Nasal administration composition for the prevention and / or treatment of viral and bacterial infections

Abstract

The present invention relates to the field of pharmaceuticals, i.e., pharmacology, and can be used for the prevention and / or treatment of nasopharyngeal and nasal cavity infections caused by viruses and bacteria, including coronavirus infection, influenza, and other acute respiratory viral infections. A nasal administration composition for the prevention and / or treatment of infectious and inflammatory diseases of the nasal cavity, comprising high molecular weight hyaluronic acid, high molecular weight polyvinylpyrrolidone (PVP), povialgol, antimicrobial polypeptide, sodium hypochlorite solution, and gerofcin, wherein these components are present in specified ratios.

Description

Detailed description of the invention

[0001] [Technical field] The present invention relates to the field of medicine, i.e., pharmacology, and can be used for the prevention and / or treatment of nasopharyngeal and nasal cavity infections caused by viruses and bacteria, including coronavirus infection, influenza, and other acute respiratory viral infections.

[0002] [Background technology] The increasing involvement of infectious viral diseases in common morbidity types within populations, along with the emergence of new and potentially dangerous viruses (including those not previously identified), is accelerating the development and implementation of promising long-acting, broad-spectrum antimicrobial compositions.

[0003] The nasal cavity acts as an antimicrobial filter, harboring a diverse flora of airborne microorganisms, including fungal spores, bacilli, micrococcuses, non-pathogenic Neisseria, Staphylococcus epidermidis, and β-hemolytic streptococci. In some cases, bacteria present in the nose include Streptococcus pneumoniae, Klebsiella pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and Candida. These species can not only cause self-infection, i.e., rhinitis, bronchitis, nasopharyngitis, and pneumonia, but can also be a source of infection for others. In children, the nasal microbiome is even more numerous and diverse.

[0004] The prototype of the solution described in the claims is an invention according to Russian Patent No. 2616523 (17 April 2017, IPC A61K9 / 06, A61K31 / 22, A61K36 / 61, A61K36 / 53, A61P31 / 04, A61P31 / 16), which is an ointment for intranasal administration for the treatment and prevention of respiratory infections. The invention is characterized by a sustained effect brought about by the synergistic action of the individual components of the drug, as described herein. The results of drug use have been shown in patients with influenza and acute respiratory viral infections.

[0005] The authors of this invention have not specified the necessary frequency and duration of use for the nasal ointment that would affect the realization of the treatment and prevention described in the claims.

[0006] Furthermore, the authors did not confirm the claimed antiviral effects resulting from the synergistic action of the individual components of the ointment through in vitro studies of those components. The authors also did not provide data on the efficacy of the nasal administration ointment against human coronaviruses, influenza viruses, and other RNA and DNA-containing viruses, nor did they claim the antibacterial effect of the ointment or its effect against microbial biofilms.

[0007] [Overview of the prefecture] The object of the present invention is to solve the problem of the lack of effective broad-spectrum compositions for the prevention and / or comprehensive treatment of infectious and inflammatory diseases of the nasal cavity when used intranasally.

[0008] The technical achievement of the present invention is the development of a broad-spectrum composition more effective than a prototype for the prevention and / or comprehensive treatment of infectious and inflammatory diseases of the nasal cavity when used intranasally, the broad-spectrum composition having sustained broad-spectrum antimicrobial activity, particularly against human coronaviruses, influenza viruses, other RNA and DNA-containing viruses, and bacteria, and can prevent the formation of microbial biofilms and disrupt already formed microbial biofilms.

[0009] The above problems are solved and the above specific technical results are achieved by developing a composition for application to the nasal cavity, wherein the composition contains an antimicrobial polypeptide which is high molecular weight hyaluronic acid, high molecular weight polyvinylpyrrolidone (PVP), gerofsin, sodium hypochlorite, povialgol, gramicidin C, or lysozyme, in the following weight % component ratios, i.e., High molecular weight hyaluronic acid -0.1~2.0 High molecular weight PVP-4.0~6.0, Povialgor -0.8~1.2, Antimicrobial polypeptides (Gramicidin C - 0.03~0.15, or Lysozyme - 0.1~18.0), Sodium hypochlorite solution with an activated chlorine content of 1566 mg / l - 49.0~51.0, Gerofsin up to -100.0 It is characterized by containing.

[0010] A synergistic effect is produced by the quantitative and qualitative combination of the constituent components in the proposed composition for patent purposes.

[0011] Another object of the present invention is the use of the composition described in the claims for the prevention and / or treatment of infectious and inflammatory diseases of the nasal cavity caused by coronaviruses, influenza viruses, other RNA and DNA-containing viruses, and bacteria, which involve the formation of microbial biofilms (particularly during periods of exacerbation of seasonal infectious diseases, epidemics and pandemics), the use characterized by the application of the composition to the nasal mucosa once daily for preventive or therapeutic purposes.

[0012] In the context of this invention, the term "high molecular weight hyaluronic acid" refers to non-sulfated glycosaminoglycans that are part of connective tissue, epithelial tissue, and nerve tissue. High molecular weight hyaluronic acid is one of the major components of the extracellular matrix found in many biological fluids (saliva, synovial fluid, etc.), plays an important role in cell proliferation and migration, and is produced by some bacteria (e.g., streptococci). The body of a person weighing 70 kg contains an average of about 15 grams of hyaluronic acid, of which one-third is converted (broken down or synthesized) every day.

