RU2178710C1 - Individual substances synthesized on base of chemical interaction of disulfide-containing peptides with purine or pyrimidine base derivatives, pharmaceutical compositions and preparations based on thereof, methods of their using for treatment of infectious diseases and prophylaxis of complications - Google Patents

Abstract

FIELD: medicine, pharmacy. SUBSTANCE: invention relates to development of medicinal agents based on active metabolites of peptide and nucleoside-nucleotide nature. Invention proposes novel compounds synthesized on the base of chemical interaction of oxidized glutathione (G-SS-G) with derivatives of purine or pyrimidine bases; pharmaceutical compositions and preparations based on thereof eliciting antiviral, antibacterial, immunomodulating and hepatoprotective effects; methods of treatment and prophylaxis of infectious diseases. EFFECT: development of novel medicinal agents and methods of their using for treatment and prophylaxis of infectious diseases of different etiology. 89 cl, 42 dwg, 60 tbl

Description

 The invention relates to medicine, in particular to anti-infectious pharmacology, in particular to the creation of new drugs based on active metabolites of peptide and nucleoside-nucleotide nature, intended for the treatment and prevention of infectious diseases, primarily viral hepatitis B and C, as well as AIDS and herpes and also caused by gram-positive and gram-negative bacteria, in addition, protozoa and fungi, in particular, for the treatment of tuberculosis, malaria, intestinal infections, chlamydial and mycoplasma infections.

 Infections caused by the hepatitis B virus (HBV) and hepatitis C virus (HCV) prevalence significantly exceed HIV infection. The rate of infection with HBV and, especially, HCV is increasing every year. In some regions of the world, hepatitis C affects more than 10% of the adult population. At the same time, HCV has the highest chroniogenic potential, being the main reason for the formation of the entire spectrum of severe chronic liver diseases. In particular, the proportion of chronic hepatitis induced by hepatitis C is more than 70%, while the proportion of liver cirrhosis is 40%, and in cases of hepatocellular carcinoma - 60% [1]. The fundamental mechanisms for the chronicity of viral hepatitis, especially hepatitis C, are the mechanisms of “escaping the virus” from under immune surveillance, as well as in connection with the existence of multiple simultaneously present variants of the virus with a modified but closely related genome - quasispecies (“quasisspecies”). Recent studies have shown the presence of virus-specific T-lymphocytes in liver cells, which confirms the immune-mediated pathogenesis of hepatitis C [2]. Moreover, the dependence of the activity of morphological changes in the liver (Knodel index) on the level of intrahepatic CD4 + cells has been established [3]. Of exceptional importance is the T-cell response, primarily the response of CD4 +, CD8 + and SD16 / 56 + to HCV NS3 protein, which is characterized as highly significant for the spontaneous resolution of hepatitis C, as well as for positive results of antiviral therapy [4, 5].

 In turn, it is known that the nature of the immune response depends on the dominant involvement of CD4 + T-lymphocyte clones - type 1 helpers (Th1) and type 2 helpers (Th2), which differ in the cytokines they produce and in the activation of the immune response in the cellular or humoral type . The activation of Th1 producing interferon gamma (IFN-γ), interleukin 2 (IL-2), tumor necrosis factor alpha and beta (TNF-α and β), leads to stimulation of the function of T-lymphocytes and resident macrophages, i.e., the development of the cellular type of immune response, which plays a decisive role in protecting against viruses. Activation of Th2, which secrete IL-4, IL-5, IL-6, IL-9, IL-10, and IL-13, triggers the humoral immunity. Hence, the imbalance in the production of Th1 / Th2 cytokines by cells plays a decisive role in the immunopathogenesis of HBV, HCV and HIV infections, and the predominant participation of Th2 type cytokines is associated with viral persistence and chronicity of the process. On the contrary, the activation of production of Th1-dependent cytokines promotes spontaneous recovery in acute viral infections and the effectiveness of therapy aimed at eliminating viruses from the body and restoring the functional capacity of affected organs [6, 7, 8, 9].

 In this regard, the function of the pleiotropic cytokine IL-12, which is one of the key "figures" in the regulation of the immune response of a macroorganism, is equally important. By regulating the balance of Th1 / Th2, IL-12 modulates the function of macrophages, especially resident macrophages (Kupffer cells) of the liver. In chronic hepatitis C, as in chronic hepatitis B, IL-12 activates the secretion of cytokines by Th1 cells, inhibiting the corresponding function of Th2. Chronization and adverse course of viral hepatitis is accompanied by a decrease in the level of IL-12 [10]. Thus, it can be argued that the determining course and outcome of HBV and HCV infection and their successful therapy - T-cell immunity suffers from functional weakness, primarily, in our opinion, in terms of the endogenous production of an adequate and consistent "team" of cytokines in response to antigenic factor. It is cytokines in conditions of constant "competition" on the speed between the formation of new "quasisspecies" and the body's immune response that determine the ability to modulate the quality and direction of the immune system response. Hence, the methods of treatment of viral hepatitis and AIDS should involve the use of immunorehabilitation agents, which are simultaneously antiviral drugs, capable of not only finely regulating the endogenous production of cytokines (including reproducing their effects when blocking helper and cytotoxic activity), but also selectively inhibiting replicative virus activity.

 All antiviral chemotherapy drugs used to treat viral hepatitis belong to 3 main groups: 1 - reverse transcriptase inhibitors, including nucleoside analogs, nucleotide analogs and non-nucleoside analogs; 2 - protease inhibitors; 3 - analogues of pyrophosphate. As shown in the scientific literature, with hepatitis B (HB) from drugs of the group of nucleoside analogs, lamivudine, azidothymidine, famciclovir is most effective [11, 12]; in hepatitis C (HS), the following are widely used therapeutic agents: ribavirin (rebetol), azidothymidine [11, 13].

 Antiviral drugs of the 1st group, namely lamivudine, ribavirin and azidothymidine, were selected as analogues in use, taking into account the data on the nature of the known antiviral agents and the basic structural formula of these substances, as well as to assess their comparative effectiveness with the new drugs proposed in this invention .

 In a number of studies [14, 15, 16], the nucleoside analogue lamivudine was called a promising drug in the treatment of chronic hepatitis B. At the same time, it is noteworthy that only 52 weeks of treatment with lamivudine in patients determine the absence of HbsAg, stable suppression of HBV DNA and stable normalization of alanine aminotransferase levels.

 Despite the lack of data on long-term monitoring of patients receiving lamivudine, numerous side effects of lamivudine are known, including: from the gastrointestinal tract (vomiting, nausea, diarrhea syndrome); hepato- and nephrotoxicity; from the hemopoietic system (neutropenia, thrombocytopenia and anemia); adverse dermatological reactions (skin rash, alopecia) occur.

 An analysis of the literature on the effectiveness of the treatment of chronic hepatitis C with ribavirin allows us to make the following main conclusion: the use of ribavirin in mono mode does not provide a satisfactory therapeutic effect; at the same time, the combination of high doses of interferon and ribavirin used for long periods of at least 12 months is the optimal therapy that provides a high percentage of a stable response to treatment [17].

 Ribavirin is also not without serious side effects, in particular such as: bronchospasm, pulmonary edema, hypotension, anemia, skin rash, asthenia. However, the main disadvantage of ribavirin is the extremely low antiviral activity and the absence of hepatoprotective effects, which is fundamentally important for the content of the methodology for treating viral hepatitis.

 The drugs of the AZT group in the recent past were the main drugs in the treatment of AIDS and herpetic infections [18]. Their widespread use was hampered by the rapid development (after 2-3 courses of treatment) of very serious complications in the form of hemo- and immunosuppression, allergic dermatoses and mucosal candidiasis, which excluded the use of AZT.

 Summing up the prior art, it should be emphasized that today the main method of treatment for hepatitis B and C are recombinant interferons and nucleoside analogues. The adopted combination therapy program (3 million ME interferon 3 times a week for 6 months and ribavirin), until recently considered the "gold standard", yesterday provided remission of the disease in more than 30% of patients, today - in 12-15% of patients. Such negative dynamics in the therapeutic efficacy of the traditional antiviral combination is due, along with other mechanisms, to the fact that interferons are high molecular weight (large) protein substances that cause the formation of antibodies. Hence, the larger and longer the similar therapy within the human population (taking into account the pace of the galloping epidemic of viral hepatitis), the lower its effectiveness every year and the higher the level of complications in the form of immunoaggression.

 In the treatment of viral hepatitis, along with the indicated drugs of direct antiviral effect, drugs with hepatoprotective effects are used. One such drug is heptral (S-adenosylmethionine). This drug is considered as an analogue of the claimed drug in terms of hepatoprotective properties.

 In this regard, moving on to the conceptual model of the present invention, we consider it possible to postulate the promise and priority of the use of low molecular weight compounds, structurally functional analogues of key cellular regulatory factors (active metabolites) that have antiviral, simultaneously immunomodulatory, as well as hepatoprotective and hemoprotective effects.

 The following terminology is used in the present invention.

 The concept of "metabolism" means the totality of all biochemical reactions that occur in the cells of living organisms [19]. "Active metabolites", in turn, are individual biochemical compounds, often short-chain peptides (from 2 to 9 amino acids), the dynamics of the content of which in the cells determines the direction and activity of certain metabolic processes.

 The term “apoptosis” refers to a morphologically recognizable form of genetically programmed cell death [20, 21, 22]. By mechanisms of apoptosis, aging cells are removed from the body; Apoptotic mechanisms induce cell death during embryogenesis, as well as the death of "spent" activated immune cells. In the classical definition, apoptosis is thus defined as the physiological suicide of cells.

 The concept of cytokines, including interleukins (IL); interferons (IFN); tumor necrosis factors (TNF) and others, involves regulatory substances of a protein nature, produced, as a rule, by immunocompetent cells and playing a key role in the development of the immune response, hematopoiesis, and apoptosis processes. Hence, the dynamics and content of endogenous cytokine production largely determines the nature and characteristics of the pathogenesis of various, including infectious diseases [20, 23].

