RU2778521C2 - Drug for treatment of bacterial infections - Google Patents

Abstract

FIELD: pharmaceutical industry.

SUBSTANCE: invention relates to the pharmaceutical industry, namely to a drug for the treatment of bacterial infections. The drug is proposed for the treatment of bacterial infections caused by bacteria selected from Mycobacterium tuberculosis, Streptococcus pyogenes, Salmonella enteritidis, Escherichia coli, Klebsiella pneumoniae, Chlamydophila pneumonia, as well as Klebsiella pneumoniae bacteria resistant to amoxicillin and clavulanic acid, Salmonella enterica bacteria resistant to streptomycin and nalidixic acid. The drug is a mixture including, in the effective amount in the form of decimal and centesimal dilutions, in the ratio of 1:1, 1:2, or 2:3, a product of technological treatment by a method for sequential multiple dilutions of initial substances of antibodies to β1 – a domain of a molecule of the main complex of histocompatibility of class 2 (HLA-DRB1), and a product of technological treatment by a method for sequential multiple dilutions of initial substance of antibodies to β2-microglobulin (β2-MG). In this case, the product of technological treatment by the method for sequential multiple dilutions of initial substance of antibodies is an aqueous or aqueous-alcohol solution obtained by multiple sequential dilution of a matrix initial solution of antibodies in combination with external mechanical impact – multiple shaking of each dilution, while a concentration of antibodies of 0.5÷5.0 mg/ml is used as the matrix solution.

EFFECT: invention provides for effective treatment of bacterial infections, reduction in antibiotic resistance, increase in the efficiency of therapeutic effect of antibiotics, including relatively to resistant bacteria.

9 cl, 2 tbl, 4 ex, 5 dwg

Description

The present invention relates to the field of pharmaceuticals, namely to agents for treating bacterial infections, increasing the effectiveness of the therapeutic action of antibiotics, including against resistant bacteria and reducing the therapeutic dose of antibiotics, as well as to a method for treating bacterial infections, a method for reducing the therapeutic dose of an antibiotic and a method increasing the therapeutic efficacy of antibiotics against bacteria resistant to them.

Bacterial infections are diseases caused by bacteria. A feature of bacterial infections is that in the process of life and after the death of bacteria, toxins are released that cause inflammation, intoxication and tissue damage. When diagnosing a bacterial infection, treatment with antibacterial drugs is prescribed. Bacteria that cause certain diseases sooner or later develop resistance (resistance) to antibiotics used in the course of treatment. This is a natural adaptation process called antimicrobial resistance. The emergence of a large number of resistant bacteria leads to an increasing number of incurable infections, resulting in increasing risks of bacterial infections, increasing medical costs for the treatment of bacterial infections, longer hospitalizations and increased mortality [WHO. Growing threat of antimicrobial resistance: possible measures. 2013].

In May 2015, the World Health Assembly approved the Global Action Plan on Antimicrobial Resistance, which includes antibiotic resistance. The Global Action Plan aims to ensure the prevention and treatment of infectious diseases with safe and effective medicines [http://www.who.int/mediacentre/factsheets/antibiotic-resistance/ru/].

Thus, the treatment of bacterial infections, as well as reducing the resistance of bacteria to antibiotics and increasing the sensitivity of bacteria to antibiotics, is a serious problem and an urgent task in the field of medicine.

From the prior art, a drug is known that contains an activated form of ultra-low doses of monoclonal, polyclonal immune or natural antibodies to the antigen of the major histocompatibility complex (HLA system) or to the complex of the antigen of the HLA system and its associated peptide [patent RU 2205025C1, 26.12.2001]. The immunotropic drug obtained in accordance with the invention is a new pharmacological preparation, which is characterized by the presence of immunotropic activity, the absence of side effects, environmental friendliness and low cost.

In the prior art, drugs for the treatment of infectious diseases are known, in particular drugs for the treatment of bacterial infections, for example, Amoxicillin, Gentamicin, Tetracycline, Levomecitin and others. Antibiotic class drugs have two main problems: an increasing number of microorganisms resistant to antibiotics, and a wide range of contraindications and side effects when using antibiotics [Belozertseva V.N. etc. Antibiotics - resistance and toxicity. New St. Petersburg Medical Gazette, 2017. - N 2. - S. 59-61]. So, for example, the drug "Amoxicillin" can cause allergic reactions (rhinitis, dermatitis, itching, etc.), nausea, diarrhea, convulsive reactions, tachycardia, joint pain, and more [https://www.rlsnet.ru/tn_iridex_id_222. htm]. When choosing the drug "Tetracycline", it is imperative to take an antibiotic sensitivity test, since over the years of using the drug, many bacteria have acquired resistance to it, in addition, when administered orally, the antibiotic can cause severe side reactions, it is considered toxic, therefore it is contraindicated for children under 8 years and pregnant women [https://www.rlsnet.ru/mnn_index_id_82.html. The drug "Gentamicin" also has a high toxicity, can cause epileptic seizures, headache, hearing loss, anemia, fever, development of superinfection and other symptoms, therefore, it is prescribed with caution, only after the final confirmation of the diagnosis [https://www. rlsnet.ru/tn_index_id_876.htm]. Modern studies have shown the danger of the drug "Levomecithin", in particular, the substance can inhibit hematopoiesis, causing aplastic anemia - a decrease or cessation of the production of blood components by the bone marrow [https://medaboutme.ru/zdorove/spravochnik/slovar-medicinskih-terminov/antibiotiki/ ; https://www.rlsnet.ru/tn_index_id_4699.htm].