[0013] One of the important functions of hyaluronic acid in connective tissue is water binding. As a result, intercellular material forms a jelly-like matrix that supports cells. Water binding and the resulting swelling determine the biological role of hyaluronic acid in regulating tissue permeability. Hyaluronic acid is a major structure-forming glycosaminoglycan because it has the ability to concentrate other glycosaminoglycans around itself and form proteoglycan aggregates that have higher hydrophilicity and elasticity compared to free proteoglycans. Hyaluronic acid creates a "buffer volume" by binding collagen fibers, other proteins and components of intercellular material, and even cells, into a single system. This "buffer volume" determines the strength and elasticity of mechanical tissues and helps them withstand temporary stresses (NNSigaeva, SVKolesov, PVNazarov, RRVLIDANOVA. Chemical modification of hyaluronic acid and application thereof in medicine / / Bulletin of the Bashkir University. 2012. Vol.17. No.3. pp.1220-1241).

[0014] In the context of this invention, the term "high molecular weight medical polyvinylpyrrolidone" (high molecular weight PVP) refers to a binder. High molecular weight PVP is a "pharmaceutically pure" pharmaceutical-grade powder with a characteristic odor and is available in the form of a loose white or yellowish-white powder with particles of varying sizes. The average molecular weight of PVP is 1,000,000 to 1,500,000 Da [VolkerBuhler. Collidon(R). Polyvinylpyrrolidone for the pharmaceutical industry / Translated from English, edited by Dr. KVAlekseev. BASF:2001.-310p.]. An aqueous solution of PVP is a viscous, clear, sterile, slightly colored liquid with a characteristic faint odor.

[0015] Although there is a wide range of materials for immobilizing pharmaceutical substances, biodegradable carriers based on high molecular weight PVP are of the greatest interest. Such biodegradable carriers allow antimicrobial substances to persist for a long time at the wound site, enabling the topical application of the biodegradable carriers for both the prevention of metal osteosynthesis and the treatment of surgical infections. In this case, the molecular weight of PVP is important. Modification of biologically active substances with high molecular weight polymers is performed to directly alter the properties of the substances, that is, complexation reduces toxicity and improves solubility, pharmacokinetics and bioavailability [Afinogenov G.E., Panarin E.F. Antimicrobial polymers, 1993. 《Hippocrates》Publishing House, St. Petersburg, 260p.]. The main function of soluble high molecular weight PVP is to sustain the action of drugs with improved bioavailability.

[0016] The authors found that high-molecular-weight PVP with adsorption and antitoxic properties as a polymer carrier provides a sustained effect of the active substance in the composition.

[0017] The term "poviargol" in the context of the present invention refers to highly dispersed metallic silver stabilized with low molecular weight medical polyvinylpyrrolidone, which is a pale greenish-gray to greenish-brown powder. In medical practice, poviargol is used as an external aqueous solution prepared immediately before use.

[0018] Poviargol is a broad-spectrum antimicrobial agent that is active against aerobic and anaerobic microflora, including those resistant to antibiotics. Poviargol inhibits the growth of most bacteria (staphylococci, streptococci, Pseudomonas aeruginosa and Escherichia coli, Proteus, Shigella, Salmonella, etc.) at concentrations up to 100 micrograms / ml.

[0019] The agent at a concentration of 1 to 3% has an anti-inflammatory effect, promotes the repair process at the epithelialization stage in wounds, has low toxicity, does not irritate skin and mucous membranes, and does not cause allergic reactions or dermatitis. Poviargol is used as an antimicrobial agent for the prevention and treatment of purulent-septic complications of wounds, ulcers, burns and pressure ulcers, including the condition that does not heal for a long time even during antibiotic treatment, for infectious diseases of the upper respiratory tract, ear, throat, nose, eye and oral cavity, and for diseases of the urogenital system and musculoskeletal system[http: / / sktb-technolog.ru / poviargolum2;“Poviargol:a new bactericidal agent for the treatment of infected wounds.(Experience of clinical use in traumatology,purulent surgery,burn therapy,gynecology,urology and ophthalmology).A reference manual for doctors / Edited by RAS member-corr.Panarina E.F.,MD,Professor.Blagitko E.M.Novosibirsk:Publishing House.1998.-66p.》].

[0020] In the context of the present invention, the term "sodium hypochlorite" refers to an effective disinfectant. The scope of its applications includes disinfection of laboratory instruments, water disinfection, local wound treatment and the like. Sodium hypochlorite exerts its biological activity through reaction with nucleic acids and amino acids. During reaction with nucleic acids, especially DNA, sodium hypochlorite chlorinates bases and prevents the formation of hydrogen bonds, which causes denaturation of the molecule and consequently results in the loss of biological activity.