 Immunocompetent cells, designated as: CD3 +; SD4 +; SD8 +; SD16 / 56 + (NK cells) are various types (differentiation markers) of T-lymphocytes that perform their functions inherent in a given type of cell, which form the body's immune response to the action of antigens or other pathogenic elements.

 Knodel's Histological Activity Index (IHA) proposed by R. G. Knodell et al. (1981), determines the nature and extent of inflammatory-necrotic lesions of the liver cells (hepatocytes), as well as the nature of the morphological changes in the liver tissue. The Knodel index is used to determine the degree of activity of damage to the liver tissue in arbitrary units - points - for various types of hepatitis, including viral hepatitis B and C. Moreover, an assessment in the range of 1-3 points is typical for minimal activity; 4-8 - for mild; 9-12 - for moderate and 13-18 - for severe, usually observed in the case of cirrhotic changes in the liver.

 Polymerase chain reaction (PCR) determines the presence (+) or absence (-) of replicative activity of DNA or RNA viruses, for example, hepatitis B virus (HBV) or hepatitis C virus (HCV), respectively. Quantitative PCR allows you to determine (measure) the number of viral copies in 1 ml of blood of the human body. Hence, for example, high replicative activity, that is, active reproduction of the virus in the body (high viral “load”), will determine the severity of the clinical course of the infectious disease and, conversely, the cessation of the replicative activity of the virus or its significant decrease indicates antiviral activity of drugs used to treat viral diseases.

 Non-structural protein of hepatitis C virus (NS3) is a key regulatory protein of the virus necessary for viral replication (therefore, activity). This protein is a multifunctional enzyme with three catalytic activities, such as protease, helicase (“untwisting” the DNA of the “host” cells for the introduction of the virus), and ATPase, that is, the ability to cleave ATP to provide energy for the virus replication process (so called "cap" synthesis) [24].

 The unique properties of oxidized glutathione (GSSG) - to stimulate the endogenous production of cytokines and hematopoietic factors - have been established and patented by the authors earlier [25].

Stabilization of the GSSG disulfide bond promotes prolongation of the residence time of exogenously introduced GSSG in biological media in the oxidized (disulfide) form and fundamentally enhances the first established biological and pharmacological effects of GSSG [25, 26]. In biological media, GSSG is metabolized by the NADP • H ⁺ -dependent glutathione reductase enzyme, which cleaves the GSSG disulfide bond to form two reduced glutathione (GSH) molecules [25, 26].

Hence, a new solution for prolonging the half-life of exogenously introduced GSSG in biological media in disulfide form was the method for producing a composite GSSG compound with inosine or with inosine monophosphate (IMP). Inosine and IMP in the composition of the named composite compound protected GSSG from the “attack” of NADP • H ⁺ -dependent glutathione reductase, providing this composite compound with a new quality of its biological and pharmacological effects, the integrative result of which is the ability to regulate cell proliferation, differentiation and apoptosis [ 26].

The above patent [26] presents a group of pharmaceutically acceptable substances containing, as an active principle, derivatives of GSSG in the form of its salts, which provide:
- a new pharmacokinetics of GSSG in the composition of these derivatives of GSSG;
- targeted delivery of GSSG to one or another organ, i.e., targeted tropism to various tissues and organs;
- the new content of the main biological and pharmacological effects of GSSG, more precisely its structurally functional analogue - hexapeptide with a stabilized disulfide bond.

 Thus, in the patent [26], as the closest analogue that we selected as a prototype of the claimed invention, the salts of oxidized glutathione are described, which are organic salts of GSSG obtained by the interaction of oxidized glutathione and inorganic bases.

 The problem to which the present invention is directed, is the creation of new individual substances, pharmaceutical compositions and preparations obtained on the basis of the interaction of disulfide-containing peptides with purines and pyrimidines, including with nitrogenous bases; nucleosides and nucleotides, as well as the development of methods for their use for the treatment of infectious diseases and the prevention of their complications.

By nature it is:
- or organic salts, in which the counterions are GSSG and nitrogenous bases; or GSSG and nucleosides of purine and pyrimidine nature;
- or new chemicals derived from the formation of a covalent bond between GSSG and nucleotide monophosphates.

 According to the IUPAC classification, not to mention the compounds obtained on the basis of the formation of a covalent bond between two substances, an organic salt formed on the basis of the formation of ionic bonds between its constituent components is also an individual substance, since the corresponding salt-forming cations and anions are strictly defined.

The present invention discloses that in the case of the preparation of a number of organic GSSG salts with nitrogen bases and / or nucleosides of purine and / or pyrimidine nature, the ionized carboxyl group of the glycine residue of the GSSG molecule acts as the anion of the formed salt, and the protonated nitrogen atom inside the heterocycle acts as the cation nitrogen base molecules (e.g. adenine, guanine or uracil); or nucleosides (e.g., inosine or thymidine with a protonated nitrogen atom N ₁ ).

At the same time, the formation of a number of organic salts based on the interaction of GSSG and nucleotides is shown, for example:
- GSSG and inosine monophosphate (GSSG-IMP);
- GSSG and uridine monophosphate (GSSG-UMF).

The present invention discloses a number of compounds obtained by forming a covalent bond between GSSG and nucleotide monophosphates, for example: inosine-5 ¹ -phosphate, uridine-5 ¹ -phosphate, cytidine-5 ¹ -phosphate, thymidine-5 ¹ -phosphate, adenosine- 5 ¹ -phosphate and guanosine-5 ¹ -phosphate. In this case, the formation of a phosphorus-amide bond between the amino group of the sodium or other inorganic salt of GSSG and the phosphoric acid residues of nucleotide monophosphates is purposefully used. The formation of a covalent bond proceeds according to one chemical reaction scheme and gives products whose formulas are shown in the corresponding figures.

One of the embodiments of the invention is the presented series of substances obtained:
a) based on the formation of ionic bonds between D-forms of GSSG and nitrogen bases, between D-forms of GSSG and nucleosides, the formulas of which are shown in the corresponding figures; and
b) based on the formation of a covalent bond between the salts of the D-forms of GSSG and nucleotide monophosphates, the formulas of which are shown in the corresponding figures.

 The invention provides organic oxidized glutathione salts (GSSG) in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and nitrogen bases, as well as nucleosides and nucleotides of purine and pyrimidine nature.

 The invention provides organic salts of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and nitrogenous bases of purine or pyrimidine nature.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and adenine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and guanine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form and thymine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and cytosine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and uracil.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and 5-methylcytosine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and 4-thiouracil.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and dihydrouracil.

 The invention provides organic salts of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and nucleosides of purine or pyrimidine nature.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and adenosine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and guanosine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form and inosine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and cytidine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and uridine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and thymidine.

 The invention provides organic salts of oxidized glutathione (GSSG), in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and nitrogenous bases, or purine nucleosides or nucleotides or pyrimidine nature.

 The invention provides organic salts of oxidized glutathione (GSSG), in the formula of which two chemically unique amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and nitrogenous bases of purine or pyrimidine nature.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and adenine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and guanine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and thymine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and cytosine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and uracil.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and 5-methylcytosine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and 4-thiouracil.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and dihydrouracil.

 The invention provides organic oxidized glutathione salts (GSSG) in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and purine or pyrimidine nucleosides.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and adenosine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and guanosine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and inosine.

 The invention provides an organic salt of oxidized glutathione in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and cytidine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and the nucleoside is uridine.

 The invention provides an organic salt of oxidized glutathione, in which the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and thymidine.

 The invention provides organic salts of oxidized glutathione (GSSG), in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, or oxidized glutathione, in the formula of which two chemically unambiguous amino acids are in D-form, and the remaining amino acids are in the L-form, and nucleosides of purine or pyrimidine nature, in the formula of which the ribose is presented in the form of β-D-ribose, or in the form of β-D-deoxyribose.

 The invention provides organic salts of oxidized glutathione (GSSG), in which the counter ions are oxidized glutathione in the L or D form, and at the same time two nitrogenous bases: one of which is purine and the other pyrimidine in nature.

 The invention provides organic salts of oxidized glutathione (GSSG), in which the counterions are oxidized glutathione in the L or D form, and two nucleosides at the same time: one of which is purine and the other pyrimidine in nature.

 The invention provides individual substances that contain organic cations represented by a series of nucleosides or nucleotides and phosphamide derivatives of oxidized glutathione (GSSG), the amino acids of which are in L-form.

 The invention provides a substance in which an adenosine monophosphate (AMP) derivative is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a guanosine monophosphate derivative (GMF) is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which an inosine monophosphate derivative (IMP) is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a cytidine monophosphate derivative (CMP) is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a uridine monophosphate (UMF) derivative is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a thymidine monophosphate derivative (TMF) is used as a phosphamide derivative of oxidized glutathione.

 The invention provides substances in which purine or pyrimidine derivatives of nucleotides are used as the phosphamide derivative of oxidized glutathione, in which the ribose is in the form of β-D-ribose or in the form of β-D-deoxyribose.

 The invention provides individual substances that contain organic cations represented by a series of nucleosides or nucleotides and phosphamide derivatives of oxidized glutathione (GSSG), in the structure of which two chemically unambiguous amino acids are represented in D-form and the remaining amino acids are represented in L-form.

 The invention provides a substance in which an adenosine monophosphate (AMP) derivative is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a guanosine monophosphate derivative (GMF) is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which an inosine monophosphate derivative (IMP) is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a cytidine monophosphate derivative (CMP) is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a uridine monophosphate (UMF) derivative is used as the phosphamide derivative of oxidized glutathione.

 The invention provides a substance in which a thymidine monophosphate derivative (TMF) is used as a phosphamide derivative of oxidized glutathione.

 The invention provides substances in which purine or pyrimidine derivatives of nucleotides are used as the phosphamide derivative of oxidized glutathione, in which the ribose is in the form of β-D-ribose or in the form of β-D-deoxyribose.

 The invention provides individual substances containing inorganic cations of alkali, alkaline earth and / or transition metals and phosphamide derivatives of oxidized glutathione in the L-form or D-form with a number of nucleosides or nucleotides of purine or pyrimidine nature, in which the ribose is in the form of β- D-ribose or in the form of β-D-deoxyribose.