Thus, incorrect and uncontrolled use of antibiotics can seriously harm health. As mentioned earlier, antibiotic treatment can be accompanied not only by adverse side effects, such as allergies, toxic complications from the liver and kidneys, disorders of the cardiovascular system and blood, nervous system, but also lead to the formation of antibiotic-resistant microbes. At the moment, the Government of the Russian Federation has approved the Strategy for preventing the spread of antimicrobial resistance in the Russian Federation for the period up to 2030. The goal of the strategy is to prevent and limit the spread of antimicrobial resistance [GOVERNMENT OF THE RUSSIAN FEDERATION, Decree of September 25, 2017 “2045-r Moscow, Strategy for preventing the spread of antimicrobial resistance in the Russian Federation for the period up to 2030]. Thus, there is a widespread, comprehensive work to reduce the level of resistance of microorganisms to antibiotics and the rational consumption of antibiotics.

A known method of overcoming the drug resistance of bacteria, including the use of antibiotics in combination with inhibitors of bacterial enzymes. For example, clavulanic acid (or clavulanate) is a beta-lactamase inhibitor whose chemical structure resembles beta-lactam antibiotics. Like other beta-lactams, clavulanic acid is able to bind to penicillin-binding proteins (PBP) of gram-positive and gram-negative bacteria and promote bacterial wall lysis. In addition, clavulanic acid has its own antibacterial activity. On this basis, combined preparations containing an aminopenicillin antibiotic and one of the beta-lactamase inhibitors (amoxicillin/clavulanate) have been created. The disadvantage of this method is the limited range of antagonistic action of the combinations of drugs used (active only against microbes whose resistance is due to the production of beta-lactamase). Also, intravenous clavulanic acid preparations are used with caution in patients with severely impaired liver function; if urticaria or an erythematous rash appears, the medication should be stopped. Clavulanic acid is contraindicated in case of individual intolerance. During pregnancy, the use of this drug with "Amoxicillin" or "Ticarcillin" is allowed only for health reasons. During lactation, the use of the drug is not recommended. Side effects of clavulanic acid include: dyspepsia, cholestatic jaundice, impaired liver function, hepatitis, pseudomembranous colitis, candidiasis, allergic reactions (erythema multiforme, Quincke's edema, exfoliative dermatitis, urticaria, anaphylactic shock) [https://www.kakprosto. ru/kak-899440-klavulanovaya-kislota-deystvie-i-svoystva-#ixzz5eejsFIpF].

In the prior art, a method is known to overcome the drug resistance of bacteria and fungi, which makes it possible to increase the efficiency

overcoming the drug resistance of microorganisms to antimicrobial drugs with different mechanisms of action, due to the combined action of low concentrations of aniline dyes and antimicrobial drugs [patent RU 2363470 C1, 10.08.2009]. It should be taken into account that aniline dyes are widely used in histological technique, they have a bactericidal, and some of them have a carcinogenic effect.

The prior art pharmaceutical combined composition for external and local use containing an active complex of bacteriolytic and proteolytic enzymes, for example, lysoamidase, a base, and at least one target additive selected from the series: an antibiotic, a synthetic antibacterial agent, including a substance that reduces the resistance of microorganisms, an antimycotic, an anesthetic, a stimulator of reparative processes. The invention provides an increase in the therapeutic activity of the pharmaceutical composition compared to known means, including strains resistant to individual components of the composition, improvement of the course of the wound process, rapid wound cleansing, reduction of convalescence time [patent RU 2655808 C2, 05/29/2018]. It should be taken into account that enzymes are sensitive to various external factors (pH, temperature, pressure, etc.) and any external influence can lead to denaturation and irreversible changes in the structure of the enzyme with the loss of its properties. For example, the enzyme lysoamidase is recommended to be stored in a dry, dark place at a temperature not exceeding +10°C, which causes additional difficulties in its production, transportation and storage, and at any of these stages may lead to a decrease in the effectiveness of the drug.

The present invention is directed to the creation of a new effective complex drug that has antibacterial, immunomodulatory activity and is also effective

against bacteria that are resistant to one or more known antibacterial agents (antibiotics), while not having a toxic or mutagenic effect on the patient's body, the invention is also aimed at creating an agent that increases the effectiveness of the pharmacological action of an antibiotic, including in relation to those resistant to it bacteria, and reducing the therapeutic dose of the antibiotic.

The solution to this problem is ensured by the fact that the claimed drug for the treatment of bacterial infections, which increases the effectiveness of the therapeutic action of antibiotics, reduces the resistance of bacteria to antibiotic therapy, and helps to reduce the therapeutic dose of antibiotics, is a product of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to β1 - the domain of the molecule major histocompatibility complex class 2 (HLA-DRB1) and β2-microglobulin (β2-MG).