[0021] In the context of this invention, the term "lysozyme" refers to enzymes in the innate immune systems of a wide variety of organisms, ranging from prokaryotes to higher eukaryotes. As a 1,4-bDN-acetylmuramidase, lysozyme exhibits chitinase activity in addition to the above, hydrolyzing glycosidic bonds of bacterial peptidoglycans, activating autolysin, and exhibiting antiviral activity, such as antiviral activity against human immunodeficiency virus (HIV). In this respect, its protein has relatively low toxicity, and no significant effects on the patient's body have been observed even when its dosage exceeds the amount sufficient to achieve a bactericidal effect.

[0022] In the context of this invention, the term "gramicidin C" refers to a molecule of organic origin that acts as a natural defense mechanism of Bacillus brevis bacteria, initiating production of this substance in response to threats to life. This antibiotic has a polypeptide structure and possesses bacteriostatic (prevents bacterial growth) and bactericidal (destroys bacteria) effects. The mechanism of action of gramicidin C is related to increased permeability of the cytoplasmic membrane of microbial cells, which disrupts the stability of microbial cells and causes cell death. Gramicidin C has a cyclic amino acid sequence, and this structure allows for the prevention of the development of microbial resistance to this antibiotic, but the development of drug dependence in various bacterial pathogens, which is important for effective treatment, is not currently described in the available literature.

[0023] In the context of this invention, the term "gerofsin" refers to a 4% solution of succinylated gelatin (also known as modified liquid gelatin) for intravenous administration, with an average molecular weight of 23,200 daltons. Gerofsin has a colloidal osmotic pressure of 34 mmHg. Its isoelectric point reaches pH 4.5. The negative charge generated within the molecule as a result of succinylation increases the molecular size, thus forming a bulkier protein chain than the non-succinylated chain while maintaining molecular weight. As a result, gerofsin has a sufficient volumetric effect over 3-4 hours. Gerofsin is used as a colloidal plasma substitute. [Brief explanation of the drawing]

[0024] [Figure 1] This shows the dynamics of the interaction between viral RNA and sodium hypochlorite. [Figure 2] This shows the time-dependent changes in the absorption spectrum of poly-AU (C=0.98×10⁻⁵ Mbp) in the presence of sodium hypochlorite (Shyp.=5×10⁻⁵ M, or 3.75 mg / l). [Figure 3] This shows the time-dependent changes in the absorption spectrum of the pAU-hypochlorite complex in the presence of povialgol at λ=260nm. [Figure 4] The absorption spectrum of the sodium hypochlorite-pobialgol mixture at λ=260nm is shown. [Figure 5] A control of human lung adenocarcinoma cell culture (ATCC® CRL-5800) is shown (x600). [Figure 6] This image (x900) shows a control of the cytotoxic effect of OC43 coronavirus on human lung adenocarcinoma cell cultures (ATCC® CRL-5800). [Figure 7] Morphological photographs (x600) of human lung adenocarcinoma cell cultures (ATCC® CRL-5800) in the presence of OC43 coronavirus and a polymer composition of three polymers are shown. [Figure 8] The absorption spectra of the DNA-hypochlorite complex are shown. CDNA = 2.2 x 10⁻⁵ Mbp (1-1 min a / a, 2-5 min a / a, 3-15 min a / a, 4-1 day a / a). a) CNaClO = 1.46 x 10⁻⁴ M, b) CNaClO = 3.5 x 10⁻⁴ M, c) CNaClO = 8.0 x 10⁻⁴ M. [Figure 9] A shows the reaction rate of the three systems of hypochlorite (Hyp)-DNA-povialgol (Pov), and B shows the time derivative of the absorption dependence of the DNA-Hyp-Pov system (black square) and DNA-Hyp (red circle) with respect to reaction time. [Figure 10] This paper shows the relationship between the bactericidal effect of a nasal administration composition and the exposure time of the nasal administration composition to various microorganisms. [Figure 11]This document demonstrates the bactericidal effect of a nasal administration composition and its antimicrobial component. [Figure 12] This shows a visualization of the nasal administration composition on the nasal mucosa 8 hours after application. [Figure 13] Eight hours after application, the absence of any visible material of the nasal administration composition based on the prototype on the nasal mucosa is demonstrated. [Figure 14(A)] A control of 0.5 MF Staphylococcus aureus is shown. [Figure 14(B)] The experimental sample of Staphylococcus aureus with a weight of 0.5 MF is shown. [Figure 14(C)] This shows a prototype of Staphylococcus aureus with a size of 0.5 MF. [Figure 15(A)] A control of 0.5 MF Pseudomonas aeruginosa is shown. [Figure 15(B)] The experimental sample of Pseudomonas aeruginosa with a density of 0.5 MF is shown. [Figure 15(C)] A prototype of Pseudomonas aeruginosa with a size of 0.5 MF is shown. [Figure 16(A)] A control of 0.5 MF Acinetobacter baumanni is shown. [Figure 16(B)] The experimental sample of Acinetobacter baumanni at 0.5 MF is shown. [Figure 16(C)] This shows a prototype of Acinetobacter baumanni at 0.5 MF. [Figure 17(A)] A control of 0.5 MF Klebsiella pneumoniae is shown. [Figure 17(B)] The experimental sample of 0.5 MF Klebsiella pneumoniae is shown. [Figure 17(C)] This shows a prototype of Klebsiella pneumoniae with a size of 0.5 MF.

