RU2301669C2 - Using compounds of antiviral class for preparing agent for treatment and prophylaxis of respiratory tract viral infection - Google Patents

Abstract

FIELD: medicine, virology, pharmacy.

SUBSTANCE: invention relates to using derivatives of dithiocarbamate of the formula: R₁R₂NCS₂H and oxidized forms of these compounds, in particular, their dimmers and their pharmaceutically compatible salts for preparing an agent used in treatment or prophylaxis of infection caused by RNA-containing viruses that damage respiratory tract and inducing disease. Also, invention relates to a disinfecting agent containing dithiocarbamate compound and a method for disinfection of surfaces, media and cell cultures.

EFFECT: valuable medicinal properties of compounds.

19 cl, 14 dwg, 14 ex

Description

FIELD OF THE INVENTION

The invention relates to the use of dithiocarbamate compounds, a disinfectant and a method for disinfecting surfaces, media and cell cultures.

State of the art

There are many viruses that cause respiratory tract diseases in humans and mammals. Although these respiratory pathogenic viruses may differ structurally and belong to different viral families, the common thing for them is that they are able to penetrate the body through the respiratory tract and infect, for example, specific cells of this tract, such as the epithelial cell layer of the respiratory tract , alveolar cells, pulmonary cells, etc. The classic symptoms of influenza infection, characterized by local inflammation and manifestations of the disease in the respiratory tract (such as runny nose, hoarseness, are common). cough, vesicles, sore throat).

Viral infections, especially in the respiratory tract, cause pathological changes in the corresponding cells, in particular in epithelial cells, due to oxidative stress. Reactive oxygen intermediates (ROIs) produced, for example, by white blood cells, pulmonary epithelial cells or xanthine oxidases, act as mediators in such cellular viral-induced disorders. During such oxidative stress, an oxidizing agent, a specific transcription factor NFkB (nuclear factor-kB), can occur. This factor NFkB was found in a wide variety of cell types and its causal relationship with gene activation in inflammatory and immune responses has been established.

Antioxidants are able to block NFkB activation by trapping reactive intermediate forms of oxygen that cause this activation. Therefore, it was proposed to use antioxidants primarily in the treatment of infections with latent viruses; however, it was found that effective treatment of such latent infections with antioxidants alone is not possible, and that, if at all, combination therapy is required using a mixture of different (i.e., having different effects) antioxidants and other virus suppressants (US 5, 686, 436) . However, inhibition of viral replication or even viral infection by antioxidants has not yet been achieved, in particular, inhibition of infections caused by influenza viruses and picornaviruses (Knobil et al., Am. J. Physiol. 274 (1) (1998) (134- 142).

On the other hand, the different antioxidants proposed for the treatment of viral infections are very different in terms of antioxidant activity. So, for example, L-ascorbic acid and vitamin E serve to protect glutathione; Vitamins K, A and E act as antagonists of peroxynitrite and other strong oxidants in the body. Anti-inflammatory steroids, non-glucocorticoid lazaroids, dithiocarbamates and N-acetyl-L-cysteine are described as inhibitors of NFkB activation.

Depending on the carrier of genetic information, viruses can be divided into DNA viruses and RNA viruses, the nucleic acid being present as a single or double strand and surrounded by a protein coat.

Moreover, the single-stranded RNA of such RNA viruses is present either as a positive chain (mRNA) or as a negative one. In addition, the genetic information of the virus can also be presented in the form of several fragments, as is the case, for example, in the influenza virus.

Human rhinoviruses (HRV) related to picornaviruses are the leading cause of the common cold. The frequent manifestation of HRV, the risk of severe secondary infections and the economic consequences of medical costs, medical visits, and the level of illness of workers make HRV an important and dangerous pathogen. Despite the frequent manifestation, an effective fight against this viral disease is not yet ensured, apart from treating the symptoms. On the other hand, the consequences, for example, from an infection caused by rhinovirus, are not so severe or even life-threatening, so that one should put up with the use of drugs that are characterized by a high risk of side effects. Therefore, the means that you want to use against such viruses should have only a minor side effect or not at all. Horse rhinitis A virus (ERAV), which, like the foot and mouth disease virus, belongs to the genus of aphthoviruses, belongs to the group of animal picornaviruses.

Other important virus families responsible for respiratory tract diseases are Orthomyxo- and Paramyxoviridae with human influenza virus as the most important representative.

The emergence of new pandemic influenza strains is usually associated with new subtypes containing new hemagglutinin or the neuraminidase gene. Such new viruses are immunologically different from pre-existing influenza viruses. In order to become infectious, influenza A and B viruses require 8 segments of RNA, influenza viruses C only 7. Influenza viruses A, B and C can form homotypic varieties in vivo, but not between types. Theoretically, 256 varieties can be formed from the corresponding 8 segments of two influenza A viruses. However, such random segregation does not occur, since at the protein level, some proteins require a partner due to its strain. This has been specifically shown for the Avian Influenza-Virus A / chicken / Germany / 34 (H7N1) FPV Rostock HA virus, which is encoded by segment 4, capable of forming a functional virus only specifically with its strain specific protein M2, encoded by segment 7 (Grambas S ., Hay AJ Maturation of Influenza A virus hemagglutinin-estimates of the pH encountered during transport and its regulation by the M2 protein. Virology 1992; 190: 11-18) (Grambas S., Bennet HS, Hay AJ Influence of amantadine resistance mutations on the pH regulatory function of the M2 protein of influenza A viruses. Virology 1992; 191: 541-549). One of the main strategic measures in the annual fight against viral influenza infection in the population is vaccination against influenza. However, despite extensive vaccination programs, influenza remains the cause of morbidity and mortality worldwide and the main cause of the disease and death of immunocompromised patients and the elderly. The antiviral activity of amantadine and rimantadine reduces the duration of symptoms of clinical flu, but important side effects and the emergence of resistant mutants are described (FIELDS et al., Virology, 3. Edition (1995) Lippincott-Raven Publ., Philadelphia, Vol.1, p.434- 436). Currently, a new group of antiviral agents has appeared on the market that suppress the influenza virus neuraminidase. Tsanamivir and oseltamivir are, for example, influenza A and B virus neuraminidase inhibitors; however, these medicines only shorten the duration of symptoms.

No. 5,686,436 describes a method for suppressing the propagation of retroviruses and latent viruses in humans and animals, such as, for example, human influenza virus (HIV), in which case a medicine is administered containing, inter alia, antioxidants and induction inhibitors of NFkB factor. In contrast, for the effective suppression of infections caused by respiratory viruses, it is necessary to effectively fight the virus already in the acute stage of infection; treatment that can be carried out only at the stage of latent viruses is unsuitable for the prevention or treatment of acute viral infections of the respiratory tract.

Thus, there remains a need for a highly effective active substance against viral infections of the respiratory tract, in particular, a person that would not have completely or only slightly side effects, would be cheap and could be produced in large quantities.

