delivering the active substance to the spleen [69] and to the lymph nodes [70], and can also be used to create dosage forms based on poly dimensional nanoparticles with a gradual (delayed) release of a drug substance [71] - [73].

Suitable for creating a pharmaceutical composition within the framework of the claimed invention, cyclodextrin may be selected from α-cyclodextrin, 2-hydroxypropyl-b-cyclodextrin, 6 monodeoxy-6-monoamine-b-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-y-cyclodextrin, sulfobutyl- β-cyclodextrin or mixtures thereof, the quantitative ratio of rifabutin: cyclodextrin in the claimed pharmaceutical composition may be from 1: 1 to 1: 5.

The composition in accordance with the claimed invention may also additionally (i.e., if necessary) contain a cryoprotectant selected from 1-50% glucose, 1-50% lactose, 1-50% mannitol or 1-50% trehalose.

Also in the framework of the present invention, we declare a method for producing a pharmaceutical composition according to the invention, characterized in that:

a) a sample of cyclodextrin or its pharmaceutically acceptable derivative (s) is dissolved in a solvent with vigorous stirring;

b) add to the resulting solution a portion of rifabutin;

c) mix;

g) homogenize;

d) add the resulting dispersion to an aqueous solution of a surfactant with vigorous stirring;

e) homogenize;

g) remove the solvent;

h) centrifuged;

i) dried.

In the framework of the present invention, drying (step (s)) can be carried out both by freeze drying and by spray drying.

Additionally, this method may also include heating the reaction mixture, and / or treating the reaction mixture with ultrasound, and / or treating the reaction mixture with microwave radiation (microwave radiation), which can be carried out in step (c).

The specified method may also (if necessary) include the addition of a cryoprotectant (preferably selected from 1-50% glucose, 1-50% lactose, 1-50% mannitol or 1-50% trehalose); in this case, the cryoprotectant is added after the centrifugation step (step (h)).

The solvent in accordance with the claimed invention is preferably removed by distillation on a rotary evaporator (under reduced pressure).

Also in accordance with the claimed invention, we declare a method of treating mycobacteriosis, characterized in that therapeutically effective amount of the pharmaceutical composition of the invention is administered intravenously and / or intramuscularly in the form of a suspension in a pharmaceutically acceptable carrier to a patient in need thereof;

a method for treating mycobacteriosis, characterized in that a therapeutically effective amount of the pharmaceutical composition of the invention is orally administered in powder form or in the form of a solid oral dosage form to a patient in need thereof;

a method for treating Helicobacter pylori infection, characterized in that a therapeutically effective amount of a pharmaceutical composition of the invention is administered intravenously and / or intramuscularly in suspension in a pharmaceutically acceptable carrier to a patient in need thereof;

A method for treating Helicobacter pylori infection, characterized in that a therapeutically effective amount of the pharmaceutical composition of the invention is orally administered in powder form or in the form of a solid oral dosage form to a patient in need thereof.

It should be borne in mind that in the framework of the present invention, mycobacteriosis may be due to

- 9 016410 with one or more pathogens selected from Muscobacterium bacilli, or Muscobacillus emanus, or Muscobacillus kaivavi, or Muscocabepillus bacillus, or Muscobacterium bacillus Helicobacter pylori infection can be caused by Hepatobacter pylori.

The following examples relating to the preparation of a pharmaceutical composition in accordance with the present invention, as well as to the confirmation of its higher effectiveness in the treatment of acute experimental mycobacterial infection caused by Mycobacterium tuberculosis, do not limit the scope of claims, but are given only to illustrate the present invention.

A person skilled in the art should also understand that the claimed pharmaceutical composition based on nanoparticles (nanocrystals) of cyclodextrin with rifabutin incorporated into them can also be successfully used to treat other infections covered by the spectrum of antibacterial activity of rifabutin [1], in particular for treatment mycobacteriosis caused by Musobacterium tuberculosis, mycobacteriosis caused by Musobacterium tuberculosis, mycobacteriosis caused by Musobacterium tuberculosis, mycobacteriosis, obu of the muscular bacillus arteriosus arthritis, or musocactus arthritis, arthropod, or arteris arterisus, or musococcus arteris, and also for the treatment of Helicobacter pylori infection; thus, the spectrum of the antibacterial activity of the composition is determined not by the new dosage form disclosed in the framework of this application, but by the spectrum of the antibacterial activity of rifabutin itself [1].

