7-АМino-[1,2,4]triazolo[1,5-А]pyrimidine derivatives and pharmaceutically acceptbale salts thereof having antiviral and antimicrobial activity

Abstract

The invention relates to pharmaceutically active viperin activator compounds having antiviral and antimicrobial activity and the use thereof, as well as to a pharmaceutical composition and a treatment method using said compounds. More particularly, the invention relates to a compound which is a 7-amino-[1,2,4]triazolo[1,5-а]pyrimidine derivative, having the general formula (I), where R1 is AlkS, AlkSO, AlkSO2, COOAlk, CON(Alk)2, substituted or unsubstituted Ph, benzyl, furyl, pyridyl; R2 is Н, Alk, Ph; R3 is ОН, OAlk, NHOAlk, NAlk2, or a saturated 6-membered nitrogen-containing heterocyclic ring bound by a nitrogen atom to a carbonyl group and optionally having a second heteroatom in the ring, selected from among X, where X is О, S, NMe; and Alk is Н, Me, Et, Pr or i-Pr, as well as to a pharmaceutically acceptable salt, solvate or hydrate of said compound. The compounds are intended for the treatment and / or prophylaxis of diseases caused by viral and / or microbial infections, such as, for example, diseases caused by influenza, ARI and SARS-CoV-2 viruses, or by ESKAPE group pathogens.

Description

[0001] IPC C07D 471 / 02; C07D 487 / 04 7-AMINO-[1,2,4]TRIAZOLO[1,5-A]PYRIMIDINE DERIVATIVES AND THEIR PHARMACEUTICALLY ACCEPTABLE SALTS HAVING ANTIVIRAL AND ANTIMICROBIAL ACTIVITY DESCRIPTION

[0002]

[0003] FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES The invention relates to new biologically active derivatives of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine of the general formula (I), their pharmaceutically acceptable salts, exhibiting activity against a wide range of viruses, primarily against the influenza virus, acute respiratory viral infections and the SARS-CoV-2 coronavirus, as well as pathogenic microorganisms, primarily from the ESCAPE group, a method for obtaining derivatives and their salts and their use as medicinal products.

[0004] LEVEL OF TECHNOLOGY

[0005] The need to find new antiviral agents is driven by the significant spread of viral infections in humans and animals, especially recently. There is every reason to believe that the spread of viral infections will continue to increase. This is due to increasing population density, weakened immune defenses in the human population, widespread viral resistance to current medications, and the emergence of new viruses. The rapid development of resistance is due to the fact that many currently used antiviral drugs are primarily derivatives of a single class of compounds, have limited efficacy, and their use in patients is unmonitored.

[0006] Influenza is an acute respiratory infectious disease caused by

[0007] An RNA-containing virus of high epidemiological and clinical significance with frequent complications, especially among individuals

[0008] Risk groups. Influenza viruses and other acute respiratory viral infections (ARVIs) cause massive outbreaks of disease that annually reach epidemic proportions. Between 27 and 41 million cases of influenza and ARVI are registered annually, with influenza affecting 5 to 15% of the Russian population annually. These diseases are considered virtually uncontrollable due to the highly variable antigenic structure of circulating influenza viruses and the heterogeneity of ARVI pathogens. Furthermore, influenza viruses and other ARVIs are capable of changing their properties and pathogenicity. The most recent example of such changes was the influenza pathogen H1N1pdmO9, which circulated during the 2009-2010 epidemic season and became known as "swine flu."From a biological point of view, influenza viruses belong to the Orthomyxoviridae family (RNA viruses with a segmented genome) and are divided into four monotypic genera: influenza A viruses (Alphainfluenza virus), influenza B viruses (Betainfluenza virus), influenza C viruses (Gammainfluenza virus), and influenza D viruses (Deltainfluenza virus), each of which is represented by only one type of virus of the same name. It is believed that only influenza A viruses have pandemic potential (Bouvier NM, Palese P. The biology of influenza viruses. Vaccine. 2008; V. 26, Suppl 4: D49-D53).Depending on the antigenic properties of hemagglutinin (a virion envelope glycoprotein that ensures recognition of target cells and binding of viral particles to the terminal sialic acid residues of glycoproteins of the plasma membrane of epithelial cells) and neuraminidase (exo-a-sialidase that catalyzes the cleavage of glycosidic bonds of the terminal sialic acid residues of oligosaccharides, glycoproteins, glycolipids and thereby ensures the release of daughter virions from infected cells), influenza A viruses are divided into subtypes.

[0009] The clinical picture of severe viral respiratory infections is often represented by a symptom complex of primary viral pneumonia. The occurrence of primary viral pneumonias in respiratory viral infections is apparently associated with the co-expression of glycoproteins and glycolipids characterized by the presence of glycans with a terminal α2,3-linked sialic acid (acting as a receptor for respiratory viruses) and transmembrane serine proteinase TMPRSS2 (proteolytically activating HA and S-protein of virions) of epithelial cells of the alveoli and bronchioles (Tortorici MA, Veesler D. Structural insights into coronavirus entry. Adv. Virus Res. 2019; 105: 93-116; Limburg H, Harbig A., Bestle D. et al. TMPRSS2 is the major activating protease of influenza A virus in primary human airway cells and influenza B virus in human type II pneumocytes. J. Virol.

[0010] 2019; 93: e00649-19). The development of predisposition to bacterial coinfection during respiratory viral pandemics is associated with many factors:

[0011] - virus-induced dysbiotic state and disruption of the barrier function of the epithelial lining of the respiratory tract (Hanada S., Pirzadeh M., Carver KY et al. Respiratory viral infection-induced microbiome alterations and secondary bacterial pneumonia. Front Immunol. 2018; 9: 2640; Sencio V, Barthelemy A., Tavares LP et al. Gut dysbiosis during influenza contributes to pulmonary pneumococcal superinfection through altered short-chain fatty acid production. Cell Rep. 2020; 30: 2934-47.e6);

[0012] - virus-induced dysfunction of effector cells of the immune system (Ghoneim HE, Thomas PG, McCullers JA. Depletion of alveolar macrophages during influenza infection facilitates bacterial superinfection. J. Immunol. 2013; 191: 1250-9; Sun K, Metzger DW. Influenza infection suppresses NADPH oxidase-dependent phagocytic bacterial clearance and enhances susceptibility to secondary methicillin-resistant Staphylococcus aureus infection. J. Immunol. 2014; 192: 3301-7) and immunosuppressive activity of cytokines relative to antibacterial immunity (Cao J., Wang D., He F. et al. Activation of IL-27 signaling promotes development of postinfluenza pneumococcal pneumonia. EMBO Mol. Med. 2014; 6: 120-40; Shepardson K, Larson K, Cho H. et al. A novel role for PDZ-binding motif of influenza A virus nonstructural protein 1 in regulation of host susceptibility to postinfluenza bacterial superinfections. Viral Immunol. 2019; 32: 131-43);

[0013] - virus-associated dysfunction of the alveolar-capillary barrier (Kamal RP, Alymova IV, York LA. Evolution and virulence of influenza A virus protein PB1-F2. Int. J. Mol. Sci. 2018; 19: E96; Short KR, Kasper J., van derAa. S. et al. Influenza virus damages the alveolar barrier by disrupting epithelial tight junctions. Eur. Respir. J.

[0014] 2016; 47: 954-66) and suppression of the activity of ion pumps that ensure fluid absorption from the alveolar lumen (Peteranderl C., Morales-Nebreda L., Selvakumar B. et al. Macrophage-epithelial paracrine crosstalk inhibits lung edema clearance during influenza infection. J. Clin. Invest. 2016; 126: 1566-80; Brand JD, Lazrak A., Trombley JE et al. Influenza-mediated reduction of lung epithelial ion channel activity leads to dysregulated pulmonary fluid homeostasis. JCI Insight. 2018; 3: 123467).

[0015] Pneumonia associated with respiratory viral infections is an independent factor in disease severity and mortality (Maruyama T, Fujisawa T, Suga S. et al. Outcomes and prognostic features of patients with influenza requiring hospitalization and receiving early antiviral therapy: a prospective multicenter cohort study. Chest. 2016; 149: 526-34; Ishiguro T, Kagiyama N., Uozumi R. et al. Clinical characteristics of influenza-associated pneumonia of adults: clinical features and factors contributing to severity and mortality. Yale J. Biol. Med. 2017; 90: 165-181). That is, to a large extent, the main problem of severe viral infections, both in the past and present, is the problem of viral, viral-bacterial and secondary bacterial pneumonia. The biology of influenza viruses and coronaviruses inevitably determines the emergence of new pandemic strains, the time of occurrence, genomic variability, and antigenic properties of which cannot be predicted.This means that pandemics of new respiratory infections will always begin in the absence of specific immune prophylaxis and therapy for these infections. This necessitates the early research and development of pathogenetic agents and methods for the prevention and treatment of respiratory viral infections based on the biological characteristics of coronaviruses and influenza A viruses.

[0016] To achieve the most complete and effective control of influenza infection, it is necessary to continually develop and introduce new antiviral drugs that are chemically unrelated to those already used clinically and that target alternative targets in the virus life cycle.

[0017] Antiviral agents for the treatment of influenza represent an extremely limited group of drugs, and resistance to them has been demonstrated for most of them. Due to the peculiarities of its genome organization (the absence of a replication error correction mechanism) and its short life cycle, the influenza virus has a high mutation rate. As a result, the virus's antigenic structure is highly susceptible to change due to the selective pressure of the host immune system. Furthermore, the use of chemotherapy drugs serves as a selection factor for the virus, which also results in the development of resistant strains. These two processes lead to the emergence of virus variants capable of evading neutralizing antibodies, thereby eluding the body's immune response, and of overcoming the effects of chemotherapy drugs targeting a specific stage of viral replication.Moreover, each type of virus has its own mechanism of adaptation to a chemical drug (Ison MG Antivirals and resistance: influenza virus. Curr. Opinion in Virol. 2011; 1: 563-573).

[0018] Currently, the most widely used group of drugs with activity against the replication of the influenza virus are neuraminidase inhibitors: Oseltamivir (Tamiflu) and Zanamivir (Relenza), registered in Russia, and Peramivir (Rapiacta) and Laninamivir (Inavir), used in the USA, which act at the stage of budding of newly synthesized influenza virions from the cell membrane, blocking the cleavage of viral progeny particles from the cell surface Ison MG Clinical use of approved influenza antivirals: therapy and prophylaxis. Influenza Other Respir. Viruses. 2013; 7 Suppl 1: 7-13). In addition, viral neuraminidase inhibitors prevent the access of virions to target cells by blocking the neuraminidase cleavage of mucopolysaccharides in the mucus of the upper respiratory tract.The practice of using neuraminidase inhibitors in the treatment of influenza has shown that the effectiveness of drugs in this group is limited by the early stage of the disease, as well as the possibility of bacterial neuraminidase viruses using concomitant infections.

[0019] Anti-influenza drugs with a different mechanism of action are also known, for example, Remantadine (a-methyl-1-adamantylmethylamine hydrochloride) and Amantadine (1-aminoadamantane) Davies WL, Grunert RR, Haff RF, McGahen JW, Neumayer EM, Paulshock M., Watts J C., Wood TR, Hermann EC, Hoffmann CE Antiviral Activity of 1 - Adamantanamine (Amantadine). Science.