SUMMARY OF THE INVENTION

The objective of the present invention is solved by the use of dithiocarbamate compounds with the structural formula R ₁ R ₂ NCS ₂ H, where R ₁ and R ₂ mean, independently from each other, unbranched or branched alkyl with 1-4 carbon atoms or form an aliphatic ring with a nitrogen atom with 4-6 carbon atoms, wherein R ₁ , R ₂ or the aliphatic ring is optionally substituted with one or more substituents selected from OH, NO ₂ , NH ₂ , COOH, SH, F, Cl, Br, I, methyl or ethyl, and oxidized forms of these compounds, in particular their dimers, as well as their pharmaceutically compatible salts for the manufacture of a medicament for treating or preventing an infection caused by RNA viruses affecting the respiratory tract and causing a disease therein. In the context of the invention, the term "respiratory tract" is understood to mean all organs and areas, starting from the openings in the body (nose, mouth, eyes (including the lacrimal canadian) and ears) and ending with pulmonary vesicles. In this case, pharmaceutically acceptable salts are, in particular, Na, K, Ca, Mg, NH ₄ , Zn. For the first time, it was unexpectedly discovered that the dithiocarbamate compounds of the invention can be effectively used against infections caused by RNA viruses that infect the respiratory tract and cause its disease, referred to herein as “respiratory RNA viruses”. Contrary to observations made by Knobel et al. (Am. J. Physiol. (1998), pp. 134-142), within the framework of the present invention, the antiviral effect of the dithiocarbamate compound of the invention against infections caused by respiratory RNA viruses, for example, HRV (human rhinoviruses) and influenza viruses, is clearly shown. This is all the more surprising because other antioxidants do not possess such an antiviral effect against respiratory RNA viruses and that a change in the redox potential is not solely responsible for the antiviral effect of the dithiocarbamate compounds according to the invention. For example, according to the invention, it can be clearly shown that antioxidants: vitamins C and E, 2-mercaptoethanol and N-acetyl-L-cysteine have absolutely no effect against respiratory RNA viruses. In addition, the effectiveness of the dithiocarbamate compounds of the invention against infections caused by respiratory RNA viruses and the propagation of these viruses is not due solely to the inhibition of NFkB activation, since it was found that using the dithiocarbamate compounds of the invention - and in the framework of this application, they should also be understood and oxidized forms, in particular, dimers, - the reproduction of respiratory RNA viruses deliberately stops.

DE 1963223 A discloses a medicament for treating viral infections of the brain, wherein the medicament should include a monoamine biosynthesis inhibitor noradrenaline, dopamine and 5-hydroxy-tryptamine. The example given in this publication indicates the effect of α-methyl tyrosine methyl ester on mice with herpes simplex infection. Thus, the mechanism of inhibition of the biosynthesis of special monoamines described in this publication is applicable only for the treatment of infections caused by DNA viruses in the brain. The treatment for respiratory RNA viruses of the invention is carried out according to a different principle and is not applicable for brain viral infections, as evidenced by the negative examples with respect to FSME (example 13) and EMC (example 14). Therefore, DE 1963223 relates to another field of application, cannot be compared with the purpose according to the invention and thus is not a close analogue.

Calvert J.G., Interferon Research 1990 (10), pp. 13-23, describes the effect of diethyldithiocarbamate (DDTC) on menoviruses, and it has been found that DDTC deactivates menovirus virions. However, Mengoviruses are causative agents of severe encephalomyocarditis, do not affect the respiratory tract and do not provoke diseases in it.

WO 95/03792 A1 discloses the use of thiol compounds for the preparation of a pharmaceutical composition for the treatment of viral diseases, the thiol compound destroying disulfide bridges in viral proteins. Among the numerous viruses, RNA viruses, in particular Picornaviridae, are also mentioned in this document. In addition, many examples of thiol compounds are given, in particular, dithiocarbamate is mentioned in a very general form. Of the many different combinational possibilities, only the following examples are given: as thiol compounds: N-acetylcysteine (NAC), cysteine, cysteine hydrochloride and N, S-diacetylcysteine-ethyl ester (DACEE), only hepatitis B and smallpox virus are listed as viral diseases a rabbit.

Therefore, not only were examples not given for most of the compounds and viral diseases, but it was also found that the treatment disclosed in this publication does not in fact fully correspond to its description: some picornaviruses (those that damage the respiratory tract and the nerves cells) by PDTC do not inhibit and their reproduction does not decrease. Therefore, not all combinations of virus-provoked diseases and thiol compounds achieve the goal, and the use of dithiocarbamate compounds selected according to the invention, which are not disclosed in WO 95/03792 A1, in the treatment or prevention of infections caused by respiratory RNA viruses, is not obvious.

GB 861043 A discloses compositions used inter alia to protect against viruses. These compositions contain, for example, dithiocarbamates, however, specific viruses are not disclosed here.

Knobil et al., Am. J.Physiol. (1998), p.134-142, report on the study of oxidants and their effects on gene expression caused by the virus. It has been unequivocally determined that neither NAC nor PDTC inhibit influenza virus infection or replication.

DE 2555730 A describes an antimicrobial agent containing a dimethyldithiocarbamate compound, the compound being an 8-hydroxyquinolinate-metal-N, N-dimethyldithiocarbamate complex. But this publication only reports antifungal and antibacterial actions.

WO 99/66918 A1 deals with the use of disulfide derivatives of dithiocarbamates for the reduction of nitrogen oxides in a patient and for the inhibition of NFkB. However, this publication provides an extremely extensive list of diseases without a detailed description of viral diseases.

Flory et al., J. Biol. Chem., March 24, 2000, 275 (12), pp. 8307-8314, report on a study of the effect of various influenza A virus proteins on activation of NFkB-dependent expression.

Tai D.I. et al., Hepatology, March 2000, 31 (3), pp. 785-787, report a study of inhibiting NFkB activation by PDTC, and it was suggested that HCV virus infection could cause antiapoptosis due to NFkB activation.

Schwarz et al., 1998 (J. Virol; Vol. 72 (7), pp. 5554-5660) investigated the effect of NFkB on the propagation of encephalomyocarditis virus (EMCV), akin to human rhinovirus. In cells without NFkB (knockout p50 - / - or p65 - / -) that were infected with the EMCV virus, although virus reproduction decreased, there was an increased cell death from apoptosis. This is in sharp contradiction with the data presented here: these examples clearly show that the compounds of the dithiocarbamate according to the invention not only inhibit the replication of related rhinovirus, but also prevent cell death caused by the virus. Therefore, inhibition of NFkB is not critical to the effectiveness of the dithiocarbamate compounds of the invention.