Additionally, it should also be noted that the method proposed for the synthesis of cyclodextrin nanoparticles with rifabutin incorporated into them allows avoiding the use of highly toxic organochlorine solvents (methylene chloride, etc.), which are used, for example, in the synthesis of nanosomal forms of rifabutin according to [53] or [62] and the residual amounts of which can be stored in the finished dosage form obtained in accordance with [53] or [62]; in the claimed synthesis method, ethanol, acetone or Ν-methylpyrrolidone can be used as organic solvents.

Example 1. Obtaining (synthesis) of nanoparticles of cyclodextrin with rifabutin incorporated into them using freeze drying, their physicochemical characteristics.

A portion of rifabutin and cyclodextrin (selected from α-cyclodextrin (α-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-b-cyclodextrin (2-GP-in-CD) or sulfobutyl-β-cyclodextrin (δΒΕ-β-CD) poured with a solvent (ethanol, acetone, dimethyl sulfoxide (DMSO) or methylpyrrolidone), left on a magnetic stirrer with vigorous stirring for 4 days, then processed in an ultrasonic bath for 15 min, an aqueous solution of P1itoshs E-68 (Poloxamer-188) was added , twice homogenized using an ultrasonic homogenizer and the solvent was distilled off at transpose pressure. Further, large agglomerates were separated by centrifugation and to the supernatant was added cryoprotectant (glucose, lactose, mannitol or trehalose), dispensed into vials (1ml / vial), frozen at -70 ° C and freeze-dried.

For each sample and for each dilution, the average nanoparticle diameter (mean ± SEM) and the polydispersity index of nanoparticles (ΡΌΙ, mean ± SEM) were determined.

During the experiment, rifabutin concentrations, types of cyclodextrin used (α-cyclodextrin, γ-cyclodextrin, 2hydroxypropyl-b-cyclodextrin, sulfobutyl-b-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin), weight ratio of rifabutin were varied (upon receipt of different batches of the preparation): cyclodextrin, the type of solvent used (ethyl alcohol, dimethyl sulfoxide (DMSO), метил-methylpyrrolidone or acetone), as well as the presence of a surfactant (preferably 0.1-1% P1itoshos E-68) or its correspondence.

Physico-chemical characteristics of the cyclodextrin (CD) nanoparticles synthesized in accordance with this example with rifabutin (Bb) incorporated therein (including the concentration of rifabutin (%), the type of cyclodextrin used (α-cyclodextrin (α-CD), sulfobutyl-β-cyclodextrin (δΒΕ- β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-in-cyclodextrin (2-GP-in-CD)) and its concentration (%), the ratio of rifabutin: cyclodextrin for each batch of nanoparticles in accordance with the invention , type of solvent used, surface used -active agent (surfactant) and concentration (%), measurement conditions, the type of cryoprotectant used and its concentration (%), average diameter and polydispersity index of nanoparticles (ΡΌΙ)) are shown in Table.

one.

- 10 016410

Table 1

Physico-chemical characteristics of the nanoparticles in accordance with example 1

No. % Type of ID; The corresponding The solution is Surfactant View Average Index about con his con- rel riter % con cryo- meter (Ζ- polydis time price- centering but- ce n- leaking Auega ^ e s1), nm persons ( tsa tra- % she- TRG Torah, ΡΒΙ) nation nie his koi- B K: copen-

one one% (1-CD; [% 1: 1 Ethanol R1ignogns P-68, 1% Mannitol one% 618.2 * 19.7 0.267 * 0.014 2 one% a-CD: 1% 1: 1 Acetone P1igop! With P-68. one% Mannitol 1% 557.9 * 9.83 0.221 + 0.073 3 one% a-CD; one% 1: 1 dmeo RShogns E-68, one% Without cryoprotectant 407.5 * 2.15 0.163 * 0.01 4 one% a-CD; 1.5% 1: 1,5 Ethanol ΡΙυΐΌπϊο P-68. one% Without cryoprotectant 353.6 * 3.87 0.120 * 0.032 5 one% a-CD; 2% 1: 2 Ethanol Rkiogns G-68, 1% Without cryoprotectant 499.7 * 7.0 0.230 * 0.010 6 2% 8BE-R-CD; 2% 1: 1 Ethanol ΡΙ Eaton ίο P-68, one% Mannitol 374.4 * 12.2 0.256 * 0.013 7 one% 5BE-r-CD; 2% 1: 1.2 5 Acetone ΡΙυΐ'οηίο E-68. one% Mannitol 451.2 * 13.4 0.265 * 0.018 8 1.5% $ BE- | 3-CD: 2% 1: 1 DMSO ΡΙιιγοπ ίο E-68. one% Without cryoprotectant 384.1 * 1 7.8 0.301 * 0.020 nine 0.5% $ BE-R-SHCHD; 2.5% 1: 1.5 Ethanol R1igo1ps E-68, one% Without cryoprotectant 276.4 * 12.3 0.259 * 0.012 10 one% δΒΕ-β-CD; 2% 1: 2 Ethanol ΡΙυΐΌπίε P-68, one% Without cryoprotectant 215.8 * 1 1.2 0.232 * 0.009 II 0.5% 2-GP-r-CD; one% 1: 1 Ethanol R1igop1S G-68, one% Without cryoprotek- 294.3 * 3.36 0.09 * 0.022