 The invention provides a substance in which a derivative with two nucleotides, one of which is purine and the other of a pyrimidine nature, is used as a phosphamide derivative of oxidized glutathione.

 The invention provides a pharmaceutical composition exhibiting antiviral, antibacterial, immunorehabilitation and hepatoprotective activity, containing as an active substance at least one of the above compounds in an effective amount, as well as a pharmaceutically acceptable carrier or excipient.

 According to the invention, there is provided a pharmaceutical preparation having antiviral, antibacterial, immunorehabilitation and hepatoprotective effects and containing at least one of the compounds described above in an effective amount as an active substance, as well as a pharmaceutically acceptable carrier or excipient.

 The invention provides a method for producing an organic salt of oxidized glutathione and inosine, in which the oxidized glutathione molecule acts as the annon, and the inosine molecule (9β-D-ribofuranosyl-hypoxanthine) acts as the cation, characterized in that to the solution of the oxidized glutathione disodium salt when while stirring and room temperature add 1 equivalent of inosine powder and leave to dissolve completely, after which the clear solution is filtered and freeze-dried.

 The invention provides a method for producing inosyl-5-phosphoryl-N-glutathione in which a phosphamide bond between the amino group of glutamic acid in an oxidized glutathione molecule and the phosphoric acid residue in an inosine-5-monophosphate molecule is obtained by reacting its N-oxysuccinimide activated ester and oxidized glutathione in dimethylformamide at pH 8.0-8.5.

 The invention provides a method of influencing infectious agents (including intracellularly localized ones) and / or stimulating T-cell and humoral anti-infectious immunity along with providing cytoprotective effects, which consists in introducing an effective amount of compounds into mammals that are based on the chemical interaction of disulfide-containing peptides with nitrogen bases, or nucleosides, or nucleotides of purine or pyrimidine nature.

 According to the invention, a method is provided for inducing apoptosis mechanisms, including by expressing an apoptosis inducer, FAS / APO-1 antigen (CD95 +) on the membranes of virus-infected cells, on the membranes of macrophages containing tuberculosis mycobacteria, on the membranes of cells infected with mycoplasmas, chlamydia, malarial plasmodium and other infectious agents, by introducing into the mammalian organism a pharmaceutical preparation containing an effective amount of an active substance selected from the group containing GSSG∞inosine, GSSG∞ uracil, GSSG∞thymn, GSSG∞ adenine, GSSG∞guanine, GSSG-inosyl monophosphate, GSSG-uracil monophosphate, GSSG-thymidine monophosphate, GSSG-cytosine monophosphate and other L-derived compounds or oxidized glutathione and its salts with nitrogenous bases or nucleosides or nucleotides of purine or pyrimidine nature.

 According to the invention, there is provided a method comprising administering to a human body of a hepatitis C virus-infected mammal a pharmaceutical preparation containing an effective amount of an active ingredient selected from the group consisting of GSSG∞inosine, GSSG-inosine monophosphate, GSSG-uracil monophosphate and other derivatives of L- or D- forms of oxidized glutathione with nucleic acid components in order to inhibit the ATP-azo / helicase activity of hepatitis C virus NS3 to stop replicative activity and eliminate hepatitis C virus, been consistent, to achieve a therapeutic effect.

 According to the invention, a method is provided which consists in predominantly inhibiting the replicative activity of RNA-containing viruses when using compounds obtained on the basis of the chemical interaction of the L- or D-forms of oxidized glutathione and its salts with nitrogen bases or nucleosides as an active substance, or nucleotides of pyrimidine nature.

 According to the invention, a method is provided for predominantly inhibiting the replicative activity of DNA-containing viruses when using compounds obtained on the basis of chemical interaction of the L- or D-forms of oxidized glutathione and its salts with nitrogen bases, or nucleosides, or nucleotides as an active substance. purine nature.

 According to the invention, there is provided a method comprising administering to a mammalian organism a pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine and other compounds of oxidized glutathione with nucleic acid components, in order to preferentially activate the functional activity of the Th1 group of T- cells, or Th2-group of T-cells, or their regulated ratio, which differentially determines the effectiveness of the immune system of a mammalian organism in wearing etiotropic factors infectious diseases.

 According to the invention, a method is provided for administering to a patient with viral hepatitis C a pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine and / or GSSG-inosine monophosphate in order to restore the normal ratio of Th1 / Th2 cells, therefore , achieving the optimal content of cytokines produced by Th1 and Th2 group of cells, including with the aim of increasing the endogenous production of IL-2, IL-12, IFN-α and γ.

 According to the invention, there is provided a method comprising administering to a patient a viral hepatitis A, B and / or C patient a pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine and other compounds of oxidized glutathione, its salts with nucleic acid components, in order to achieve hepatoprotective effects, including the prevention and / or treatment of complications of the aforementioned chronic viral hepatitis, namely the prevention and treatment of cirrhosis of the liver and hepatocytes llyulyarnyh carcinomas.

 The invention provides a method in which a disease is selected from all types of infectious diseases, including viral and bacterial, including anaerobic infections; chlamydial and mycoplasma infections; mycoses (candidiasis); protozoal infections.

 The invention provides a method in which substances derived from the chemical interaction of disulfide-containing peptides with nitrogen bases, nucleosides and nucleotides are administered parenterally, orally, in the form of inhalation solutions, solutions for local instillations, ocular or intranasal drops, ointments, creams or gels for skin application, as well as in the form of suppositories - until the therapeutic effect is achieved kta and / or prevention of complications caused by infectious agents during the development of an infectious disease.

 The invention provides a method of treating a subject in need of terminating the replicative activity of viruses, bacteria or other infectious agents, inducing apoptosis mechanisms of infected cells, restoring and stimulating anti-infective capable immunity: T-cell, and / or humoral, systemic cytoprotective effects, comprising administering to the subject a pharmaceutical a preparation containing a biologically active substance selected from the group consisting of biologically active compounds, semi based on the chemical interaction of the L- or D-forms of oxidized glutathione, its salts with nitrogen bases, or nucleosides or nucleotides of purine or pyrimidine nature in an amount sufficient to achieve a therapeutic effect.

 According to the invention, the method includes the treatment of acute, prolonged course of viral hepatitis B, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine and GSSG-inosine monophosphate.

 According to the invention, the method includes the treatment of acute viral hepatitis C, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞uracil and GSSG-inosine monophosphate.

 According to the invention, the method includes the treatment of chronic viral hepatitis B, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞guanosine, GSSG∞adenosine, GSSG-inosine monophosphate and GSSG-thymidine monophosphate.

 According to the invention, the method comprises the treatment of chronic viral hepatitis C, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞uracil, GSSG∞cytosine, GSSG∞dihydrouracil, GSSG-uracilyl monophosphate, GSSG-cytosine monophosphate, uracil-GSSG-ynose.

According to the invention, the method comprises treating chronic viral hepatitis in a cirrhotic stage, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞uridine, GSSG∞thymidine, Li ₂ -GSSG-inosine monophosphate and Na ₂ -GSSG-thymidine monophosphate.

According to the invention, the method comprises the treatment of pulmonary tuberculosis, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞cytosine, GSSG-5-methylcytosine, Li ₂ -GSSG-inosine monophosphate.

According to the invention, the method comprises treating tuberculosis of the genitourinary system, and the pharmaceutical preparation is selected from the group consisting of GSSG∞ thymine, Na ₂ -GSSG-gyneosine monophosphate and uracil-Li ₂ -GSSG-guanosine monophosphate.

According to the invention, the method includes the treatment of AIDS, as well as cytomegalovirus infection and infection caused by the Epstein-Barr virus and / or pneumocystis, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞dihydrouracil, GSSG∞4-thiouracil, as well as Zn ₂ -GSSG-thymidine monophosphate and Ag ₂ -GSSG-yracilyl monophosphate and uridine∞GSSG∞inosine.
According to the invention, the method comprises treating infections caused by herpes viruses, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, Li ₂ -GSSG-guanosine monophosphate, and also D-form Na ₂ -GSSG-cytosine monophosphate and D-form GSSG∞-uracil.
According to the invention, the method involves the treatment of mycoses (candidiasis), and the pharmaceutical preparation is selected from the group consisting of GSSG∞uridine, GSSG∞4-thiouracil and Ag ₂ -GSSG-yrapylmonophosphate.

According to the invention, the method comprises the treatment of mycoplasma infection, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞adenosine, and also Na ₂ -GSSG-adenosine monophosphate.

According to the invention, the method comprises treating chlamydial infection, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞thymine, GSSG∞uridine, GSSG∞guanosine, and also Na ₂ -GSSG-guanosine monophosphate.

и GSSG∞5-метилцитозин.
Согласно изобретению способ включает лечение анаэробной инфекции, а фармацевтический препарат выбирают из группы, включающей D-форму GSSG∞инозина (D-цистеин) и D-форму GSSG∞урацила (D-глутаминовая кислота).According to the invention, the method includes the treatment of malaria, leishmaniasis, and the pharmaceutical preparation is selected from the group comprising GSSG∞inosine,

and GSSG∞5-methylcytosine.
According to the invention, the method comprises treating anaerobic infection, and the pharmaceutical preparation is selected from the group comprising the D-form of GSSG∞inosine (D-cysteine) and the D-form of GSSG∞uracil (D-glutamic acid).

According to the invention, the method includes the treatment of viral hepatitis A, dysentery and cholera, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, Li ₂ -GSSG-inosine monophosphate and Li ₂ -GSSG-guanosine monophosphate, as well as the D-form of GSSG∞ uracil (D- glutamic acid).

According to the invention, the method includes the treatment of infectious meningitis, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, Li ₂ -GSSG-inosine monophosphate, D-form GSSG∞uracil (D-glutamic acid), GSSG∞5-methylcytosine, Ag ₂ -GSSG -yracilyl monophosphate.