According to the invention, the antiviral effect of the dithiocarbamate compounds is also independent of the combination of certain substances. The dithiocarbamate compounds according to the invention can be used completely independently, regardless of other additives, in particular antioxidants, which is absolutely necessary in US 5686436 to achieve the antiretroviral effect of NFkB activation inhibitors, since it was unexpectedly found that the antiviral effect not only helped to overcome oxidative stress, but also thanks to the dithiocarbamate compounds of the invention, it has become possible to suppress infection / replication.

The dithiocarbamate compounds of the invention have been shown to induce genes that act as transcription factors inducing antioxidants (Meyer et al., EMBO J. 12, 2005-2015, 1993). The heterodimeric transcription factor API can be induced with N-acylcysteine (NAC) and the dithiocarbamate compounds of the invention, resulting in DNA binding and transactivation. Activation of AP1 by the dithiocarbamate compounds of the invention is dependent on protein synthesis and includes transcription of the c-jun and c-fos genes. However, only activation of AP1 does not provide reliable inhibition of the viruses according to the invention.

Pyrrolidine dithiocarbamate (PDTC) is already known as a pro- and antioxidant, an inhibitor of the activation of transcription factor NFkB, zinc ionophore and a metal chelating agent. Sherman et al., Biochem. Biophys. Res. Comm., 191 (3): 1301-1308, 1993, described PDTC as an inhibitor of NFkB factor activation and NO synthesis. WO 01/00193 A2 reports compositions with a diethyl dithiocarbamate content in the pico- and nanomolar ranges, characterized by a pronounced anti-apoptosis effect.

It was shown that the compounds of the dithiocarbamate according to the invention have a pronounced effect against RNA viruses - respiratory RNA viruses that affect the respiratory tract and provoke a disease in it, both in vitro and in vivo.

Thus, according to the invention, infection is prevented already at a very early stage, even before significant cell destruction or even cell death occurs.

In the framework of the present invention, respiratory RNA viruses are understood to mean any human and mammalian viruses that infect the body through the respiratory tract, i.e. airways and lungs, and penetrating into it, while they cause disease in the respiratory tract. Obviously, the biological processes that occur during such an infection are so similar to each other that the action of the dithiocarbamate compounds according to the invention is equally effective despite the biological heterogeneity of this group of viruses. However, the opposite result was also noted, according to which, for other viruses that have entered the body through other infectious pathways and have gone through other biological cycles (for example, that can integrate into the host genome as a latent virus) or cause the disease in other organs, for example, in the brain, the action only the dithiocarbamate compounds of the invention are not sufficient to treat a viral infection.

Meanwhile, in this regard, it has been shown that in treating patients with AIDS with dithiocarbamates, improvement of the condition of patients or their cure is not achieved (Multicenter, randomized, placebo-controlled study of dithiocarb (Imuthiol) in human immunodeficienty virus-infected asymptomatic and minimally syptomatic patients The HIV87 Study Group. AIDS Res. Hum. Retroviruses 1993 Jan .; 9 (1): 83-9). Based on these results, subsequent clinical studies of latent outbreaks have not been conducted (see US 5686436).

The compounds of the dithiocarbamate according to the invention show their particular effect primarily at an early stage of a viral infection or, if they are taken before the infection begins. Thus, dithiocarbamate compounds can prevent the onset of a viral infection if they are taken prophylactically, for example, in areas and at times when there is a risk or even an increased risk of respiratory viral infections, for example, in areas where there are epidemics or long-term colds are sustained. It is preferable to use the dithiocarbamate compounds according to the invention for the prevention of viral infection.

However, it is most preferable to inhibit the virus according to the invention, especially at an early stage of an already begun infection caused by a respiratory RNA virus. Moreover, the dithiocarbamate compounds according to the invention are purposefully used to inhibit virus replication, i.e. at the time when the infected individual has not yet suffered significant damage. As a result, not only the consequences of a progressive viral infection in the individual can be prevented, but also the subsequent spread of infectious viruses among other individuals; the risk of further infection is minimized, which has a great healing effect and is of great importance especially with respect to the human influenza virus.

In particular, the following are considered respiratory RNA viruses within the scope of the present invention: rhinoviruses, Coxsackie viruses, echoviruses, coronaviruses, enteroviruses, human orthomyxoviruses (e.g. influenza virus (A, B and C), paramyxoviruses (e.g. parainfluenza virus and pneumovirus), respiratory virus synthesis (RSV) and other RNA viruses as long as they are capable of causing disease (at least) in the respiratory tract Viruses or their strains causing the disease not in the respiratory tract, but in another organ, for example, in the brain, for example, viruses of mining, encephalus Iocarditis, polyvirus, cardiovirus, etc. are not the subject of this application. Respiratory RNA viruses according to the invention are characterized by an infection of the epithelial cells of the upper and lower respiratory tract. In addition, other organs can be additionally affected. The resulting local inflammation of the respiratory tract is the main cause the appearance of typical symptoms of a flu infection, such as a runny nose, sore throat, hoarseness, cough, vesicles, and often fever. Also, an infection caused by the indicated respiratory viruses contributes to the frequent manifestation of secondary infections in an immunologically weakened individual.

In the framework of the present invention, picornaviruses are understood to mean any “real” picornaviruses in accordance with the currently accepted classification of Picornaviridae based on King et al. (“Picornaviridae” in “Vims Taxonomy, Seventh Report of the International Committee for the Taxonomy of Viruses” (2000), Eds. Van Regenmortel et al., Academic Press 657-673) if they affect the respiratory tract and cause disease in it , i.e. genus: enterovirus, rhinovirus, aftovirus, parechovirus, erbovirus, cobuvirus and tesovirus. These viruses of the Picornaviridae family are distinguished by: similar genetic structure, protein composition, cultivation characteristics and resistance to temperature and antiviral agents.

It turned out that the present invention is especially applicable for the defeat of pathogenic for humans and animals representatives of the genuine respiratory Picornaviridae, in particular, from the family of enterovirus (entervirus 70, 71, Coxsackie virus), rhinovirus (e.g. human rhinovirus) and aftovirus (e.g. foot and mouth disease virus) in contrast to other viruses, including picornaviruses that cause latent infections, such as HAV, the inherent advantages of the invention have not been confirmed. This is probably due to the fact that the group of "real" respiratory viruses is in itself so homogeneous and the pathophysiological processes in infections are so similar that the dithiocarbamate compounds act in a similar way.

Under the "viral infection" in the framework of this application refers to any attack of a respiratory virus on cells, containing, for example, one of the following: fixing the viral particle on the cell, introducing the virus genetic information into the cell at a subsequent stage, producing new viral particles and expressing the infectious viral particles.