12 0.5% 2-GP-r-CD; 1.5% 1: 1,5 Ethanol ΡΙιι gote P-68, one% Without cryoprotectant 326.5 * 1.90 0.086 * 0.013 thirteen 0.5% 2-GP-11-CD; 2% 1: 2 Ethanol ΡΙϋΓοηία G-68, one% Without cryoprotectant 310.6 * 2.06 0.106 * 0.011 14 one% 2-GP-y-CD; 1.5% 1: 1.5 Ethanol ΡΙιττοιιΐο P-68, 1% Mannitol one% 146.5 * 5.73 0.374 * 0.036 fifteen 1.5% 2-GP-y-CD: 2.25% 1: 1,5 Ethanol Ρΐιιτοπΐε P-68, ί% Without cryoprotectant 141.7 * 5.73 0.236 * 0.012 sixteen 0.5% 2-GP- in CD; 0.5% 1: 1 Ethanol Ρ) ιι ιοηίε R-68. one% Mannitol one% 247.55 * 3.29 0.419 * 0.023 17 0.5% 2-ΓΓΙ- y - CD 0.5% 1: 1 Ethanol R1igO1YS G-68, 1% Without cryoprotectant 1235.0 * 25.11 0.419 * 0.023 eighteen 0.33 % 2-GP- y - CD 0.5% 1: 1,5 Ethanol ΡΙιιγοπ) with P-68, one% Without cryoprotectant 774.1 * 31.87 0.217 * 0.041 nineteen 0.33% 2-GP- y - CD 0.5% 1: 1.5 Ethanol P1igol1S P-68. one% Without cryoprotectant 263.7 * 12.34 0.19 * 0.034

rifabuti incorporated into them. Example 2. Obtaining (synthesis) of nanoparticles of cyclodetetrin using spray drying, their physicochemical characteristics.

The synthesis of cyclodextrin nanoparticles with rifabutin incorporated into them was carried out analogously to Example 1, however, at the final stage of the synthesis, the particles were subjected to spray drying rather than lyophilic drying.

Upon receipt of various batches of the drug, the concentrations of rifabutin and the types of cyclodextrin used (α-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-b-cyclodextrin,

- 11 016410 sulfobutyl-b-cyclodextrin, 2-hydroxypropyl-y-cyclodextrin), the weight ratio of rifabutin: cyclodextrin, the type of solvent used (ethyl alcohol, Ν-methylpyrrolidone or acetone), as well as the presence of surfactants (preferably 0.1-1% P1igoshs P-68) or its correspondence. For each sample, the average nanoparticle diameter (mean ± standard deviation) and the polydispersity index of nanoparticles (ΡΌΙ, mean ± standard deviation) were determined.

During the experiment, the rifabutin concentrations, types of cyclodextrin used (α-cyclodextrin, γ-cyclodextrin, 2-hydroxypropylβ-cyclodextrin. Sulfobutyl-b-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin), weight ratio of rifabutin were varied (upon receipt of different batches of the preparation): cyclodextrin, the type of solvent used (ethyl alcohol, dimethyl sulfoxide (DMSO), метил-methylpyrrolidone or acetone), as well as the presence of a surfactant (preferably 0.1-1% P1itoshos P-68) or its correspondence.