[0025] [Modes for carrying out the invention] A composition according to the claim, for use in the nasal cavity solely for the prevention and / or treatment of infectious and inflammatory diseases of the nasal cavity, or as part of a comprehensive treatment of such diseases, having sustained antimicrobial activity against human coronaviruses, influenza viruses, other RNA and DNA-containing viruses, and bacteria, having the ability to prevent the formation of microbial biofilms and to disrupt already formed microbial biofilms, wherein the components are in the following weight % ratios, i.e., High molecular weight hyaluronic acid -0.1~2.0, High molecular weight PVP-4.0~6.0, Povialgor -0.8~1.2, Antimicrobial polypeptides (Gramicidin C - 0.03~0.15, or Lysozyme - 0.1~18.0), Sodium hypochlorite solution with an activated chlorine content (AH) of 1566 mg / l - 49.0~51.0 Gerofsin up to 100.0 The composition according to the claim, comprising:

[0026] The amount of high molecular weight hyaluronic acid may also be 0.5–2% by weight, 0.5–1% by weight, or 1–2% by weight.

[0027] The amount of high molecular weight PVP can also be 4.5–5.5% by weight.

[0028] The amount of Povialgol can also be 0.8–1% by weight.

[0029] The amount of lysozyme can also be 1 to 16.0% by weight.

[0030] The compositions described in the claims may be used, by application to the nasal mucosa, solely for the prevention of infectious and inflammatory diseases of the nasal cavity caused by coronaviruses, influenza viruses, other RNA and DNA-containing viruses, and bacteria, or as part of a comprehensive treatment for such diseases.

[0031] To obtain this composition, all powdered components (hyaluronic acid, polyvinylpyrrolidone, povialgol, gramicidin C (or lysozyme)) are successively dissolved in a sodium hypochlorite solution according to the quantitative ratios described above. Then, gerofsin solution is added up to 100.0% by weight and mixed until a homogeneous mixture is obtained.

[0032] A formulation of the claimed composition based on gerofcin, high molecular weight hyaluronic acid, and polyvinylpyrrolidone provides a sustained effect resulting from the formation of a protective barrier film on the mucosal surface of the nasal cavity, thereby enhancing the long-term antimicrobial effects of povialgol and sodium hypochlorite, moistening the nasal passages, and reducing the risk of irritation, dryness of the nasal mucosa, pain, and bleeding.

[0033] The proportions of the components of the composition described in the claim are the result of extensive experimental research and are optimally suited to achieve the above results. In this regard, an important advantage of the composition described in the claim is that it has a sustained antimicrobial effect against a wide range of RNA and DNA-containing viruses, as well as bacteria and microbial biofilms, and that it has no side effects.

[0034] The composition described in the claim is characterized by forming a thin polymer barrier film on the mucosal surface of the nasal cavity with a single application, which provides high adsorption of viral and bacterial particles for one hour. Furthermore, the presence of povidolgol for the first 50 seconds accelerates the denaturing effect of sodium hypochlorite on viral RNA or viral and bacterial DNA. [Examples]

[0035] As a non-limiting example, the following experimental data is provided.

[0036] Example 1 The following components by weight percentage, namely, Sodium hypochlorite solution (AH 1566 mg / L) - 49.0 Hyaluronic acid -1.0 PVP-5.0 Povialgol -1.0 Gramicidin C-0.03 Gerofsin -43.97 The antiviral activity of a proposed intranasal administration composition containing [the specified ingredient] against RNA-containing viruses was studied.

[0037] The following virus strains, namely, Human coronavirus, OC43 strain, Influenza A / Puerto Rico / 8 / 34(H1N1) virus I used it.

[0038] To analyze antiviral activity, 1 ml of the proposed nasal administration composition sample was mixed in a 1.5 ml vial with 0.1 ml of virus-containing liquid (initial titer: 6.5 g of OC43 coronavirus and 7.5 g of influenza virus). Instead of the composition, 0.1 ml of cell medium was added to a control vial. Each vial was incubated at room temperature for 3 minutes, after which the infectivity activity of the corresponding viruses in the virus-containing liquid was determined. Three parallel samples were used for analysis in each study group. [Table 1]

[0039] Based on the data above, the proposed nasal administration composition moderately reduced the titers of influenza virus and coronavirus OC43 in cell cultures.

[0040] The effectiveness of the antiviral effect against RNA-containing viruses was at least 99.9% during a 3-minute exposure.

[0041] Example 2 The following components by weight percentage, namely, Sodium hypochlorite solution (AH 1566 mg / L) - 49.0 Hyaluronic acid -1.0 PVP-5.0 Povialgol -1.0 Lysozyme -16.0 Gerofsin -28.0 The antiviral activity of a proposed intranasal administration composition containing [the specified ingredient] against RNA-containing viruses was studied.

[0042] The following virus strains, namely, Human coronavirus, OC43 strain, Influenza A / Puerto Rico / 8 / 34(H1N1) virus I used it.