In addition, it is also possible to use oxidized forms of such compounds, in particular, their dimers, since the latter are known to be rapidly converted by metabolism in the body to form reduced forms. In the framework of this application, such compounds should be understood as "oxidized forms", the remainder S of which is oxidized. A preferred example of such an oxidized dimeric form is disulfiram-tetraethylthiuram disulfide (C ₁₀ H ₂₀ N ₂ S ₄ ), also known as "Antabus" or "Abstinyl." Tetraethylthiuramdisulfide is known per se, and it is used in particular in the treatment of alcoholism: it serves as a mediator in oxidation-reduction and deactivates aldehyde dehydrogenase. When ethanol is taken, acetaldehyde is concentrated in the body, which has an extremely negative effect on overall well-being: while taking disulfiram and alcohol, fears appear, nausea occurs, vision loss occurs, pains in the chest, head, etc. appear, and these symptoms last for 3-4 days and even weeks. Therefore, disulfiram is prescribed to alcoholics as a therapeutic agent, since any subsequent drinking should be prevented by such strong negative effects.

Other known actions of disulfiram are inhibition of enzymes such as, for example, fructose, 1,6-diphosphate dehydrogenase, xanthine oxidase, hexokinase, aldehyde oxidase and dopamine β-hydroxylase.

In addition, disulfiram is used in the treatment of pediculosis, scabies and nickel dermatitis.

The metabolism of disulfiram has already been analyzed in detail, see, for example, Dollery C., 1999, Therapeutic Drugs, Second Edition, Vol. 1, Churchill Livingstone, Edinburgh. It is described that diethyldithiocarbamate is the main metabolite of tetraethylthiuramdisulfide in the body, and such a conversion occurs very quickly.

The oxidized form of the dithiocarbamate compound of the invention is highly soluble in oil and can be used, for example, as an oral agent, and the compounds are absorbed in the stomach. The compounds can be used, in particular, in the form of a powder aerosol. In addition, the oxidized forms of the compounds according to the invention can be provided with hydroxyl groups to increase water solubility and obtain an aerosol form. If the compounds of the dithiocarbamate according to the invention are prepared in the form of an oxidized, in particular, dimeric, preparation, such an agent will have a depot effect, i.e. the dithiocarbamate compounds according to the invention will be deposited gradually in the body over time and then absorbed from it. To obtain such a depot, oxidized dithiocarbamate compounds can be implanted in a known manner into the body to be treated.

A preferred compound according to the invention is characterized in that R ₁ and R ₂ are, independently of one another, alkyl with 1 to 3 carbon atoms or form together with a nitrogen atom an aliphatic ring with 4 to 6 carbon atoms. It has proven itself particularly positively in the treatment and prevention of infections caused by the respiratory RNA virus.

It is preferable to select a dithiocarbamate compound from pyrrolidine dithiocarbamate (PDTC) and N, N-diethyldithiocarbamate (DDTC). These compounds are characterized by pronounced activity against infections caused by respiratory RNA viruses.

Especially preferred for this is an infection caused by picornavirus, orthomyxovirus or parmixovirus. The dithiocarbamate compounds of the invention are particularly active against infections caused by these viruses.

Preferably, the orthomyxovirus is a human influenza virus, derived in particular from the group consisting of viruses A, B, C of influenza, paramyxovirus, parainfluenza virus or pneumovirus. Preferably, the orthomyxovirus is a mammalian influenza A virus, and the picornavirus is a rhinovirus, in particular human or horse rhinovirus, enterovirus, in particular enterovirus 70, 71, or Coxsackie virus or aftovirus, in particular, foot-and-mouth virus and horse rhinitis virus A. Moreover, as already explained above, such viruses and their strains that cause diseases not in the respiratory tract, but, for example, in the brain or in nerve cells, for example, cardioviruses or polioviruses related to picornaviruses, are excluded. Such viral infections are particularly effectively treated or prevented by the compound of the invention.

The Picornaviridae family covers a number of small RNA viruses, including rhinoviruses (e.g. human rhinovirus), enteroviruses (e.g. enterovirus 70, 71, Coxsackie virus, which is, for example, the causative agent of diseases of the hands, legs and mouth), aftoviruses (e.g., foot and mouth disease virus , and in this case, the disease is caused in the mouth, as well as horse rhinitis virus A). Regarding, for example, foot-and-mouth disease, it is particularly interesting to note that using the compound according to the invention it is necessary to bridge the time interval between the moment of vaccination and the moment the vaccine is effective (about 10-12 days), since during this time the vaccinated animals remain unprotected and probably after infection secrete a large number of viruses, without being ill themselves. Therefore, the compound of the invention could be administered to animals at the same time as vaccination, and possibly for a further 2 weeks afterwards.

The names orthomyxo- and paramyxoviruses are assigned after separation of the former collective designation of influenza viruses and other similar viruses. In humans, paramyxoviruses cause measles, mumps, and respiratory and neurological diseases. Paramyxoviruses include, but are not limited to parainfluenza virus, mumps virus, avian pseudo-plague virus, syncytium respiratory virus (RSV), measles virus and bovine plague virus. Orthomyxoviruses include, but are not limited to viruses A, B and C, which are the causative agents of human influenza.

Type A influenza viruses are responsible for more influenza epidemics and all pandemics. Although influenza A viruses are found among horses and pigs, as well as among birds, for example, as a causative agent of classical bird plague, only human and mammalian influenza viruses, such as horses, can be considered respiratory viruses in the truest sense of the word, since biology avian influenza virus (avian influenza virus (AI)) is completely different from the biology of human influenza virus. Therefore, just the AI virus cannot be considered as a respiratory virus. In ducks, the AI virus primarily multiplies in the intestinal tract, which does not occur in humans. As a result, AI viruses can be isolated from feces of birds (Hinshaw et al., 1980, Canad. J. Microbiol. 26, 622-9). In addition, the rate of change of the nucleotides of the AI viruses is lower than the same rate of viruses that can be isolated from mammals. The evolution of viral proteins in organisms other than the bird organism, usually indicates a rapid accumulation of mutations that are not found in AI viruses (Gorman et al., 1991, J. Virol. 65: 3704-14; Ludwig et al., 1995, Virology 212: 555-61). Receptor specificity varies between different influenza viruses. Most AI viruses are associated primarily with the alpha2-3-galactose-sialic acid receptor. In contrast, human influenza viruses are primarily associated with the alpha 2-6-galactose-sialic acid receptor (Rogers + Paulson, 1983, Virology 127, 361-73; Baum + Paulson, 1990, Acta Histochem. Suppl. 40: 35-8).

Since the viruses of the invention cause a number of widespread diseases in humans and mammals, the antiviral effect of the dithiocarbamate compound of the invention is of particular importance in light of these viruses. Given that the compounds of the dithiocarbamate according to the invention have a pronounced effect against these viruses, these compounds are particularly suitable for the preparation of a number of agents for the treatment and prevention of such viral infections. The dithiocarbamate compounds of the invention can be prepared in large quantities easily and cheaply and even at elevated concentrations have almost no toxic effect on the cells being treated.