Physico-chemical characteristics of some cyclodextrin (CD) nanoparticles synthesized in accordance with this example with rifabutin (Kb) incorporated therein (including the concentration of rifabutin (%), type of cyclodextrin used (α-cyclodextrin (α-CD), sulfobutyl-b-cyclodextrin (βΒ-β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-b-cyclodextrin (2-GP-WCD)) and its concentration (%), the ratio of rifabutin: cyclodextrin for each batch of nanoparticles in accordance with invention, type of solvent used, used The surface-active substance (SAS) and its concentration (%), measurement conditions, the type of cryoprotectant used and its concentration (%), average nanoparticle diameter and polydispersity index (ΡΌΙ)) are presented in Table. 2.

table 2

Physico-chemical characteristics of the nanoparticles in accordance with example 2

Sample No. % concentration B View 11D; its concentration,% The ratio r <b: KD Solvent Surfactant. View % krnoprotek- KOI-torus price- tra- Cia Average diameter (кAkega ^ e ά), nm Index of half-finishedness (PO1) one one% o + CD: one% 1: 1 Acetone ΡΙυΐΌη- Mannitol 1C E- 68.1% 605.94 9.2 0.324 + 0.08 2 one% a-CD: 2% 1: 2 Ethanol ΡΙιίΓοη- Without cryopro: with G-tector 68.1% 721.7 + 8.0 0.300 + 0.050 3 one% 5BE.-R- CP: 2% 1: 1.25 Acetone ΡΙοϊΌη- Mannitol 1s E- 68.1% 323.8 + 14.7 0.291 + 0.0.34 4 0.5% 8BE-R- CP: 2.5% 1: 1,5 Ethanol ΡΙαιοπ- Without cryopro- 1s E-Tector 68, 1% 254.9 + 12.1 0.318 + 0.048 5 0.5% 2-Γ'Π-β- CD one % 1: 1 Ethanol P1igop * Without cryopro- 1s E-Tector 68, 1% 297.1 ± 4.67 0.110 + 0.012 6 0.5% 2-ΓΠ-β- CD 2% 1: 2 This, yul ΡΙιιΐοη- Bezkriopro- 1C G-Tector 68, 1% 340.9 + 6.15 0.305 + 0.241 7 1.5% 2-ΓΠ-γID; 2.25% 1: 1.5 Ethanol P1igop- Without cryoproject G-tector 68, 1% 209.3 + 7.36 0.296 + 0.032 8 0.5% 2-GP-rCD; 0.5% 1: 1 Ethanol P1igop- Mannitol E · 68.1% 327.42 + 5.44 0.521 + 0.294 nine 0.33% 2-GP-r- CD 0.5% 1: 1.5 Ethanol ΡΙηγοιί- Without cryoproίο G-tector 68, 1% 578.2 + 28.13 0.262 + 0.054

As a result of a series of experiments (examples 1, 2), it was found that both spray and freeze drying are suitable for preparing a pharmaceutical composition in accordance with the invention (a composition based on cyclodextrin nanoparticles with rifabutin incorporated therein).

It was also found that the optimal ratio for the production of nanoparticles (nanocrystals) in accordance with the invention containing rifabutin is rifabutin: cyclodextrin, equal to 1: 1-1: 1.5; for α-cyclodextrin, this ratio is 1: 1.25, for 2-hydroxypropyl-cyclodextrin, approximately 1: 1.

As the result of the experiment, dimethyl sulfoxide (DMSO) was recognized as the most optimal solvent, however, since DMSO is difficult to completely remove from the finished preparation (pharmaceutical composition) in accordance with the invention, it was decided to use ethanol as an organic solvent.

The most preferred cyclodextrins for use in the pharmaceutical composition of the invention in accordance with the results obtained in Example 1 were

- 12 016410 recognized 2-hydroxypropyl-b-cyclodextrin and sulfobutyl-b-cyclodextrin.

One of the samples of the claimed pharmaceutical composition of rifabutin, synthesized in the framework of examples 1, 2 using 2-hydroxypropyl-b-cyclodextrin nanoparticles with rifabutin incorporated in them, was investigated as part of the present work on the efficacy in the treatment of experimental tuberculosis in mice caused by Musobacillus tuberculinum818. The methodology for studying the effectiveness and results are presented in example 3 (intravenous administration) and example 4 (oral administration).

Example 3. The study of the efficacy of the nanosomal pharmaceutical composition of rifabutin in accordance with the invention in the treatment of acute experimental tuberculosis in mice caused by infection with Musoacobacter pylori infection strain (S37V), when administered intravenously.

The purpose of the experiment was to compare the therapeutic effect of the claimed pharmaceutical composition of rifabutin (2-hydroxypropyl-b-cyclodextrin nanoparticles with rifabutin incorporated in them according to the present invention) when administered intravenously and the substance of rifabutin (when administered orally) in the treatment of acute experimental tuberculosis in mice.

The experiments were carried out on experimental animals — 100 mice (n = 100) —bait Ba1b / c females aged 7–8 weeks, weighing 20–22 g. Animals taken into the study were infected with Mycobacterium tuberculosis (Musobacepit Ibegi1-818, strain Η37Βν) at a dose of 5 , 6x10 ⁷ CFU / mouse by introducing a suspension of mycobacteria into the lateral tail vein of animals.