[0043] To analyze antiviral activity, 1 ml of the proposed nasal administration composition was mixed in a 1.5 ml vial with 0.1 ml of virus-containing liquid (initial titer: 4.7 g of OC43 coronavirus and 5.5 g of influenza virus). Instead of the composition, 0.1 ml of cell medium was added to a control vial. Each vial was incubated at room temperature for 3 minutes, after which the infectivity activity of the corresponding viruses in the virus-containing liquid was determined. Three parallel samples were used for analysis in each study group. [Table 2]

[0044] Based on the data above, the proposed intranasal administration composition moderately reduced the titers of influenza virus and coronavirus OC43 in cell cultures. The effectiveness of the antiviral activity against RNA-containing viruses was at least 99% during a 3-minute exposure.

[0045] Example 3 To demonstrate that the antiviral effect of the components of the claimed composition is enhanced, the antiviral activity of the components of the proposed nasal administration composition against RNA-containing viruses (coronaviruses) at the following wt% concentrations was studied. Sample number 1 Gramicidin (polypeptide antibiotic) - 0.03 Sample number 2 Pobialgol (silver nanoparticles)-1.0 Sample number 3 Gramicidin (polypeptide antibiotic) - 0.03 Pobialgol (silver nanoparticles)-1.0 Sample number 4 Lysozyme (antimicrobial polypeptide)-16.0 Pobialgol (silver nanoparticles)-1.0 Sample number 5 Lysozyme (antimicrobial polypeptide)-16.0 Gramicidin (polypeptide antibiotic) - 0.03 Sample number 6 Lysozyme (antimicrobial polypeptide)-16.0.

[0046] The following virus strains, namely, Human coronavirus, OC43 strain I used it.

[0047] To analyze antiviral activity, 1 ml of each sample was mixed in a 1.5 ml vial with 0.1 ml of virus-containing liquid (initial titer of coronavirus OC43, 5.5 g). Instead of the corresponding sample, 0.1 ml of cell medium (complete RPMI-1640 medium containing 2 mM L-glutamine, 250 mg / l gentamicin, and 10% fetal bovine serum) was added to a control vial. Each vial was incubated at room temperature for 3 minutes, after which the infectivity activity of the corresponding virus in the virus-containing liquid was determined. Three parallel samples were used for analysis in each study group. [Table 3]

[0048] Since there is no data on the antiviral effect of gramicidin C in the available literature, this is the first time that the antiviral activity of gramicidin C alone has been demonstrated.

[0049] In this case, an additive effect (a distinctive feature) was observed in the interaction between gramicidine C and povialgol, and between lysozyme and povialgol, against the OC43 coronavirus strain. This interaction provides the advantage of sustained action in nasal administration compositions as the initial concentration of antimicrobial components gradually decreases.

[0050] Example 4 We studied the ability of Povialgol (silver nanoparticles) to accelerate the destruction of viruses and synthetic RNA by sodium hypochlorite.

[0051] This study used human coronavirus OC43 from virus strains collected at the St. Petersburg Pasteur Research Institute of Epidemiology and Microbiology (St. Petersburg). The initial viral RNA sample (C=3×10) was collected until it reached the minimum volume (600 μl) required for measurement in a special spectrophotometer cell. -4 M bp The solution was diluted with 0.001 M NaCl. The concentration of viral RNA used in the experiment was C = 1 × 10⁻⁶. -5 M bp That was the case.

[0052] In all experiments, the concentration (moles) of sodium hypochlorite was 5 times the concentration of nucleotides in the solution, i.e., S hyp. = 5 × 10 -5 The concentration was M, or 3.75 mg / l.

[0053] Figure 1 shows the results of interactions between viral RNA and sodium hypochlorite at various concentrations.

[0054] The various time-dependent behaviors in different regions of the spectrum indicate the interaction between sodium hypochlorite and various RNA components present in the solution. Specifically, the 260 nm wavelength region is where the interaction between sodium hypochlorite and the nucleic acid bases of polynucleotides can be reflected.

[0055] A similar experiment was conducted using synthetic RNA (poly AU). The results are shown in Figure 2.

[0056] The pattern of time dependence of the relative change in absorption and its rate in the region of λ = 260 nm that characterizes the state of nucleic acid bases in polynucleotides is similar between viral RNA and synthetic RNA. This indicates that the interaction process between polynucleotide and hypochlorite has at least two stages. The first stage is fast, and usually manifests as an increase in absorption intensity associated with denaturation of the polynucleotide or destruction of its secondary structure. Nucleic acid bases are also chlorinated in this stage. In the second stage, gradual destruction of nucleic acid bases occurs, which is caused by products formed in the first stage of interaction.

[0057] The time-dependent change in the absorption spectrum of the "poly AU + sodium hypochlorite" complex in the presence of povargol is shown at the following component concentrations: C pAU = 7.6 × 10 -5 M, C NaOCl = 3.8 × 10 -4 M, C poviargol = 0.002%. The results are shown in Figure 3.

[0058] Nucleotide destruction occurring in the first two minutes of the interaction between sodium hypochlorite and polynucleotide is significantly enhanced in the presence of povargol.

[0059] This mechanism of action is that sodium hypochlorite interacts with povargol. This is suggested by the absorption spectrum of the "sodium hypochlorite-povargol" mixture (Figure 4).

[0060] The product of this interaction contributes to the destruction of nucleotides in RNA.

