RU2747550C1 - Means for prevention of interstitial pneumonia - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: present invention relates to the field of medicine, namely to the use of an agent for the prevention of interstitial pneumonia, comprising oxidized dextran with M.M. from 10 to 50 kDa in the form of an aqueous solution with a concentration of 0.5-10.0 wt. %.EFFECT: present invention provides a decrease in the severity of pathological morphological changes in the lung tissue, characteristic of interstitial pneumonia.1 cl, 5 ex, 6 tbl

Description

The invention relates to experimental biology, pharmacology, medicine and can be used to create medicines for the prevention of interstitial pneumonia, including those accompanied by acute respiratory distress syndrome.

Currently, there is a steady increase in viral interstitial pneumonia, developing as a complication of acute respiratory viral infections.

Interstitial pneumonia is a progressive inflammatory process affecting the walls of the alveoli and the connective tissue of the parenchyma, with possible secondary inside alveolar exudation and the outcome in fibrous remodeling of the pulmonary structures.

Modern medicine has a very extensive arsenal of traditional and promising drugs for the drug therapy of pneumonia, but the COVID-19 pandemic has highlighted the development of effective means of preventing viral interstitial pneumonia. It is known that a feature of coronavirus pneumonia is the prevalence of acute respiratory distress syndrome in its pathogenesis and the early development of fibrotic complications.

For the treatment and prevention of acute respiratory distress syndrome and pneumonia, the use of derivatives of 4- (4-cyano-2-thioaryl) -dihydropyrimidinones, which are inhibitors of neutrophil elastase, has been described. In the pathogenesis of lung tissue damage in interstitial pneumonia, neutrophil elastase can act as one of the agents damaging lung tissue, but its role is ambiguous and there are no convincing in vivo data confirming the high therapeutic and prophylactic efficacy of neutrophil elastase inhibitors in pneumonia and acute respiratory distress syndrome. [RF patent for invention No. 2497813 "4- (cyano-2-thioaryl) -dihydropyrimidinones and their use", publ. 10.11.2013, Bul. No. 31].

To delay the development of pulmonary complications, in particular bronchitis and pneumonia, it is proposed to use an alpha-1 proteinase inhibitor (A1PI) by inhalation [RF patent No. 2635482 "Alpha-1 proteinase inhibitor for delaying the onset or progression of pulmonary exacerbations," publ. 13.11.2017, Bul. No. 31]. A1PI inhibits trypsin, chymotrypsin and various forms of elastases, cutaneous collagenase, renin, urokinase, and polymorphonuclear lymphocyte proteases. One of the endogenous roles of A1PI is to regulate the activity of neutrophil elastase, which degrades foreign proteins and damages native tissue present in the lungs. The main disadvantage of the claimed invention is the limited contingent of patients in whom the A1PI content is reduced, and they are subject to the potential risk of pulmonary complications, including pneumonia. The patent also lacks any data confirming the effectiveness of A1PI for the prevention of interstitial pneumonia, especially of viral etiology.

For the treatment of influenza and pneumonia, it is proposed to use a combination of phillirin and phylligenin. Both compounds are the main biologically active substances of the fruits of Forzitia drooping (of the Olive family) and have antiviral, antibacterial, antioxidant, antitumor and anti-inflammatory effects. According to the authors of the patent, the combination of phillirin and philligenin has a more pronounced antiviral effect in in vivo and in vitro models than traditional antiviral agents such as oseltamivir and ribavirin, while the claimed composition has an anti-inflammatory, antipyretic pharmacological effect that can potentially relieve influenza symptoms and pneumonia and can be considered as a promising drug [patent for invention of the Russian Federation No. 2655616 "The use of a composition of phillirin / philligenin in the preparation of a drug or medical product for the relief and / or prescription for the treatment of viral diseases, and a drug or medical product for the treatment of viral diseases ", publ. 05/29/2018, Bull. No. 16]. The disadvantage of the present invention is that the claimed composition does not have a prophylactic effect in viral interstitial pneumonia. While at present the arsenal of anti-inflammatory, antipyretic drugs for the treatment of interstitial viral pneumonia is quite extensive. However, despite a wide range of medications for the treatment of interstitial pneumonia, especially of viral etiology, early prevention of this serious disease is a priority and subsequently determines its easier clinical course.