The drug of the present invention, which is a product of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to the β1 - domain of the molecule of the major histocompatibility complex class 2 (HLA-DRB1) and β2-microglobulin (β2-MG), is effective for the prevention and treatment of bacterial infections, caused by different groups of bacteria. While these groups of bacteria are, for example: spirochetes (eg Treponema, Borrelia, Leptospira); aerobic and microaerophilic, motile, spiral and curved Gram-negative bacteria (eg Campylobacter, Helicobacter, Spirillum); gram-negative aerobic and microaerophilic rods and cocci (eg Achromobacter, Bordetella, Kingella, Neisseria); facultative anaerobic Gram-negative rods (for example,

Cedecea, Escherihia, Klebsiella, Plesiomona, Haemophilus, Streptobacillus (coli infections)); Gram-negative anaerobic straight, curved and spiral bacteria (eg Anaerobiospirrilum, Bacteroides, Porphyromonas); anaerobic Gram-negative cocci (eg Veillonella); rickettsia and chlamydia (eg Ehrlichia, Chlamydophila); Gram-positive cocci (eg Aerococcus, Staphylococcus, Streptococcus); gram-positive rods and cocci that form endospores (for example, Bacillus, Clostridium); regular non-spore-forming Gram-positive rods (eg Erysipelothrix, Listeria); irregularly shaped non-spore-forming Gram-positive rods (eg Bifidobacterium, Corinebacterium, Rothia); mycobacteria (eg Mycobacterium); actinomycetes (eg Actinomadura, Nocardia, Streptomyces); mycoplasmas (for example, Mycoplasma, Ureaplasma) and others [I.V. Smirnov. Causative agents of human bacterial infections. Clinical Microbiology and Antimicrobial Chemotherapy, No. 2, Volume 2, 2000, p. 4-11].

The claimed drug is also effective against bacteria that are resistant to one or more antibiotics.

Bacterial infections, for which the claimed agent is effective, can be systematized not only by the type of bacterium causing the disease, but also by the mechanism of transmission:

Bacterial infections of the digestive system / intestinal bacterial infections, can be caused by shigella, staphylococci, vibrio cholerae, typhoid bacillus, salmonellosis - mainly fecal-oral transmission (salmonellosis, typhoid fever, dysentery, food poisoning, campylobacteriosis, dysentery, enteritis, eshirichiosis, campylobacteriosis , colitis, food poisoning, gastritis, peptic ulcers, botulism, dysentery).

Bacterial infections of the respiratory tract - aspiration route of transmission, can be caused by staphylococci, pneumococci, streptococci, whooping cough, meningococci, chlamydia, mycobacteria, mycoplasmas and others (sinusitis, rhinitis, tonsillitis, laryngitis, tracheitis, tonsillitis, epiglottitis, sinusitis, pneumonia, bronchitis, tuberculosis, etc.).

Bacterial infections of the genitourinary system - bacterial vaginosis (gardnerellosis), chlamydia, cystitis, dysbacteriosis, pyelonephritis, glomerulonephritis, urethritis, bacterial balanitis, prostatitis, bacterial cystitis, pyelonephritis, urethritis, bacterial prostatitis, vesiculitis, etc.

Bacterial infections of the skin and soft tissues, caused, for example, by staphylococci, streptococci - a contact route of transmission (erysipelas, impetigo, cellulitis, phlegmon, furunculosis, hidradenitis, etc.).

Blood bacterial infections caused, for example, by rickettsia, yersinia, coccobacteria - a transmissible transmission mechanism (tularemia, plague, typhus, trench fever, etc.).

Infections of the lymphatic system, for example, lymphadenitis, lymphangitis can be caused by streptococcus, staphylococcus aureus.

Infection of the nervous system, for example, meningitis, encephalitis, myelitis and others caused by meningococci, pneumococci, Haemophilus influenzae type b, Borrelia, mycoplasmas, fungi of the genus Candida, etc. can be transmitted by airborne droplets, and there may also be a lymphogenous transmission route.

Infection of bones and joints can be caused by Lyme disease bacteria, staphylococci, streptococci, mycobacteria, gonococcal bacteria - the mechanism of transmission is through the bloodstream, through direct penetration, for example, wounds, surgical interventions, etc., through infections in nearby structures (infectious arthritis, osteomyelitis, Reiter's disease, etc.).

Ear infection, infection of the mastoid process and sinuses, can be caused by streptococci and staphylococci, fungi of the genus Candida, Haemophilus influenzae, etc. - the infection is transmitted through the Eustachian tube, from the external auditory canal, or through the blood (labyrinthitis, sensorineural hearing loss, mastoiditis and other).

The pharmaceutical combination of the present invention is a set of active ingredients for the treatment of bacterial infections, that is, a drug that is a product of technological processing by the method of sequential multiple dilutions of the original substances of antibodies to HLA-DRB1 and antibodies to β2-MG and the corresponding antibiotic, while both are administered to the patient by co-administration. The co-administration of a medicinal product and an appropriate antibiotic can be carried out by means of simultaneous administration of drugs independently of each other (in separate dosage forms) at the same time or sequentially within time intervals, especially when such time intervals allow drugs in combination to show an improved effect. The claimed drug is used in an effective dose, while the antibiotic should be taken according to the instructions or as prescribed by the doctor. In this case, the specified antibiotic can be selected from the group, for example: beta-lactam (penicillins (amoxicillin, methicillin, oxacillin, benzylpenicillin, etc.); cephalosporins (cephalexin, cefazolin, ceftriaxone, cefepime, etc.); carbapenems (meropenem, doripenem, biapenem, etc.)); macrolides (ketolides, erythromycin, roxithromycin, clarithromycin, etc.); tetracyclines (tetracycline, oxytetracycline, doxycycline, metacycline, etc.); aminoglycosides (streptomycin, kanamycin, amikacin, gentamicin, etc.); chloramphenicol; oxazolidinones; glycopeptides (targocid, daptomycin, vancomycin, etc.); lincosamides (delacin C); fluoroquinolones (levofloxacin, gemifloxacin, sparfloxacin, etc.); antifungal

antibiotics (nystatin, amphotericin B, etc.); anti-tuberculosis antibiotics (ftivazide, metazid, saluzide, etc.); antileprosy drugs (solusulfon, diucifon, etc.); anticancer drugs (doxorubicin, rubomycin, carminomycin, etc.); various antibiotics (fosfomycin, fusidin, rifampicin, etc.) [https://ru.wikipedia.org/wiki/Antibiotics].