[0020] 1964; 144: 862). The mechanism of action of these drugs has been studied quite thoroughly {Cady SD, Schmidt-Rohr K., Wang J., Soto CS, DeGrado WF, Hong M. H. Structure of the amantadine binding site of influenza M2 proton channels in lipid bilayers. Nature. 2010; 463: 689-692). Currently, as a result of the widespread use of adamantane drugs, their antiviral properties against influenza A viruses have been significantly lost; it has been shown that in Western Europe today there are no strains sensitive to this group of drugs. In the USA, the drug baloxavir marboxil (Xofluza) is registered - an inhibitor of the endonuclease activity of the polymerase complex of the influenza virus {Yang T. Baloxavir Marboxil: The First Cap-dependent Endonuclease inhibitor for the treatment of influenza. Ann. Pharmacother. 2019; 53: 754-759).In Russia and a number of other countries, Arbidol (umifenovir) is used to control influenza. It blocks the fusogenic activity of viral hemagglutinin and thus prevents the fusion of viral and cellular membranes {Blaising J., Polyak SJ, Pecheur EI. Arbidol as a broadspectrum antiviral: an update. Antiviral Res. 2014. doi: http: / / dx.d0i.0rg / l 0.1016 / j.antiviral.2014.04.006). In addition to direct antiviral activity, it has interferonogenic properties, so it can be used for both therapy and prevention of influenza infection. Influenza viruses and other RNA-containing viruses demonstrate the ability to develop resistance to direct-acting antiviral drugs, regardless of their mechanism of activity. One way to overcome this resistance is considered to be the simultaneous use of several drugs aimed at different viral targets.In this case, the probability of selection of virus variants resistant to two or more drugs is greatly reduced. Despite this, a number of studies have demonstrated the formation of double mutants resistant to both adamantane drugs and neuraminidase inhibitors (Sheu TG, Fry AM, Garten RJ. Dual Resistance to Adamantanes and. Oseltamivir Among Seasonal Influenza A (H1N1) Viruses: 2008-2010. J. Infect. Dis., 2011; 203 (1): 13-17).

[0021] One of the main challenges in modern healthcare is the development of test systems that can quickly identify the viral pathogen. This is because symptoms such as fever, cough, runny nose, and malaise are characteristic of infections with all respiratory viruses, and it is often difficult to make a correct diagnosis without clearly identifying the pathogen causing the illness. However, patients with upper respiratory tract infections often do not seek medical care and do not identify the specific virus causing their illness. Therefore, the search for and development of broad-spectrum antiviral agents is recognized as a major challenge.

[0022] The development of new approaches to treating infectious diseases remains a priority for research in chemistry and infectious biology in Russia and internationally. One of the most important areas of social significance involves identifying original, promising compounds with antimicrobial activity, with the goal of subsequently developing new, effective and safe antimicrobial drugs based on these compounds, primarily for combating ESCAPE microorganisms, such as Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter.

[0023] The objective of the present invention is to find new pharmacologically active compounds against a wide range of viral infections, primarily against influenza, coronavirus, respiratory syncytial virus, including strains resistant to currently existing drugs, and active against microorganisms of the ESCAPE group, which would have low toxicity and would not cause side effects in warm-blooded living organisms.

[0024] The World Health Organization (WHO) has identified antimicrobial resistance as a threat to global stability and, at the country level, a threat to national security. According to a WHO systemic analysis published in 2023, 541,000 deaths in the European Region in 2019 were attributed to bacterial infections caused by problematic resistant strains of pathogens. Seven leading pathogens—Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Enterococcus faecium, Streptococcus pneumoniae, and Acinetobacter baumannii—were responsible for more than 80% of resistance-related deaths.

[0025] According to forecasts from the Organization for Economic Cooperation and Development, by 2035 we expect a twofold increase in bacterial resistance to last-line antibiotics compared to 2005 levels, which, according to WHO experts, highlights the urgent need to apply effective methods for managing antimicrobial therapy and expand the coverage of epidemiological surveillance for antibiotic-resistant strains of microorganisms worldwide.

[0026] Despite more than five years of active attention to the problem of antibiotic resistance worldwide and a range of measures to control it, the WHO has noted the insufficient real impact of measures taken both globally and in individual countries (draft document "WHO strategic and operational priorities to address drug-resistant bacterial infections, 2025-2035"). The reason for this situation is obvious: the problem of antibiotic resistance has been growing for decades due to insufficient attention to it both at the practical level of antibiotic use in healthcare, agriculture, and veterinary medicine, and due to the lack of intensive scientific research in this area. It is precisely because of the insufficient development of the scientific base that humanity has proven unprepared for this challenge.Antibiotic resistance is a natural, ongoing, evolutionary process. Therefore, there is no single solution to this problem. Control requires the simultaneous application of various approaches. This is impossible without establishing a scientific foundation, developing highly competent scientific personnel, and developing technological capabilities.

[0027] This application implements the original idea of ​​searching for next-generation drugs that activate a unique factor of innate immunity, viperin. These drugs, therefore, possess both antiviral and antimicrobial activity and are resistant to the development of resistance by both viruses and bacteria, due to the fact that the claimed compounds affect host susceptibility rather than the pathogenicity of the infection.

[0028] Viperin protein, also known as RSAD2 (Radical S-adenosyl methionine domain-containing 2) and cig5 (cytomegalovirus-induced gene 5), is involved in many cellular processes, among which the most studied are the immune response and mitochondrial metabolism (Rivera-Serrano EE, Gizzi A. S., Arnold J. J, Grove TL, Almo SC, Cameron CE. Viperin reveals its true function. An. Rev. Virol. 2020; 7 (1): 421-446; Wang X., Hinson ER, Cresswell P. The interferon-inducible protein viperin inhibits influenza virus release by perturbing lipid rafts. Cell Host Microbe. 2007; 2 (2): 96-105; Gizzi A. S., Grove TL, Arnold JJ, Jose J., Jangra RK, Garforth SJ A naturally occurring antiviral ribonucleotide encoded by the human genome. Nature. 2018. 558 (7711): 610-614).

[0029] In humans, the RSAD2 gene is located at 2p25.2 and is 2858 bp in length. The 1083-nucleotide-long transcript encodes a 361-amino-acid protein weighing approximately 42.17 kDa. RSAD2 has three domains: an N-terminal α-helical domain, a central S-adenosyl methionine domain (SAM), and a conserved C-terminal domain. The N-terminal α-helical domain localizes viperin to the outer (cytosolic) lipid layer of the endoplasmic reticulum (ER) membrane, the Golgi apparatus (GA), and lipid droplets. SAM contains four conserved domains, the first of which contains three cysteine ​​residues, CXXCCXC, responsible for Fe-S binding. The C-terminal domain is required for viperin dimerization and is also important for interaction with the cytosolic iron-binding cofactor CIAO1.

[0030] Viperin mRNA was first identified among transcripts whose levels increased in fibroblasts during human cytomegalovirus (HCMV) infection. It was later confirmed that viperin, as an interferon-inducible factor, is also involved in the interferon-independent immune response.

[0031] One of the proven mechanisms of antiviral defense involving viperin is the blocking of the formation of newly synthesized virions. Viperin binds and inhibits the enzyme farnesyl diphosphate synthase (FDPS), which is necessary for the synthesis of lipids, particularly cholesterol. This results in decreased plasma membrane fluidity and disruption of PCT / RU2025 / 000306.

[0032] 9

[0033] the formation of lipid rafts—the sites of viral budding. This protective mechanism has been shown to be effective against many viral infections, such as influenza and HIV.

[0034] Another mechanism of viperin's antiviral defense is the specific inhibition of viral RNA synthesis. SAM contains an active site for redox reactions that convert cytidine triphosphate (CTP) to 3'-deoxy-3'4'-didehydro-CTP (ddgCTP). This is a derivative of one of the four nucleotides from which RNA polymerases construct RNA. Viral RNA polymerases identify ddgCTP and incorporate it into RNA, but due to the lack of a 3'-terminal hydroxyl, the enzyme cannot continue synthesis, leading to chain termination. This does not affect the activity of eukaryotic RNA polymerase or host RNA synthesis.

[0035] Interestingly, in humans, the RSAD2 gene is located adjacent to the oppositely directed CMPK2 gene, which encodes cytidylate monophosphate kinase 2, which is involved in the synthesis of CTP and UTP in mitochondria. Both RSAD2 and CMPK2 are coexpressed and function upon interferon induction, but are inhibited by lncRNA-CMPK2 (multi-exonic nuclear-localized long noncoding RNA (IncRNA)), whose gene is located adjacent to CMPK2.

[0036] Viperin plays various roles during pathogenic infection, sometimes detrimental to the host, as is typical for infections with DNA-containing viruses of the Herpesviridae family. For example, at the onset of HCMV infection, viperin functions as a protective antiviral protein, but in later stages, the viral protein vMia transports viperin to the mitochondria, where it facilitates various processes that facilitate viral replication (reconfiguration of mitochondrial metabolism, destruction of the actin cytoskeleton, etc.). Also, herpes simplex virus type 8 (Kaposi's sarcoma-associated herpesvirus, KSHV) uses viperin to stabilize its ORF44 helicase, which enhances its replication.

[0037] Viperin also blocks the replication of RNA-containing viruses of the Flaviviridae family (HCV, West Nile virus, Zika virus, Dengue virus, Tick-borne encephalitis virus, Japanese encephalitis virus, etc.) when it forms a complex with their proteins (e.g., NS3 or NS5A). The complex enters the mitochondria, triggering reactions that inhibit viral RNA synthesis. The structure of the C-terminal domain of viperin is important for complex formation in these cases. A similar reduction in replication has been shown for other families of RNA viruses: Reoviridae (viperin binds Rotavirus via the N-terminal protein NSP4) and Picornaviridae (the 2C protein of Enterovirus A71 interacts with viperin via the N-terminal domain).

[0038] Viperin synthesis is activated both directly through pathogenic RNA and DNA receptors and through type I and II interferon responses. Viperin expression is minimal in most cells, with the exception of some liver and heart tissues, adipose tissue, and immune cells. Increased viperin expression is caused by various DNA- and RNA-containing viruses, synthetic analogs of viral nucleic acids, and bacterial lipopolysaccharides.

[0039] The direct pathway for RSAD2 gene induction is mediated by the activation of RLR receptors, which directly recognize foreign RNA molecules and, via the mitochondrial antiviral sensor MAVS, lead to the translocation of IRF1 and IRF3 factors to the RSAD2 promoter and other genes. Viperin expression is also stimulated by pathogenic DNA via the cGAS-STING signaling pathway.

[0040] Viperin expression increases under the influence of a number of interferons: IFN-γ, IFN-I (α / [3]), and IFN-III (λ), whose activity increases upon antigen binding to cellular receptors. TLR receptors, which function in both innate and adaptive immunity, also activate RSAD2 expression.

[0041] Viperin itself modulates signaling pathways that include its expression, as well as those involved in parallel immune cascades. This is mediated by its ability to directly interact with, translocate, and participate in the activation of key factors in these cascades: MAVS, TRAF6, IRAKI, TBK1, and STING. Viperin plays a key role in the differentiation, maturation, and function of immune cells. In dendritic cells, it modulates IFN-I secretion induced by TLR7 and TLR9 receptors and is a factor in the differentiation and maturation of dendritic cells, as well as in the maturation of T lymphocytes mediated by them.

[0042] Viperin is important for both macrophage polarization (differentiation) and cytokine synthesis. Disruptions in RSAD2 expression in the M1 macrophage population have been observed in chronic inflammation, including autoimmune diseases.

[0043] High levels of RSAD2 gene expression in neutrophils have been observed during processes associated with IFN-I induction, such as lymphocytic choriomeningitis virus infection, rheumatoid arthritis, and asthma. The role of viperin in NETosis (the formation of neutrophil traps—the release of chromatin fibrils from neutrophils containing adherent bactericidal components of granules, nuclei, and cytoplasm) remains unclear.

[0044] RSAD2 is essential for B-cell activation via the NF-κB signaling pathway. It is produced in elevated quantities by all T-cell subsets, but its levels increase more significantly in CD4+ T-cells during immune disorders. RSAD2 influences IFN-I-mediated differentiation of CD4+ naive T-cells into Th17 and Tfh cells. Furthermore, viperin is essential for an optimal Th2 response and the activation of signals mediated by T-cell receptors, NF-κB, and AP-1.