According to the invention, the method includes the treatment of especially dangerous infections of plague, tularemia and anthrax, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞adenine, GSSG∞thymine, GSSG∞5-methylcytosine, GSSG∞4-thiouracil, D-form GSSG∞ uracil (D-glutamic acid), D-form GSSG∞inosine (D-cysten), adenine∞GSSG∞thymine.
According to the invention, the method comprises treating prion infections (“core proteins”), and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞uridine, GSSG∞dihydrouracil, Ag ₂ -GSSG-uracilyl monophosphate, Ag ₂ -GSSG-thymidine monophosphate, as well as uracil monophosphate-Li ₂ -GSSG-inosine monophosphate.

According to the invention, the method includes the treatment of influenza and acute respiratory viral infections, and the pharmaceutical preparation is selected from
the group comprising GSSG∞inosine, GSSG∞adenosine, GSSG∞racil and GSSG∞thymine,
The invention is illustrated by drawings, which are schematic and were made to represent the structural formulas of new substances. However, not every element is indicated on each drawing and not every component for each embodiment of the idea of the invention is given if the illustration is not necessary in order to allow an ordinary person skilled in the art to understand the invention. The figures shown respectively:
In FIG. 1 is the structural formula of the organic salt of bis- (γ-L-glutamyl) -L-cysteinyl-bis-glycine (GSSG) with adenine.

 In FIG. 2 is a structural formula of an organic salt of GSSG with guanine.

 In FIG. 3 is a structural formula of an organic salt of GSSG with thymine.

 In FIG. 4 is a structural formula of the organic salt of GSSG with uracil.

 In FIG. 5 is a structural formula of an organic salt of GSSG with cytosine.

 In FIG. 6 is a structural formula of an organic salt of GSSG with 5-methylcytosine.

 In FIG. 7 is a structural formula of an organic salt of GSSG with 4-thiouracil.

 In FIG. 8 is a structural formula of an organic salt of GSSG with dihydrowracil.

 In FIG. 9 is the structural formula of a substance (organic salt), the counterions of which are GSSG and inosine (9-β-D-ribofuranosylhypoxanthine).

 In FIG. 10 is the structural formula of a substance (organic salt), the counterions of which are GSSG and thymidine (3-β-D-2-deoxyribofuranosylthymine).

 In FIG. 11 is the structural formula of a substance (organic salt), the counterions of which are GSSG and uridine (3-β-D-ribofuranosyluracil).

 In FIG. 12 is the structural formula of a substance (organic salt), the counterions of which are GSSG and guanosine, where R is the ribose residue.

 In FIG. 13 is the structural formula of a substance (organic salt), the counterions of which are GSSG and adenosine, where R is the ribose residue.

In FIG. 14 is a structural formula of a substance obtained on the basis of the formation of a covalent bond between Na ₂ -GSSG and uridine-5 ¹ -phosphate (UMF).

In FIG. 15 is a structural formula of a substance derived from the formation of a covalent bond between Na ₂ -GSSG and cytidine-5 ¹ -phosphate (CMF).

In FIG. 16 is a structural formula of a substance derived from the formation of a covalent bond between Na ₂ -GSSG and thymidine-5 ¹ -phosphate (TMP).

In FIG. 17 is a structural formula of a substance obtained on the basis of the formation of a covalent bond between Na ₂ -GSSG and adenosine-5 ¹ -phosphate (AMP).

In FIG. 18 is a structural formula of a substance derived from the formation of a covalent bond between Na ₂ -GSSG and guanosine-5 ¹ -phosphate (GMF).

In FIG. 19 is a structural formula of a substance derived from the formation of a covalent bond between Zn ₂ -GSSG and thymidine-5 ¹ -phosphate.

In FIG. 20 is a structural formula of a substance derived from the formation of a covalent bond between Ag ₂ -GSSG and uridine-5 ¹ -phosphate.

In FIG. 21 is a structural formula of a substance derived from the formation of a covalent bond between Li ₂ -GSSG and guanosine-5 ¹ -phosphate.

In FIG. 22 is a structural formula of a substance derived from the formation of a covalent bond between the D form of Na ₂ —GSSG (D-glutamic acid) and UMF.

In FIG. 23 is a structural formula of a substance derived from the formation of a covalent bond between the D form of Na ₂ —GSSG (D-cysteine) and CMP.

 In FIG. 24 is the structural formula of a substance (organic salt), the counterions of which are D-form GSSG and uracil.

 In FIG. 25 is the structural formula of a substance (organic salt), the counterions of which are D-form GSSG and inosine.

 In FIG. 26 is a structural formula of a combined organic salt, the counterions of which are GSSG and nitrogenous bases of purine and pyrimidine nature.

 In FIG. 27 is a structural formula of a combined organic salt, the counterions of which are GSSG and nucleosides of purine and pyrimidine nature.

 In FIG. 28 is a structural formula of a combined compound formed on the basis of covalent bonds between the amino groups of the inorganic salt GSSG and the phosphamide groups of purine and pyrimidine nucleotides.

FIG. 29 is the structural formula of 9-β-D-ribofuranosyl-5'-phosphoryl-N-bis- (γ-L-glutamyl) -L-cystinyl-bis-glycine disodium (dilithium) salt, obtained on the basis of the formation of a covalent bond between Na ₂ (Li ₂ ) -GSSG and inosine-5'-monophosphate (IMP).

FIG. 30 - the dynamics of clinical, laboratory and morphological parameters of patient K., who received therapy with GSSG∞inosine.
FIG. 31 - the dynamics of clinical, laboratory and morphological parameters of patient Z., who received therapy with GSSG∞inosin.
FIG. 32 - chest x-ray, left lung. Before treatment (October 1999) (example 16).

 FIG. 33 - chest x-ray, left lung. After treatment (January 2000) (example 16).

 FIG. 34 - therapeutic efficacy of the studied hepatoprotective compounds in experimental dichloroethane hepatitis (Example 17).

 FIG. 35 - therapeutic efficacy of the studied hepatoprotective compounds in experimental acetaminophen hepatitis (Example 17).

 FIG. 36 - therapeutic efficacy of the studied hepatoprotective compounds with combined (dichloroethane + acetaminophen) experimental hepatitis (Example 17).

 FIG. 37 - change in indicators of cytolytic syndrome (ALT, ACT) with combined experimental hepatitis (dichloroethane + acetaminophen) (Example 17).

 FIG. 38 - change in the level of bilirubin in blood plasma with combined experimental hepatitis (dichloroethane + acetaminophen) (Example 17).

FIG. 39 - the dynamics of immunological parameters in patients with chronic viral hepatitis C before and during treatment with GSSG∞inosine.
FIG. 40 - the dynamics of immunological parameters in the treatment of acute viral hepatitis B with GSSG∞inosine.
FIG. 41 - the level of cytokines in the blood serum of patients with chronic viral hepatitis C before and during treatment with the drug GSSG∞inosin.
FIG. 42 is a synthesis scheme for inosyl-5′-phosphoryl-GSSG-Na ₂ .

 As a preferred embodiment, the invention provides a new compound prepared as an organic salt of oxidized glutathione in which 9-β-D-ribofuranosylhypoxanthine (inosine) is a cation. The procedure for producing GSSG∞inosine is presented below, its structural formula in FIG. 9.

The method of obtaining organic salt GSSG∞inosine
200 g (0.65 M) of reduced glutathione (GSH) is taken and dissolved in 700 ml of distilled water with constant stirring and a temperature of 12-15 ^o C (ice bath). 163 ml (0.65 M) of 4N NaOH solution are added. After complete dissolution, 295 ml of a 3% solution of H ₂ O ₂ is slowly added, while the solution is in an ice bath and the temperature remains at the same level. Mixing continues for another 3-4 hours (this time was determined empirically by working with platinum as a catalyst), while the temperature of the solution is maintained below 20 ^o C. After completion of the reaction, HPLC is carried out (final content of GSSG - 98% or more). The pH of the solution is checked and, if necessary, a 4N NaOH solution is added, the pH is adjusted to 5.5.

Then 174.2 g (0.65 M) of inosine (in the form of a homogeneous dry powder) is added and stirring is continued at 23-25 ^{° C. for} another 10-12 hours until it is completely dissolved.

 The resulting solution is filtered through a filter with a pore size of not more than 0.7 microns. The filtered solution should be transparent, colorless, not opalescent, pH 5.3 +/- 0.2. The pH is adjusted to 6.0 by adding a 1-4N NaOH solution. Control - HPLC: peaks of GSSG and inosine (Nucleosil C 18, MeCN - 0.1% TFA).

The resulting solution is lyophilized in accordance with the following recommendations (see also the attached lyophilization scheme):
- shelves are cooled to -20 +/- 0.2 ^o C;
- place the containers with the solution on the shelves;
- freeze the solution to -35 +/- 0.2 ^o C;
- the frozen solution is kept at the indicated temperature for 3 +/- 0.5 hours;
- disable the shelves;
- turn on the condenser for cooling to -54 +/- 2 ^o С for 15 +/- 3 min;
- vacuum treatment for 30 +/- 5 min until 4-6 Pa;
- the material is left for 2 hours;
- the temperature on the shelves gradually rises until a constant temperature on the shelves and the material temperature are reached.

 An important element of the proposed new technical solution for obtaining the named compound (GSSG∞inosine) is the established fact that inosine, poorly soluble in aqueous solutions in the composition of the organic salt with GSSG, becomes a highly soluble compound, which is additional evidence of the formation of ionic bonds between GSSG and inosine, i.e., the formation of a special type of organic salt.

 A further preferred embodiment of the invention is the preparation of the D form of GSSG∞inosine. In this case, in the GSSG formula, two chemically unambiguous amino acids (cysteines) are presented in the D-form, and the remaining amino acids in the L-form (Fig. 25). At the same time, in FIG. 24 shows the formula D-GSSG-uracil in which the glutamic acid is in the D form.

 In turn, in a further embodiment of the invention, it is disclosed that a pharmaceutical preparation based on GSSG∞inosin has a unique combination of biological and pharmacological effects providing a completely new solution to the problem of treating viral diseases, such as: viral hepatitis B and C and their complications; AIDS (see examples of the implementation of the invention 5-8, 12-13); herpetic and urogenital infections (see examples of the invention 14-15).