Also, the present invention is aimed at increasing the effectiveness of the pharmacological action of an antibiotic, including against bacteria resistant to it, by the simultaneous or sequential administration of the corresponding antibiotic and the claimed medicinal product, which is a product of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to HLA-DRB1 and β2-MG. Thus, with the additional introduction of the claimed medicinal product while taking antibiotics, the therapeutic dose of the antibiotic decreases.

The technical result of the present invention consists in expanding the arsenal of medicinal antibacterial agents, creating a broad-spectrum antibacterial agent that is effective against, among other things, bacteria resistant to one or more antibiotics, has low toxicity and mutagenicity, which makes it possible to reduce the burden on the patient's body during antibacterial therapy; also, the claimed agent increases the effectiveness of the pharmacological action of the antibiotic, including against resistant bacteria, and reduces the therapeutic dose of the antibiotic, which leads to a decrease in side effects during their administration and after a course of antibiotics.

The products of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to the β1 - domain of the molecule of the major histocompatibility complex class 2 (HLA-DRB1) and β2-microglobulin (β2-MG) are aqueous or aqueous

alcohol dilutions of antibodies (or a combination of such dilutions) obtained by sequential multiple dilutions (potentiation) of the matrix stock solution of antibodies in combination with external influence - shaking each dilution.

In previous works, for example, in patent RU 2205025 C1, December 26, 2001, the applicant used the terms “activated-potentiated form of antibodies”, “ultra-low doses of antibodies” to describe the products of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies. At the same time, the applicant draws attention to the fact that the term "product of technological processing by the method of successive multiple dilutions of the initial substances of antibodies" and the term "activated - potentiated form of antibodies", "ultra-low doses of antibodies", "ultra-high dilutions of antibodies", "potentiated antibodies", " release-active antibodies” are completely interchangeable [Epshtein O.I. Release activity (modern view of homeopathy and non-homeopathy). M.: RAMN Publishing House, 2017. 48 p. // Epstein O.I. Ultra-low doses (history of one study). M.: Publishing house of the Russian Academy of Medical Sciences, 2008. 336 p.].

In this case, the dilution of antibodies to HLA-DRB1 and antibodies to β2-MG should be understood as any, including decimal, hundredths, thousandths, dilutions of the matrix solution of antibodies, starting from C2 (i.e., from a dilution of the matrix solution by 100 ² times), as well as any combinations thereof (for example, C2+D34+M45, D20+C4, etc.) and ratios (for example, 1:1:2, 2:3, etc.).

For the preparation of the claimed medicinal product, natural, monoclonal or polyclonal antibodies are used as the initial substance of antibodies, which can be obtained using known technologies - methods described, for example, in: Myagkova M.A., Morozova B.C. Immunochemical properties of natural antibodies to physiologically active compounds, Basic Research No. 11, 2014, p. 1066-1070 and Immunological methods, under

ed. G. Frimelya, M., "Medicine", 1987, p. 9-33; or, for example, in the article Laffly E., Sodoyer R. Hum. Antibodies. Monoclonal and recombinant antibodies, 30 years after. - 2005 - Vol.14. - N 1-2. P.33-55 and specific for antigens - HLA-DRB1 and β2-MG.

Natural antibodies are immunoglobulins that are produced by the body in strictly defined quantities in the complete absence of any external antigenic stimulation, so these antibodies, as a rule, circulate in the bloodstream even of healthy people. Natural autoantibodies are usually polyreactive and have a fairly low affinity for a specific set of autoantigens.

Monoclonal antibodies are heterogeneous antibodies to a specific antigen epitope derived from a single clone of antibody-producing B cells [Lipman NS1, Jackson LR, Trudel LJ, Weis-Garcia F. Monoclonal versus polyclonal antibodies: distinguishing characteristics, applications, and information resources. ILAR J. 2005; 46(3):258-68. // IHC Staining Methods. Fifth edition. Preface Chapter 1. Thomas Boenisch. Antibodies. p:1-9]. Monoclonal antibodies are obtained, for example, using hybridoma technology. Moreover, the initial stage of the process includes immunization based on the principles already developed in the preparation of polyclonal antisera. Further stages of the work involve obtaining hybrid cells producing clones of antibodies with the same specificity. Their isolation in an individual form is carried out by the same methods as in the case of polyclonal antisera.

Polyclonal antibodies are a collection of immunoglobulins that react with different epitopes of a specific antigen and are secreted by B cells of various body lines [Lipman NS1, Jackson LR, Trudel LJ, Weis-Garcia F. Monoclonal versus polyclonal antibodies: distinguishing characteristics, applications, and information resources. ILAR J. 2005; 46(3):258-68. // IHC Staining Methods. Fifth edition.

Preface Chapter 1. Thomas Boenisch. Antibodies. p:1-9]. Polyclonal antibodies can be obtained by active immunization of animals. To do this, according to a specially developed scheme, a series of injections of the antigen substance required in accordance with the invention is given to animals. As a result of this procedure, monospecific antisera with a high content of antibodies are obtained, which are used in the technological processing of the initial substances by the method of sequential multiple dilutions to obtain the final product. If necessary, the antibodies present in the antiserum are purified, for example, by affinity chromatography, by the use of fractionation by salt precipitation or by ion exchange chromatography.