[0061] We have for the first time identified and explained the effect of sodium hypochlorite accelerating the RNA disruption process in the presence of povialgol. This mechanism involves the interaction of sodium hypochlorite with povialgol. The products of this interaction contribute to the faster disruption of nucleotides in RNA.

[0062] Example 5 The sorption activity of polymer combinations of the following weight percent compositions against RNA-containing viruses was studied. Sample number 1 High molecular weight PVP-5.0 Cell culture medium - up to 100.0°C. Sample number 2 High molecular weight hyaluronic acid - 1.0 Gerofsin (4% liquid gelatin) - up to 100.0. Sample number 3 High molecular weight hyaluronic acid - 1.0 High molecular weight PVP-5.0 Gerofsin (4% liquid gelatin) - up to 100.0.

[0063] The following virus strains, namely, Human coronavirus, OC43 strain, Influenza A / Puerto Rico / 8 / 34(H1N1) virus I used it.

[0064] 100 μl of virus-containing liquid (initial titer: 6.5 g of OC43 coronavirus and 7.5 g of influenza virus) was added to 1 ml of the corresponding polymer combination, and the resulting mixture was incubated at room temperature for 1 hour. The sample was then centrifuged at 3000 rpm for 20 minutes, and the viral infectivity was detected in the supernatant. Cell culture medium was used as a control sample instead of the polymer combination. [Table 4]

[0065] The above data shows that, during a one-hour simultaneous pre-incubation, high molecular weight PVP had the ability to reduce the titer of both test viruses by 1 lb, a mixture of high molecular weight hyaluronic acid and gelatin by 2.5 lb, and a mixture of three polymers, namely high molecular weight PVP and hyaluronic acid on gerofusin, by 3.5 lb.

[0066] The polymer composition has been found to have exceptionally superior antiviral effects, which are attributed to the high ability of the polymers and their combinations to adsorb RNA-containing viral particles. This antiviral effect leads to the ability of the studied polymer combination to prevent viral particles from adhering to the nasal mucosa, to retain the viral particles within the polymer matrix, and to enhance the action of antimicrobial agents in the antimicrobial nasal administration composition.

[0067] Figures 5-7 show morphological images of the cell culture state of the experimental OC43 coronavirus in the presence of three polymer compositions, namely high molecular weight PVP and hyaluronic acid on gerofsin.

[0068] As can be seen in Figure 6, OC43 coronavirus has a clear cytotoxic effect and completely destroys the monolayer of the cell culture.

[0069] As can be seen from Figure 7, the polymer composition has a protective effect on the monolayer of the cell culture, preventing its destruction due to the cytotoxic effects of the coronavirus.

[0070] Example 6 We studied the ability of Povialgol (silver nanoparticles) to accelerate the destruction of microbial plasmid DNA by sodium hypochlorite.

[0071] Plasmid DNA from E. coli DH5-alpha strain, with a size of 5,500-6,000 nucleotide pairs, was used in this experiment. The concentration used was C = 2.2 × 10⁻¹⁴. -5 M bp That was the case.

[0072] Three plasmid DNA and hypochlorite complexes were prepared for spectral analysis. Three different solutions containing a constant concentration of DNA and different concentrations of sodium hypochlorite were used, i.e., 1) C NaOCl = 1.46 × 10 -4 M, 2) C NaOCl = 3.5 × 10 -4 M, 3) C NaOCl = 8.0 × 10 -4 M These were prepared for each complex.

[0073] The complex was prepared by direct mixing. A sodium chloride (NaCl) aqueous solution with an ionic strength of 0.015 M was used as the solvent.

[0074] Figure 8 shows the absorption spectra of the E. coli plasmid DNA complex with sodium hypochlorite at various concentrations of NaOCl.

[0075] As can be seen from the obtained spectrum, regardless of the concentration of sodium hypochlorite, the absorption of the solution increases across the entire DNA absorption region during the first minute after mixing. The overall spectrum is close to the sum of the spectra of DNA and sodium hypochlorite at the concentrations used.

[0076] After 10-15 minutes, significant changes were observed in the spectral shape and absorption intensity at a wavelength of 260 nm.

[0077] After one day, the DNA absorption band (λmax=260nm) has essentially disappeared. NaClO =8.0x10 -4 At concentration M, the absorption zone of sodium hypochlorite persisted throughout the day. This indicates that, at a given concentration, this disinfectant was incorporated in excess and remained in the solution even after interaction.

[0078] From the above, it was demonstrated that interaction with sodium hypochlorite results in both the destruction (denaturation) of the secondary structure of microbial DNA and chemical modification of nucleic acid bases, which is presumed to be chlorination. The presence of secondary structure delays the chemical reaction between sodium hypochlorite and the nucleic acid bases of DNA.

[0079] A solution of povidolgol was prepared from a dry powder weighed substance. The solvent was an aqueous solution of NaCl salt with an ionic strength of I = 0.001 M. In addition, an equal volume of reagent C was added. Pov To achieve a 0.5% mixing condition, the concentration of the prepared solution was halved.