[0045] In mitochondria, viperin inhibits the protein HADHB (the P-subunit of the mitochondrial trifunctional protein), which influences fatty acid metabolism. Viperin-mediated increases in mitochondrial AMP levels induce AMP-activated protein kinase (AMPK), which activates glucose transporter 4 (GLUT4), increasing the amount of cytoplasmic glucose available for glycolysis and lipid synthesis, as well as translocation of ChREBP to the nucleus, which triggers the synthesis of metabolic enzymes and induces de novo lipogenesis.

[0046] Thus, viperin is involved in numerous immune responses and metabolic regulation processes, which occur not only during immune defense but also in inflammation, autoimmune disorders, and cancer. Understanding the complex mechanisms involved in viperin-mediated immune responses has important therapeutic implications.

[0047] From the perspective of the claimed invention, it must be said that today in the world not only are there no antiviral and / or antimicrobial drugs whose action is mediated by the activation of viperin, but there is also a lack of scientific and patent information on any xenobiotics that activate its production.

[0048] DISCLOSURE OF THE INVENTION

[0049] This application implements the original idea of ​​searching for next-generation drugs that activate an important factor of innate immunity, viperin, thereby acquiring their antiviral and antimicrobial activity. PCT / RU2025 / 000306

[0050] 12

[0051] This problem can be solved by using derivatives of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine of general formula I:

[0052]

[0053] (I)

[0054] Where

[0055] R 1represents AlkS, AlkSO, AlkSO2, COOAlk, CON(Alk)2, substituted or unsubstituted Ph, benzyl, furyl, pyridyl;

[0056] R 2 represents H, Aik, Ph;

[0057] R 3 is OH, OAlk, NHOAlk, NAlk2, or a saturated 6-membered nitrogen-containing heterocyclic ring attached via a nitrogen atom in a carbonyl group, and optionally containing a 2nd heteroatom in the ring selected from X, where X is O, S, NMe;

[0058] Aik is H, Me, Et, Pr, or i-Pr.

[0059] The present invention also applies to pharmaceutically acceptable salts, solvates and hydrates of the compounds of formula I.

[0060] The proposed compounds have antiviral and antimicrobial activity and can be used for the treatment and / or prevention of diseases caused by viral and / or microbial infections.

[0061] In a preferred embodiment, the invention relates to compounds of formula (1) selected from the group consisting of

[0062] 7-amino-2-(methylthio)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid and its pharmaceutically acceptable salts, such as

[0063] sodium 7-amino-2-(methylthio)[1,2,4]trizolo[1,5-a]pyrimidine-6-carboxylate, potassium 7-amino-2-(methylthio)[1,2,4]trizolo[1,5-a]pyrimidine-6-carboxylate, or 7-amino-2^ 1 -[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid and its salts or

[0064] 7-amino-2-R 1 -6-hydroxycarboxamide- [1,2,4]trizolo [1, 5 -a] pyrimidine.

[0065] The term "pharmaceutically acceptable salts of a compound of formula (I)" means salts of an inorganic or organic base that possess the required pharmacological activity of the parent compound. These salts may be obtained in situ during the synthesis, isolation or purification of the compound of formula (I) or prepared specifically.

[0066] Pharmaceutically acceptable salts with bases are characterized by the fact that they contain therapeutically active non-toxic addition salt forms with bases, which compounds of formula I are capable of forming. The said addition salts with bases can be obtained by treating compounds represented by the general formula I with suitable bases, alkalis, for example, inorganic alkalis: sodium, calcium or potassium hydroxide; organic bases, for example: compounds containing a guanidine fragment, ammonia, dimethylamine, triethylamine, compounds having a strongly basic amino group.

[0067] The compounds or salts of the present invention may exist as solvates, where the solvent may be water (i.e., hydrates) or organic solvents (e.g., methanol, ethanol, or acetonitrile, forming methanolate, ethanolate, and acetonitrile, respectively). A detailed description of the properties of such salts is provided in the article by Berge SM et al. Pharmaceutical Salts. / . Pharm. Sci. 1977; 66: 1-19.

[0068] The term "pharmaceutically acceptable" means that the noun it defines is suitable for use as a pharmaceutical product or as part of a pharmaceutical product.

[0069] The term "therapeutically effective amount" refers to the total amount of each active substance sufficient to demonstrate a significant benefit to the patient, such as a reduction in viral or microbial load.

[0070] The term "substituted" refers to 1-3 substituents selected from halogen atoms, Aik and AlkO, where Aik is selected from Me, Et, Pr or i-Pr.

[0071] The term "saturated nitrogen-containing heterocyclic ring attached via a nitrogen atom in a carbonyl group and optionally containing a 2-membered heteroatom in the ring selected from X, where X is O, S, NMe" refers to a 3-6-membered heterocyclic ring, preferably piperidine, morpholine, thiomorpholine, piperazine, which may also be substituted, for example, as indicated above.

[0072] The compounds can be obtained and used in crystalline form. Compounds of general formula I and their pharmaceutically acceptable salts have been studied against pathogenic viruses and can be used to produce medicinal products for the treatment or prevention of viral diseases caused by respiratory viruses or coronaviruses, in particular, for example, those caused by influenza, acute respiratory viral infections (ARVI) viruses (rhinovirus), or the SARS-CoV-2 coronavirus, as well as hepatitis B or C viruses.

[0073] Compounds of general formula I and their pharmaceutically acceptable salts can be used to obtain medicinal products based on them for the treatment or prevention of diseases caused by pathogenic microbes of the ESCAPE group, in particular, for example, Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter, mycobacterium tuberculosis.

[0074] The invention also relates to a pharmaceutical composition containing an effective amount of a compound of formula I or a pharmaceutically acceptable salt, solvate or hydrate thereof as an active component and pharmaceutically acceptable excipients.

[0075] Pharmaceutically acceptable excipients include diluents, auxiliary agents and / or pharmaceutical carriers used in the pharmaceutical industry.

[0076] Pharmaceutical carriers include those traditionally used in pharmaceutical formulations for medicinal products. When preparing compositions for oral administration, binders, lubricants, disintegrants, solvents, diluents, stabilizers, suspending agents, colorless agents, and flavoring agents are used; in injection formulations, antiseptic agents, solubilizers, and stabilizers are used; and in topical formulations, bases, diluents, lubricants, and antiseptic agents are used.

[0077] The pharmaceutical composition can be used as a medicinal product in the form of a tablet, capsule, pill, powder, or in a form suitable for topical application. If the compounds of the claimed structure are water-soluble, they can be used to prepare parenteral administration forms and effervescent tablets.

[0078] The pharmaceutical composition can be obtained by mixing the active component in an effective amount and an appropriate excipient.

[0079] The subject of the invention also includes a method for preventing and / or treating a disease caused by a viral or microbial infection, comprising administering or applying to a subject (patient) in need thereof a compound of formula I, a pharmaceutically acceptable salt or a pharmaceutical composition based on them in a therapeutically effective amount.

[0080] The method of treatment using the compounds, pharmaceutical composition or medicinal product based on them claimed in the invention is also effective against strains resistant to currently existing drugs.

[0081] In vitro experiments have shown the ability of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula I to activate the production of viperin in vivo, achieving a sanitizing effect against a number of viruses and pathogenic microorganisms.

[0082] Taking this into account, compounds of general formula (I), their pharmaceutically acceptable salts, solvates, and hydrates can be used as activators of viperin production, as a method of treating humans, including infectious diseases whose dynamics are sensitive to the level of viperin in the body.

[0083] The compounds, as studies have shown, are suitable for the combined treatment of viral and microbial infections, as well as for combination therapy.

[0084] Examples of the preparation of compounds according to the invention are shown below.

[0085] The examples include specific embodiments of the present invention, but do not limit the present invention. It is understood that the examples are intended to illustrate some embodiments, reproducibility and suitability of the invention for practical use. They are presented as the most appropriate and understandable descriptions of its methods and conceptual aspects. General method for the synthesis of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidines of general

[0086] formulas!

[0087]

[0088] The first stage of the synthesis involves the production of ethyl 2-cyano-3-ethoxy-2-propenoate (2) from cyanoacetic ester (1) and triethyl orthoformate in the presence of acetic anhydride with a yield of at least 57%. The quality of the resulting product allows it to be used in the next stage without further purification.

[0089] The second stage of the synthesis involves the closure of the pyrimidine ring and consists of the reaction of 2-cyano-3-ethoxyacrylate (2) obtained in the previous stage with 3-amino-1H-1,2,4-triazole (3) in acetic acid. In this case, both stages of the chemical process, the addition of 2-cyano-3-ethoxyacrylate at the amino group with the elimination of ethyl alcohol and the closure of the pyrimidine ring, occur without the isolation of an intermediate product. Ethyl ester of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid (4) is obtained in 74% yield and the resulting substance is of sufficient purity to be used in the next stage without further purification.

[0090] The third stage of the synthesis is the alkaline hydrolysis of the ester group of ethyl 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate (4) and the formation of the corresponding acid (5). The yield of the resulting acid, the key scaffold, is 91% and it is used without further purification in the synthesis of the target 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of formulas (6), (7), (8), or (9) or the corresponding salt.

[0091] EXAMPLES

[0092] Example 1: Synthesis of ethyl 2-cyano-3-ethoxy-2-propenoate (2) A 1000 ml round-bottomed single-neck flask equipped with a magnetic stirrer and reflux condenser is charged with 60 ml of cyanoacetic ester, 106 ml of triethyl orthoformate and 160 ml of acetic anhydride. The reaction mixture is boiled for 4 hours, cooled to room temperature and the ethyl alcohol and the remaining unreacted reagents are distilled under vacuum until a straw-colored liquid oil is formed. 40 ml of isopropanol is added to it, the mixture is stirred and left for 16 hours at -18 °C. The resulting white crystalline precipitate is then filtered off and washed on the filter with very cold isopropanol (2 ^ 50 ml). 38.3 grams of ethyl 2-cyano-3-ethoxyacrylate were obtained (yield 43%). The combined mother liquors were evaporated, 15 ml of isopropanol was added, and the mixture was cooled for 18 hours at -18°C. The resulting precipitate was filtered, yielding an additional 12.6 grams of ethyl 2-cyano-3-ethoxyacrylate. The mixture was dried at room temperature for 24 hours.The total yield of product 2 is 50.9 grams (57.2%). Tm = 50-52 °C.

[0093] Synthesis of ethyl 7-amino-2-(methylthio)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate (4)

[0094] In a 500 ml round-bottomed single-necked flask equipped with a magnetic stirrer and a reflux condenser, 30 grams of ethyl 2-cyano-3-ethoxyacrylate (2) (0.177 mol) and 23 grams of 3-amino-5-methylthio-1H-1,2,4-triazole (3) (0.177 mol) are suspended in 90 ml of glacial acetic acid. Bring to a boil and the resulting solution is boiled for 3.5 h. During the reaction (after about 2 h), a suspension is obtained. The reaction is monitored by TLC (hexane: acetone 2:1 and then chloroform: methanol 10:1). The reaction mass is cooled to 50 °C and acetic acid is distilled under vacuum to a dry residue. 400 ml of water are added to it and left for 16 hours at +4-6 °C. The light yellow precipitate is filtered off, washed with cold water (3x200 ml), squeezed out and dried at 100 °C for 10 hours. 33.1 grams of ethyl 7-amino-2-(methylthio)[1,2,4]-triazolo-[1,5-a]pyrimidine-6-carboxylate (74.02%) are obtained, of sufficiently good quality to be used in the next step without purification. Purity 91% (HPLC). Tm = 211-213 °C.

[0095] -Amino-2-(methylthio)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid (5) In a 1000 ml flat-bottomed flask equipped with a magnetic stirrer, 28.0 g of ethyl 7-amino-2-(methylthio)[1,2,4]-triazolo-[1,5-a]pyrimidine-6-carboxylate (4) (0.11 mol) are suspended in 300 ml of water and 11.7 g of solid sodium carbonate (11 mmol) are sprinkled. The reaction mixture is boiled until a clear solution is formed (~ 4.5 hours), cooled to room temperature and 20 ml of acetic acid are slowly added to it in portions. This gives a white suspension, to which 700 ml of water are then added. The mixture is cooled at +4°C for 16 hours and the amorphous white precipitate is filtered off. The mixture is washed with cold water (2 x 200 ml), squeezed out, and dried at 110°C to yield 22.72 grams of the target 7-amino-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid (5).

