RU2728938C1 - Use of dalargin for production of drug preparations for preventing pneumonia - Google Patents

Abstract

FIELD: medicine; pharmacology.SUBSTANCE: invention refers to pharmacology and medicine and aims at prevention of pneumonia. For preventing pneumonia, a medicinal agent containing a hexapeptide of formula H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH or its pharmaceutically acceptable salt are used.EFFECT: use of the invention provides prevention of developing pneumonia and reduced risk of adverse consequences of developing pneumonia, including a reduced risk of mortality from pneumonia.1 cl, 2 dwg, 4 tbl, 5 ex

Description

Scope of invention

The invention relates to the field of pharmacology and medicine, in particular to the use of dalargin hexapeptide in the production of medicines intended for the prevention of pneumonia.

Background Art

Pneumonia is an infectious disease characterized by focal lesions of the respiratory parts of the lungs with the obligatory presence of intraalveolar exudation. According to the International Classification of Diseases (ICD-10), pneumonia is classified as a respiratory disease (codes J10-J18) caused by infectious agents such as viruses, bacteria, mycoplasma, fungi, chlamydia, or unspecified pathogens. Pneumonia is usually divided into community-acquired and nosocomial pneumonia. According to statistics, the incidence of community-acquired pneumonia in Russia among all age groups is 10-15 cases per 1000 population, and the mortality rate from this nosology is about 5% among middle-aged people and 30% among the elderly. Methodical instructions "Respiratory tract infections" MU 3.1.2.3047-13 of 2013 . Antibiotic therapy is one of the main approaches to the treatment of pneumonia, the effectiveness of which, however, decreases due to the development of resistance to existing antibiotics. A meta-analysis of 26 studies (5659 patients) showed that 11 to 22% of patients with pneumonia did not respond to antimicrobial therapy. van Hecke et l. Implications of Antibiotic Resistance for Patients' Recovery From Common Infections in the Community: A Systematic Review and Meta-analysis. Clin Infect Dis. 2017, 65 (3): 371-382 . Vaccine prophylaxis is one of the preventive approaches to reducing the incidence of pneumonia. However, vaccination can cause a protective effect only against a specific pathogen, while pneumonia can be caused by many, including undefined, infectious pathogens. Thus, there is an urgent need for new approaches to the prevention of pneumonia using non-specific means of prevention, especially those that reduce mortality in severe forms of pneumonia.

Dalargin is a regulatory hexapeptide with the chemical formula:

corresponding to the amino acid sequence H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH (YdAGFLR). Dalargin is a strong base and is used in the form of pharmaceutically acceptable, i.e. non-toxic salts, almost exclusively in the form of diacetate.

Dalargin is part of commercially available agents for the treatment of gastric ulcer and duodenal ulcer, pancreatitis, and pancreatic necrosis. The use of dalargin in medicinal products is disclosed in the following RF patents (Nos.): 2032422, 2241488, 2198641, 2104717, 2270025, 2351334, 2343885, 2405534, 2363455, 2515550, 2180598, 2635083, 2436588, 2473325, 2144831, 2646565 2146530, 2155608, 2185176, 2139725, 2266130, 2326661, 2299742, 2218896, 2416398, 2318503, 2430753, 2299438, 2258529, 2099077, 2122415, 2261713, 2006039, 2366416, 22287642, 2167849, 2017, 2284192, 2290203, 2286793, 2366432, 2299065, 2429002, 2196603, 2142814, 2285522, 2203693, 2230549, 2180591, 2142736, 2113856, 2217139, 2266752, 2261722, 2362580, 222376941, 22281814 2672888, 2416398, 2318503, 2430753, 2299438, 2258529, 2099077, 2122415, 2261713, 2006039, 2366416.

The effectiveness of dalargin in the treatment of pneumonia has already been shown earlier. Intramuscular administration of dalargin led to the normalization of the mucous membrane of the lobar bronchus in patients with community-acquired pneumonia. Borovskaya and others. Bulletin of physiology and pathology of respiration. 2004, 17: 85-88 . However, the effectiveness of dalargin in the prevention of pneumonia is unknown in the art.

Description of the invention

The present invention relates to the use of a hexapeptide of formula H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention of pneumonia.

Further, the present invention relates to the use of a medicament containing a hexapeptide of formula H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH (I) or a pharmaceutically acceptable salt thereof for the prevention of pneumonia.

The technical result of the present invention is that the drug containing the hexapeptide of formula (I) as an active substance, when used prophylactically, prevents the development of pneumonia and reduces the risk of adverse consequences of pneumonia, including reducing the risk of death from pneumonia.