Both a suspension of rifabutin substance (BB) and a solution of its pharmaceutical composition (BB-CD NP) in accordance with the invention (sample No. 12 in Table 1) were prepared no more than 1 hour before the start of therapy (in accordance with passport recommendations and with the synthesis method disclosed in example 1). To prepare a suspension of rifabutin substance, 100 μl of 90 vol.% Ethanol was added to 10 mg of rifabutin substance, diluted with sterile water (up to 4 ml) to a final concentration of 2.5 mg / ml. The rifabutin composition of the present invention (BB-CD NP) was diluted in physiological saline (0.9% sodium chloride solution) containing a small amount of ascorbic acid, and diluted with water to a final concentration of 2.5 mg / ml.

Treatment was started on day 7 after infection and continued for 4 weeks.

During this period, 3 times a week, the animals of the experimental group were injected intravenously (ί.ν.) into the lateral tail vein of the pharmaceutical composition of rifabutin in accordance with the invention (B-CD LF), at a dose of 100 μg / mouse (20 animals) or 500 mcg / mouse (20 animals) (intravenously). Animals included in the comparison group were administered orally (reg oz) (in a volume of 0.2 ml) a solution of rifabutin substance at a dose of 100 μg / mouse (20 animals) or 500 μg / mouse (20 animals). Animals of the control group (20 animals) were injected intravenously (into the lateral tail vein) with 0.2 ml of physiological saline.

The day after the end of treatment, the animals were euthanized, their autopsy, removal and homogenization of internal organs (lungs, spleen); whole homogenates (0th dilution) and the first ten-fold dilution of homogenates (for animals that received an intravenous pharmaceutical composition of rifabutin in accordance with the invention and for animals that received an oral solution of rifabutin) were seeded on medium Brook 7-11 agar; for animals of the control group, Middlebrook 7–11 agar was sown on the first (tenfold) and second (hundredfold) dilutions of the homogenate. Counting the number of CFU was made 21 days after sowing; decimal logarithm (1d (CFU)) was taken for each value; the results for each group of animals, for each of the organs (lungs, spleen) are presented in table. 3 (CFU / organ, as: mean ± SD) and in FIG. 3.

TABLE 3 Seeding of organs of mice infected with M. luyegsi818 Η37Βν after intravenous treatment with the rifabutin composition in accordance with the invention

A drug The dose of treatment, mcg / mouse Lungs Spleen kj-cd ich 500 0.35 ± 0.29 2.53 ± 0.23 (sample 12 one hundred 3.22 ± 0.20 4.02 ± 0.15 according to Table 1) ί.ν. K, reg about " 500 1.62 ± 0.19 2.90 ± 0.07 one hundred 4.32 ± 0.10 4.52 1 0.07 Control group - 6.37 ± 0.06 5.86 ± 0.12

Thus, the intravenously injected cyclodextrin form of rifabutin (B-CD LF), created in the framework of the present invention, as can be seen from the data presented in table. 3 and in FIG. 3 was more effective than the rifabutin substance administered orally in the treatment of experimental mouse tuberculosis. Deviations between groups were significant at both doses used (100 and 500 μg / mouse). The greatest difference between the groups was observed when seeding from lung tissue.

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Example 4. The study of the effectiveness of the nanosomal pharmaceutical composition of rifabutin in accordance with the invention in the treatment of acute experimental tuberculosis in mice caused by infection with Musoacobacterium tuberculosis infection strain (Η37Βν), when administered orally.

By analogy with example 2, the purpose of the experiment was to compare the therapeutic effect of the claimed pharmaceutical composition of rifabutin (nanoparticles of 2-hydroxypropyl-b-cyclodextrin with rifabutin incorporated in them in accordance with the present invention), by oral administration of the composition (in powder form) and the substance rifabutin (also when administered orally) in the treatment of acute experimental tuberculosis in mice.

The characteristics and number of animals (type of animals, average body weight), the method of their infection with mycobacterium tuberculosis (Musoacantepyllobi1188, strain 3737), as well as the experimental design, were consistent with Example 3.

Analogously to example 3, treatment of animals began on the 7th day after infection, but continued for 8 weeks. During this period, 3 times a week, animals were injected orally, in a volume of 0.2 ml, at doses of 500 and 100 μg / mouse or a suspension of rifabutin (BB), or a rifabutin composition in accordance with the invention (BB-CD NP, reg о§, sample No. 12 in Table 1) (20 animals for each dose, for each form of rifabutin); the control group (20 animals) was administered orally with 0.2 ml of physiological saline.