According to the above, the term "antibodies" means immunoglobulins of any origin (natural, polyclonal or monoclonal antibodies) that specifically bind to a target molecule (antigen). At the same time, the ability of antibodies to recognize a certain type of epitopes of an antigen molecule and interact with it - specificity - is a key property of antibodies that determines the full range of effects of a particular antibody [Boyd WC. Fundamentals of immunology. fundam. Immunol. 1946; Langman RE. The specificity of immunological reactions. Mol. Immunol. 2000; 37: 555-561]. The specificity does not depend on the nature of origin and the method of obtaining the latter: natural, polyclonal or monoclonal antibodies with the same specificity will affect the same target [A. Roit et al., Immunology. Per. from English. - M.: Mir, 2000, p. 149-161, 527-544], and therefore can be used to obtain the claimed funds.

An option for the preparation of the claimed medicinal product is the use of polyclonal antibodies, which, as a matrix (primary) solution with a concentration of 0.5÷5.0 mg/ml,

used for technological processing by the method of successive multiple dilutions to obtain the final product.

Technological processing by the method of successive multiple dilutions of the initial substances of antibodies is a uniform decrease in concentration as a result of serial dilution of 1 part of the said matrix solution in 9 parts (for decimal dilution D) or in 99 parts (for centesimal dilution C) or in 999 parts (for thousandth dilution M) a neutral solvent in combination with an external influence (for example, in the form of shaking) for each dilution obtained and the use of separate containers for each subsequent dilution [B. Shvabe "Homeopathic medicines", M., 1967, p. 14-29].

It was found that the outwardly simple procedure of successive multiple reduction in the concentration of substances is a complex technology, the products of which acquire unique properties. Historically, potentization technology products have been referred to as "small doses", "potentiated preparations", "high dilutions".

The final product can be presented in various dosage forms [OFS.1.6.2.001.18 State Pharmacopoeia of the Russian Federation, Edition XIV] and contain pharmaceutically acceptable additives.

So, for example, the claimed medicinal product can be made in a solid dosage form and contain a technologically necessary (effective) amount of a pharmaceutically acceptable additive, which is a neutral carrier (for example, lactose), saturated with a mixture of technological processing products by the method of sequential multiple dilutions of the initial substances of antibodies to HLA -DRB1 and β2-MG, and pharmaceutically acceptable excipients (excipients), which include, for example,

hypromellose, maltitol, glycerol, potassium sorbate, anhydrous limanic acid, isomalt, silicon dioxide, sodium cyclamate, sodium saccharin, microcrystalline cellulose, magnesium stearate and others.

Pilotlab» производства компании

GmbH) производят орошение до насыщения вводимых в псевдоожиженный - кипящий слой гранул нейтрального носителя - лактозы (молочного сахара), предварительно полученным водным или водно-спиртовым раствором (концентрация подбирается экспериментальным путем,To obtain a solid oral form of the claimed medicinal product in a fluidized bed installation (for example, the “

Pilotlab" produced by the company

GmbH) produce irrigation until saturation of the granules of a neutral carrier - lactose (milk sugar) introduced into the fluidized - fluidized bed, with a previously obtained aqueous or aqueous-alcoholic solution (the concentration is selected experimentally,

OFS.1.6.2.0010.18 State Pharmacopoeia) of products of technological processing by the method of successive multiple dilutions of the initial substances of antibodies with simultaneous drying in a stream of heated air supplied under the grate at a temperature not exceeding 40°C. The resulting tablet mass is uniformly mixed and tableted by direct dry pressing (for example, in a tablet press Korsch - XL 400) [WO 2007105981(A1), 20.09.2007]. After tableting, tablets weighing 300 mg are obtained, impregnated with an aqueous or aqueous-alcoholic solution of technological processing products by the method of sequential multiple dilutions of the initial substances of antibodies to HLA-DRB1 and β2-MG.

Also, the claimed drug, together with a known antibiotic, can be a fixed or non-fixed pharmaceutical combination used to treat bacterial infections. The term "fixed combination" means that the active ingredients, the claimed drug and the combination partner - an antibiotic, are both administered to the patient simultaneously in a single dosage form or dosage. The terms "non-fixed combination" or "kit" mean that the active ingredients, the claimed medicinal product and the partner in

combinations - an antibiotic, both administered to the patient as separate elements or simultaneously or sequentially without a specific time frame, where such administration provides therapeutically effective levels of the two active ingredients in the patient's body. The claimed combination of drug and antibiotic can be used for simultaneous or sequential use for the treatment of bacterial infections. In this case, the antibiotic is administered in a known therapeutic dose.

The present invention is illustrated by the following examples, together with the accompanying drawings:

Fig. 1. Bladder weight 7 days after infection.

Fig. 2. Bacterial density in bladder tissue in mice 7 days after infection.

Fig. 3. Kidney weight at 7 days after infection.

Fig. 4. Bacterial density in kidney tissue in mice 7 days after infection.

Fig. 5. Bacterial density of urine 1, 3 and 6 days after infection.

Example 1

Anti-tuberculosis activity of the claimed drug against Mycobacterium tuberculosis caused in guinea pigs.