[0096] Yield: 91%. Mp = 294-296 °C. Mass (El), m / z I relat.(%)): 224.2260 [M-N]' (50). C7H7N5O2S. *H NMR (300 MHz, DMSO-de) 5 2.74 (s, 3H, CH3S), 8.46 (br s, 2H, NH2), 8.75 (s, 1H, CH).

[0097] Elemental analysis of C7H7N5O2S. Calculated: C, 37.33; H, 3.13; N, 31.09. Found: C, 37.41; H, 3.02; N, 31.12.

[0098] The analytical data for compound 5 correspond to those described in Reiter J, Pongo L, Dvortsak P. Triazoles. VIII. The reaction of 5-amino-l,2,4-triazoles with ethyl 2-cyano-3-ethoxyacrylate and 2-cyano-3-ethoxyacrylonitrile. J. Heterocyclic Chem. 1987; 24 (4): 1149–54.

[0099] Example 2: Sodium 7-amino-2-(methylthio)[1,2,4]trizolo[1,5-a]pyrimidine-6-carboxylate (6a).

[0100] In a 1000 ml flat-bottomed flask equipped with a magnetic stirrer, 22.72 grams (0.1 mol) of 7-amino-6-carboxy-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine (5) are suspended in 300 ml of methanol and a freshly prepared solution of sodium methoxide is added (2.32 grams of sodium (0.1 mol) in 80 ml of methanol). At the moment of addition of the sodium methoxide solution, a slight liquefaction of the suspension is achieved, it is kept at room temperature for 4 hours and during this time it thickens greatly. The resulting suspension is treated with 500 ml of PCT / RU2025 / 000306

[0101] 19

[0102] diethyl ether and mix well and leave at minus 18 °C for 16 hours. The resulting white amorphous precipitate is filtered and washed with 50 ml of diethyl ether. The mixture is squeezed out and dried first in air for 3 hours and then at 110 °C for 6 hours. 22.11 grams of sodium 7-amino-2-(methylthio) [1,2,4]trisolo [1,5-a]pyrimidine-6-carboxylate are obtained.

[0103] Output: 86%. T пл= 312-314 °C. Mass (El), m / z Ireiai J) 224.2260 [M-N]' (52). C7H7N5O2S. 'H NMR (300 MHz, DMSO-d6) 52.68 (s, 3H, CH3S), 8.21 (br s, 1H, NH), 8.73 (s, 1H, CH), 10.23 (br s, 1H, NH).

[0104] Elemental analysis: C7H6NsNaO2S. Calculated: C, 34.01; H, 2.45; N, 28.33. Found: C, 33.97; H, 2.43; N, 28.42.

[0105] The analytical data for compound 6a correspond to those described in Reiter J, Pongo L, Dvortsak P. Triazoles. VIII. The reaction of 5-amino-l,2,4-triazoles with ethyl 2-cyano-3-ethoxyacrylate and 2-cyano-3-ethoxyacrylonitrile. J. Heterocyclic Chem. 1987; 24 (4): 1149–54.

[0106] Example 3: Calcium di-7-amino-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate dihydrate (6b)

[0107] To a solution of 4.0 grams of sodium 7-amino-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate in 60 ml of water, 10 ml of an aqueous solution of calcium chloride (0.9 g) are added, immediately producing a fine white suspension. The reaction mixture is kept at room temperature for 24 hours and then filtered, washed with water, alcohol, and acetone. Dry at 100°C for 48 hours.

[0108] Yield 92%. Mass (EI), m / z I relat .(%)): 224.2260 [MH]⁻ (14). C7H7N5O2S. l H NMR (300 MHz, DMSO-d6) 6 NMR (300 MHz, D2O) 82.68 (s, 3H, CH3S), 8.23 ​​(br s, 1H, NH), 8.74 (s, 1H, CH), 10.21 (br s, 1H, NH).

[0109] Elemental analysis: Ci4CaHi2Nio04S2 x 2H20. Calculated: C, 32.06; H, 3.07; N, 26.70. Found: C, 31.89; H, 2.85; N, 26.54

[0110] Example 4: Argininium 7-amino-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate hydrate (6c)

[0111] To a solution of 4.5 grams of sodium 7-amino-2-(methylthio)[1,2,4]triazolo[1,5-p]pyrimidine-6-carboxylate in 60 ml of water, add 25 ml of an aqueous solution of arginine hydrochloride (3.8 g), the reaction mixture in the form of a solution is boiled for 10 minutes, left for 24 hours at room temperature. The resulting solution is evaporated under vacuum to dryness and the residue is recrystallized twice from water, washed with ice-cold ethanol and acetone. Dry at 100 °C for 48 hours. Yield 84%. T П l = 276-279 °C. Mass (El), m / z I re iai.(° / >)') 224.2260 [MH]' (79). C7H7N5O2S. 'H NMR (300 MHz, D2O) 5 1.5-1.8 (dm, 4H, CH2CH2), 2.68 (s, 3H, CH3S), 3.19 (t, 3H, CH2), 3.76 (t, 1H, CH), 8.69 (s, 1H, CH).

[0112] Elemental analysis: C13H21N9O4S. Calculated: C, 37.40; H, 5.55; N, 30.20. Found: C, 37.44; H, 5.53; N, 30.39.

[0113] Example 5: Ethanol amine 7-amino-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate (6d)

[0114] To a suspension of 3.8 grams of the previously obtained 7-amino-2-(methylthio)[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylate in 80 ml of ethanol, 1 ml of ethanol is added with stirring. The reaction mixture initially goes into solution, and then a white precipitate quickly forms. The mixture is maintained with stirring at room temperature for 4 hours. The suspension is cooled to 4 °C for 3 hours, and the finely crystalline white precipitate is filtered, washed with ice-cold methanol and acetone. The mixture is crystallized from an ethanol / water mixture. Dry at 100 °C for 48 hours.

[0115] Yield 94%. T пл = 239-241 °C. Mass (El), m / z

[0116]

[0117] 224.2260 [MH]' (43). C7H7N5O2S. 'H NMR (300 MHz, D2O) 52.65 (s, 3H, CH3S), 3.17 (t, 2H, CH2), 3.62 (t, 2H, CH2), 8.67 (s, 1H, CH).

[0118] Elemental analysis of C9H14N6O3S. Calculated: C, 37.75; H, 4.93; N, 29.35. Found: C, 37.72; H, 4.90; N, 29.43.

[0119] Example 6: 7-Amino-2-(methylsulfinyl)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid (be)

[0120] To a nitrating mixture consisting of 0.086 ml of fuming nitric acid (D = 1.46) and 0.4 ml of concentrated sulfuric acid, 0.15 g of dry 7-amino-2-(methylthio)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid is added in small portions with vigorous stirring so that the temperature does not rise above 20 °C. The reaction mixture is stirred at the specified temperature for 8 hours and diluted with cold water (2 ml). The formed precipitate is filtered and washed with a minimum amount of cold water, then with alcohol. The resulting product is recrystallized from an alcohol-water mixture. Yield 0.100 grams.

[0121] Yield 56%. Mp = 288-290 °C. Mass (El), m / z I reial .(%)) 240.2284 [M-N]' (48). C7H7N5O3S. H NMR (300 MHz, DMSO-d6) 53.29 (s, 3H, CH3S), 8.34 (br s, 1H, NH), 8.78 (s, 1H, CH), 9.32 (br s, 1H, NH). PCI7RU2025 / 000306

[0122] 21

[0123] Elemental analysis: C7H7N5O3S. Calculated: C, 34.85; N, 2.92; N, 29.03. Found: C, 34.81; N, 2.87; N, 29.12.

[0124] Example 7: Sodium 7-amino-5-methyl-2-(methylthio)[1,2,4]trizolo[1,5-a]pyrimidine-6-carboxylate (6f)

[0125] The synthesis of the compound was carried out according to Example 1-2 using triethyl orthoacetate instead of orthoformic ether.

[0126] Yield 74%. Mp = 264-266 °C. Mass (El), m / z

[0127]

[0128] 239.2556 [M-N]' (61). C8H8N5O2S. 'H NMR (300 MHz, DMSO-d6) δ 2.25 (s, 3H, CH3S), 2.79 (s, 1H, CH), 8.34 (br s, 1H, NH), 8.78 (s, 1H, CH), 9.32 (br s, 1H, NH).

[0129] Elemental analysis of Cs^NsNaChS. Calculated: C, 36.78; H, 3.09; N, 26.81. Found: C, 36.67; H, 3.07; N, 26.70.

[0130] Example 8: Sodium 7-amino-2-phenyl-[1,2,4]trizolo[1,5-a]pyrimidine-6-carboxylate (6g)

[0131] The compound was synthesized according to Example 1-2 using 3-amino-5-phenyl-1H-1,2,4-triazole (CAS 4922-98-9) instead of 3-amino-5-methylthio-1H-1,2,4-triazole, obtained according to the method of Dolzhenko A. V, Pastorin G., Dolzhenko A. V, Chui WK Tetrahedron Lett., 2009; 59: 2124. https: / / doi.org / 10.1016 / j.tetlet.2009.02.172.

[0132] Yield 63%. T пл = 248-250 °C. Mass (El), m / z

[0133]

[0134] 254.2324 [MH]' (32). C12H9N5O2. 'H NMR (300 MHz, DMSO-d6) δ 7.73 and 8.18 (m, 5H, Ph), 8.26 (br s, 1H, NH), 8.79 (s, 1H, CH), 9.53 (br s, 1H, NH).

[0135] Elemental analysis of CnHsNsNaCh. Calculated: C, 51.99; H, 2.91; N, 25.26. Found: C, 52.13; H, 2.96; N, 25.32.

[0136] Example 9: 7-Amino-2-methylthio-6-methoxycarbonyl[1,2,4]trizolo[1,5-a]pyrimidine (7)

[0137] To a suspension of 2 grams of 7-amino-2-(methylthio)-[1,2,4]triazolo[1,5-a]pyrimidine-6-carboxylic acid (5), obtained according to Example 1, in 50 ml of dichloromethane are added dropwise 2 ml of thionyl chloride and the mixture is boiled for 3-4 hours until the precipitate is completely dissolved. The resulting solution is evaporated under vacuum to dryness and the resulting 7-amino-2-methylthio-[1,2,4]triazolo[1,5-a]pyrimidine-5-carbonyl chloride is used in further synthesis without additional purification.

[0138] A solution of 100 mg of 7-amino-2-methylthio[1,2,4]trizolo[1,5-a]pyrimidine-5-carbonyl chloride in 3 ml of dry acetone is added dropwise to 20 ml of methanol while cooling to 0-5 °C. The resulting solution is left for 2 hours at room temperature, evaporated under vacuum, and the residue is crystallized from methanol. PCT / RU2025 / 000306

[0139] 22

[0140] Yield 78%. T П l = 143-145 °C. Mass (El), m / z ( / re / aZ .(%): 240.2556 [M+H] +(49). C8H9N5O2S. 'HNMR (300 MHz, DMSO-d6) δ 2.71 (s, 3H, CH3S), 3.74 (s, 3H, OCH3), 8.56 (br s, 2H, NH2), 8.81 (s, 1H, CH).

[0141] Elemental analysis of C8H9N5O2S. Calculated: C, 40.16; H, 3.79; N, 29.27. Found: C, 40.19; H, 3.70; N, 29.21.

[0142] Example 10: 7-Amino-2-methylthio-6-hydroxycarboxamide-[1,2,4]trizolo[1,5-a]pyrimidine (8a)

[0143] To a solution of 100 mg of 7-amino-2-methylthio-[1,2,4]trizolo[1,5-a]pyrimidine-5-carbonyl chloride, obtained according to Example 9, in 3 ml of dry acetone, 58 mg of dry hydroxylamine hydrochloride are added, and then, while cooling to 0-5 °C, 0.9 ml of triethylamine is added dropwise. The resulting solution is left for 4 hours at room temperature, evaporated under vacuum, the residue is triturated with water, filtered, and the resulting white solid is crystallized from ethanol.