The day after the end of treatment, the animals were euthanized, their autopsy, removal and homogenization of internal organs (lungs, spleen); whole homogenates (0-fold dilution) and the first ten-fold dilution of homogenates (for animals that received the antibiotic rifabutin in one form or another) were seeded on Middlebrook 7–11 agar medium; for animals of the control group (20 animals), 7-11 agar of the first (tenfold) and second (hundredfold) dilutions of the homogenate was sown on Middlebrook.

Counting the number of CFU was made 21 days after sowing; decimal logarithm (1d (CFU)) was taken for each value; the results for each group of animals, for each of the organs (lungs, spleen) are presented in table. 4 (CFU / organ, as: mean ± SD) and in FIG. 4.

Table 4

Insemination of organs of mice infected with M. luyegsi818 Η37Βν after oral treatment with rifabutin composition in accordance with the invention

A drug The dose of treatment, mcg / mouse Lungs Spleen K-vdnch (ob- 500> 0.00 ± 0.00 0.71 ± 0.32 zets 12 according Table 1), reg o $ one hundred 3.45 ± 0.27 3.66 t 0.18 K, reg 08 500 0.41 g 0.27 1.85 g 0.14 one hundred 4.40 t 0.05 4.25 ± 0.19 Control - 6.02 ± 0.10 6.16 ± 0.06 ..................

Of the presented in example 4, in table. 4 and in FIG. From the data in Figure 4, it can be seen that the orally administered rifabutin pharmaceutical composition of the present invention was found to be more effective than the rifabutin substance administered in the same manner in the treatment of acute experimental tuberculosis in mice. Deviations between groups were significant at the two doses used (100 and 500 μg / mouse). The greatest difference between the groups was observed when seeding from lung tissue. With the oral administration of the pharmaceutical composition of rifabutin in accordance with the invention at a dose of 500 μg / mouse, the effect (phenomenon) of sterilization in the lung tissue was observed.

Thus, in the framework of the present invention, it was possible to create a soluble dosage form of the rifabutin antibiotic, suitable for both parenteral and oral administration to a patient in need thereof, based on cyclodextrin nanoparticles containing rifabutin in a concentration of 4 ± 1 mg / ml; it was shown that the effectiveness of such a soluble form of rifabutin is not inferior to the effectiveness of the substance rifabutin administered orally (at the same doses), and even surpasses it in relation to the elimination of M. luyegsi-D8 in the spleen, which constitutes an unexpected technical result of the present invention.

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Claims (46)