According to a preferred embodiment, the dithiocarbamate compounds according to the invention are contained in the agent in a concentration of 0.01-5000 mM, preferably 1-300 mM, particularly preferably 10-100 mM. At such concentrations, the dithiocarbamate compounds of the invention are particularly effective against infections caused by respiratory RNA viruses and have almost no side effects. The concentration used is selected depending on the viral infection being treated, its intensity, the organism being treated, for example, a person or animal, and age.

It is particularly optimal that the concentration of the dithiocarbamate compounds of the invention in the agent is 10 mM-1 M. In this case, the dithiocarbamate compounds of the invention are present in a highly concentrated form and the agent may be diluted to the desired concentration before treatment.

Preferably, the agent also contains a pharmaceutically compatible carrier. In this case, any medium known to the specialist in the field of pharmacy can be used, for example, physiological saline with phosphate buffer (PBS) or otherwise buffered solutions of sodium chloride or composition containing liposomes, and in this case, the medium is selected taking into account the type of treatment, viral infection and the body to be treated.

Preferably, the agent can be administered orally, through the nose, intravenously, parenterally, rectally or in the form of eye or ear drops, rinse or aerosol. Moreover, the route of administration depends in particular on the viral infection to be treated. An infection in the respiratory tract is treated, for example, by administering the agent through the nose, for example, in the form of an aerosol containing dithiocarbamate compounds, since the treatment of the viral infection is carried out at the site of the virus attack. Depending on the type of application, a certain concentration of dithiocarbamate compounds is selected or the product contains additional substances that are effective in this form of application. It is also possible to use the product in dry form, and before treatment it is diluted with an appropriate solvent.

Particularly effective use is achieved when the product contains additional antiviral substances. Thus, it becomes possible to suppress a viral infection in the respiratory tract from several sides and at the same time it is possible to weaken or completely destroy a whole set of different viruses. Such additional antiviral substances are, for example, substances that suppress replication, immunostimulating substances, neutralizing antibodies, etc., as well as, if necessary, substances that can stimulate the immune system as a whole.

Preferably, the agent comprises a combination of at least two different dithiocarbamates according to the invention, in particular a mixture of pyrrolidine dithiocarbamate and N, N-diethyl dithiocarbamate. The dithiocarbamate compounds of the invention perform pro- and antioxidant functions in cells. Their antioxidant effects include the removal of hydrogen peroxide, superoxide radicals, peroxynitrites, hydroxyl radicals and lipid peroxidation products. Due to this removal, dithiocarbamates are oxidized to thiuram disulfide. Thiuram disulfides are responsible for the prooxidative effects of dithiocarbamates, and in some cases the formation of thiurams depends on the presence of metals. It is described that the antiapoptotic activity of dithiocarbamates is probably determined by the deactivation of caspases caused by the oxidation of thiol.

It is particularly preferred that the agent according to the invention additionally contains substances selected from antibiotics, vaccines, immunosuppressants, stabilizers, immunostimulating substances, blood preparations or mixtures thereof. If antibiotics are additionally used, then other infectious bacteria can be destroyed along with respiratory viruses. If the product additionally contains vaccines, which are understood as both passive and active vaccines, then at the same time as treating or preventing a viral infection according to the invention, some other additional viral infections are also possible that can easily infect a weakened organism. Stabilizers can be added to increase shelf life or to extend shelf life. Blood preparations are, for example, plasma, blood cells, coagulation factors, etc., depending on the course of treatment of the patient.

It is advisable to use a tool to inhibit the growth of viruses. This ensures the non-proliferation of an infection that has already occurred, or, otherwise, its very quick treatment.

In accordance with another aspect of the invention, the latter relates to a disinfectant containing at least one dithiocarbamate compound of the invention, as described above. Under the "disinfectant" in the framework of the present invention refers to any tool used outside the body of a person or animal to fight viruses, for example, on surfaces, inside tools, in particular in media, or for cell cultures. Such disinfectants can be used, in particular, in the case when the processed material is sensitive to other aggressive antiviral substances. For example, a disinfectant containing the dithiocarbamate compounds of the invention can be used as an additive to media or to treat cells or cell cultures that are sensitive to other, more aggressive disinfectants or antiviral agents. The dithiocarbamate compounds of the invention have been particularly effective in the treatment and prophylaxis of respiratory cell infections and cell cultures caused by respiratory RNA viruses.

A disinfectant is particularly effective when it contains the dithiocarbamate compounds of the invention at a concentration of 10 μM-5 M, in particular 30 μM-1 M. Such concentrations are effective, on the one hand, against infections caused by respiratory RNA viruses, and on the other hand, such a disinfectant is extremely gentle, for example, when it is used as an antiviral substance for cell cultures. The concentration is determined by the type and stage of development of the viral infection used in the treatment of the substance, for example, on the type and sensitivity of the cells. It goes without saying that it is also possible to prepare a sanitizing agent in the form of a concentrate, and before use it is adjusted with an appropriate solvent to the desired concentration of the dithiocarbamate compound according to the invention.

Preferably, the disinfectant contains additional disinfectants, in particular antiviral agents. Such substances, known to any specialist in the field of microbiology, are added in the case when it is necessary to simultaneously destroy other viruses, for example, DNA viruses. It goes without saying that antibacterial substances, in particular antibiotics, can also be added.

Another aspect of the present invention is a method of disinfecting surfaces, media and cell cultures, the disinfectant according to the invention, as described above, is applied to the surface or culture of cells or introduced into the medium. For disinfecting a surface, it is sufficient, for example, to clean it with a disinfectant. In the case of cell media and cultures, the disinfectant can be exposed for a longer time, and, as described above, the concentration of the disinfectant can vary depending on the purpose.

According to another aspect of the invention, the latter relates to the treatment and prophylaxis of an infection caused by respiratory RNA viruses using the dithiocarbamate compounds of the invention. In this case, the agent containing the dithiocarbamate compounds according to the invention, as described above, is administered to the patient or animal in an appropriate form and concentration.

The present invention is explained in more detail below with the aid of examples and drawings, to which it is not limited.

Brief Description of the Drawings

Figure 1 is a graphical representation of the inhibitory effect of pyrrolidine dithiocarbamate (PDTC) on human rhinovirus replication in cell culture;

Figure 2 represents the inhibition of pyrrolidine-dithiocarbamate cytopathic effects induced by rhinoviruses;

Figure 3 shows an increase in cell viability using pyrrolidine dithiocarbamate and N, N-diethyl dithiocarbamate (DDTC);

Figure 4 shows the effect of pyrrolidine-dithiocarbamate in treatment, depending on the duration of use;

Figure 5 shows the cleavage of eIF4GI;

6 illustrates a Western Blot assay for determining expression of rhinovirus capsid proteins;

7 shows the effect of other antioxidants on human rhinovirus infection (HRV);

Figures 8 and 9 show the effect of pyrrolidine dithiocarbamate on influenza virus replication;

Figure 10 illustrates the dependence of the action of pyrrolidine dithiocarbamate on Vero cells infected with influenza viruses on its concentration;

11A and 11B illustrate the effect of pyrrolidine dithiocarbamate on mice infected with influenza virus;

12 shows the efficacy of pyrrolidine dithiocarbamate against horse rhinitis virus A (ERAV);

13A and 13B illustrate the efficacy of pyrrolidine dithiocarbamate against MKS;

Fig. 14 shows the absence of the action of pyrrolidine dithiocarbamate on encephalomyocarditis virus (EMCV).