 It is emphasized that a new level of therapeutic efficacy of GSSG∞inosin, especially in relation to viral hepatitis, is due to the fact that the drug has three types of activity: antiviral, immunorehabilitation and hepatoprotective.

 It was found that each of the three types of pharmacological activity of GSSG∞inosine is unique and, in this regard, the most important embodiment of the invention is the discovery of the mechanisms of the antiviral, antibacterial, immunorehabilitation and hepatoprotective activity of GSSG∞inosine (its D-forms) and other compounds based on interactions of disulfide-containing peptides and purines / pyrimidines.

With respect to antiviral activity, these compounds have been shown to possess two systems of antiviral mechanisms of action. The first, a kind of nonspecific system of mechanisms is due to the general property of drugs to induce apoptosis mechanisms of virus-infected cells, in particular, due to increased expression of the apoptosis inducer FAS / APO-1 receptor (CD95 +) (see, example of implementation of the invention 3). It is this mechanism that determines the high efficiency of GSSG∞inosine and a number of other compounds (GSSG-IMP, GSSG-UMP, Li ₂ -GSSG-IMP, Zn ₂ -GSSG-TMP) when used as a treatment for infectious diseases caused by DNA and RNA viruses.

 In particular, the antiviral activity of GSSG∞inosin has been established in a number of viral infection models, namely: Rift Valley Fever (LDR); generalized forms of herpes infection; Venezuelan equine encephalomyelitis (VEL); influenza (A virus, H3N2) (see examples of the invention 2). The principal result of these studies was the establishment of the fact that GSSG∞inosine was more effective than traditional antiviral drugs.

 In turn, the unique therapeutic efficacy of GSSG∞inosine and the other compounds under consideration has been demonstrated in the clinic of infectious diseases when using these compounds for the treatment of tuberculosis patients (see examples of the invention 16); urogenital infections (see examples of the implementation of the invention 14-15); AIDS (see examples of the implementation of the invention 12-13); acute and chronic hepatitis B (see examples of the implementation of the invention 5-8, tab. 12-21, Fig. 30 and 31).

 Thus, the revealed ability of GSSG∞inosine and other compounds under consideration to induce apoptosis of virus-infected cells also extends to cells infected with mycobacterium tuberculosis; chlamydia, mycoplasma, ureaplasma and other infect agents.

In most of the given examples of the invention, which demonstrate a new quality of therapeutic solutions to these socially significant diseases, the effectiveness of the drugs proposed in the invention is compared with the effectiveness recommended by current medical practice, therapeutic and prophylactic agents. At the same time, it was shown that therapeutic efficacy, for example, GSSG∞inosine (the patented name in the Russian Federation is Moliksan ^® ) in the case of acute and chronic hepatitis B, is determined by the normalization time for such objective indicators of the course of the infectious process as: transaminase activity (ALT and ACT ); bilirubin content; prothrombin content; the content of HbsAg is significantly higher than the therapeutic efficacy of conventional therapy.

 So, in the case of acute hepatitis B, traditional therapy provides a positive dynamics of transaminase activity, the content of bilirubin and HbsAg, as a rule, by 50-60 days of treatment. The use of GSSG∞inosine in mono mode - to 14-17 days of treatment (see examples of the invention 5 and 6, tab. 12-17, Fig. 40).

 In the case of chronic hepatitis B, the key indicators of the effectiveness of the therapy are, again, the activity of transaminases (an indicator of the degree of cytolysis of hepatocytes) and the replicative activity of the virus, determined using the PCR test.

According to this main indicator of the effectiveness of the treatment of chronic hepatitis B, traditional therapy (recombinant interferons plus nucleoside analogues) provide a decrease in the percentage of positive PCR of HBV DNA from 100% by no more than 30-40%; i.e., in 60-70% of patients, treatment is ineffective. The use of GSSG∞inosin provides the conversion of HBV DNA PCR from 100% positive to negative in 70-80% of patients (see examples of the invention 7 and 8, tables 17-21, Figs. 30 and 31). The restoration of the functional capacity of the liver in the case of viral and toxic hepatitis, which is determined by such indicators as protein synthesizing and detoxification functions of the liver, is achieved with GSSG∞inosine 2–3 times shorter than with conventional therapy, for example, heptral ^® (S α-adenosyl-methionine) or Essential ^® (see examples of the invention 11; 14).

A further preferred embodiment of the invention is to disclose a specific, ie. E. Direct antiviral activity GSSG∞inozina (Moliksana ^®), GSSG-IMP and GSSG-UMP, as well as their salts, with respect to hepatitis C virus (RNA HCV), m. E. RNA -containing virus as opposed to hepatitis B DNA-containing virus (HBV DNA). This embodiment of the invention claims the unique property of GSSG∞inosine (Molixan ^® ), as well as GSSG-IMP and GSSG-UMP - the ability to inhibit the ATP-azo / helicase activity of the regulatory non-structural protein-enzyme (NS3) of hepatitis C virus (see example implementation of the invention 4 ) Our data indicate that Zn ₂ -GSSG-IMP, as well as other salts of GSSG-UMP, for example Ag ₂ -GSSG-UMP, have the ability to inhibit the activity of non-structural (regulatory) enzyme proteins not only of HCV, but also of HIV .

 Therefore, the convincing therapeutic activity of GSSG∞inosine with respect to acute and chronic viral hepatitis; herpetic infection and AIDS (see examples of implementation of the invention 5-14) is due to a combination of its mediated (induction of apoptosis of virus-infected cells) and direct antiviral activity (inhibition of the ATP-azo / helicase activity of NS3 HCV), which defines a new solution to treatment and prophylactic measures regarding chronic hepatitis C.

 At the same time, it has been shown that with respect to viral infections, and especially viral hepatitis, as well as AIDS, the D-form of GSSG∞inosine has greater antiviral activity, ensuring the termination of the high replicative activity of viruses and their elimination from the body (see, for example, implementation of the invention 12 )

 Extremely relevant for the treatment of chronic viral hepatitis and, especially for the treatment of hepatitis C, was the vector of immunoreactive activity of GSSG∞inosine, as the second type of pharmacological activity of this and its related compounds.

The characteristic features of the immunorehabilitation efficacy of GSSG∞inosine are:
- normalization of the ratio of production of Th1 / Th2 cytokines by cell types with predominant activation of T-cell immunity;
- predominant stimulation of endogenous production of IL-2, IL-12, as well as IFN and TNF.

As follows from clinical examples of the implementation of the invention 9-11, which is characteristic of a severe course and chronicity of the infectious process (chronic adverse development of the hepatitis C pattern), the imbalance in the production of Th1 / Th2 cytokines by groups of T-helpers is positively corrected by the use of GSSG∞inosine. Moreover, the prevalence of activity of Th2 group of cytokines observed in patients with HCV infection: IL-4, IL-10, IL-6 and IL-13 under the influence of GSSG∞inosine is replaced by the predominant predominance of activity of Th1 group with increased secretion of IL-2; IL-12; IFN-α and γ, as well as TNF-α and β. At the same time, the T-cell immunity is significantly activated (increase in blood CD4 ⁺ ; CD8 ⁺ ; CD16 ⁺ / 56 ⁺ ; CD25 ⁺ ; increased activity of resident macrophages), thereby ensuring that HBV and HCV (hepatitis B and C viruses) get under proper immunological surveillance, and therefore the elimination of viruses from the body and a favorable therapeutic outcome.

 One of the most important mechanisms of the therapeutic effect of GSSG∞inosine is the induction of IL-12 secretion, which helps to stabilize the predominance of activity of the Th1 group, therefore, the active production of endogenous (intrinsic to the body) interferons (see examples of the invention 5-7, 12).

The third type of biological and pharmacological activity of GSSG∞inosin, as a drug that preferably implements an embodiment of the invention, is its hepatoprotective activity, which purposefully determines the unique anti-cirrhotic effects of GSSG∞inosin (Molixan ^® ).
The very formulation of this issue is organically connected with one of the main conceptual provisions of the invention - the creation of individual substances capable of realizing a fundamentally new solution to the problem of treating such infectious diseases as viral hepatitis, especially hepatitis C.

 Taking into account what was said in the introduction that more than 40% of liver cirrhosis and 60% of liver cancer are caused by chronic hepatitis C, this means that any constructive solution to the treatment of chronic hepatitis C involves both the elimination of the actual virus cytopathic lesion of the liver tissue and the prevention of fibrosis liver, along with the prevention of malignant transformation of hepatocytes.

полученных на экспериментальной модели цирроза печени и в клинике.The invention provides the rationale for anti-cirrhotic effects

obtained on an experimental model of liver cirrhosis and in the clinic.

 In particular, on the model of cirrhosis obtained as a result of chronic administration of dimethylnitrosoamine (DMNA) to experimental animals, it was found that the use of the drug GSSG∞inosine at a dose of 10 mg / kg 3 times a week for 6 weeks ensured a decrease in the amount of connective tissue in the liver by 64% and contributed to the restoration of damaged hepatocytes. In this case, the comparison drug Heptral (S-adenosyl-methionine) had a significantly lower therapeutic effect, reducing the degree of liver fibrosis of experimental animals by only 35% and without significantly affecting the restoration of the functional capacity of the liver (protein synthesizing and others) (see implementation example inventions 29).

 Equally convincing results were obtained in the clinic when treating patients with a diagnosis of GSSG∞inosin with a diagnosis of: chronic viral hepatitis B (and / or chronic hepatitis C); cirrhotic stage (PCR HBV +; PCR HCV +), ascites, portal hypertension. The use of the drug GSSG∞inosine in mono mode during repeated courses provided a unique therapeutic effect, confirmed by the positive dynamics of the clinical condition and special methods for examining patients (see Examples of the invention 8-11).

Thus, the treatment of patients with chronic viral hepatitis, especially hepatitis C, with the use of dosage forms of the drug GSSG∞inosin (Molixan ^® ) provides not only the elimination of the actual infection process, but also the prevention of complications of viral hepatitis, especially cirrhosis and liver cancer. In addition, it is revealed that in the case of the formed toxic cirrhosis, which is absolutely analogous to alcoholic cirrhosis, the use of GSSG∞inosine in the experiment and clinic demonstrates therapeutic efficacy that is unparalleled in terms of the corresponding use of known hepatoprotective drugs.