CLAIM 1. Pharmaceutical composition for the treatment of mycobacteriosis and helicobacter infection based on dispersed nanoparticles of cyclodextrin in size dispersed in a pharmaceutically acceptable carrier - 17 016410 rum from 100 to 1000 nm, free of polymer and containing a therapeutically effective amount of rifabutin dispersed in them, as well as pharmaceutically acceptable excipients. 2. The pharmaceutical composition according to claim 1, characterized in that it is a lyophilisate, and when dispersed in a pharmaceutically acceptable carrier, a suspension for intravenous and / or intramuscular administration in a therapeutically effective amount to a patient in need thereof. 3. The pharmaceutical composition according to claim 1, characterized in that it is a powder for the manufacture of solid oral dosage forms. 4. The pharmaceutical composition according to claim 3, characterized in that the solid oral dosage forms are tablets, capsules or dragees. 5. The pharmaceutical composition according to any one of claims 1 to 4, characterized in that the surface of said nanoparticles is additionally modified with one or more surfactants selected from the group consisting of poloxamer-101, poloxamer-105, poloxamer-108, poloxamer-122 Poloxamer-123, Poloxamer-124, Poloxamer-181, Poloxamer-182, Poloxamer-183, Poloxamer-184, Poloxamer-185, Poloxamer-188, Poloxamer-212, Poloxamer-215, Poloxamer-217, Poloxamer-231, Poloxamer-231, Poloxamer-231 -238, poloxamer-282, poloxamer-288, poloxamer-331, poloxamer-401, poloxamer-407, sorbi en monostearate (8ΡΆΝ 60), sorbitan tristearate (8ΡΆΝ 65) sorbitan monolaurate (8ΡΆΝ 20), sorbitan monooleate (8ΡΆΝ 80), sorbitan monopalmitate (8ΡΆΝ 40), sorbitan trioleate (8ΡΆΝ 85) or mixtures thereof. 6. The pharmaceutical composition according to claim 1, characterized in that the cyclodextrin or its pharmaceutically acceptable derivative (s) is selected from α-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, sulfobutyl-β-cyclodextrin, 6-monodeoxy-6 -monoamine-β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, or mixtures thereof. 7. The pharmaceutical composition according to claim 1, characterized in that the quantitative ratio (rifabutin: cyclodextrin) is from 1: 1 to 1: 5. 8. The pharmaceutical composition according to claim 1, characterized in that it further comprises a cryoprotectant selected from 1-50% glucose, 1-50% lactose, 1-50% mannitol or 1-50% trehalose. 9. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutically acceptable carrier is a 0.9% aqueous solution of sodium chloride, or 0.01-0.1% aqueous solution of polysorbate-80, or 0.05-5% aqueous poloxamer-188 solution, or 0.05-5% aqueous poloxamer solution 407. 10. The pharmaceutical composition according to claim 9, characterized in that the pharmaceutically acceptable carrier further comprises ascorbic acid. 11. The pharmaceutical composition according to any one of claims 1 to 10, characterized in that the mycobacteriosis is caused by one or more pathogens selected from Muso bacillus lacticulosum, or Muscobacillus capvavi, or ^ um - ^ ηΐ ^ е 11 11 11 и 11 сот р ех ех, или или или или или или или или или или или или или или или или или или или или или или или или или или или или или или М ус ус ус 5 5 5 5 5. 12. The pharmaceutical composition according to claim 11, characterized in that the mycobacteriosis is HIV / AIDS-associated mycobacteriosis. 13. The pharmaceutical composition according to p. 12, characterized in that the HIV / AIDS-associated mycobacteriosis is due to Musobacillitis amphibylacterium tuberculosis. 14. The pharmaceutical composition according to claim 12, wherein the HIV / AIDS-associated mycobacteriosis is due to Musobacillus tuberculosis. 15. A method of obtaining a pharmaceutical composition according to any one of claims 1 to 14, characterized in that: a) a sample of cyclodextrin or its pharmaceutically acceptable derivative (s) is dissolved in a solvent with vigorous stirring; b) add to the resulting solution a portion of rifabutin; c) mix; g) homogenize; d) add the resulting dispersion to an aqueous solution of a surfactant with vigorous stirring; e) homogenize; g) remove the solvent; h) centrifuged; i) dried. 16. The method according to clause 15, wherein in step c), the reaction mixture is further heated and / or sonicated by ultrasound and / or microwave treatment of the reaction mixture. 17. The method according to p. 15, characterized in that as cyclodextrin or its pharmaceutically acceptable (acceptable) derivative (s) using α-cyclodextrin, β 18 016410 cyclodextrin, 2-hydroxypropyl-b-cyclodextrin, sulfobutyl-b-cyclodextrin, 6-monodeoxy-6-monoamine-b-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, or mixtures thereof. 18. The method according to clause 15, wherein the solvent used is acetone, or ethanol, or dimethyl sulfoxide, or Ν-methylpyrrolidone. 19. The method according to clause 15, wherein the solvent in step g) is removed by distillation on a rotary evaporator (under reduced pressure). 20. The method according to clause 15, wherein the homogenization is carried out using ultrasonic treatment or using a high pressure homogenizer. 21. The method according to clause 15, characterized in that after centrifugation (step h)) add a cryoprotectant. 