For the treatment and prevention of influenza and pneumonia caused by the influenza virus, the search for effective neuraminidase inhibitors is still relevant. Thus, fluorine-substituted (3r, 4r, 5s) -5-guanidino-4-acylamino-3- (pentan-3-yloxy) cyclohexene-1-carboxylic acids and their esters [patent for invention RF No. 2489422, "Fluoro-substituted (3R, 4R, 5S) -5-guanidino-4-acylamino-3- (pentan-3-yloxy) cyclohexene-1-carboxylic acids, their esters and method of application", published 10.08.2013 Bul ... No. 22]. These compounds can be used for the treatment and prevention of influenza and pneumonia caused by this virus. The disadvantage of the claimed compounds is their narrow specificity with respect to the influenza virus, and the prophylactic effect of these compounds does not apply to other viruses that do not have neuraminidase activity.

Known immunomodulators, which also have a complex pharmacological effect (anti-inflammatory, antioxidant, immunostimulating, interferon-inducing, etc.), in the treatment of acute respiratory infections. Such compounds include azoxymer bromide (polyoxidonium) [Bulgakov V.A. Immunomodulators for the prevention and treatment of acute respiratory infections: the effectiveness of Azoximer bromide // Therapeutic archive. 2014. T. 86. No. 12, S. 92-97] and cycloferon [patent for invention of the Russian Federation No. 2281097 "Medicinal product, including cycloferon, method of its preparation and use", publ. 10.08.2006 Bul. No. 22]. The disadvantage of polyoxidonium is the lack of in vivo data confirming its effectiveness for the prevention of interstitial pneumonia.

Known use for the treatment of viral pneumonia of a combination of antiviral agents with different mechanisms of action of the components, for example, a combination for the treatment of viral influenza pneumonia, including riamilovir and oseltamivir [RF patent No. 2703535 "Combination of antiviral agents for the treatment of viral influenza pneumonia and its use", publ ... 2019, Bull. No. 30]. The disadvantage of the claimed antiviral composition is that it is focused only on the treatment of influenza pneumonia, as well as the lack of data on the prevention of interstitial pneumonia using the claimed composition of antiviral agents, especially for pneumonia caused by viruses, without pronounced neuraminidase activity.

The COVID-19 pandemic differs from all previously known acute respiratory viral infections primarily by the high incidence of severe forms of viral interstitial pneumonia, the treatment for which is still in the intensive search stage, as well as massive fibrotic complications after coronavirus pneumonia. To this should also be added the very high mortality rate in coronavirus pneumonia. Given the severity of this disease, the absence of specific medications for its treatment, the most promising direction is the prevention of viral interstitial pneumonia with the use of nonspecific medications aimed at different links of pathogenesis, in particular, acute respiratory distress syndrome, which is characteristic, among other things, of pneumonia caused by the COVID-19 virus. ... Sepsis and acute respiratory distress syndrome (ARDS) are common elements in the pathogenesis of severe COVID-19.