The purpose of the study was to study the anti-tuberculosis activity of the claimed drug against Mycobacterium tuberculosis in a model of experimental tuberculosis in guinea pigs in comparison with the anti-tuberculosis drug Isoniazid. The preparation of the present invention contains, in a ratio of 1:2 by volume, the product of technological processing by the method of successive multiple dilutions of the initial substance of antibodies to HLA-DRB1 (in aqueous-alcoholic

dilution of C12C150 in a ratio of 2:3, monoclonal antibodies to HLA-DRB1) and the product of technological processing by the method of sequential multiple dilutions of the initial substance of antibodies to β2-MG (in aqueous dilution of C12C150 in a ratio of 2:3, monoclonal antibodies to β2-MG). Experimental groups:

Group 1 - intact control, uninfected animals (n=7);

Group 2 - negative control (infected, untreated animals) (n=7);

Group 3 - animals treated with placebo (purified water) orally 2 times a day for 5 days before infection and 6 days a week for two months two weeks after infection in a volume of 4 ml/kg (n=14);

Group 4 - animals treated with placebo (purified water) orally 2 times a day for 5 days before infection and 6 days a week for two months two weeks after infection in a volume of 4 ml/kg and Isoniazid at a dose of 10 mg/kg in a volume of 0.4 ml 6 days a week for two months two weeks after infection (n=14);

Group 5 - animals treated orally 2 times a day for 5 days before infection and 6 days a week for two months two weeks after infection in a volume of 4 ml/kg (n=14);

Study design:

The duration of this study was (196 days, 6.5 months): September-December - observation of experimental animals and their treatment; January-March - study of the diagnostic material selected during necropsy by microbiological and histological methods. Clinical examination, assessment of appearance, general health, detection of mortality were performed daily before feeding the animals.

Study drugs were administered orally to guinea pigs using a syringe without a needle in a total volume of 8 ml/kg twice daily for

5 days before infection and six days a week (daily except Sundays) for 2 months after infection. The dose of drugs was adjusted once a week based on the results of weighing the animals. An aqueous starch suspension of isoniazid was administered to the animals at the same time and in the same manner as the test samples. The dose of isoniazid is 10 mg/kg of body weight in 0.4 ml of water-starch gruel.

During the study, in all groups of guinea pigs, the survival rate, weight, growth rate of M.tuberculosis in crops, histological changes in the organs of animals were evaluated.

Research results:

Conclusion:

An analysis of the data obtained during the study allows us to conclude that a positive therapeutic effect was equally noted in the group in the treatment of laboratory animals with Isoniazid and in the group in the treatment of the claimed Drug. Histological examination of the organs of animals of these two groups showed the presence of signs of therapeutic pathomorphosis. The efficacy of the claimed drug against lung infections has also been demonstrated.

Example 2

Antibacterial activity of the claimed drug against Streptococcus pyogenes in mice in combination with and without an antibiotic.

This study is a blind, placebo-controlled study. The purpose of the study was to evaluate the effectiveness of the claimed drug with linezolid (a synthetic antibiotic) or telithromycin (an antibiotic of the ketolide class) or without antibiotics against Streptococcus pyogenes in a mouse soft tissue neutropenic infection model. The preparation of the present invention contains, in a ratio of 2:3 by volume, the product of technological processing by the method of sequential multiple dilutions of the initial substance of antibodies to HLA-DRB1 (in a water-alcohol dilution of C4D20 in a ratio of 3:2; monoclonal antibodies) and the product of technological processing by the method of sequential multiple dilutions. dilutions of the initial substance of antibodies to β2-MG (in water-alcohol dilution of C10C150 in a ratio of 1:2; natural antibodies).

Experimental groups:

Group 1 - negative control, female mice were infected with S. pyogenes and did not receive drug or antibiotic administration during the study period; n=3.

Group 2 - female mice received placebo (glycine buffer) orally daily in a volume of 20 ml/kg 5 days before and on the day of infection with S. pyogenes; n=6.

Group 3 - female mice received placebo (glycine buffer) orally daily in a volume of 20 ml/kg 5 days before and on the day of infection with S. pyogenes; in combination with linezolid (50 mg/kg) on the day of infection 2 and 14 hours after infection; n=6.

Group 4 - female mice received placebo (glycine buffer) orally daily in a volume of 20 ml/kg 5 days before and on the day of infection with S. pyogenes; in combination with telithromycin (10 mg/kg) on the day of infection 2 and 14 hours after infection; n=6.

Group 5 - female mice received the drug orally daily in a volume of 20 ml/kg 5 days before and on the day of infection with S. pyogenes; n=6.

Group 6 - female mice received the drug orally daily in a volume of 20 ml/kg 5 days before and on the day of infection with S. pyogenes; in combination with linezolid (50 mg/kg) on the day of infection 2 and 14 hours after infection; n=6.

Group 7 - female mice received the drug orally daily in a volume of 20 ml/kg 5 days before and on the day of infection with S. pyogenes; in combination with telithromycin (10 mg/kg) on the day of infection 2 and 14 hours after infection; n=6.

Study Design:

The duration of the study was 7 days. Neutropenia in mice was induced by two intraperitoneal doses of cyclophosphamide (4 days prior to challenge, 150 mg/kg and 1 day, 100 mg/kg). Mice received no further immunosuppression throughout the study.