22. The method according to item 21, wherein the cryoprotectant is selected from 1-50% glucose, 1-50% lactose, 1-50% mannitol or 1-50% trehalose. 23. The method according to clause 15, wherein the aqueous solution of a surfactant has a concentration of 0.05-5%. 24. The method according to p. 15, characterized in that the surfactant is selected from the group comprising poloxamer-101, poloxamer-105, poloxamer-108, poloxamer-122, poloxamer-123, poloxamer-124, poloxamer-181, poloxamer -182, Poloxamer-183, Poloxamer-184, Poloxamer-185, Poloxamer-188, Poloxamer-212, Poloxamer-215, Poloxamer-217, Poloxamer-231, Poloxamer-238, Poloxamer-282, Poloxamer-288, Poloxamer-331 , poloxamer-401, poloxamer-407, sorbitan monostearate (δΡΑΝ 60), sorbitan tristearate (δΡΑΝ 65), sorbitan monolaurate (δΡΑΝ 20), sorbitan monooleate (δΡΑΝ 80), sorbitan monopal mitate (δΡΑΝ 40), sorbitan trioleate (δΡΑΝ 85) or mixtures thereof. 25. The method according to clause 15, wherein the centrifugation is carried out at a speed of 10005000 rpm 26. The method according to clause 15, wherein the drying in step i) is carried out using lyophilic or spray drying. 27. The method according to clause 15, wherein the product obtained in step i) is mixed with excipients and other excipients and the resulting mixture is used to make a solid oral dosage form. 28. A method of treating mycobacteriosis, characterized in that a therapeutically effective amount of a pharmaceutical composition according to any one of claims 1 to 2, 614 is administered intravenously and / or intramuscularly in a pharmaceutically acceptable carrier to a patient in need thereof. 29. The method according to p. 28, wherein the pharmaceutically acceptable carrier is a 0.9% aqueous solution of sodium chloride, or 0.01-0.1% aqueous solution of polysorbate-80, or 0.05-5% aqueous solution poloxamer-188, or 0.05-5% aqueous solution of poloxamer-407. 30. The method according to clause 29, wherein the pharmaceutically acceptable carrier further comprises ascorbic acid. 31. The method according to claim 28, characterized in that the MAI caused by one or more pathogens selected from MueoaeYupit 1iegsi1o818 or MueoaeYupit Βονίδ, or MueoaeYupit kapzazi or MueoaeYupit zegoShaeeit or MueoaeYupit au1it-1p1gaee11i1age eotr1eh or MueoaeYupit 1ergae or MueoaeYupit raga1iegei1o818 . 32. The method according to p. 28, wherein the mycobacteriosis is HIV / AIDS-associated mycobacteriosis. 33. The method according to p, characterized in that the HIV / AIDS-associated mycobacteriosis is due to MueoLaeJupit and Leohei1o818. 34. The method according to p. 32, characterized in that the HIV / AIDS-associated mycobacteriosis is due to Mueo-ea-ц ц пп пп 1 1 ит ит-ее ее 11 11 е е е е е е е е ех е. 35. A method of treating mycobacteriosis, characterized in that a therapeutically effective amount of a pharmaceutical composition according to any one of claims 1, 3-14 is orally administered in powder form or in the form of a solid oral dosage form to a patient in need thereof. 36. The method according to clause 35, wherein the mycobacteriosis is caused by one or more pathogens selected from MueoLaeupitus and Egeeio818, or MueoBaeiupitus L0 \ '15. either MueoAeIUpit kapzazi, or MueoAeIeUpit zaShaeyeit, or MueoAeIyupit kapzazi, or MueoAeIUpit kapzazi, or MueoAeIUpit kapzazi, or MueoAeIUpit kapzazi, or MueoAeIupit kapzazi, or MueoAeIupit kapzazi. 37. The method according to any one of claims 35, 36, wherein the mycobacteriosis is HIV / AIDS-associated mycobacteriosis. 38. The method according to clause 37, wherein the HIV / AIDS-associated mycobacteriosis is due to Mueo-ejupit 1 and egei1o518. 39. The method according to clause 37, wherein the HIV / AIDS-associated mycobacteriosis is caused by MueoAe1e1a1aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa. 40. The method according to clause 35, wherein the solid oral dosage forms are tablets, capsules or dragees. - 19 016410 41. A method of treating Helicobacter pylori infection, characterized in that therapeutically effective amount of the pharmaceutical composition according to any one of claims 1 to 3, 6-14 is administered intravenously and / or intramuscularly in a pharmaceutically acceptable carrier to a patient in need thereof. 42. The method according to paragraph 41, wherein the pharmaceutically acceptable carrier is a 0.9% aqueous solution of sodium chloride, or 0.01-0.1% aqueous solution of polysorbate-80, or 0.05-5% aqueous solution poloxamer-188, or 0.05-5% aqueous solution of poloxamer-407. 43. The method according to § 42, wherein the pharmaceutically acceptable carrier further comprises ascorbic acid. 44. A method of treating helicobacter infection, characterized in that a therapeutically effective amount of a pharmaceutical composition according to any one of claims 1, 3-14 is orally administered in powder form or in the form of a solid oral dosage form to a patient in need thereof. 45. The method according to item 44, wherein the solid oral dosage forms are tablets, capsules or dragees. 46. The method according to any of paragraphs 44, 45, characterized in that the patient is additionally administered a therapeutically effective amount of one or more drugs selected from amoxicillin, pantoprazole, esomeprazole, rabeprazole, omeprazole, metronidazole, clarithromycin, bismuth citrate or tetracycline.