 Therefore, the conceptual platform of the claimed invention is the presented group of original substances (possible name - thionucleins), obtained (synthesized) as organic salts of GSSG with nitrogen bases and / or nucleosides; and / or as compounds based on the formation of a covalent bond between GSSG and nucleotides of purine and pyrimidine nature.

 These individual substances (formulas in Figs. 1-29) have a unique combination of biological and pharmacological effects, namely: a) antiviral effect - direct (inhibition of the ATP-azo / helicase activity of NS3 HCV) and indirect (through the induction of apoptosis of virus-infected cells, in including due to the expression of the apoptosis inducer FAS / APO-1 receptor); b) immunorehabilitation activity; and iv systemic cytoprotective and, especially, specific hepatoprotective effects with the ability to prevent cirrhotic changes in the liver and even reverse the development of liver fibrosis.

 Pharmaceutical preparations containing the compounds of the invention as biologically active substances can be used in various dosage forms: a) for parenteral administration; b) for per os administration in capsules and tablets; c) in the form of suppositories - rectal and vaginal; g) in the form of eye drops; and e) in the form of creams, gels and ointments for external use.

 In this regard, it seems appropriate to use the presented substances as medicines for the treatment of a wide group of infectious diseases and the prevention of their complications. First of all, this refers to the group of infectious diseases for which the principal factors of the etiopathogenesis of the chronicity and progression of the disease process are: a) the presence of an intracellular persistent persistent infect agent; b) the quasivariance of the genetic apparatus of the infectagent, which allows it to "escape" from the immunological surveillance of the affected organisms; c) a violation of the rational, from the standpoint of protecting the body, ratio of the production of Th1 / Th2 cytokines by groups of helper lymphocytes, i.e., a violation of the orientation of the body's effective immune response; d) systemic cytopathic, especially hepato-, nephro and hemocytopathic effects.

 In turn, the performed experimental and clinical studies (see examples of the implementation of the invention 2-18) showed that GSSG∞inosine and other drugs presented have high therapeutic efficacy precisely in connection with their ability to correct all of these factors of the etiopathogenesis of infectious processes, which allows formulate the following medical indications of their use.

 First of all, for the treatment of infectious diseases in respect of which the intracellular pathogen is the etiotropic factor, for example: DNA and RNA viruses; chlamydia, mycoplasma and ureaplasma, hence the treatment of acute and chronic viral hepatitis; AIDS herpetic, mycoplasma and chlamydial infections.

 For the treatment of protozoal infections (malaria, leishmaniasis); for the treatment of infectious diseases caused by fungi (mycoses) and pneumocysts, in combination with traditional antibacterial therapy in the acute period and in mono mode as part of maintenance therapy.

 For the treatment of infectious diseases caused by gram-positive and gram-negative bacteria, as well as non-spore-forming anaerobes, which contain the stage of intracellular persistence of the pathogen during etiopathogenesis, for example, as is typical for tuberculosis and persistence of mycobacteria in the macrophages of the body. In this case, the induction of apoptosis of infected cells (including macrophages) by drugs of the GSSG∞inosin group ensures mycobacterial affection by specific chemotherapy and the T-cell immunity system.

Moreover, for the treatment of acute viral hepatitis B and C, as well as mixed hepatitis, it is preferable to use GSSG∞inosine, as well as GSSG-inosine monophosphate, Li ₂ -GSSG-inosine monophosphate.

 For the treatment of chronic hepatitis B and C, it is preferable to use GSSG∞inosine, as well as GSSG-inosine monophosphate (GSSG-IMP), as well as GSSG-uracil monophosphate (GSSG-UMF).

For the treatment of chronic hepatitis in the cirrhotic stage, it is preferable to use GSSG∞inosine, as well as GSSG∞adenosine, GSSG∞uridine, GSSG∞thymidine and Na ₂ - GSSG-thymidine monophosphate (Na ₂ - GSSG-ТМФ), depending on the characteristics of morphological changes in the liver and the degree of violation of its functional capacity.

 For the treatment of pulmonary tuberculosis, the use of GSSG∞inosin, GSSG∞cytosine and GSSG-5-methylcytosine is preferred.

In the case of tuberculosis of the genitourinary system, it is preferable to use Na ₂ -GSSG-guanosine monophosphate (Na ₂ -GSSG-FMP) and / or uracil-Li ₂ -GSSG-guanosine monophosphate (UMF-Li ₂ -GSSG-GMF).

For the treatment of AIDS, as well as CMV infection; infections caused by the Epstein-Barr virus and / or pneumocysts, it is preferable to use Pyyp∞inosine, GSSG∞-dihydrouracil, as well as Zn ₂ -GSSG-TMF, Ag ₂ -GSSG-UMF and uridine∞GSSG∞inosine, depending on the stage of the disease, the nature of opportunistic infections and the presence of Kaposi’s sarcoma.

For the treatment of herpetic infections, it is preferable to use Li ₂ -GSSG-GMF, as well as the D-form of Na ₂ -GSSG-cytosine monophosphate (D-form Na ₂ -GSSG-CMF) and the D-form of GSSG∞ uracil.
For the treatment of mycoses, it is preferable to use GSSG∞uridine, GSSG∞4-thiouracil and Ag ₂ -GSSG-yuracyl monophosphate.

For the treatment of mycoplasma infection, it is preferable to use GSSG∞inosine, GSSG∞adenosine, and also Na ₂ -GSSG-adenosine monophosphate (Na ₂ -GSSG-AMP).

For the treatment of chlamydial infection, it is preferable to use GSSG∞inosine, GSSG∞guanosine, GSSG∞thymine, and Na ₂ -GSSG-GMF.

For the treatment of infections caused by herpes viruses, it is preferable to use GSSG∞inosine, Li ₂ -GSSG-guanosine monophosphate, as well as the D-form of Na ₂ -GSSG-cytosine monophosphate and the D-form of GSSG∞uracil.
For the treatment of viral hepatitis A, as well as intestinal infections (dysentery, cholera), it is preferable to use Li ₂ -GSSG-GMF, GSSG∞inosine, and also the D-form GSSG∞uracil (D-glutamic acid).

 For the treatment of influenza, as well as other acute respiratory viral infections (SARS), it is preferable to use GSSG∞inosine, GSSG∞adenosine, GSSG∞uracil and GSSG∞thymine, depending on the type of influenza virus and the viruses that cause SARS.

 The results of a preclinical study of the general toxic effects of the claimed pharmaceutical preparations make it possible to attribute the claimed dosage forms to practically non-toxic medicinal substances - class 5 [28, 29].

 The following examples of the invention demonstrate the possibility of obtaining the above substances, methods of their medical use, as well as the therapeutic efficacy of the family of developed new pharmaceutical preparations (obtained on the basis of the presented new individual substances) taking into account their testing on a wide palette of infectious diseases (Examples of the invention 1- 18).

Example 1
The method of obtaining (synthesis) inosyl-5'-phosphoryl-GSSG-Na ₂
I. General characteristics of the drug.

 1. Name: 9-β-D-ribofuranosyl-5'-phosphoryl-N-bis- (γ-L-glutamyl) -L-cystinyl-bis-glycine disodium (dilithium) salt.

 2. The structural formula is shown in FIG. 29.

3. Gross formula: C ₃₆ H ₅₅ N ₁₀ O ₁₉ Na ₂ S ₂ P.

 4. Molecular weight: 1072.96 (for disodium salt).

 5. Appearance: odorless white powder.

 6. Solubility: soluble in water, 0.9% isotonic sodium chloride solution for injection; insoluble in 95% alcohol, chloroform, ether and other organic solvents.

 7. The transparency and color of the solution: a solution of 0.05 g of the drug in 10 ml of water is transparent and colorless.

 8. pH of 0.1% solution: 4.5-5.5 potentiometrically.

9. Authenticity:
a) amino acid analysis (6 n HCl, 110 ^o C, 20 h), (margin of error 20%, for cysteine 35%), in accordance with: glycine - 2.0; glutamic acid - 2.0; cysteine - 2.0;
b) NMR ( ¹ H) - spectroscopy, according to "BRUKER" AM 500, 500 MHz, D ₂ O (see table. A);
c) according to the output time, it corresponds to the standard.

10. Purity (content of the main substance):
a) HPLC: not less than 97%
BECKMAN instrument "Gold Nouveau Chromatography Data System" Version 1.0, Diod Array Detector Module 126. Sample 20 μl of a 0.1% drug solution, chromatography on an ULTRASPERE ODS 250x4.6 mm column with acetonitrile reversed C ₁₈ phase in isocratic mode - 0.1 % trifluoroacetic acid (2: 98); flow rate 1 ml / min, detection at 220 nm, scanning 190-600 nm, PDA functions - Contour Plot, 3D;
b) according to the amino acid analysis of 85% (analysis according to p. 9a with an exact weight);
c) TLC - homogeneous, analysis is carried out when applying in a strip of 5 μl of a 0.1% solution of the drug; Kieselgel plates 60 _f (Merck) 10x5 cm, system: n. butanol - acetic acid - water (4: 1: 1); manifestation according to standard methods - ninhydrin and chlorine / benzidine; R _f = 0.15;
g) the content of sodium (Na) according to the emission spectral method of 4.0-4.8%.

11. The found element content, mcg / g:
Silver (Ag) - <1.0 (less than 0.0001%)
Aluminum (Al) - 2.0
Arsenic (As) - <1.0
Barium (Ba) - <0.50
Beryllium (Be) - <0.05
Calcium (Ca) - 7.0
Cadmium (Cd) - <0.05
Cobalt (Co) - <0.5
Chrome (Cr) - 1.7
Copper (Cu) - <0.5
Iron (Fe) - <1.0
Potassium (K) - <2.5
Selenium (Se) - <2.0
Magnesium (Mg) - <2.5
Manganese (Mn) - <0.2
Molybdenum (Mo) - <0.2
Sodium (Na) - 48 mg / g
Nickel (Ni) - <0.5
Lead (Pb) - <0.40
Strontium (Sr) - 1.9
Titanium (Ti) - <0.5
Vanadium (V) - <0.5
Zinc (Zn) - 0.65
Antimony (Sb) - <0.5
Determination method
An exact sample weight (about 50 mg) was dissolved in 50 ml bidistilled water and the solution was used for analysis.