The term "pneumonia" means an acute infectious disease of any etiology, characterized by focal lesions of the respiratory parts of the lungs with the obligatory presence of intraalveolar exudation. Non-exclusive examples of pneumonia include pneumonias listed in the International Classification of Diseases (ICD-10) codes J10-J18.

The term "drug" in the present invention means a substance or combinations of substances that come into contact with the human or animal body, penetrate the organs, tissues of the human or animal body, used to prevent disease. Medicines include pharmaceutical substances and medicinal products.

The term "pharmaceutical substance" means an active substance with pharmacological activity, which is intended for the production, manufacture of medicinal products and determines their effectiveness

The term "drug" means a drug in the form of a dosage form used to prevent disease.

The term "dosage form" means the state of the drug, corresponding to the methods of its administration and use, and ensuring the achievement of the desired therapeutic effect.

The term "dose" means the amount of active ingredient that must be taken at one time.

The term "prevention" means an intervention aimed at preventing a disease, reducing the risk of developing one or more symptoms of the disease, preventing the progression of one or more symptoms of the disease, and reducing the adverse consequences of the disease.

In one embodiment of the present invention, the medicament of the present invention is administered to a person who has a high risk of pneumonia due to epidemiological grounds. Such epidemiological features include, not exclusively, the presence of an epidemic of an infectious disease associated with a pathogen that increases the risk of pneumonia. Non-exclusive examples of such pathogens include influenza viruses Alphainfluenzavirus (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.

In another embodiment of the present invention, the medicament of the present invention is administered to prevent pneumonia in a person who has a high risk of developing pneumonia due to the early signs of an infectious disease that has already appeared in that person and who is at a high risk of developing pneumonia.

In another embodiment of the present invention, the medicament of the present invention is administered to a person at high risk of developing pneumonia to prevent complications associated with developing pneumonia, including disability and premature mortality.

The hexapeptide of formula (I) has the gross formula C ₃₅ H ₅₁ N ₉ O ₈ , molecular weight 725.84 daltons and CAS registration number 81733-79-1 as an identifier.

The present invention relates to the hexapeptide of formula (I) and all pharmaceutically acceptable salts thereof, as well as solvates. A preferred pharmaceutically acceptable salt of the hexapeptide of formula (I) is hexapeptide diacetate (I) having the formula C ₃₅ H ₅₁ N ₉ O ₈ x 2C ₂ H ₄ O ₂ .

The hexapeptide of formula (I) can be obtained by any method known in the art, in particular by solid phase synthesis. The hexapeptide of formula (I) and its salts and solvates are commercially available, for example, in Bachem catalog No. 4030569.0100 (https://shop.bachem.com/4030569.html). In addition, hexapeptide diacetate of formula (I) is produced as a commercially available pharmaceutical substance in the Russian Federation by several manufacturers.

The drug of the present invention contains the hexapeptide of the formula (I) as a pharmaceutical substance in an amount of 0.1 to 50 mg per dose.

In one embodiment of the present invention, the medicament is dosed and contains from 1 to 10 mg of hexapeptide diacetate of formula (I) in one dose.

The medicament of the present invention may contain excipients.

The term "excipient" means a substance of inorganic or organic origin used in the production process, manufacture of a medicinal product to give it the necessary physicochemical properties.

Non-exclusive examples of adjuvants for use in the dosage form of the present invention include purified water, water for injection, propellants, solvents, antimicrobial preservatives, surfactants, stabilizing agents, emulsifiers, buffering agents to maintain the pH at 3.0-8, five.

In some embodiments of the present invention, the medicament of the present invention may contain from 0.001 to 99.999% of an excipient.

The medicament of the present invention can be produced in both liquid and solid dosage forms.

In one embodiment of the present invention, the liquid dosage form is formulated as a suspension, emulsion, or solution for subsequent administration as injection, infusion, nasal spray, or inhalation nebulizer spray.

In another embodiment of the present invention, a solid dosage form, for example, a lyophilisate is dissolved or dispersed in a liquid medium prior to use and the resulting suspension, emulsion, or solution is administered by injection, infusion, nasal spray, or inhaled with a nebulizer.

In another embodiment of the present invention, the dosage form is produced as a suspension, emulsion or solution under pressure in a package with a metering valve-spray system (aerosols and sprays) for subsequent nebulization.