[0144] Yield 64%. Mp = 231-234 °C. Mass (El), m / z I re iat.(%y) 241.2437 [M+H] +(66). C7H8N6O2S. 'H NMR (300 MHz, DMSO-d6) 8 2.70 (s, 3H, CH3S), 8.34 (br s, 2H, NH2), 8.56 (br d, 1H, NH), 8.81 (s, 1H, CH), 9.12 (br d, 1H, OH).

[0145] Elemental analysis of C7H8N6O2S. Calculated: C, 35.00; H, 3.36; N, 34.98. Found: C, 35.13; H, 3.30; N, 35.01.

[0146] Example 11: 7-Amino-2-ethoxycarbonyl-6-hydroxycarboxamide-[1,2,4]trizolo[1,5-a]pyrimidine (8b)

[0147] The compound was synthesized according to Example 10 using 3-amino-5-ethoxycarbonyl-1H-1,2,4-triazole (CAS 63666-11-5) as the starting material, obtained according to the method of Wang H., Lee M., Peng Z., Blazquez B., Lastochkin E., Kumarasiri M., Bouley R., Chang M., Mobashery SJ Med. Chem. 2015; 58: 4194. doi: 10.1021 / jm501831g.

[0148] Yield 38%. Tm = 128-131 °C. Mass (El), m / z (1^%): 267.2137 [M+H] + (23). C9H10N6O4. 'HNMR (300 MHz, DMSO-d6) 8 1.54 (t, 3H, CH3), 6.63 (q, 2H, CH2), 8.34 (br s, 2H, NH2), 8.81 (s, 1H, CH), 9.07 (br d, 1H, OH).

[0149] Elemental analysis of C9H10N6O4. Calculated: C, 40.61; H, 3.79; N, 31.57. Found: C, 40.55; H, 3.65; N, 31.63.

[0150] Example 12: 7-Amino-2-methylthio-6-methylcarboxamide-[1,2,4]trizolo[1,5-a]pyrimidine (9)

[0151] The compound was synthesized according to Example 10 using a solution of methylamine in isopropyl alcohol. Yield 76%. Tm = 193-195 °C. Mass (El), m / z I re iat.( )) 239.2708 [M+H] + (39). C8HION6OS. 'H NMR (300 MHz, DMSO-d6) δ 2.71 (s, 3H, CH3S), 2.88 (s, 3H, NCH3), 8.32 (br s, 2H, NH2), 8.81 (s, 1H, CH), 9.34 (br s, 1H, NH).

[0152] Elemental analysis of CsHioNeOS. Calculated: C, 40.33; H, 4.23; N, 35.27. Found: C, 40.27; H, 4.29; N, 35.54.

[0153] Example 13: Activation of viperin production by 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I)

[0154] High-throughput cDNA sequencing (RNA-seq) was used to identify changes in the transcriptome of canine epithelial cells, the Madin-Darby Canine Kidney (MDCK) cell line, induced by compound 6a. The MDCK cell line was treated with compound 6a for 3 h at a concentration of 10 μg / ml; untreated cells were used as a control. Each experiment was performed in three biologically independent replicates. Cells were harvested (3-4 million cells per sample) and resuspended in Trizol (Invitrogen). Total RNA was isolated from the samples by extraction with Trizol reagent (Invitrogen) according to the manufacturer's standard protocol. Preparation of cDNA libraries consisted of the selection of poly-A mRNA molecules from total RNA using magnetic beads bearing poly-T oligonucleotides; mRNA fragmentation; synthesis of the first and second strands of cDNA; adapter ligation and PCR enrichment.Sample preparation for sequencing was performed using the NEBNext® Poly(A) mRNA Magnetic Isolation Module and NEBNext® Ultra™ RNA Library Prep Kit for Illumina®. Six cDNA libraries were obtained. Sequencing was performed using an Illumina Novaseq6000 with a read length of 150 nucleotides, and a minimum of 20 million reads per library. Sequencing results were analyzed using standard procedures for read set quality assessment, filtering, and normalization.To process the RNA-seq data, we used a number of software packages, including HISAT2 for mapping the obtained nucleotide reads to the reference genome; samtools for manipulating the mapped data, obtaining mapping statistics, determining the degree of overamplification of cDNA libraries, and other information; and a number of R programming language packages (DESeq2, edgeR) for intersecting genomic coordinates with the mapped reads, determining the statistical significance of the expression difference for each gene, and so on. Nucleotide reads were aligned to the Canis lupus familiaris Ensembl CanFam3.1 genome assembly. Genes with a combined expression of at least 100 reads across all samples were selected for analysis.

[0155] Among the differentially expressed genes, the RSAD2 (Radical S-adenosyl methionine domain containing 2) gene, which encodes the antiviral protein viperin, is of particular interest. Viperin is a multifunctional, nonspecific antiviral defense protein that inhibits many viruses, such as CHIKV, HCMV, HCV, DENV, WNV, SINV, influenza virus, and others (Mattijssen, Pruijn, 2012). The mechanisms of viperin's antiviral activity are currently being extensively studied (Rivera-Serrano et al., 2020), including the induction of type I interferons and the destruction of lipid rafts. Viperin synthesis is activated by type I interferons and gamma interferons; non-interferon induction of viperin has also been described (Rivera-Serrano et al., 2020). The identified transcriptome change was validated by quantitative RT-PCR on MDCK cell culture.

[0156] The ability of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) to induce viperin expression was compared. To determine the level of RSAD2 gene expression, MDCK cells were treated with compound 6a, 6b, or 8a at a concentration of 10 μg / ml for 3 hours; untreated cells served as a control. The experiment was performed in a blinded manner; samples were refrigerated, including the control. Total RNA was isolated using the RNAEasy reagent kit (Qiagen) according to the manufacturer's protocol. RNA samples were treated with Turbo DNase (Life Technologies) to remove traces of genomic DNA. RNA quantity and purity were determined spectrophotometrically and by electrophoresis in a 1% agarose gel. cDNA synthesis was performed from 1 mg of total RNA using random hexanucleotides (random primers, Evrogen) and SuperScript III reverse transcriptase (Life Technologies) as primers.The obtained cDNA samples were used as a template for quantitative RT-PCR (qRT-PCR) performed using qPCRmix-HS SYBR (Evrogen) on a LightCycler 480 (Roche). Gene-specific primers were selected using the Primer3 program (https: / / www.bioinformatics.nl / cgi-bin / primer3plus / primer3plus.cgi) for the canine viperin gene mRNA sequence annotated in the databases (ENSCAFT00000005279.4). The following program was used: 95°C, 20 sec; 61°C, 20 sec, 72°C, 30 sec, 40 cycles. To ensure statistical significance of the results, technical triplicates for each sample were used. The obtained data were analyzed using the LinRegPCR v2014.6 program. Data were normalized to GAPDH mRNA.

[0157] The results of the experiment are shown in Fig. 1A. An increase in the number of RSAD2 transcripts was observed for all three derivatives of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine of general formula (I): ~30 times for compound 6a, ~5 times for compound 6a, and ~50 times for compound 8a.

[0158] Western blotting with anti-viperin antibodies (Viperin Polyclonal Antibody, Invitrogen) was used to determine protein synthesis levels. Actin, detected with the Anti-Actin antibody (Sigma-Aldrich), served as an internal control. In addition to exposing MDCK cells to the test substances, an experiment was also conducted on MDCK cells infected with the influenza A virus. Cells unexposed to the substances, infected with the virus, and uninfected cells served as controls. The exposure time was 3 hours, and the concentration was 10 μg / ml. The experimental results are shown in Fig. 1B.

[0159] Influenza virus infection leads to a ~4-fold increase in viperin levels in all cases. When using compound 6a, viperin levels increase ~6-7-fold and are virtually independent of the presence of infectious agents. Compounds 6a and 8a increase viperin levels ~4-fold, and in combination with viral infection, ~5-6-fold (see Fig. 1).

[0160] Thus, we have shown that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) activate viperin synthesis in epithelial cells at both the RNA and protein levels. The induction of viperin synthesis by these substances is comparable to that observed during cell infection with the influenza virus; upon exposure of infected cells to the substances, the amount of viperin increases even further.

[0161] Example 14: Therapeutic and prophylactic antiviral action of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against the influenza virus

[0162] Influenza is a respiratory infection that causes significant harm to the human population, causing annual epidemics during the cold season. In the past, this disease has caused several global pandemics spanning the globe, including the "Spanish flu" of 1918, which caused the deaths of over 50 million people. A comparative study of the efficacy of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) was conducted in accordance with the Guidelines for the Study of Specific Antiviral Activity of Pharmacological Substances (2005), Order of the Ministry of Health of Russia dated August 23, 2015.2010 No. 708n "On approval of the rules of laboratory practice in the Russian Federation", and also taking into account the recommendations of international regulatory authorities (World Health Organization, Center for Expertise and Study of Drugs of the Food and Drug Administration of the US Department of Health and Human Services).

[0163] Currently available medications for the treatment of influenza are divided into two broad groups: symptomatic agents, which alleviate the clinical manifestations of the disease, and etiotropic agents, which directly block viral replication. The latter include M2 ​​channel blockers (amantadine and rimantadine), neuraminidase inhibitors (oseltamivir and zanamivir), and viral polymerase inhibitors (baloxavir marboxil and favipiravir).

[0164] Remantadine and amantadine have lost their role as influenza preventatives and treatments, as most currently circulating strains carry resistance mutations. Neuraminidase inhibitors remain relevant, but cases of the emergence and spread of resistant strains in the population were already noted in 2008, and a recurrence cannot be ruled out. Baloxavir promotes the emergence of resistant strains even after a single dose, and favipiravir has been associated with numerous side effects, primarily due to teratogenicity.

[0165] A study of the efficacy of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) was conducted on pure female Balb / c mice aged 5-7 weeks and weighing 18-20 g. The selection of this strain fully complies with the guidelines. Twenty-seven mice per group were used to determine protective activity, reduce mortality and viral load in the lungs, and evaluate the histological pattern of lung lesions.

[0166] Animals were obtained from the Federal State Budgetary Scientific Institution "Scientific Center for Biomedical Technologies of the Federal Medical and Biological Agency of Russia," Stolbovaya Branch. Animals were maintained under standard conditions in accordance with SOP B-P-004 / 03-24 "Routine Procedures for the Care of Laboratory Animals." During the acclimatization and experimental periods, mice were housed in polycarbonate cages with a diameter of 1200 cm. 2, in groups of 15 individuals intended for determining body weight and mortality dynamics, and separately in groups of 12 individuals intended for organ harvesting, on a bedding. The cages used are covered with steel mesh lids with a feeding recess. Complete granulated feed (OOO "Laboratorkorm", complete compound feed PK-120 for keeping laboratory animals (mice, rats, hamsters) was given ad libitum in the feeding cavity of the steel mesh cover of the cage. The animals were given water obtained by a water treatment system that meets the standards of SanPiN 1.2.3685-21 "Hygienic standards and requirements for ensuring the safety and (or) harmlessness of environmental factors for humans" in standard drinkers with steel spouts ad libitum. Dust-free corn litter for laboratory animals (medium fraction) No. 2 was used as bedding.After receipt and prior to the study, laboratory animals were kept in quarantine for 14 days and then acclimated for an additional 10 days in group housing in cages. During this period, the animals' clinical condition was monitored daily by visual inspection by veterinarians. Animals with abnormalities detected during the examination were not included in the experimental groups. Animals were randomly assigned to groups. The drugs, in the form of an aqueous solution prepared ex tempore, were administered intragastrically through a gavage tube at a dose of 0.5 ml daily from the first to the sixth day after infection. The negative control group consisted of animals that received the solvent (water) in an equivalent volume, while the positive control group consisted of animals that received ribavirin at a dose of 10 mg / kg. After drug administration and non-invasive testing, the animals were routinely euthanized using a CO2 chamber.