An experimental approach is described in determining the prospects for the use of medications, in particular, the hexapeptide of the formula H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH (I) or its pharmaceutically acceptable salt, for the prevention of pneumonia [RF invention patent No. 2728938 "The use of dalargin for the production of drugs for the prevention of pneumonia", publ. 08/03/2020, Bulletin No. 22], including caused by COVID-19 [patent for invention of the Russian Federation No. 2728939 "The use of dalargin for the production of drugs for the treatment of coronavirus infection Covid-19", publ. 08/03/2020 Bul. No. 22]. Taking into account the complexity of modeling this disease in vivo, it was proposed to use a model of pneumonia induced by intratracheal administration of lipopolysaccharide (LPS-induced pneumonia), which, like coronavirus pneumonia, develops in the form of interstitial pneumonia with a predominance of acute respiratory distress syndrome. A similar experimental model is described in other sources with different options for technical performance and criteria for assessing the severity of the pathological process, mainly morphological and immunological [Rybakova A.V., Makarova M.N., Sokolov V.D., Kryshen K.L., Khodko S. .V., Makarov VG Guidelines for experimental modeling of LPS-induced acute pneumonia in rats // International veterinary bulletin, 2013. No. 4, pp. 106-116; Katelnikova A.E. et al. Experimental models of acute bronchitis in animals // Laboratory animals for scientific research, 2019, No. 1, pp. 127-150; Golubev A.M., Kuzovlev A.N., Sundukov D.V., Golubev M.A. Morphological characteristics of the lungs during inhalation of lipopolysaccharide and perfluorane // General Reanimatology, 2015, Volume 11, No. 1, pp. 7-13].

Closest to the claimed technical solution (prototype) is the use of a non-specific drug containing a hexapeptide of the formula H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH (I) or a pharmaceutically acceptable salt thereof for the prevention of interstitial pneumonia [patent for the invention of the Russian Federation No. 2728938 "The use of dalargin for the production of drugs for the prevention of pneumonia", publ. 08/03/2020, Bul. No. 22]. In the claimed medicinal product, the hexapeptide of formula (I) is contained in an amount of 0.01 to 1.0 mass. %, the rest is pharmaceutical fillers and excipients. Examples of excipients: purified water, water for injection, propellants, solvents, antimicrobial preservatives, surfactants, stabilizing agents, emulsifiers, buffer components to maintain the pH value at 3.0-8.5. According to the prototype, the hexapeptide of formula (I) is administered intranasally, intramuscularly or by inhalation. The claimed medicinal product is used in an amount of 0.1 to 50 mg per dose. Non-exclusive examples of pneumonia pathogens in which the hexapeptide of formula (I) prevents pneumonia are Alphainfluenzavirus influenza viruses (serotypes H1N1, H1N2, H2N2, H3N2, H5N1, H7N9) and Betainfluenzavirus; coronaviruses SARS-CoV, SARS-CoV-2, MERS; the human respiratory syncytial virus Human orthopneumovirus; bacteria Streptococcus pneumoniae, Haemophilus influenzae, Legionella pneumophila, Escherichia coli; fungi Aspergillus, Mucor, Candida, Blastomyces, Coccidioides, Histoplasma, Pneumocystis, C. albicans, P. Carinii. The claimed agent was tested in a model of pneumonia in mice, induced intratracheal administration of lipopolysaccharide. After the administration of lipopolysaccharide and the induction of pneumonia, the hexapeptide of formula (I) was administered to the mice of the experimental groups intranasally, intramuscularly and inhaled at a dose of 10 μg / kg for 5 days. 120 hours after the induction of pneumonia, the lungs were removed from the animals, dried to constant weight, and the ratio of dry weight to wet weight of the lung was determined (assessment criterion - pulmonary edema), prophylactic administration of hexapeptide (I) reliably prevented pulmonary edema, one of the symptoms of pneumonia, in the experimental models of pneumonia. The disadvantage of the prototype tool is its lack of effectiveness.

The problem to be solved by the claimed invention is to increase the effectiveness of the prevention of interstitial pneumonia.

The technical result achieved with the implementation of the invention is to reduce the severity of pathological morphological changes in the lung tissue, characteristic of interstitial pneumonia.

The solution to this problem is achieved by the fact that oxidized dextran with M.M. is used as a prophylactic agent. from 10 to 50 kDa in the form of an aqueous solution with a concentration of 0.5 to 10.0 wt. %.