Examples

Example 1. Reducing the production of infectious particles of rhinovirus using pyrrolidine dithiocarbamate (PDTC)

To determine the effectiveness of human rhinovirus (HRV) infections, PDTC supplementation was introduced after cell infection with different HRV serotypes.

HeLa cells were infected with serotypes 1A, 2, 14 and 16 of the HRV virus at TCID ₅₀ (dose 50% of tissue culture infection) per cell. At the same time, 125 μM PDTC was added to the medium. After 4 hours of infection, excess virus was removed and PDTC was again added to fresh medium. After 24 hours (Fig. 1, top) and 48 hours (Fig. 1, bottom) after infection (R.I.), the supernatant was collected and the number of viral offspring was determined using TCID ₅₀ tests. Processing of the cells with PDTC decreased after 24 hours the virus titer by 10 ³ (FIG. 1, top). The supernatant of PDTC-treated cells, which were collected within 48 hours after infection, also showed a significant decrease in virus titer (FIG. 1, bottom). These tests suggest that PDTC has a strong antiviral effect on different HRV serotypes.

Example 2. Inhibition of cytopathic effects induced by HRV virus and increasing the viability of infected cells using PDTC

In the late stages of rhinovirus infection, morphological changes in the cells, known as “cytopathic actions” (CPE), occur. These HRV-induced “cytopathic actions” are characterized by cell rounding, wrinkling, nuclear deformation, and chromatin condensation. In HeLa cell infections caused by the HRV2, HRV14, HRV1A, and HRV16 viruses, a clear cytopathic effect was observed at 100 TCID ₅₀ per cell 8 hours after infection. PDTC supplementation during infection inhibited the appearance of these cytopathic effects. Figure 2 shows that the cell morphology 8 hours after infection in the presence of 125 μM PDTC (see the lower right figure) did not differ from the morphology of uninfected cells (see the upper two figures).

Thus, the use of PDTC leads to the cure of a viral infection at a very early stage, as a result of which significant cell damage is prevented.

Example 3. Increasing the viability of infected cells using pyrrolidine dithiocarbamate (PDTC) and N, N-diethyl dithiocarbamate (DDTC)

A rapid cell multiplication experiment was conducted to test the effects of PDTC and DDTC on cell viability during viral infection.

For this cell titer was determined by an instrument 96 ^® Aq _ueous Non-radioactive test for rapid cell proliferation (Promega; Madison, Wisconsin, USA ) according to the manufacturer's instructions. The day before the infection, the cells were placed on plates with 96 corrugations. These cells were infected with different HRV serotypes at 20 TCID ₅₀ / cell. Cell viability was determined by adding tetrazolium, incubation was performed for 2 hours at 37 ° C, and absorbance was measured at 492 nm.

Twenty-four hours after HRV2 infection, ZeLa cells showed a complete loss of their metabolic activity compared to uninfected HeLa cells (mock infection medium = MIM false infectious medium). In FIG. 3 (top: DDTC; bottom: PDTC), it can be seen that the viability of the cells themselves increased significantly after the addition of PDTC or DDTC in small concentrations. Only PDTC or only DDTC affects uninfected cells only slightly. However, the concentrations used to inhibit viral infections did not cause toxicity (“-” means “without”).

Example 4. The effectiveness of PDTC depending on the time of action

To determine the stage in the life cycle of the virus at which PDTC begins to act, the latter was added at different points in time after the viral infection at 20 TCID ₅₀ / cell and proliferation tests were performed. It was found that the addition of PDTC (125 μM) during the first six hours after infection (“-” means “without”) provides the best protection against virus-induced proliferation loss (FIG. 4A). This effect is not typical of a single serotype, since serotypes 1A, 2, 14 and 16 of the HRV virus were also used. Only treatment of cells with PDTC (25 μM) more than 8 hours after infection reduced the protective effect.

Similar results were obtained when the titers of the virus were determined in the supernatant of infected and PDTC-treated cells (see FIG. 4B). The addition of PDTC for up to four hours after infection reduced the titers of HRV2 produced by 10 ³ . Even if PDTC treatment started 6 hours after infection, the produced virus titers were significantly underestimated. These results indicate that PDTC supplementation also has an antiviral effect when it occurs at a too late stage of infection, whereby cell viability increases and the number of infectious viruses decreases sharply.

This indicates that the antiviral effect is manifested not only in the early stages of the life cycle of the virus, for example, during the adhesion of the receptor or its introduction into the cell.

Example 5. The study of the process of infection caused by HRV in the presence of PDTC

To study the process of rhinovirus infection in the presence and absence of PDTC, various proteolytic activities were analyzed. A characteristic proteolytic activity manifested during infections caused by rhino- and enteroviruses is the enzymatic cleavage of cellular translational-initiating factors 4GI (eIF4GI) and 4GII viral 2A-proteinase, resulting in the completion of the synthesis of the protein of the host cells. Depending on the HRV serotype, eIF4G proteins are cleaved at an early stage of infection. An additional, only briefly described cleavage activity for rhinovirus infection is the cleavage of the intermediate filament protein cytokeratin 8. Also, this cleavage depends on the 2A protease, but occurs at a late stage of infection. To determine the effect of PDTC treatment on the course of a viral infection, a Western Blot assay was performed in which 2A protease substrates were analyzed.

The Western Blot analysis removes the medium at appropriate times. Cells were lysed by adding 100 μl of protein buffer (8% sodium dodecyl sulfate, 20% β-mercaptoethanol, 20% glycerol, 0.04% bromophenol blue). 20 μl of the protein extract was separated per track using SDS-PAGE and blotting was performed on a PVDF membrane. Antibody incubation is carried out at 0.1% Tween 20 and 5% skim milk powder in TBS. When determining the immunity used polyclonal rabbit antibodies against eIF4GI. Rabbit anti-immunoglobulin conjugated with alkaline phosphatase was used as secondary antibodies. Alkaline phosphatase staining, molecular sizes were determined using a pre-stained marker SDS-7B (Sigma).

In HeLa cells infected with HRV2 (100 TCID ₅₀ per cell), eIF4GI cleavage was detected 4 hours after infection (“cp” means cleavage product, “cleavige product”), complete cleavage occurred 8 hours after infection (FIG. 5 ) At this point, no eIF4GI cleavage was observed in infected cells in the presence of PDTC. In later periods, approximately 24 hours after infection, a slight cleavage of eIF4GI could also be seen in PDTC treated cells.