[0167] To conduct the study, animals were divided into 5 experimental groups:

[0168] Group 1 - negative control, placebo - individuals received 0.5 ml of water.

[0169] 2nd group - treatment with compound ba at a dose of 100 mg / kg in 0.5 ml of water,

[0170] 3rd group - treatment with compound be at a dose of 100 mg / kg in 0.5 ml of water,

[0171] Group 4 - treatment with compound 8a at a dose of 100 mg / kg in 0.5 ml of water,

[0172] Group 5—positive control, treatment with Tamiflu at a dose of 10 mg / kg in 0.5 ml of water. Influenza virus infection was shown to cause manifestation and progression of the pathological process in animals. External signs of the disease included limited mobility, increased respiratory rate, and decreased feed and water consumption, leading to weight loss and death. These signs are typical of influenza pneumonia. Data on the dynamics of animal mortality during influenza pneumonia are presented in Table 1, and virus titer values ​​are grouped in Table 2.

[0173] Table 1. Dynamics of mortality in Balb / c mice and indicators of protective activity of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) during experimental lethal influenza pneumonia caused by the influenza virus A / Aichi / 2 / 68 (H3N2) _ _ _

[0174] Animal mortality Index Reliability Drug, by days after infection Mortality,

[0175] protection, differences from dose, mg / kg %

[0176] 6 7 8 9 10 I 12 13 14 % placebo, p Distilled

[0177] 1 2 1 1 1 2 57.1 0 - water (placebo)

[0178] Compound 6a, 100 mg / kg 1 1 20 65.0 0.994 Compound 6a, 100 mg / kg 1 2 1 26.7 53.3 0.206 Compound 8a, 100 mg / kg 1 1 20 65.0 0.151

[0179]

[0180] Tamiflu, 20 mg / kg 1 6.7 84.1 0.005* Note: * - differences with placebo characteristics are significant at p<0.05

[0181] Table 2. Values ​​of A / Aichi / 2 / 68 (H3N2) virus titer in the lungs of mice after administration of drugs

[0182] Virus titer in the lungs, 1 gTCIDso

[0183] Drug, dose, mg / kg Mean Standard Value (M) Deviation (SD) from the placebo group (p) Distilled

[0184] water (placebo) 5.792 0.459 —

[0185] Conn. 6a, 100 mg / kg 3.500 2.356 0.8113 Comp. be, 100 mg / kg 4.708 0.510 0.0002* Comp. 8a, 100 mg / kg 3.625 0.905 0.0014* Tamiflu, 20 mg / kg 2.388 0.656 0.0457*

[0186]

[0187] Note: * - differences with placebo characteristics are significant at p<0.05

[0188] The presented data indicate that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) have antiviral activity against the influenza virus, which is confirmed by an increase in survival and a decrease in viral titers in the lungs of mice compared to mice in the control group that received a placebo (statistical significance level p<0.05).

[0189] Example 15: Therapeutic and prophylactic antiviral activity of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against mouse-adapted human respiratory syncytial virus. Respiratory syncytial infection is an acute viral disease that primarily affects the lower respiratory tract, caused by an RNA-containing virus (RS virus). Currently, two serological strains of the RS virus are distinguished, but they both belong to the same serotype. This virus has low stability in the external environment and is sensitive to disinfectants and boiling. Infection with this disease occurs through airborne droplets from an infected person. The virus enters the body through the pharynx and nasopharynx. Then, after multiplying in the mucous membrane, it spreads to the lower respiratory tract, causing inflammation at the site of the lesion.

[0190] A comparative study of the effectiveness of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) was carried out in accordance with regulatory documents and harmonized requirements of regulatory authorities similar to example 14.

[0191] The experiment was conducted on 50 albino juvenile mice of both sexes weighing 10-12 g, randomly divided into five groups. Five animals were housed in temperature-controlled boxes (averaging 21°C) with a 12-hour light / dark cycle and ample food and water. The study utilized the PC virus, previously adapted to growth in mouse lungs at the Pasteur Research Institute of Epidemiology and Microbiology in St. Petersburg. Mice were infected intranasally with the PC virus under light ether anesthesia at a dose of 5.0 logTCID50, in a volume of 0.05 ml for each mouse.The efficacy of the new 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) according to the present invention was characterized by determining the viral titer in the lungs of mice from the experimental groups and statistically comparing them with the control group on the 5th and 7th days of infection by titrating the isolated mouse lung suspension in the Hep-2 cell culture by the cytopathic effect (CPE) Markushin S. G., Kost V. Yu., Rtishchev A. A., Akopova I. I., Koptyaeva I. B., Lisovskaya K. V. Epidemiology and Vaccine Prevention, 2016; 5 (90): 79-85). The preparations in the form of an aqueous solution prepared ex tempore were administered PCT / RU2025 / 000306.

[0192] 30

[0193] The animals were administered 0.5 ml intragastrically through a tube daily from the first to the sixth day after infection. To compare parameters and assess the validity of the model, a negative control group was formed consisting of animals that received the solvent (water) in an equivalent volume, and a positive control group consisting of animals that received ribavirin at a dose of 10 mg / kg. After drug administration and noninvasive testing, the animals were euthanized using a CO2 chamber.

[0194] To conduct the study, the animals were divided into 5 experimental groups of 10 individuals:

[0195] 1st group - negative control, placebo - individuals received 0.5 ml of water.

[0196] 2nd group - treatment with compound 6a at a dose of 100 mg / kg in 0.5 ml of water,

[0197] 3rd group - treatment with compound be at a dose of 100 mg / kg in 0.5 ml of water,

[0198] Group 4 - treatment with compound 8a at a dose of 100 mg / kg in 0.5 ml of water,

[0199] Group 5 - positive control, treatment with ribavirin at a dose of 10 mg / kg in 0.5 ml of water.

[0200] The characteristics of the antiviral activity of compounds with the general formula (I) against the RS virus are presented in Table 3.

[0201] Table 3. Antiviral action of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) in the model of experimental respiratory syncytial virus infection in mice

[0202] The test substance, Infectious titer of the virus in the lungs of mice IgTCIDso dose 5 days after infection 7 days after infection Distilled water

[0203] 4.97±0.37 3.54±0.43 (placebo)

[0204] Compound 6a, 100 mg / kg 3.23±0.31* 1.42±0.26* Compound be, 100 mg / kg 3.56±0.42* 2.14±0.37* Compound 8a, 100 mg / kg 3.12±0.29* 1.57±0.34* Ribavirin, 10 mg / kg 3.04±0.33* 1.23±0.25* Note: * - differences with placebo characteristics are significant at p<0.05

[0205]

[0206] The presented data indicate that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) have antiviral activity against the RS virus, which is confirmed by a decrease in viral titers in the lungs of mice compared to mice in the control group that received a placebo (statistical significance level p < 0.05). Example 16: Therapeutic and prophylactic antiviral effect of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) against 985 rhinovirus

[0207] The experiment was conducted on white mongrel mice of both sexes weighing 9-10 g, which were randomly divided into five groups of ten animals each. Five animals were housed per group in temperature-controlled boxes (averaging 21°C) with a 12-hour light / dark cycle and sufficient water and food. Human rhinovirus, strain HRV-1 (deposited in GKV2730), previously titrated in mice, was used for the study. Mice were infected intranasally with rhinovirus under light ether anesthesia at a dose of 5.0 lgTCID50, in a volume of 0.05 ml for each mouse. To evaluate the antiviral efficacy of new compounds of general formula (I) according to the present invention, measurements of the viral titer were carried out in the lungs of mice of the experimental groups compared to the control group on the 3rd day after infection by titrating the isolated lung suspension of mice in a HeLa cell culture for cytopathic effect (CPE).Animals were treated with experimental formulations based on the compound of formula (I) 1000 intragastrically using a tube once a day for 5 days (two days before infection and for 3 days after infection), similarly, animals of the control group received a placebo.

[0208] A comparative study of the effectiveness of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) was carried out in accordance with 1005 regulatory documents and harmonized requirements of regulatory authorities similar to examples 14 and 15.

[0209] Experimental groups:

[0210] Group 1 - negative control, placebo - individuals receiving 0.5 ml of water, Group 2 - treatment with compound 6a at a dose of 100 mg / kg in 0.5 ml of water,

[0211] 1010 3rd group - treatment with compound be at a dose of 100 mg / kg in 0.5 ml of water,

[0212] Group 4 - treatment with compound 8a at a dose of 100 mg / kg in 0.5 ml of water,

[0213] Group 5 - positive control, treatment with ribavirin at a dose of 10 mg / kg in 0.5 ml of water.

[0214] The characteristics of the antiviral activity of compounds with the general formula (I) 1015 against rhinovirus are presented in Table 4. Table 4. Antiviral action of new compounds of the general formula (I) according to the present invention in the model of experimental rhinovirus infection in mice _ _

[0215] Drug, Infectious titer of the virus in the lungs of mice, dose mg / kg IgTCIDso 3 days after infection

[0216] Distilled water (placebo) 2.73±0.37

[0217] Compound 6a, 100 mg / kg 0.92±0.41*

[0218] Conn. be, 100 mg / kg 1.44±0.33*

[0219] Compound 8a, 100 mg / kg 1.12±0.42* Ribavirin, 10 mg / kg 2.53±0.29 Note: * - differences with placebo characteristics are significant at p<0.05

[0220]

[0221] 1020

[0222] The presented data indicate that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) have antiviral activity against rhinovirus type 1, which is confirmed by a decrease in viral titers in the lungs of mice compared to mice in the control group that received placebo (statistical significance level p<0.05).

[0223] Example 17: Therapeutic antiviral activity of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against the SARS-CoV-2 virus

[0224] To evaluate the efficacy of 7-amino-[1,2,4]triazolo[1,5-1030 a]pyrimidine derivatives of general formula (I), a SARS-CoV-2 infection model was reproduced in Syrian hamsters using compound 6a as an example. The virus was titrated in Vero-B cell culture by the number of plaque-forming units and administered to animals intranasally at a dose of 4x10 at a dose of 26 μl / hamster. 4SARS-CoV-2 (strain PV 10734) TCID50. Several groups of animals from the same litter (1035) were formed.

[0225] Experimental groups:

[0226] Group 1 - animals that received compound 6a at a dose of 15 mg / kg daily intraperitoneally for 5 days without infection, infection control

[0227] Group 2 included animals infected with SARS-CoV-2 and given 0.5 ml of 1040 water (negative control, placebo). Group 3 included animals given compound 6a at a dose of 15 mg / kg intraperitoneally daily for 5 days after infection, from days 3 to 7 of SARS-CoV-2 infection, during the onset of its manifest signs.

[0228] The study of the effectiveness of the 7-amino-[1,2,4]triazolo[1,5- 1045 a]pyrimidine derivative of general formula (I) was carried out in accordance with regulatory documents and harmonized requirements of regulatory authorities similar to example 14-16.

[0229] During observation of infected animals, manifestations of disease symptoms (sneezing, nasal discharge) were assessed; the frequency of these symptoms did not significantly differ between infected animals. Infected animals experienced weight loss.

[0230] Animals were sacrificed on day 7. At necropsy, the lungs and spleen were removed, weighed, and their specific gravity calculated as a percentage of body weight. The right lung was placed in Petri dishes containing saline and transilluminated to count the number of tissue compactions and 1,055 foci of hyperemia with hemorrhages in the lung parenchyma.