Disclosure of the essence of the invention

The agent for the prevention of interstitial pneumonia includes oxidized dextran with M.M. from 10 to 50 kDa in the form of an aqueous solution with a concentration of 0.5 to 10.0 wt. %.

Oxidized dextran is a dextran containing carbonyl groups. It can be obtained by known methods based on the chemical or radiation oxidation of dextran. In particular, oxidized dextran can be obtained chemically, for example, by oxidizing dextran ("Fluka", Switzerland) with sodium periodate and precipitating oxidized dextran with ethanol [RF patent No. 2125451 "Method for producing isoniazid-dextran conjugate", publication date 27.01. 1999].

Oxidized dextran can also be obtained by oxidizing dextran with potassium permanganate in an acidic medium with heating (80-100 ° C), after which the impurities are removed by separating the liquid from the precipitate that precipitated at the stage of dextran oxidation and containing manganese dioxide; oxidized dextran is precipitated from the supernatant with ethanol [Eurasian patent for invention No. 011718 "Method for producing dialdehyde dextran", publication date 04/28/2009].

Dextran with a molecular weight of 10-50 kDa in the form of an aqueous or aqueous-salt solution with a concentration of 1-10% is used as a starting product for obtaining the claimed agent.

Water for injection can be used as a pharmaceutically acceptable solvent.

For a comparative assessment of the prophylactic effect of the claimed agent and the hexapeptide in the form of the pharmacopoeial drug "Dalargin" described in the prototype, a histological study of the lungs of ACR mice was carried out in the prevention of LPS-induced interstitial pneumonia by inhalation. The study was carried out on 30 male ACR mice, which were injected intranasally with 20 μl of a solution (concentration 20 mg / ml) of Escherichia coli lipopolysaccharide (LPS), followed by inhalation in experimental groups of oxidized dextran or hexapeptide of formula (I) for 5 minutes, followed by withdrawal from the experiment after 24 hours. All animals in the study were divided into 5 groups: group 1 - control - male ACR mice with intranasal administration of 20 μl of LPS solution (2 mg / ml) and subsequent withdrawal from the experiment after 24 hours; Group 2 - male ACR mice with intranasal injection of 20 μl of LPS solution (2 mg / ml) followed by inhalation of a solution of hexapeptide of formula (I) with a concentration of 1 mg / ml (pharmacopoeial drug "Dalargin") for 5 min and excretion from experiment after 24 hours; Group 3 - male ACR mice with intranasal injection of 20 μl of LPS solution (2 mg / ml) followed by inhalation of 2% oxidized dextran solution with M.M. 40 kDa for 5 min and removal from the experiment after 24 hours; Group 4 - male ACR mice with intranasal injection of 20 μl of LPS solution (2 mg / ml) followed by inhalation of 0.5% solution of oxidized dextran with M.M. 50 kDa for 5 min and removal from the experiment after 24 hours; Group 5 - male ACR mice with intranasal injection of 20 μl of LPS solution (2 mg / ml) followed by inhalation of 10% oxidized dextran solution with M.M. 10 kDa for 5 min and removal from the experiment after 24 hours. The material for the histological examination was samples of lung tissue (lower right lobe), which were subjected to standard histological processing followed by staining with hematoxylin and eosin for visualization by direct light microscopy. Histological studies were performed using an AxioImager A1 microscope with an AxioCam MRc camera (Carl Zeiss, Germany). At the same time, the following pathological changes were assessed in sections of lung tissue samples from mice: the severity of interstitial and alveolar edema, hemocirculatory disorders, cellular inflammatory infiltration, destructive changes (areas of emphysema, atelectasis, dystrophy and necrosis of bronchial epithelium). The morphological criteria for assessing the preventive action of the claimed agent on the LPS-induced pneumonia model have been expanded in comparison with the prototype, for a more complete account of the acute respiratory distress syndrome developing on this model, which is an important pathophysiological link in severe interstitial pneumonia, including in COVID-19 ...

The results of a comparative study of the preventive action of the claimed agent and the prototype agent are presented in Table 1.