 The platinum content was quantified by inductively coupled plasma mass spectrometry using a PQe model instrument, VG Elemental, England. The relative accuracy of the analysis is 5%.

 The content of other elements is quantified by inductively coupled plasma atomic emission spectrometry using a TRACE 61E model, Thermo Jarrell Ash, USA. The relative accuracy of the analysis is 5%.

12. Mass loss during drying: 10% upon drying to constant weight at 100 ^{° C.} in vacuo (1 mm Hg) over CaCl ₂ and P ₂ O ₅ .

 II. Description of the synthesis procedure.

 13. The chemical synthesis scheme (see Fig. 42).

I - inosine-5-monophosphate,
HOSu - oxysuccinimide,
DCC - dicyclohexylcarbodiimide,
II - inosine-5-monophosphate oxysuchninmide activated ether,
III - inosine-5-monophosphoryl-N-glutathione.

14. Description of the method
Inosine-5-monophosphate (I) is dissolved in dimethylformamide and 1 equivalent of N-oxysuccinimide and 1.2 equiv. Are added with stirring and cooling to 0-5 ^{° C.} N, N-Dicyclohexnlcarbodiimide. Stirred under cooling for 1 h, then 12 hours at room temperature. The precipitated dicyclohexylurea is filtered off and 3 equivalents of the disodium salt of oxidized glutathione are added to the residue. Stirring at room temperature for 24 hours. Then dimethylformamide is evaporated and by preparative HPLC in the mode described above, the product is purified. UV control over the absorption band of a purine base of 260 nm.

Example 2
Antiviral activity of the drug GSSG∞inosine
1) Rift Valley Fever (LDR) is an acute febrile disease of viral etiology that affects domestic animals and people. A person’s disease is characterized by acute onset, rapid development of febrile symptoms, pain in the joints and limbs, eye damage, and symptoms of hemorrhagic diathesis. One of the characteristic signs of LDR in humans and animals is the defeat of the liver virus, as a result of which hemorrhages and massive necrosis of the parenchymal tissue are observed in this organ. The disease is also accompanied by leukopenia.

Existing means and methods of etiotropic and pathogenetic therapy of LDR are not effective enough. Given the peculiarities of the pathogenesis of LDR, in particular, the high tropism of the pathogen to hepatocytes, it seemed advisable to evaluate the antiviral activity of the drug GSSG∞inosin in the experimental model of this infection.
1. Materials and research methods.

 1.1. Animals. The experiments used adult randomly bred white male mice weighing 18-21 g, sourced from the Rappolovo nursery of the Russian Academy of Medical Sciences.

1.2. The causative agent of infection. A virus strain isolated in Uzbekistan in 1987 from a person with hemorrhagic fever from the Museum of Viral Cultures of the Research Institute of Military Medicine was used to simulate the infection of LDR. A brain suspension of newborn mice infected with an intracerebrally indicated strain of the pathogen served as an inoculum. Infectious doses of the virus in the present study were in the range of 1-20 LD ₅₀ , the pathogen was administered intraperitoneally. The period of observation of animals was 14 days.

1.3. Preparations. GSSG∞inosine - dosage form for injection (1% and 3%) - was used in doses of 3, 10 and 30 mg / kg per administration. The drug was administered intraperitoneally 1 time per day for 6-7 days. The beginning of administration is 1-2 days before infection, then the drug was administered on the day of infection (4 hours before infection) and continued for another 4 days. Thus, the course of treatment consisted of 6-7 injections of GSSG∞inosin.
Ribamidil is a comparison drug (analogue of imp. Virazole, ribavnrin), manufactured by Olaine (Latvia), injectable dosage form. Ribamidil is currently one of the most effective antiviral chemotherapy drugs, in particular for hemorrhagic fevers. The drug was administered subcutaneously according to the optimal, previously worked out scheme: a single (daily) dose was 100 mg / kg, the timing of administration corresponded to the time of administration of GSSG∞inosine.
1.4. Evaluation of the effectiveness of drugs. The protective efficacy of the drugs was evaluated by the level of survival (%) and average life expectancy (days) of the animals of the experimental and control groups.

2. The results of the study
2.1. The effect of GSSG∞inosine monotherapy on the outcome of LDR infection
In the first series of experiments, a study was made of the effectiveness of GSSG∞inosine when administered in monotherapy. The drug was started to be introduced before the animals were infected with the pathogen; in one scheme - 48 hours before infection, with the other two - 4 hours before infection. In general, the courses of treatment consisted of 6-7 injections of GSSG∞inosine intraperitoneally in single doses of 3, 10 and 30 mg / kg. Ribamidil was administered subcutaneously at a subtherapeutic dose of 100 mg / kg for 5 days.

 In animals of the control group, the incubation period was 6 days. At the same time, in the animals of the experimental groups, signs of infection appeared 2 days later. The final results of this experiment are presented in table 1.

As follows from the data presented, the mortality of animals in the control group infected with the virus in doses of 10-20 LD ₅₀ was 100%. At an infectious dose of 1-2 LD ₅₀ , 67% of infected mice died. Comparison drug ribamidil, taken as a positive control, protected from death about one third of the animals infected with a high dose of the virus and up to 60% of the animals infected with low doses of the pathogen. The drug GSSG∞inosin in this model of acute generalized viral infection also had a certain protective effect, increasing the survival of animals by 22-59% relative to control depending on the therapeutic dose and severity of the infection.

 In the second series of experiments, the protective efficacy of the combined use of ribamidyl and GSSG∞inosine was evaluated. The drugs were used in the same doses and according to the same schemes as in the first series. The results of this experiment are presented in table. 2. The data obtained indicate that the antiviral effect of the combined use of GSSG∞inosine and ribamidil in experimental infections caused by the LDR virus in white mice is higher than each drug separately.

 In the third series of experiments, the effect of GSSG∞inosine on the course of LDR infection was studied by survival rates and the presence of hemorrhage on the surface of the liver in experimental and control mice exposed during the development of this infection.

In the experiment, we used 40 randomly bred male mice weighing 18 g, sourced from the Rappolovo nursery RAMS. For infection, the virus was taken at a dose of 4 LD ₅₀ , infection was carried out intraperitoneally.

 GSSG∞inosine was also administered intraperitoneally in two doses - 3 and 30 mg / kg for 7 days. The beginning of therapy is immediately after infection of the animals, and then daily (1 time per day). Observation was carried out for 10 days (table. 3).

It was found that with this administration scheme, GSSG∞inosine at a dose of 3 mg / kg had a protective effect against experimental infection in white mice caused by intraperitoneal infection of 4 LD ₅₀ of the Rift Valley fever virus, increasing the survival rate of animals by about 2 times (66% against 31 % in control). It should be noted that the general condition of the animals of the experimental groups was better than the control ones, in particular, they consumed food much better during the entire observation period, while anorexia was observed in mice of the control group from 5 days after infection.

 The autopsy of live mice of the experimental and control groups was performed at 5 days (the beginning of death) and 10 days (completion of the experiment) of observation. Apparently healthy mice were taken for autopsy. A study of the fallen mice was not performed. On the 5th day, 1 animal from each group was opened, on 10 - 2 from the experimental groups and 3 animals from the control group. An external examination of the liver of animals showed that the number of hemorrhagic foci in the animals of the experimental groups is less than in the control. In the latter case, there were more than 15-20 of them on each liver, and in animals of the experimental groups - 1-3, with a better indicator observed in animals receiving a higher dose of GSSG∞inosine - 30 mg / kg (no foci were detected).

Thus, the new systemic cytotoprotector and immunomodulator GSSG∞inosin has antiviral activity and increases the resistance of white mice to the causative agent of Rift Valley fever, which causes a fatal generalized infection in these animals with damage to the liver and other organs. The introduction of GSSG∞inosine intraperitoneally in single doses of 3, 10, 30 mg / kg for 6 days with experimental LDR in white mice infected with 1 -20 LD ₅₀ virus, allows to increase the survival rate of animals by 30-60%, and also significantly ( 2 times or more) increase their life expectancy. The administration of GSSG∞inosin during experimental rift valley fever in white mice together with the antiviral drug ribamidil allows increasing the protective effect of the latter, as evidenced by an increase in the survival rate (by 20%) and life expectancy (by 2 times) of infected mice.

2.2. The effect of GSSG∞inosine on the course of generalized herpetic infection in white mice
The experiments used white random-bred male mice weighing 12-14 g, sourced from the Rappolovo nursery RAMS.

 To simulate a herpes infection, herpes simplex virus, strain L-2 from the museum of the Research Institute of Military Medicine was used. Infection of animals was carried out intraperitoneally with an inoculum, which was an emulsion of the brain of infected and diseased newborn mice.

 GSSG∞inosine was administered daily, 1 time per day at a dose of 30 mg / kg for 5 days; beginning of administration - 2 hours before infection.

 Herpetic infection proceeded against the background of immunodeficiency caused by cyclophosphamide (CFA) - 1 group, hydrocortisone (HA) - 2 group, irradiation (Obl) - 3 group.

 The comparison drug is cycloferon (once, 2 hours before infection at a dose of 100 mg / kg).

 The observation period is 14 days.

 Results: The experiment showed that GSSG∞inosin also has a protective effect against DNA-containing viruses, which include the herpes simplex virus. Even in conditions of lethal infecting doses, GSSG∞inosine contributed to the survival of up to 30% of animals with complete death of animals in the control groups (Table 4).

2.3. The effect of the drug GSSG∞inosin on the course of infection caused by equine Venezuelan encephalomyelitis virus (VEL) in experimental animals
In the experiments we used white non-inbred male mice weighing 18-20 g, sourced from the Rappolovo nursery RAMS. In total, 144 animals were used in the experiment.