[0231] Compound 6a was found to inhibit weight loss in animals, with significant increases in the placebo group. The lungs of infected animals showed viral pneumonia with multiple foci of irregular consolidation with unclear boundaries of varying sizes, but usually not very large and tending to merge, especially in the lower parts of the lungs. Spots on the lung surface varied in color, from light gray and gray-pink to light red and brown. Discolored lung areas had a granular surface when sectioned and bulged slightly above the surrounding tissue. In the negative control group (placebo), virtually no areas of normal, light pink lung tissue were detected. The lungs appeared compacted, tightly elastic, and edematous. The surface of the lung section was mottled and unevenly filled with blood. Fluid was virtually not squeezed out of the compacted areas when sectioned.Compound 6a, when administered therapeutically, reduces the number of compactions in lung tissue and the number of foci of hyperemia and hemorrhage: lung tissue 1070 is represented by isolated foci of compaction with fairly clear boundaries of varying sizes, but usually not very large and without confluence, most of the surface has the appearance of normal lung tissue of light pink and light red color. According to gravimetric parameters of organs, compound 6a significantly affects the development of viral pneumonia - 1075 reliable differences were observed between treated animals and the negative control (placebo) for the controlled parameters. Figure 2 shows the results of a study of the antiviral effect of compound 6a on the SARS-CoV-2 coronavirus in a disease model: the relative weight of the lung (Fig. 2A) and spleen (Fig. 2B) on the day of autopsy.

[0232] 1080 A decrease in spleen specific gravity was observed in infected animals, apparently reflecting the developing immunodeficiency. It was shown that therapeutic use of compound 6a, according to formula (I), maintained organ parameters at intact levels, and these characteristics significantly differed from those in the control group.

[0233] Viral infectivity was determined by titrating organ suspensions in Veto (B) cell culture, followed by evaluation of the virus's cytopathic effect. To prepare the suspension, a maintenance medium (EMEM with 2% serum and 1% antibiotic) was added to a test tube containing the test material (apical and cardiac lobes of the right lung or nasal turbinate tissue fragments) at a 1:10 (weight / volume) ratio and homogenized in a FastPrep-24 5G homogenizer at 4.0 m / s for 30 s. The homogenate was centrifuged at 3500 rpm and 4°C for 10 min, 100 μl of the supernatant was transferred to an Eppendorf tube containing 900.0 μl of the maintenance medium and filtered through a 1095 syringe filter with a 0.22 μm pore diameter. Serial tenfold dilutions were then prepared in the maintenance medium from 10' 2 up to 10' 8The nutrient medium was removed from a 96-well Veto cell culture plate (B), 100.0 µl of maintenance medium was added to each well, and then 100.0 µl of the appropriate virus dilution was added to each row (4 replicates for each dilution). The last row was filled with 200.0 µl of maintenance medium as a cell culture control. The plate was incubated for 3 days at 37°C and 5% CO2 in a CO2 incubator (ICO240med (Memmert, Germany). The cytopathic effect was assessed visually under an inverted microscope (AE31 Trinocular inverted microscope (Moys, China). The TCID50 was calculated using the method of Reed and Mench (1938) in decimal logarithms of the fifty percent infectious dose per 1 g of lung tissue or nasal turbinate tissue (1 g TCID50 / G).

[0234] Figure 3 shows the data on the SARS-CoV-2 virus titer in the lungs on the day of autopsy, as characteristics of the antiviral effect of compound 6a on the SARS-CoV-2 coronavirus in an experimental model of the disease. 1110 The presented data indicate that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) have antiviral activity against rhinovirus type 1, which is confirmed by a decrease in virus titers in the lungs of mice compared to mice in the control group receiving a placebo (statistical significance level p < 0.05).

[0235] 1115 Example 18: Protective and therapeutic antimicrobial action of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against Staphylococcus aureus

[0236] The experiment was conducted on 40 female white SHK mice weighing 14-16 g, which were randomly divided into 4 groups, each group of 10 animals. 1120 Animals were kept five individuals per thermoregulated box (with an average temperature of 21 °C) with a 12-hour light / night cycle and a sufficient amount of water and food. A live virulent culture of S. aureus 1986, previously titrated on mice, was used for the study. Mice were infected intravenously in the infraorbital vein under light ether anesthesia at a dose of 5.0 1125 IgTCIDso, in a volume of 0.05 ml for each mouse. Animals received treatment with experimental formulations based on compounds according to formula (1) intragastrically once a day two days before infection and for 5 days after infection; animals of the control group received a placebo in a similar manner.To evaluate the antimicrobial efficacy of new ISO derivatives of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine of general formula (I), animal survival measurements were carried out.

[0237] Experimental groups:

[0238] 1st group - treatment with compound 6a at a dose of 125 mg / kg in 0.5 ml of water, 2nd group - treatment with compound 8a at a dose of 250 mg / kg in 0.5 ml of water, 1135 3rd group - treatment with ciprofloxacin at a dose of 5 mg / kg in 0.5 ml of water,

[0239] Group 4 – control (placebo) – individuals received 0.5 ml of water. The antibacterial activity characteristics of compounds with the general formula (I) against Staphylococcus aureus are presented in Table 5.

[0240] Table 5. Protective and therapeutic antimicrobial action of 7-1140 amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against Staphylococcus aureus in mice _

[0241] Drug Number of mice Dose, regimen Death / in group survived Compound 6a 10 125 mg / mouse, 2 days before and 5 days 0 / 10*

[0242]

[0243] after infection, 7 days in total Compound 8a 10 250 mg / mouse, 2 days before and 5 days 2 / 8* after infection, 7 days in total Ciprofloxacin 10 5 mg / kg, 5 days after infection 0 / 10* Placebo 10 - 10 / 0

[0244]

[0245] Note: * - differences with placebo characteristics are significant at p<0.05

[0246] The presented data indicate that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives have a protective and therapeutic antimicrobial effect 1145 against Staphylococcus aureus, which is confirmed by a decrease in the mortality of mice compared to mice in the control group that received a placebo (statistical significance level p<0.05).

[0247] Example 19: Protective and therapeutic antimicrobial action of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against 1150 Klebsiella pneumoniae

[0248] The experiment was conducted on 50 female white SHK mice weighing 14-16 g, randomly divided into 5 groups of 10 animals each. Five animals were housed per group in temperature-controlled boxes (with an average temperature of 21 °C) with a 12-hour light / dark cycle and an adequate supply of water and food. A live virulent culture of Klebsiella pneumonia 204, previously titrated in mice, was used for the study. Mice were infected intravenously in the infraorbital vein under light ether anesthesia at a dose of 5.0 IgTCIDso, in a volume of 0.05 ml for each mouse. Animals received treatment with the compounds intragastrically through a gavage once daily two days before infection and for 5 days after infection. Animals of the control group similarly received a placebo. To evaluate the antimicrobial efficacy of new 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I), animal survival was studied.

[0249] The following experimental groups were formed:

[0250] 1165 Group 1 - positive control 1, treatment with ciprofloxacin at a dose of 10 mg / kg in 0.5 ml of water.

[0251] 2nd group - positive control 2, treatment with cefotaxime at a dose of 15 mg / kg in 0.5 ml of water.

[0252] Group 3 - treatment with compound 6a at a dose of 150 mg / kg in 0.5 ml of water, 1170 Group 4 - treatment with compound 8a at a dose of 150 mg / kg in 0.5 ml of water,

[0253] Group 5—negative control (placebo)—individuals received 0.5 ml of water. The antibacterial activity characteristics of compounds with the general formula (I) against virulent Klebsiella are presented in Table 6.

[0254] 1175 Table 6. Protective and therapeutic antimicrobial action of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against Klebsiella pneumonia in mice

[0255] Number of mice in Palo / Drug Dose, regimen

[0256] group survived Ciprofloxacin 10 5 mg / kg, start on the day of infection, 9 / 1

[0257] 5 days

[0258] Cefotaxime 10 15 mg / kg, start on the day of infection, 8 / 2

[0259] 5 days

[0260] Compound 6a 10 150 mg / mouse, 2 days before and 5 days 0 / 10* after, total 7 days

[0261] Compound 8a 10 150 mg / mouse, 5 days before and 5 days after, 7 days in total

[0262] Placebo 10 - 9 / 1 Note: * - differences with placebo characteristics are significant at p<0.05

[0263]

[0264] The presented data indicate that 7-amino-1180 [1,2,4]triazolo[1,5-a]pyrimidine derivatives have a protective and therapeutic antimicrobial effect against Klebsiella pneumoniae, which is confirmed by a decrease in the mortality of mice compared to mice in the control group that received a placebo (statistical significance level p<0.05).

[0265] Example 20: Therapeutic antimicrobial action of 7-amino-1185 [1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) against Mycobacterium tuberculosis

[0266] An experiment to identify anti-tuberculosis activity was carried out in vivo using microbiological and biological methods. Using the classical model of hematogenous disseminated tuberculosis in laboratory animals 1190, the chemotherapeutic activity of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) and the features of the course of the tuberculosis process were studied. To determine the chemotherapeutic efficacy of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of the general formula (I) in vivo, experimental reproduction of hematogenous disseminated tuberculosis 1195 was carried out on 40 BALB / c mice weighing 18-20 g. Infection was carried out with a two-week virulent culture of M. tuberculosis H37Rv by introducing a suspension of mycobacteria into the ophthalmic venous sinus at a dose of 5×107 CFU (colony forming units). All experimental animals were divided into groups.

[0267] 1200

[0268] 1st group - positive control treatment with isoniazid at a dose of 25 mg / kg in 0.5 ml of water.

[0269] 2nd group - treatment with compound ba at a dose of 50 mg / kg in 0.5 ml of water, 3rd group - treatment with compound 6a at a dose of 150 mg / kg in 0.5 ml of water, 1205 4th group - treatment with compound 8a at a dose of 150 mg / kg in 0.5 ml of water,

[0270] Group 5 – negative control (placebo) – individuals received 0.5 ml of water.

[0271] Animals were treated per os, starting on the day after infection, daily (except Sunday) for 28 days. Animals 1210 were euthanized using ether anesthesia. To determine the effectiveness of each chemotherapy regimen, macroscopic changes in the parenchymatous organs of animals and the index of Mycobacterium tuberculosis isolation from pathological material were taken into account. Microbiological examination included culture. The right lung and spleen (separately) were subjected to special processing: 1215 were homogenized in a porcelain mortar with 6% sulfuric acid, then centrifuged for 15 minutes at 3000 rpm. The resulting dense sediment was washed first with distilled water, then with saline. The supernatant liquid was decanted. To the sediment (0.5 ml) was added 1.0 ml of saline. solution, homogenized, and seeded into test tubes (0.2–0.4 ml) on a dense 1220 Finn-2 nutrient medium. The cultures were incubated in a thermostat at 37°C for 1 month from the time of seeding.The growth intensity of the cultures was taken into account based on the number of MBT colonies grown on the nutrient medium.

[0272] I. During daily observation of mice receiving the studied drugs during the first month, no pronounced features in behavioral reactions were noted.

[0273] After 23-26 days from infection, mass mortality of mice in the placebo group was observed. On the 28th day of isoniazid treatment, all mice died; in the group of animals treated with compound 6a at a dose of 50 mg / kg, three died. In groups 3-4, survival was 100%.

[0274] 1230 Macroscopic examination of the internal organs of mice from group pl revealed pronounced signs of tuberculous inflammation: 15-20 large confluent foci of inflammation were determined in the lungs, the spleen was enlarged by 3.0-3.5 times. The pattern of damage to the internal organs of mice from groups 2 and 4 was similar and included the presence of up to 10 single foci in the lungs, 1235 located over the entire surface of the organ; the spleen was enlarged by approximately 1.5-2.0 times. Macroscopic examination of the parenchymatous organs of mice from group 3 revealed about 5 small foci of infection in the lungs. The spleen was enlarged by 1.5 times. In the lungs of mice from group 1, no foci of tuberculous infection visible through a magnifying glass were observed. The spleen was either not enlarged or enlarged only slightly.