[0080] The material used in this study has a characteristic absorption spectrum in the wavelength range of 220–500 nm. The spectrum was measured using a Shimadzu 1800 spectrophotometer in a quartz cuvette with optical path lengths d1=1 cm and d2=0.101 cm. The aforementioned spectrophotometer operates in a double-beam configuration. In all measurements, the cuvette for the background solution was left empty; that is, the measurements were performed against the atmosphere, and its absorption in the range examined was D Air The values ​​did not exceed ≤0.005. After each measurement, the cuvette containing the solution was rinsed and used to measure the solvent used in the previous measurement. Using the Origin software package, the solvent spectrum was subtracted during the data processing stage. Solutions were prepared using a laboratory electronic balance (accuracy 0.0002 g) and a dispenser, and stirred using a vortex mixer.

[0081] Differential analysis of the time-dependent absorption change at a wavelength of 260 nm (Figure 9) reveals that the first step of the DNA disruption reaction is much faster. This fact is consistent with the fact that in the presence of sodium hypochlorite, povidolgol (silver nanoparticles) induces denaturation of microbial DNA, thereby making nucleic acid bases more susceptible to chlorination.

[0082] Therefore, the disruption of microbial DNA molecules by sodium hypochlorite occurs more rapidly in the presence of povidol in the initial stages of the interaction. The reaction product of silver nanoparticles and sodium hypochlorite promotes the denaturation of the microbial DNA double helix, which accelerates the chlorination reaction between nucleic acid bases and sodium hypochlorite.

[0083] Therefore, the proposed nasal administration composition destroys RNA and DNA-containing viruses, as well as bacteria.

[0084] Example 7 The following components by weight percentage, namely, Sodium hypochlorite solution (AH 1566 mg / l) - 49.0 Hyaluronic acid -2.0 PVP-5.0 Povialgol -1.0 Gramicidin C-0.03 Gerofsin -42.97 The dependence of the bactericidal effect of a proposed intranasal administration composition containing [ingredient name] on exposure time was studied.

[0085] Bacterial isolates were obtained from infection sites of patients in various departments of an interdisciplinary medical institution, and the proposed antimicrobial compositions were tested on these bacterial isolates using the "checkerboard" method.

[0086] Figure 10 shows the evaluation results of the dependence of the bactericidal effect on exposure time.

[0087] It was observed that the growth of the tested microorganisms completely ceased after only 1.5 hours of exposure to the nasal administration composition. In this case, it should be noted that the disinfectant components of the nasal administration composition—sodium hypochlorite and povidol—were used at bactericidal doses that were several hundredths and several thousandths of the permissible dose of a single agent for clinical topical use.

[0088] Example 8 The following components by weight percentage, namely, Sodium hypochlorite solution (AH 1566 mg / L) - 49.0 Hyaluronic acid -1.0 PVP-5.0 Povialgol -1.0 Lysozyme -16.0 Gerofsin -28.0 The bactericidal effect of a proposed nasal administration composition containing [ingredient name] was studied.

[0089] Lysozyme is known to have very weak antibacterial activity against Gram-negative microorganisms [Renata Cegielska-Radziejewska, Grzegorz Lesnierowski, Tomasz Szablewski, Jacek Kijowski. Physico-chemical properties and antibacterial activity of modiWedeggwhite-lysozyme / / Eur Food Res Technol(2010)231:959-964;DOI:10.1007 / s00217-010-1347-y;Goncharova AI, Okulich VK, Zemko V.Yu., Senkovich SA Antimicrobial activity of lysozyme as a factor of nonspecific resistance / / Bulletin of Vitebsk State Medical University.-2019.-volume] 18,No.4.-pp.40-45;DOI:https: / / doi.org / 10.22263 / 2312-4156.2019.4.40].

[0090] Clinically multi-resistant Staphylococcus epidermidis 8242008 (resistant to more than five antibiotic groups, including gentamicin), isolated from patient blood cultures, was used in the study.

[0091] We used the Klebsiella pneumoniae 5299639 clinical strain isolated from wounds in patients with maxillofacial profiles. This isolate was characterized by resistance to eight antimicrobial agents, including carbapenems.

[0092] The proposed intranasal administration compositions were tested using the "checkerboard" method after 1.5 hours of exposure.

[0093] Figure 11 shows the results of the evaluation of the bactericidal effect of the nasal administration composition and its antimicrobial component.

[0094] Regarding Gram-negative Klebsiella pneumoniae, lysozyme was found to have no antimicrobial activity. Povialgol reduced the microbial contamination level by 1 lb immediately after 1.5 hours of exposure. The nasal administration composition was found to reduce the number of Klebsiella pneumoniae by 2 lb during application.

[0095] Regarding Gram-positive Staphylococcus epidermidis, lysozyme was found to reduce contamination by 1 lb after 1.5 hours of exposure. Povialgol reduced the microbial contamination level of Staphylococcus by 2 lb. The nasal administration composition was found to reduce the number of Staphylococcus epidermidis by 3 lb during application.

[0096] The above results indicate that the bactericidal effect of lysozyme against pathogenic microorganisms is increased in the presence of povialgol.

[0097] Example 9 The following components by weight percentage, namely, Sodium hypochlorite solution (AH 1566 mg / l) - 49.0 Hyaluronic acid -1.0 PVP-5.0 Povialgol -1.0 Gramicidin C-0.03 Gerofsin -43.97 Evaluation of the sustained effect of a nasal administration composition containing the above.