Experimental samples were administered within 5 days before infection, as well as on the day of infection (after 2 and 14 hours) orally at a dose of 20 ml/kg. On the day of infection, the mice were injected into the thigh muscle with a strain of S. pyogenes (ATCC BAA-2469) at a dose of 5.4 lg CFU. Also 2 and 14 hours after infection

the animals were injected with antibiotics - linezolid (50 mg/kg), to which this strain of S. pyogenes is sensitive, and telithromycin (10 mg/kg), to which the strain is resistant.

In the course of the study, survival, clinical manifestations of infection, and body weight were evaluated in all groups of mice.

Research results:

2 hours after challenge, untreated mice (Group 1) were harvested with thigh muscle to assess bacterial density (CFU/g, log ₁₀ ). In the remaining groups, the thigh muscle was taken to assess the bacterial density 24 hours after infection. The data obtained are presented in table 2.

Conclusion:

At 24 hours post-challenge, the test groups (Group 3 and 6), when administered in combination with linezolid, showed a statistically significant reduction in mean bacterial load. Thus, the claimed drug is effective against infections of the musculoskeletal system, and also in combination with antibiotics, increases their effectiveness.

Example 3

Efficacy of the claimed medicinal product alone and in combination with an antibiotic against E. coli in a model of urinary tract infection in mice.

The purpose of the study was to evaluate the effect of the test drug under conditions of experimental infection in mice caused by E. coli in combination with the antibiotic Norfloxacin and without it. The preparation of the present invention contains, in a ratio of 1:1 by volume, the products of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to HLA-DRB1 (in aqueous dilution of С12С30С50 in a ratio of 1:1:1; polyclonal antibodies) and antibodies to β2-MG (in aqueous dilution of C12C30C50 in a ratio of 1:1:1; monoclonal antibodies).

Experimental groups:

Group 1 - mice of both sexes (n=10) received the claimed drug (20 ml/kg) daily via oral tube for 5 days before infection and 7 days after infection and 1 hour after infection intraperitoneally (volume 10 ml/kg).

Group 2 - mice of both sexes (n=10) received the claimed drug (20 ml/kg) daily via oral gavage for 5 days before infection and 7 days after infection. Norfloxacin 5 mg/kg (volume 10 ml/kg) was intraperitoneally administered 1 hour after infection.

Group 3 - control, mice of both sexes (n=10) received glycine buffer (20 ml/kg) daily via oral gavage for 5 days before infection and 7 days after infection and 1 hour after infection intraperitoneally (volume 10 ml/kg ).

Group 4 - mice of both sexes (n=10) received glycine buffer (20 ml/kg) daily by oral gavage for 5 days before infection and 7 days after infection. Norfloxacin 5 mg/kg (volume 10 ml/kg) was intraperitoneally administered 1 hour after infection.

Study Design:

Mice were infected with E. coli UTI89 (2.107 cfu/mouse) by intraurethral bladder inoculation (day 0).

At 1, 3 and 6 days after infection, urine was collected for CFU determination. Seven days after challenge, 10 animals per group were sacrificed and kidneys and bladders were harvested for bacterial load analysis. Body weight was recorded every 2 days throughout the experiment.

Research results:

The dynamics of body weight in all groups was positive, which indicates a good tolerance of experimental samples by animals.

Bladder weight was statistically significantly lower in Group 2 (drug + norfloxacin) compared to Group 4 (global buffer + norfloxacin). This may indicate that the claimed drug, in combination with antibiotics, can reduce bladder edema.

The claimed drug in combination with norfloxacin (Group 2) statistically significantly reduced the bacterial load in the bladder. Thus, this combination may be promising as a therapeutic regimen (Fig. 1, 2).

There were no statistically significant differences between groups in kidney weight.

The claimed drug in combination with norfloxacin (Group 2) significantly reduced the bacterial load in the kidneys compared with glycine buffer (Group 3). Thus, this combination may be promising as a therapeutic regimen (FIGS. 3, 4).

Starting on day 3, the Group 2 sample (drug + norfloxacin) had a reduced bacterial density in the urine compared to Group 4 (global buffer + norfloxacin). Group 3 sample (main buffer) had higher

CFU in urine at all time points compared to the sample of Group 1 (preparation) (Fig. 5). Conclusion:

Thus, the claimed drug in combination with the antibiotic Norfloxacin exhibits antibacterial activity against E. coli in a model of urinary tract infection in mice.

Example 4 Toxicity

Toxicological studies were carried out in accordance with the principles of good laboratory practice (GOST 33044-2014) and the "Guidelines for conducting preclinical studies of drugs" [Mironov A.N., Bunatyan N.D. Guidelines for conducting preclinical studies of drugs. M. Grif and K. 2012; 944 p.].

Due to the fact that the usual parallels of pharmacokinetics and pharmacodynamics (dose-concentration-effect relationship), accepted for most drugs, to the claimed drug (product of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to HLA-DRB1 and β2-MG) are not applicable (the existing physicochemical and immunological methods do not allow recording drug concentrations in the biological media of the body) when selecting doses for toxicological studies, we were guided by physiologically acceptable volumes of the carrier (purified water in experimental studies) in which the drug is introduced.

Acute Toxicity:

The study was performed on non-linear mature white mice of both sexes (weight 22-26 g, age 2.5 months) and mature Wistar rats of both sexes (weight 225-250 g, age 4 months) with intragastric

administered. The study drug (n=18) or purified water (control, n=18) was administered twice with an interval of 2 hours in the maximum allowable volumes: intragastrically - 25 ml/kg to mice and 20 ml/kg to rats. The intact group did not receive any substances (n=18). After administration of the substances, the animals were observed for 2 weeks.