This suggests that either the protease function was blocked, or the amount of viral protein decreased sharply.

Example 6. Detection of integumentary proteins of viruses

To determine the effect of PDTC on viral protein expression, HRV2 capsid proteins were determined in protein extracts of HeLa cells infected with HRV2 virus (100 TCID ₅₀ ) by Western Blot analysis (see FIG. 6). Western Blot analysis was performed as described above in Example 5, using rabbit polyclonal anti-HRV2 antiserum. Significant amounts of rhinovirus proteins VP1, VP2 and VP3 were detected in untreated cells 6 hours after infection. PDTC treatment prevents the expression of these capsid proteins for the first 8 hours after infection. Weak expression of VP1, VP2, and VP3 was detected only later, approximately 24 hours after infection.

As a result, it was shown that PDTC causes the late production of viral proteins, which explains the anti-rhinoviral effect of PDTC and its cell defense.

Example 7. Determination of the dependence of the antiviral effect of redox potential

The inhibitory effect of other antioxidants during the infection of HeLa cells caused by the HRV2 virus was investigated (Fig. 7). HeLa cells were infected on a 6-well plate with HVR2 virus (100 TCID ₅₀ / cell) (ni means “not infected”). One hour after infection, the medium is removed and fresh medium or medium with PDTC or NAC in different concentrations is added. After 24 hours, the cell layer was washed with saline with phosphate buffer (PBS) and stained with crystal violet. On figa you can see that the cell layer was destroyed by the infection caused by the HRV2 virus, this effect can be prevented using PDTC, but not NAC. The mechanism of action of vitamin C, Trolox and β-mercaptoethanol (2-ME) is determined as described in example 3 (figures 7B, C, D). It was unexpectedly discovered that all of these antioxidant substances did not exert any protective effect during the propagation of viruses. For control purposes, the toxic effects of substances in the absence of viruses were verified. A large dose of vitamin C (100 μg / ml) strongly inhibited cell growth.

Thus, it was shown that the antiviral effect of PDTC is not explained by its antioxidant effect, but is obviously associated with other properties.

Example 8. The effect of PDTC on the replication of influenza virus

Influenza virus A / PR8 / 34 or Vienna / 47/96 at 0.01 multiplicity of infection (moi = multiplicity of infection) infects 5 × 10 ⁵ Vero cells and incubates for 1 h at room temperature. The inoculum is then removed and 5 μg / ml trypsin and 600 μM PDTC are added to the infectious medium. After 48 hours and 72 hours, the supernatant was removed and the titer of the virus was measured using a standard plate assay. As shown in FIG. 8, the titer of the A / PR8 / 34 virus decreased in the presence of PDTC by more than 2 log steps compared to the control sample (c).

To determine the effectiveness of PDTC, a TCID ₅₀ test (50% dose of tissue culture infection) is performed using and without PDTC. TCID _{50 was} calculated by the Kaerber method for any concentration: 96-well microplates were infected with the virus at double dilution, 1 hour after infection, the supernatant was removed and the medium containing PDTC at the indicated concentrations was added. The number of infected wells is determined after 4 days. Figure 9 shows how, at a concentration of 300 μM PDTC, TCID ₅₀ decreased by 76.4% and 82.2% for A / PR8 / 34 and A / Vienna / 47/96.

Inhibition of more than 99.9% for both viruses was achieved at a concentration of 1200 μM.

Example 9. Determination of the effective concentration of PDTC

The effective concentration of PDTC is determined by cytopathic reduction assay (CPE Reduktion-Assay) (CPE = cytopathic effect): Vero cells are cultured on 96-well microplates and infected at 5 TCID ₅₀ and 50 TCID ₅₀ per well with influenza virus A / PR8 / 34 and at 5 TCID ₅₀ per well, influenza virus A / Vienna / 47/96. 1 h after infection, the supernatant is removed and medium with 5 μg / ml trypsin and two-fold diluted PDTC with an initial concentration of 1200 μM is added. In the next four days, the tablets are visually examined for cytopathic effect. The manifestation of the cytopathic effect is calculated in relation to the control sample for each concentration. Positive 100% means complete lysis in all wells. In Fig. 10 it can be seen that a 50% decrease in the positive wells was obtained at a concentration of from 50 to 100 μM PDTC, a complete cessation of the cytopathic effect for all viruses occurred at a concentration of 600 μM PDTC.

Example 10. Determination of the effect of PDTC in vivo

10 C57 / BL6 mice were inoculated through the nose with a lethal dose (50 μl of 10 ⁷ pfU) of A / PR8 / 34 virus. One hour after this, they were injected through the nose with 25 μl of PDTC 600 mM and 25 μl of saline with phosphate buffer. Mice are examined and treated for the first 48 hours every 12 hours, then every 24 hours. The weight of the mice is measured and expressed in% of the initial weight (% w, Fig. 11A). On figv it can be seen that all treated with PDTC mice (square) suffered a viral infection and 7 days after infection gained weight. All mice treated with saline with phosphate buffer (rhombuses) died within 12 days after infection (% s =% survivors).

Thus, it was shown that already one PDTC has a strong effect against infections caused by influenza virus, in vitro and in vivo.

Example 11. Antiviral activity of PDTC against horse rhinitis virus A (ERAV)

A decrease in the propagation of horse rhinitis A virus by PDTC supplementation is examined as follows: Vero cells infect 10 TCID ₅₀ per cell with ERAV virus. At the same time, PDTC at various concentrations (1 mM-50 μm) was added. 4 hours after infection, the inoculum is removed and fresh medium containing PDTC is added. 24 hours after infection, the supernatant is removed and the titer of the virus is determined using a standard plate assay.

12 shows how the titer of the virus (vt) in the supernatant is reduced compared to untreated cells.

Example 12. The effect of PDTC on the propagation of foot and mouth disease virus (MKS) in cell culture

Using a cytopathic reduction assay (CPU Reduktions-Assay), the effective concentration of PDTC was determined when exposed to cells destroyed by foot-and-mouth disease virus.

Cells of IB-RS-2 were cultured with 96-well microplates, infected with foot-and-mouth disease virus O-Manisa at 0.1 TCID ₅₀ per cell and incubated for 1 h at 37 ° C. Then the inoculum is removed and the infectious medium with the indicated concentration of PDTC (10 μM-1200 μM) is added. After 24 hours, the number of infected wells was determined by observing through a microscope the cytopathic effect.

On figa shows that the number of infected wells (% pos.) After 24 hours depended on the concentration of PDTC. 600 μM PDTCs protect cells 100% of the cytopathic effect caused by viruses. A 50% reduction in infected wells is achieved at a concentration in the range of 75-150 μM.