[0275] Table 7. Results of microbiological examination of parenchymatous organs of mice after chemotherapy with the studied drugs (29th day) Group Drug, dose Log CFU growth after 21 days of cultivation Right lung Spleen 1 Isoniazid, 25 mg / kg 3.96* 3.42* 2 Compound 6a, 50 mg / kg 8.08 8.20 3 Compound 6a, 150 mg / kg 5.49* 4.81* 4 Compound 8a, 150 mg / kg 6.36 5.29* 5 Distilled water (placebo) 8.16 8.52

[0276]

[0277] Note: * - differences with placebo characteristics are significant at p<0.05

[0278]

[0279] 1245 The presented data indicate that 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives have a therapeutic antimicrobial effect against Mycobacterium tuberculosis, which is confirmed by a decrease in the mortality of mice compared to mice in the control group that received a placebo (statistical significance level p<0.05) and the number of CFU in the lungs and 1250 spleen.

[0280] Example 21: Acute toxicity of 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivatives of general formula (I) in mice upon intragastric administration The experiments were performed on male and female Balb / c mice with an initial weight of 20±0.7 g (2 months old) provided by the Andreevka Branch of the Federal State Budgetary Institution of Science 1255 “National Center for Biomedical Technologies” of the Federal Medical and Biological Agency of Russia, where they were maintained as an inbred line. The animals were kept under standard experimental vivarium conditions corresponding to the current requirements for experimental biological vivariums (“Laboratory Animals” Regulations and Guidelines, Moscow, 2003) in plastic cages measuring 42×25×15 cm. Fine PCI7RU2025 / 000306 was used as bedding

[0281] 40

[0282] 1260 shavings from environmentally friendly deciduous wood (TU 5313-001-1897639-92). The mice had free access to food and water. The animals received complete extruded compound feed for laboratory animals - mice, rats, hamsters; standard ROOS PR98 B01220. Quality certificate dated 10.07.08. The daylight period was 12 hours (from 8:00 to 20:00). Illumination during the light period of the 1265 cycle was 70-90 lux. The temperature in the constant maintenance room was 22 °C. The experiments were conducted in the spring.

[0283] All manipulations with animals were carried out in compliance with the rules for the humane treatment of laboratory animals. In carrying out this work, the general requirements were met: Interstate standard GOST 33044-2014 1270 "Principles of Good Laboratory Practice", Guidelines for the conduct of preclinical studies of drugs, edited by Mironov A.N., Bunatyan N.D., Vasiliev A.N., Moscow: Grif i K., 2012: 944; Guide for the care and use of laboratory animals. National Academy press. Washington, DC, 2011 and "Guidelines for the maintenance and breeding of laboratory animals in nurseries and experimental biological clinics (vivaria) and their use for scientific, educational and industrial purposes", Moscow, 2003. After delivery to the vivarium and before the start of the experiment, the animals were kept in quarantine for 14 days for adaptation.

[0284] The studied 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivative of general formula (I), using compound 6a as an example, was administered as a solution in saline 1280 (0.9% aqueous sodium chloride solution) in a single dose in the maximum permissible volume of 0.5 ml per mouse. The behavior and condition of the animals were observed for 14 days. Each group consisted of 6 males and 6 females.

[0285] The general condition of the test animals was determined based on their appearance, behavioral responses, clinical signs of intoxication (pattern of motor activity and gait, presence and nature of seizures, skeletal muscle tone, responses to various stimuli, respiratory rate and depth, heart rate, condition of hair and skin, tail position, frequency of urination, and urine color), feeding activity, and body weight dynamics. Changes in these parameters and possible death of the test animals served as criteria for acute toxicity.

[0286] Males. Male mice were administered intragastrically a 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivative of the general formula (I) using compound 6a as an example in a dose range from 300 to 1000 mg / kg. Four doses of the drug were tested to determine acute toxicity. The dynamics of animal mortality are shown in Table 8. The integral 1295 indices of surviving experimental male mice after a single intragastric administration of FS corresponded to the control ones.

[0287] Found: LD16 - >1000 mg / kg, LD50 = >1000 mg / kg, LD84 = >1000 mg / kg, which indicates that substance 6a is a low-toxicity compound. Table 8. Dynamics of mortality in male mice after a single intragastric administration of FS

[0288] Dose, No. Animal death (day) Died Survived Frequency mg / kg animal death, % in 1 2 3 4 5-14

[0289] group

[0290] 1000 6 0 0 0 0 0 0 6 0 800 6 0 0 0 0 0 0 6 0 500 6 0 0 0 0 0 0 6 0

[0291]

[0292] 300 6 0 0 0 0 0 0 6 0

[0293] Females. Female mice were administered intragastrically a 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivative of the general formula (I) using compound 6a as an example in a dose range from 300 to 1000 mg / kg. A total of 1305 doses of the drug were tested to determine acute toxicity. The dynamics of animal mortality are shown in Table 9. The integral indices of surviving female experimental mice after a single intragastric administration of FS corresponded to the control ones.

[0294] Found: LDie = >1000 mg / kg, LD50 = >1000 mg / kg, LD84 = >1000 mg / kg, which indicates that substance 6a is a low-toxicity compound. 1310 Table 9. Dynamics of mortality in female mice after a single intragastric administration of FS _

[0295] Dose, Number of animals killed (day) Died Survived % diemg / kg animals in 1 2 3 4 5-14

[0296] group

[0297] 1000 6 0 0 0 0 0 0 6 0 800 6 0 0 0 0 0 0 6 0 500 6 0 0 0 0 0 0 6 0

[0298]

[0299] 300 6 0 0 0 0 0 0 6 0

[0300] Thus, when administering the 7-amino-[1,2,4]triazolo[1,5-a]pyrimidine derivative of general formula (I), using compound ba as an example, even at very high doses (up to 1000 mg / kg), no deaths were observed in either the female or male groups. Moreover, all animals remained active and mobile throughout the entire observation period, and did not disrupt their feeding or drinking regimens.

Claims

CLAUSES OF THE INVENTION 1. A compound corresponding to the general structural formula I. (I) Where R 1 represents AlkS, AlkSO, AlkSO2, COO Aik, CON(Alk)2, substituted or unsubstituted Ph, benzyl, furyl, pyridyl; R 2 represents H, Aik, Ph; R 3 is OH, OAlk, NHOAlk, NAlk2, or a saturated 6-membered nitrogen-containing heterocyclic ring attached via a nitrogen atom in a carbonyl group, and optionally containing a 2nd heteroatom in the ring selected from X, where X is O, S, NMe; Aik is H, Me, Et, Pr or i-Pr, as well as their pharmaceutically acceptable salts, solvates or hydrates, wherein R 1 cannot mean MeS if R 2 means H, R 3 means OH or OEt or its sodium salt.

2. A compound according to formula (I) according to claim 1, where R 1= SMe, R 2=H, R 3 =OH, which is a potassium salt.

3. A compound according to formula (I) according to claim 1, where R 2 =H, R 3 =NH0H.

4. Use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate or hydrate thereof Where R 1 represents Aik, AlkO, AlkS, AlkSO, AlkSO, COOAlk, C0N(Alk)2, substituted or unsubstituted Ph, benzyl, furyl, pyridyl; R 2 represents H, Aik, Ph; R 3 is OH, OAlk, NHOAlk, NAlki, or a saturated 6-membered nitrogen-containing heterocyclic ring attached via a nitrogen atom to a carbonyl group, and optionally containing a 2nd heteroatom in the ring selected from X, where X is O, S, NMe; Aik is H, Me, Et, Pr or i-Pr, possessing antiviral and antimicrobial activity for the treatment and / or prevention of diseases caused by viral and / or microbial infections.

5. The use of a pharmaceutically acceptable salt of a compound, its solvate or hydrate according to formula (I) according to claim 4, where R 1 =SMe, R 2 =H, R 3 =OH, possessing antiviral and antimicrobial activity for the prevention or treatment of diseases caused by viral and / or microbial infections.

6. Use of a potassium or sodium salt of a compound according to formula (I) according to claim 4 or 5, where R 1 =SMe, R 2 =H, R 3 =OH, which has antiviral and antimicrobial activity for the prevention and / or treatment of diseases caused by viral and / or microbial infections.

7. Use of a compound according to formula (I) according to claim 4 where R 1 =SMe, R 2 =H, R 3=NHOH, a pharmaceutically acceptable salt, solvate or hydrate thereof, having antiviral and antimicrobial activity for the prevention and / or treatment of diseases caused by viral and / or microbial infection 8. Use of a compound of formula (I) according to any one of claims 4-7 or a pharmaceutically acceptable salt, solvate or hydrate thereof, according to which the compound activates the production of viperin in the treatment of a human with an infectious disease, including an infectious disease, the dynamics of which is sensitive to the level of viperin in the body.

9. The use of a compound of formula (I) according to any of claims 4 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof for the prevention and / or treatment of viral infection in humans and animals caused by viruses selected from influenza, acute respiratory viral infections and / or SARS-CoV-2.

10. The use of a compound of formula (I) according to any one of claims 4 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof for the prevention and / or treatment of a viral infection in humans caused by the hepatitis B or C virus.

11. The use of a compound of formula (I) according to any one of claims 4 to 8, or a pharmaceutically acceptable salt, solvate or hydrate thereof, for the prevention and / or PCI7RU2025 / 000306 44 treatment of infection in humans or animals caused by pathogenic microbes of the ESCAPE group, selected from Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, 12. The use of a compound of formula (I) according to any one of claims 4 to 8 or a pharmaceutically acceptable salt, solvate or hydrate thereof for the prevention and / or treatment of infection caused by Mycobacterium tuberculosis.

13. The use according to claim 11 or 12 of a compound of formula (I), where R 1= SMe, R 2 =H, R 3=OH, in the form of potassium or sodium salt for the prevention and / or treatment of infection in humans and animals caused by pathogenic microbes of the ESCAPE group or Mycobacterium tuberculosis.

14. The use of a compound of formula (I), its pharmaceutically acceptable salt, solvate or hydrate according to any of paragraphs 4-10, for the prevention and / or treatment of a viral infection in humans or animals, as part of combination therapy.

15. The use of a compound of formula (I), its pharmaceutically acceptable salt, solvate or hydrate according to any of claims 4 or 11-13, for the prevention and / or treatment of a microbial infection in a human or animal, as part of combination therapy.

16. The use of a compound of formula (I), a pharmaceutically acceptable salt, solvate or hydrate thereof according to any of paragraphs 4-8 or 13 for the prevention and / or treatment of a combined viral and microbial infection in humans or animals.

17. A pharmaceutical composition having antiviral and antimicrobial activity for the treatment and / or prevention of diseases caused by viral and / or microbial infection, containing a compound of general formula I in a therapeutically effective amount as an active component. 2 2 where R 1 represents Aik, AlkO, AlkS, AlkSO, AlkSO2, COOAlk, CON(Alk)2, substituted or unsubstituted Ph, benzyl, furyl, pyridyl; R 2 represents H, Aik, Ph; R 3 represents OH, OAlk, NHOAlk, NAlk2, or saturated 6- a membered nitrogen-containing heterocyclic ring attached via a nitrogen atom to a carbonyl group and optionally containing a 2-membered heteroatom in the ring selected from X, where X is O, S, NMe; Aik is H, Me, Et, Pr or i-Pr; a pharmaceutically acceptable salt, solvate or hydrate thereof, and pharmaceutically acceptable excipients.

18. A method for the treatment and / or prevention of diseases caused by viral and / or microbial infection, comprising administering or applying to a patient in need thereof a therapeutically effective amount of a compound of general formula I NH2O 2 where R 1 represents Aik, AlkO, AlkS, AlkSO, AlkSO2, COOAlk, C0N(Alk)2, substituted or unsubstituted Ph, benzyl, furyl, pyridyl; R 2 represents H, Aik, Ph; R 3 is OH, OAlk, NHOAlk, NAlki, or a saturated 6-membered nitrogen-containing heterocyclic ring attached via a nitrogen atom to a carbonyl group, and optionally containing a 2nd heteroatom in the ring selected from X, where X is O, S, NMe; Aik is H, Me, Et, Pr or i-Pr; a pharmaceutically acceptable salt, solvate or hydrate thereof, or a pharmaceutical composition according to claim 16.

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