The data obtained were analyzed using multivariate analysis of variance (two-way ANOVA) with subsequent comparison of all groups with each other by Tukey's post-hoc test. Differences were considered statistically significant at p<0.05.

The introduction of the claimed preparation containing the product of technological processing by the method of successive multiple dilutions of the initial substances of antibodies to HLA-DRB1 (in aqueous dilution of C12C30C50 in a ratio of 2:1:3; polyclonal antibodies) and antibodies to β2-MG (in aqueous dilution of C12C30C50 in a ratio of 2: 1:3; polyclonal antibodies) intragastrically in the maximum allowable volumes to sexually mature mice and rats did not change the general condition of the animals (there were no signs of anxiety, changes in appetite, secretions, the state of mucous membranes, wool, skin, etc.) and did not affect the increase in the total weight of rats and mice (both females and males).

The absence of mortality in animals treated with the drug did not allow us to determine the LD50 index. Conventionally, the LD50 was taken as a dose exceeding the maximum amount of the drug administered to the animals.

Considering that the double intragastric administration of the claimed drug in the maximum allowable volumes did not have a toxic effect on the animal organism, it can be concluded that it is harmless and the reason is attributed to the class of low toxicity or to category 5 according to the GHS classification.

Repeated dose toxicity:

The study was carried out on 200 mature white conventional rats of both sexes (weight 180-242 g, age 3.5-4 months) and

24 mature rabbits of the Chinchilla breed of both sexes (weight 1.9 kg, age 2.5-3 months). Since it was not possible to determine the LD50 value for the drug, in the study of chronic toxicity in rats, the drug or purified water was administered intragastrically in the maximum allowable volume for intragastric administration and 1/4 of the maximum allowable volume. Rabbits received the drug in a volume close to the daily water intake.

The duration of administration was 6 months, the condition of rats and rabbits was assessed 3 and 6 months after the start of the drug administration, as well as after a month of withdrawal.

Chronic intragastric administration of the drug according to the present invention to adult rats of both sexes at doses exceeding more than 100 times the recommended daily doses for humans did not cause the death of animals within 6 months of administration, significant changes compared to the control group in behavioral activity, morphological and functional indicators of peripheral blood - after 3 and 6 months of administration, as well as 1 month after discontinuation of the drug

Chronic administration of the drug at the indicated doses was also not accompanied by significant changes in the macro- and micromorphological state and histological architectonics of the internal organs of rats.

In a study on rabbits, the entire period of the study also showed no death of animals. Chronic administration of the drug did not have a significant effect on the general condition, behavior, body weight of animals, as well as on indicators of peripheral blood and bone marrow hematopoiesis. According to the results of biochemical studies of blood plasma in rabbits, no significant changes were found. There were also no changes in the examination of urine in rabbits treated with the drug for 6 months. Macroscopic examination of the internal organs of rabbits treated with the drug for 6 months, pathological

no changes were found. Histological examination of organ tissues also showed no pathological changes.

Thus, on the basis of the conducted studies, it can be concluded that the claimed drug, when administered six months intragastrically to mature rats at a dose of 10 ml/kg and to rabbits at a dose of 50 ml/kg (more than 100 times higher than the average daily dose for humans), does not have a significant effect on the general condition and behavior of animals; does not cause pathological and morphofunctional changes in the cardiovascular, digestive and excretory systems.

Claims (9)

1. A drug for the treatment of bacterial infections caused by bacteria selected from Mycobacterium tuberculosis, Streptococcus pyogenes, Salmonella enteritidis, Escherichia coli, Klebsiella pneumoniae, Chlamydophila pneumonia, as well as Klebsiella pneumoniae bacteria resistant to amoxicillin and clavulanic acid, Salmonella enterica bacteria resistant to streptomycin or nalidixic acid, which is a mixture that includes in an effective amount in the form of decimal and hundredth dilutions in a ratio of 1:1, 1:2 or 2:3 the product of technological processing by the method of sequential multiple dilutions of the initial substances of antibodies to β1 - the domain of the molecule of the main complex class 2 histocompatibility (HLA-DRB1) and the product of technological processing by the method of sequential multiple dilutions of the initial substance of antibodies to β2-microglobulin (β2-MG), where the product of technological processing by the method of sequential multiple dilutions of the initial substance ounce of antibodies is an aqueous or water-alcohol solution obtained by repeated sequential dilution of the matrix stock solution of antibodies in combination with external mechanical action - repeated shaking of each dilution, while the concentration of antibodies is used as a matrix solution of 0.5 ÷ 5.0 mg / ml. 2. The drug according to claim 1, wherein the bacterial infection is tuberculosis. 3. The drug according to claim 1, wherein the bacterial infection is salmonellosis. 4. The medicament according to claim 1, wherein the bacterial infection is a coli infection. 5. The drug according to claim 1, wherein the bacterial infection is a streptococcal infection. 6. The drug according to claim 1, where the antibody is a monoclonal, polyclonal or natural antibody to HLA-DRB1. 7. The drug according to claim 1, where the antibody is a monoclonal, polyclonal or natural antibody to β2-microglobulin. 8. A medicament according to claim 1, further comprising pharmaceutically acceptable additives. 9. The drug according to claim 1, made in a solid dosage form and containing an effective amount of neutral carrier granules saturated with a mixture of technological processing products by the method of sequential multiple dilutions of the initial substances of antibodies to HLA-DRB1 and antibodies to β2-microglobulin, and pharmaceutically acceptable excipients .

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