Note: +++ - pronounced changes, ++ - moderate changes, + slight changes. When there is a mixed version, for example, "+ / ++", it means that the changes are worn depending on the visual fields on the preparation under the microscope from mild to moderate changes.

As can be seen from the presented results, according to the morphological evaluation criteria, the claimed agent, in comparison with the prototype agent, has a more pronounced prophylactic effect on the LPS-induced pneumonia model.

The range of concentrations of oxidized dextran selected in the claimed agent was selected experimentally. When the concentration of the oxidized dextran solution is less than 0.5 wt. % the preventive effect of solutions of oxidized dextran against interstitial pneumonia is reduced, and the use of solutions of oxidized dextran over 10 wt. % is impractical due to the high viscosity, which reduces the formation of stable aerosols when inhaled with standard nebulizers.

In addition, the range of molecular weights used for oxidized dextrans is determined by the availability and cost of primary raw materials (dextrans). The use of dextrans with molecular weights of less than 10 kDa and over 50 kDa is limited by the availability of their raw material base and these dextrans have a high cost, which ultimately leads to a significant increase in the cost of the final product for widespread medical use in the prevention of viral interstitial pneumonia.

Implementation of the invention

The claimed technical solution is illustrated by the following examples of specific implementation.

Example 1. Preparation of the claimed means for inhalation.

For the preparation of the claimed means for inhalation, oxidized dextran with M. 10 kDa is dissolved in purified water in the proportions indicated in Table 2 and the resulting solution is sterilized by filtration through microporous filters with a pore diameter of 0.2 μm, then poured into sterile vials or ampoules under nitrogen atmosphere. For the prevention of pneumonia, the resulting solution is administered to a subject in need of it by inhalation in the form of an aerosol using a nebulizer.

Example 2. Preparation of the claimed means for inhalation.

For the preparation of the claimed means for inhalation, oxidized dextran with M. 50 kDa is dissolved in purified water in the proportions indicated in Table 3 and the resulting solution is sterilized by filtration through microporous filters with a pore diameter of 0.2 μm, then poured into sterile vials or ampoules under nitrogen atmosphere. For the prevention of pneumonia, the resulting solution is administered to a subject in need of it by inhalation in the form of an aerosol using a nebulizer.

Example 3

Preparation of the declared means for inhalation.

For the preparation of the claimed means for inhalation, oxidized dextran with M. 40 kDa is dissolved in purified water in the proportions indicated in Table 4 and the resulting solution is sterilized by filtration through microporous filters with a pore diameter of 0.2 μm, then poured into sterile vials or ampoules under nitrogen atmosphere. For the prevention of pneumonia, the resulting solution is administered to a subject in need of it by inhalation in the form of an aerosol using a nebulizer.

Example 4

Preparation of the declared means for inhalation.

For the preparation of the claimed means for inhalation, oxidized dextran with M. 10 kDa is dissolved in purified water in the ratios indicated in Table 5 and the resulting solution is sterilized by filtration through microporous filters with a pore diameter of 0.2 μm, then poured into sterile vials or ampoules under nitrogen atmosphere. For the prevention of pneumonia, the resulting solution is administered to a subject in need of it by inhalation in the form of an aerosol using a nebulizer.

Example 5

Preparation of the declared means for inhalation.

For the preparation of the claimed means for inhalation, oxidized dextran with M. 30 kDa is dissolved in purified water in the proportions indicated in Table 6 and the resulting solution is sterilized by filtration through microporous filters with a pore diameter of 0.2 μm, then poured into sterile vials or ampoules under nitrogen atmosphere. For the prevention of pneumonia, the resulting solution is administered to a subject in need of it by inhalation in the form of an aerosol using a nebulizer.

Claims (1)

The use of an agent for the prevention of interstitial pneumonia, including oxidized dextran with M.M. from 10 to 50 kDa in the form of an aqueous solution with a concentration of 0.5-10.0 wt. %.