To determine the effect of PDTC on foot-and-mouth disease virus replication, IB-RS-2 cells are infected in T25 cm ² flasks with FMD virus O-Manisa at 0.001 TCID ₅₀ per cell and incubated for 1 h at 37 ° C. The inoculum is then removed and the infectious medium containing PDTC at the indicated concentrations (0 μM-200 μM) is added. 24 hours after infection, the supernatant is removed and the virus titer (TCID) is determined in a standard plate assay.

As shown in FIG. 13B, the titer of 0-Manisa foot and mouth disease virus decreased in the presence of PDTC by more than 2 log steps compared to control (0). No PDTC was added to the control sample.

Example 13. The effect of PDTC on the propagation of FSME viruses in cell culture

FSME viruses are not picornaviruses and do not cause diseases in the respiratory tract.

The monolayers of BHK-21 cells (ATCC) are infected with the FSME virus at 10 pfu / cell (Neudörfel) in the presence of PDTC at the following concentrations: 1000 μM, 500 μM, 250 μM, 125 μM, 62.5 μM, 31.25 μM, 15 6 μM, 7.8 μM, 3.9 μM, 1.95 μM, 0.975 μM.

Then the cells are incubated for 4 days at 37 ° C and the monolayer of cells is analyzed under a microscope. Virus propagation is determined by enzyme immunoassay. Microscopic analysis showed no evidence of toxic effects of PDTC. A quantitative assessment of virus reproduction indicates that PDTC in any (of the studied) concentration is not able to reduce virus reproduction.

A similar study was conducted to identify the effect of PDTC on the propagation of the virus in cells in suspension: the same concentrations were tested as above. And in this experiment, there was also no evidence of a decrease in virus reproduction under the influence of PDTC.

Example 14. The effect of PDTC on mice infected with EMC virus

C57B16 mice were intraperitoneally infected with EMC virus at 10 TCID ₅₀ . The control group was injected intraperitoneally once a day with 50 μl of physiological saline with phosphate buffer. Two other groups were injected with 50 μl of 50 mM PDTC per day, with treatment being started simultaneously with infection (PDTC) in one group and 24 hours after infection in the other group (PDTC, 24 hours after infection).

On Fig shows the weight change of mice after the onset of infection. It is obvious that the processed and unprocessed mice do not differ from each other. Among infected mice of all groups, death occurred on average after 5.5 days.

Thus, it was shown that PDTC has no effect on the infection caused by the EMC virus, which although it belongs to picornaviruses, however, the disease does not cause in the respiratory tract, but in the nerve cells.

Claims (19)

1. The use of dithiocarbamate compounds of the structural formula R ₁ R ₂ NCS ₂ H, in which R ₁ and R ₂ mean, independently from each other, unbranched or branched (C ₁ -C ₄ ) alkyl or form an aliphatic ring with 4- 6 carbon atoms, wherein R ₁ , R ₂ or the aliphatic ring is optionally substituted with one or more substituents selected from OH, NO ₂ , NH ₂ , COOH, SH, F, Cl, Br, I, methyl or ethyl, and oxidized forms of these compounds, in particular their dimers, as well as their pharmaceutically compatible salts, for the preparation of a treatment or prevention of infection caused by RNA viruses that affect the respiratory tract of humans and animals and cause disease in it. 2. The use according to claim 1, characterized in that R ₁ and R ₂ mean, independently of each other, (C ₁ -C ₃ ) alkyl or form together with the nitrogen atom an aliphatic ring with 4-6 carbon atoms. 3. The use according to claim 1 or 2, characterized in that the dithiocarbamate compound is selected from pyrrolidindithiocarbamate and N, N-diethyldithiocarbamate. 4. The use according to claim 1, characterized in that the viral infection is an infection caused by picornavirus, orthomyxovirus or paramyxovirus. 5. The use according to claim 4, characterized in that the orthomyxovirus is a human influenza virus, in particular, originating from the group of influenza viruses A, B and C, and paramyxovirus is a parainfluenza virus or pneumovirus. 6. The use according to claim 4, characterized in that the orthomyxovirus is a mammalian influenza virus A. 7. The use according to claim 4, characterized in that the picornavirus is a rhinovirus, in particular a human or horse rhinovirus, an enterovirus, in particular an enterovirus 70, 71, or a Coxsackie virus or an aftovirus, in particular foot-and-mouth disease virus, or horse virus A rhinitis. 8. The use according to claim 1, characterized in that the tool contains a compound of dithiocarbamate in a concentration of 0.01-5000 mm, preferably 1-300 mm, particularly preferably 10-100 mm. 9. The use according to claim 1, characterized in that the agent contains dithiocarbamate compounds at a concentration of 10 mM-1 M. 10. The use according to claim 1, characterized in that the tool further comprises a pharmaceutically compatible carrier. 11. The use according to claim 1, characterized in that the tool is intended for use orally, through the nose, intravenously, rectally, parenterally or in the form of drops for the eyes or ears or as a gargle for a throat or aerosol. 12. The use according to claim 1, characterized in that the tool additionally contains antiviral substances. 13. The use according to claim 1, characterized in that the tool contains a combination of at least two different compounds of dithiocarbamate. 14. The use according to claim 1, characterized in that the agent additionally contains substances selected from antibiotics, vaccines, immune suppressors, stabilizers, immunostimulating substances, blood products or mixtures thereof. 15. The use according to claim 1, characterized in that the tool is used to inhibit the growth of viruses. 16. A disinfectant for eliminating RNA viruses that infect the respiratory tract of humans and animals, containing at least one dithiocarbamate compound described in any one of claims 1-15. 17. The disinfectant according to clause 16, characterized in that it contains a compound of dithiocarbamate at a concentration of 10 μm-5 M, in particular 30 μm-1 M. 18. A disinfectant according to claim 16 or 17, characterized in that it further comprises disinfectants, in particular antiviral substances. 19. A method of disinfecting surfaces, media and cell cultures, characterized in that the disinfectant according to claim 17 or 18 is applied to the surface or culture of cells or introduced into the medium. Priority on points and signs: According to paragraphs 1, 4, 7 - in terms of the use of dithiocarbamates with the exception of dimers (oxidized forms) for the treatment or prevention of picornavirus infection, Convention priority is set on July 16, 2001 according to application AT 1102/01. Regarding the use of dithiocarbamates, with the exception of dimers (oxidized forms) for the treatment or prevention of viral infection in the respiratory tract, priority has been set for all items on the date July 16, 2001 according to application AT 1103/01. Regarding the use of dithiocarbamates, including oxidized forms, in particular their dimers for the treatment and prevention of viral infection in the respiratory tract, priority is set for all claims by date of December 17, 2001 according to AT 1973/01. Regarding the use of dithiocarbamates, as well as their oxidized forms, in particular dimers for the treatment or prophylaxis of picornavirus infection, according to all claims, priority is set to date 17.12.2001 according to application AT 1972/01.

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