[0098] This study was conducted with 10 male and female volunteers aged 18 to 83 years. Prior to the study, pulverized activated carbon was added to the composition to improve visualization. The proposed nasal administration composition was applied once to both nasal canals for all subjects, and then the subjects compressed their nasal wings with their fingers to evenly distribute the composition across the mucosal surface. The distribution of the nasal administration composition along the nasal mucosa was evaluated 8 hours after application under prenatal rhinoscopy conditions using a hardware technique (STORZ Karl Storz Endoscope tele pack XLEDTP100 device).

[0099] The nasal administration composition was observed to be continuously and uniformly distributed on the anterior nasal cavity mucosa in all volunteers throughout the entire period, indicating the presence of the proposed nasal administration composition during the 8-hour observation period (Figure 12).

[0100] Similar tests were conducted using a nasal administration composition based on a prototype for nasal mucosa, and no visible material was observed 8 hours after application (Figure 13).

[0101] Example 10 Comparative test of a nasal administration composition (hydrogel) and a prototype. When the antiviral activity was evaluated according to the method described above, it was found that the viral titer decreased to 1.3 lb TCID50 during a 3-minute co-incubation of OC43 coronavirus and the prototype ointment (the viral titer in the control was 5.3 lb).

[0102] For comparison, Example 1 showed a decrease in the titer of OC43 coronavirus during co-incubation with the nasal administration composition for 3 minutes, with this decrease reaching 3 lb (the viral titer in the control was 6.5 lb). In Example 2, the decrease in the titer of OC43 virus during incubation with the nasal administration composition containing lysozyme was 1.5 lb (in this case, the viral titer in the control was 4.7 lb).

[0103] Therefore, the proposed hydrogel has higher efficiency compared to the prototype.

[0104] Example 11 Dosage 10 during 1 hour of exposure 5 Comparative evaluation of the bactericidal activity of the nasal administration composition (hydrogel) and prototype based on the dosage of test cultures of microorganisms at CFU / ml (Staphylococcus aureus (Figure 14 (A~C)), Pseudomonas aeruginosa (Figure 15 (A~B)), Acinetobacter baumanni (Figure 16 (A~B)), Klebsiella pneumoniae (Figure 17 (A~B)) and co-incubation time (according to the method of Example 7, this time-kill assay is an evaluation of the time-lethal effect relationship).

[0105] The following results were obtained. The prototype drug had no bactericidal effect on the test cultures of microorganisms. Microbial growth was observed in the petri dishes (Figures 14(C), 15(C), 16(C), and 17(C)).

[0106] In this respect, the nasal administration composition according to Example 7 ultimately reduces the microbial population by 4-5 g for all Gram-positive and Gram-negative microorganisms.

[0107] Based on the above, the composition described in the claim is more effective than the prototype agent in both virucidal activity and bactericidal activity.

Claims

1. Hyaluronic acid, polyvinylpyrrolidone, silver nanoparticles, antibacterial polypeptide, sodium hypochlorite solution, and gelatin are used in the following weight percentages of components, i.e., Hyaluronic acid -0.1 to 2.0 Polyvinylpyrrolidone -4.0 to 6.0 Silver nanoparticles - 0.8 to 1.2 Antimicrobial polypeptide - 0.03 to 18.0 Sodium hypochlorite solution - 49.0 to 51.0 Liquid gelatin - up to 100.0 A composition for intranasal administration for the prevention and / or treatment of infectious and inflammatory diseases of the nasal cavity, comprising [a specific compound / component].

2. The nasal administration composition according to claim 1, characterized in that the amount of hyaluronic acid is 0.5 to 2% by weight.

3. The nasal administration composition according to claim 1, characterized in that the amount of hyaluronic acid is 1 to 2% by weight.

4. The nasal administration composition according to claim 1, characterized in that the amount of polyvinylpyrrolidone is 4.5 to 5.5% by weight.

5. The nasal administration composition according to claim 1, characterized in that the amount of silver nanoparticles is 0.8 to 1% by weight.

6. The nasal administration composition according to claim 1, characterized in that the antimicrobial peptide is gramicidin C or lysozyme.

7. The nasal administration composition according to claim 6, characterized in that the amount of gramicidin C is 0.03 to 0.15% by weight.

8. The nasal administration composition according to claim 6, characterized in that the amount of lysozyme is 0.1 to 18.0% by weight.

9. The nasal administration composition according to claim 6, characterized in that the amount of lysozyme is 1 to 16.0% by weight.

10. The nasal administration composition according to claim 1, characterized in that infectious and inflammatory diseases of the nasal cavity are caused by coronaviruses, influenza viruses, other RNA and DNA-containing viruses, and bacteria, and are accompanied by the formation of microbial biofilms.

11. Use of the nasal administration composition according to claim 1 for the prevention and / or treatment of infectious and inflammatory diseases of the nasal cavity caused by coronaviruses, influenza viruses, other RNA and DNA-containing viruses, and bacteria, which involve the formation of microbial biofilms, wherein the nasal administration composition is applied to the nasal mucosa once daily.

12. The use according to claim 11, characterized in that the nasal administration composition is used alone or as part of a combination therapy.

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