In another embodiment of the present invention, the solid dosage form is produced in the form of one or more doses of powder (lyophilisate) for subsequent spraying using a powder inhaler adapted for use with single-dose capsules, blisters or other suitable forms containing a powder for inhalation, or having a reservoir for powder. In the latter case, the dose is metered out using the dosing device of the inhaler.

The term "nebulizer" refers to an inhalation device that converts a liquid drug for nebulization into a dispersion in a gaseous medium for delivery of the active substance to the lungs. Examples of nebulizers include compressor, ultrasonic, or other types of nebulizers.

In one embodiment of the present invention, the drug is a stable aqueous solution containing the hexapeptide of formula (I) in water in the following ratios, wt%:

Hexapeptide of formula (I), diacetate 0.01 - 1.00% Water for injection or purified water up to 100%

In another embodiment of the present invention, the drug is a stable aqueous solution containing hexapeptide diacetate of formula (I) in water in the following ratios, wt%:

Hexapeptide of formula (I), diacetate 0.01 - 1.00% Sodium chloride 0.40 - 0.90 Water for injection or purified water up to 100%

The medicament of the present invention can be manufactured by methods well known in the art and in accordance with recognized pharmaceutical procedures, for example, as described in the Remington Pharmaceutical Science Directory, 17th edition, ed. Alfonso R. Gennaro, Mack Publishing Company, Easton, PA, 18th edition (Remington's Pharmaceutical Sciences, seventeenth edition, ed. Alfonso R. Gennaro, Mack Publishing Company, Easton, Pa., Eighteenth edition, 1990).

The present invention further provides a method for preventing pneumonia, characterized in that a therapeutically effective amount of a hexapeptide of formula H-Tyr-D-Ala-Gly-Phe-Leu-Arg-OH (I) or a pharmaceutically acceptable salt thereof is administered to a subject in need thereof.

The term "therapeutically effective amount" means an amount of a hexapeptide of formula (I) that is sufficient to provide the desired prophylactic effect. In a preferred embodiment, the therapeutically effective amount of hexapeptide (I) is from 0.01 to 1 mg / kg body weight of the subject.

The term "subject" means a mammal, including a human.

In the method for preventing pneumonia of the present invention, a therapeutically effective amount of a hexapeptide of formula (I) can be administered to a subject in need thereof by injection (intravenous, intramuscular, or subcutaneous), inhalation, or intranasal as drops, gel, or spray.

The following examples demonstrate the invention. The examples illustrate the invention and are not intended to limit the scope of the invention in any way.

Example 1.

Preparation of a medicinal product for inhalation.

Table 1. Composition of the medicinal product.

Composition Content, wt. % Hexapeptide (I) diacetate 0.01-1.00 Purified water up to 100

To prepare a medicament for inhalation, the lyophilisate of hexapeptide diacetate (I) is dissolved in purified water in the proportions indicated in Table 1 and the resulting solution is poured into vials or ampoules under nitrogen atmosphere. In the method of preventing pneumonia, the resulting solution is administered to a subject in need thereof by inhalation in the form of an aerosol using a nebulizer.

Example 2.

Preparation of a medicinal product for injection.

Table 2. Composition of the medicinal product.

Composition Content, wt. % Hexapeptide (I) diacetate 0.01-1.00 Sodium chloride 0.90 Water for injections up to 100

For the preparation of an injectable drug, the lyophilisate of hexapeptide diacetate (I) is dissolved in an isotonic aqueous solution of sodium chloride in the proportions indicated in Table 2. In the method of preventing pneumonia, the resulting solution is administered to a subject in need of this by injection (subcutaneously, intramuscularly, or intravenously).

Example 3.

Preparation of a medicinal product for intranasal administration.

Table 3. Composition of the medicinal product.

Composition Content, wt. % Hexapeptide (I) diacetate 0.01-1.00 Purified water up to 100

To prepare a drug for intranasal administration, the lyophilisate of hexapeptide diacetate (I) is dissolved in purified water in the ratios indicated in Table 3, the resulting solution is poured into vials under nitrogen atmosphere and the vials are equipped with a pump to obtain a coarse aerosol when pressed. In the method for preventing pneumonia, the resulting solution is administered to a subject in need thereof intranasally in the form of an aerosol.

Example 4.

The effectiveness of the drug in the prevention of pneumonia.

The efficacy of a drug containing a hexapeptide of formula (I) in the prevention of pneumonia was studied in a pneumonia model caused by intratracheal administration of lipopolysaccharide (LPS), a major component of the cell wall of gram-negative bacteria, as described in D'Alessio FR. Mouse Models of Acute Lung Injury and ARDS. Methods Mol Biol. 2018, 1809: 341-350. Severe fatal pneumonia was induced by the addition of the immunosensitizer alpha-galactosylceramide as described in Aoyagi T et al., Shock. 2019, 52 (1): 83-91 . Male mice of the C57Bl line were selected by randomization into two groups of 20 animals each. To induce pneumonia, all mice were injected intratracheally with alpha-galactosylceramide (1 μg / mouse) and 24 hours later with LPS (E. coli; 300 μg / mouse) with the addition of a complex of immunosensitizers muramyldipeptide G2 and complete Freund's adjuvant. LPS caused pneumonia. Figure 1 shows a histological specimen (hematoxylin-eosin stain) of healthy lung tissue (Figure 1A) and a lung specimen obtained from an animal with LPS-induced pneumonia as indicated above (Figure 1B). A preparation of healthy tissue is characterized by moderate blood circulation, absence of pathological changes in the stroma and parenchyma, as well as in the walls of the bronchi and bronchioles. The preparation of animal tissue induced by intratracheal administration of LPS is characterized by a pronounced picture of bronchopneumonia with areas of destruction of the walls of the bronchus and bronchioles, desquamated epithelium over a greater length, and with pronounced inflammatory infiltration extending to the adjacent alveoli, in the center of which there is an accumulation of inflammatory infiltrate. Thus, this experimental model of pneumonia reproduces the essential signs of pneumonia, characteristic of pneumonia in humans, and can be used to test substances suitable for the prevention of pneumonia.

Dosage regimen: The study design is shown in Figure 2A. In the first group (group HP (I) + LPS), animals received hexapeptide (I) at a dose of 10 μg / kg in 50 μL of physical. solution intramuscularly, once a day for five days. In the second group (LPS group, control), animals received 50 μl of physical. solution intramuscularly, once a day for five days. One day after the last injection of hexapeptide (I) or physical. solution, pneumonia was induced in all animals by sequential administration of alpha-galactosylceramide (1 μg / mouse) and 24 hours later with LPS (E. coli; 300 μg / mouse) with the addition of a complex of immunosensitizers as described above. The results are shown in Figure 2B as Kaplan-Meier curves describing animal survival. The figure shows that 80% of animals in the control group (LPS) died within 5 days after the induction of pneumonia with lipopolysaccharide versus 15% in the group of animals that received prophylactic hexapeptide (I) (group HP (I) + LPS). Analysis of the curves using the Mantel-Cox log rank test showed that the prophylactic administration of hexapeptide (I) reliably prevented the death of animals from pneumonia (p <0.0001) compared to the control. The hazard ratio of death from pneumonia for the group of animals receiving prophylactic hexapeptide (I) was 0.148 (95% confidence interval 0.060-0.365) relative to control animals. Thus, prophylactic administration of a drug containing hexapeptide (I) significantly reduced the risk of death from pneumonia.

Example 5.

The effectiveness of the drug in the prevention of pneumonia.

Male C57Bl mice received hexapeptide (I) at a dose of 10 μg / kg in 50 μl of physical. solution once a day for five days intramuscularly (group 1), intranasally (group 2), or inhalation (group 3). One day after the last administration, pneumonia was induced in the animals as described in Example 4. 120 hours after the induction of pneumonia, lungs were removed from the animals, dried to constant weight, and the ratio of dry weight to wet weight of the lung was determined. The results are presented in table 4 as the mean ± error of the mean of the indicated ratio in the experimental groups 1-3, as well as in the control group consisting of intact animals.

Table 4. The ratio of dry weight of the lung to the wet weight of the lung.

Group Ratio,% Control (n = 4) 26.6 ± 4.8 Group 1 (n = 4) 28.2 ± 4.0 Group 2 (n = 4) 21.3 ± 4.8 Group 3 (n = 4) 24.2 ± 4.1

One-way analysis (one-way ANOVA) with Tukey's test showed that there were no significant differences in the ratio of dry lung weight to wet weight of the lung between animals from experimental groups 1-3 and intact animals from the control group (F = 0.467; p = 0.71). Thus, prophylactic administration of hexapeptide (I) reliably prevented pulmonary edema, one of the symptoms of pneumonia, in an experimental model of pneumonia.

Claims (2)

1. Use of a medicament 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 pneumonia. 2. Use according to claim 1, characterized in that the pharmaceutically acceptable salt is a hexapeptide diacetate of formula (I).

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