The causative agent of infection: VEL virus, strain "TRINIDAD", infectious doses - 1 and 2 LD ₅₀ . The virus was administered subcutaneously.

 Test subject: GSSG∞inosine. The drug was administered intraperitoneally at doses of 3 and 30 mg / kg; dry samples were dissolved in an isotonic sodium chloride solution so that the required single dose was contained in a volume of 0.5 ml.

 Schemes of drug administration: the drug in the indicated doses was administered according to 2 schemes - prophylactic (-72 h, -48 h, -24 h, 0) and emergency prophylactic (+2, +24, +48, +72, +96, + 120 h).

 Comparison drug (positive control): inducer of interferon Cycloferon (CF) at a dose of 50 mg / kg, which was administered 4 hours before infection.

 Virus control: animals were infected with virus-containing material, there was no treatment.

 Combined use of drugs: in 2 experimental groups, the combined prophylactic administration of GSSG∞inosine at a dose of 3 mg / kg and Cycloferon at a dose of 50 mg / kg was used.

 Observation of animals: carried out within 14 days after infection.

 Assessment of the effectiveness of drugs: by the difference in survival rates (%) and average life expectancy (T, day) of animals from experimental and control groups.

 The results are presented in table. 5.

As follows from the data obtained, the infecting dose of the VEL virus in this experiment turned out to be even slightly lower than the calculated one - death from 2 LD _{50 of the} pathogen occurred in 50% of the animals of the control group, and from 1 LD ₅₀ in 33%.

 In groups 1, 4 and 9 on the 3rd and 4th day after infection, nonspecific death of 1-2 animals was noted, which, as a rule, is caused either by toxic drugs or injuries. Specific mortality in VEL in white mice is usually recorded from 5 days after infection.

Comparison drug Cycloferon, taken in a subtherapeutic dose of 50 mg / kg, did not have a protective effect at 2LD ₅₀ , and at 1 LD _{50 the} protective effect was 33%.

 The drug GSSG∞inosine when used at a dose of 30 mg / kg exerted an antiviral effect. In the case of the use of a preventive regimen: in this case, its protective effectiveness was equal to the effectiveness of cycloferon (increase in survival by 33%, a significant increase in life expectancy).

 In experiments with combined prophylaxis, a clear positive effect of the drug GSSG∞inosin was noted both in terms of survival (increasing it by 17-30%) and in the value of T - average life expectancy (3 times increase).

2.4. Antiviral activity of the drug GSSG∞inosine
In the chemotherapy laboratory of the Influenza Institute of the Ministry of Health of the Russian Federation (St. Petersburg), the antiviral activity of GSSG∞inosine against influenza A virus (NZ 2) was studied. Antiviral activity was determined by the ability of the test drug to inhibit the reproduction of influenza virus on the model of surviving fragments of the chorionallantoic membrane of the chicken embryo (CAO).

Drugs that caused a decrease in the infectious titer in the experiment compared with the control (neutralization index) by more than 2.0 lg ID ₅₀ . considered active; from 1.0 to 2.0 lg ID ₅₀ - moderately active and less than 1.0 lg ID ₅₀ - inactive.

 A preliminary study of the toxicity of the drug GSSG∞inosin showed that even at a concentration of 1 mg / 0.5 ml, it did not cause a damaging effect on the cells of ChAO. Thus, the maximum tolerated dose (MTD) is ≥ 1000 μg / 0.5 ml.

 The results of experiments to study the antiviral activity of GSSG∞inosin samples against model influenza virus A / Hong Kong / 1/68 (NC 2) are presented in table. 6.

 As can be seen from the data presented, the GSSG∞inosin samples have a pronounced antiviral activity (CTI = 5) and reduce the infectious activity of the virus.

 Thus, the data presented justify the choice of experimental models of toxic hepatitis for studying the specific pharmacological activity of GSSG∞inosin, followed by extrapolation of the results to viral lesions of the liver.

повышает устойчивость белых мышей к возбудителю лихорадки долины Рифт, вызывающего у этих животных летальную генерализованную инфекцию с поражением печени и других органов.CONCLUSIONS
1. New systemic cytotoprotector and immunomodulator

increases the resistance of white mice to the causative agent of Rift Valley fever, which causes a fatal generalized infection in these animals with damage to the liver and other organs.

2. The introduction of GSSG∞inosine intraperitoneally in single doses of 3, 10, 30 mg / kg for 6 days with experimental LDR in white mice infected with 1-20 LD ₅₀ virus, allows to increase the survival rate of animals by 30-60%, and significantly (2 times or more) increase their life expectancy.

 3. The administration of GSSG∞inosine during experimental rift valley fever in white mice together with the antiviral drug ribamidil allows increasing the protective effect of the latter, as evidenced by an increase in the survival rate (by 20%) and life expectancy (by 2 times) of infected mice.

 4. GSSG∞inosine at a dose of 30 mg / kg when administered for 5 days increases the resistance of immunocompromised white mice to the DNA-containing herpes simplex virus.

 5. On a model of neuroviral infection caused by the causative agent of Venezuelan encephalomyelitis in horses (VEL). revealed the antiviral effect of the drug GSSG∞inosin. The prophylactic regimen of GSSG∞inosine was more effective than the emergency prophylactic.

 6. The study of antiviral activity against influenza A virus showed that the drug GSSG∞inosine has a pronounced antiviral activity (CTI = 5) and reduce the infectious activity of the virus.

 Thus, the unique property of GSSG∞inosine to induce apoptosis mechanisms of virus-infected cells and to activate the proliferation and differentiation of normal cells ensures the high efficiency of GSSG∞inosine against a wide range of viral infections.

Example 3
Enhanced expression of the apoptosis inducer - Fas / APO-I - receptor (CD95) in virus-infected hepatocytes with GSSG∞inosin
Fas receptors, also referred to as CD95 antigen, are the starting point of the receptor-mediated apoptotic cascade. Liver tissue is rich in these receptors and therefore apoptosis of liver cells is usually realized through a Fas-dependent mechanism. Activation of Fas membrane receptors in defective hepatocytes leads to the death of genetically modified or virus-infected cells. In the case of a favorable course of the disease caused by the hepatitis C virus, the accumulation of Fas receptors in the cells infected with the virus is shown, which contributes to their programmed death and elimination of, thus, infected cells from the tissue. The presence of domains rich in reactive cysteines in the extracellular part of the Fas receptor determines its activation upon a sharp change in the state of SH groups in the intercellular medium. This could happen when the ratio of oxidized and reduced glutathione changes in the treatment of patients with Glutoxim drugs. In this study, the effect of treatment with GSSG∞inosine on the expression of Fas receptors in liver biopsy samples of such patients was studied.

The formation of patient groups for examination using a liver biopsy was carried out depending on the clinical course of the disease, the type of virus detected, and the use of GSSG∞inosine instead of standard medications. A biopsy was performed only for patients with a chronic course of hepatitis B or C. The first biopsy was performed to assess liver damage before treatment. A second biopsy was performed 3 (hepatitis B) or 6 months (hepatitis C) after the start of standard therapy or therapy with GSSG∞inosine injections.
A total of 84 patients with hepatitis B and 63 patients with hepatitis C were examined. During the morphological study of biopsy samples, they were divided in accordance with Knodell classification. Thus, the protocol involved patients with a moderate degree of inflammatory process in the liver, as well as with moderate manifestations of fibrotic substitution of the parenchyma. In the blood of patients at the beginning of the course of therapy by PCR, from 500,000 to 1,000,000 copies / ml of viral particles were detected. A total of 78 patients with hepatitis B and 54 patients with hepatitis C were selected.

 In addition to the morphological assessment of fibrotic processes and the detection of viral particles, immunomorphological analysis of cells containing Fas receptors was carried out, and the content of Fas receptors in freshly prepared homogenate from biopsy samples was quantitatively determined by the immunochemical method. For immunomorphological analyzes, reagents and antibodies from DAKO (DENMARK) were used, and immunoassays were performed with a kit from Oncogene (USA).

 Fixed tissue samples from paraffin blocks are prepared in the form of sections with a thickness of 5 microns, which are dewaxed and dehydrated, as described in the protocol of the company "DAKO". Monoclonal antibodies against Fas / Apo1 receptors recognize it on the surface. Antibodies were diluted in accordance with the instructions (1: 150) and applied to a glass slide with a slice. Incubated in a humid chamber for 30 minutes or 1 hour in accordance with the attached prescription. Unbound antibodies were washed twice in 50 ml of PBS buffer for 2 minutes and then the bound primary antibodies were detected by secondary antibodies containing a biotin tag using a DAKO kit (DAKO LSAB kit k675). Bound biotin is detected by streptavidin-peroxidase conjugate in the presence of a chromogenic substrate, for example diaminobenzidine (DAB), as described in the protocol. Relevant components and reagents are included in the DAKO kit. The ratio of cells with expressed Fas receptors to other cells was evaluated in accordance with the protocol.

Immunoassay of the Fas antigen was performed with a cell lysate of a freshly biopsy sample. Enzyme immunoassays were performed by the sandwich method using murine monoclonal antibodies to human Fas protein immobilized in the wells of a 96-well Oncogene plate (USA). Incubation of the lysate for one hour causes the binding of the Fas antigen, and upon subsequent washing, the bound Fas antigen remains in the wells. Other specific biotinylated antibodies to the Fas antigen were added to the washed wells, and after binding to the Fas antigen, the wells were washed again. The conjugate of streptavidin with horseradish peroxidase was then added. Streptavidin with horseradish peroxidase sewn to it specifically binds to the biotin of the second antibodies, and after washing the whole complex remains in the wells. A chromogenic substrate of tetramethyl benzidine (TMB) was added to the wells, which under the action of peroxidase changes from a colorless state to a bright blue colored product, which is recorded spectrophotometrically. Used ready-made analysis kits Fas-APO-1 company Oncogene ^TM Research Products (USA).

Lysing buffer was added to the biopsy sample at a ratio of 10: 1 and homogenized. Incubate 30 minutes on ice, shaking occasionally. Cell debris was removed by centrifugation for 5 minutes, 12,000 rpm in an Eppendorf centrifuge. The supernatant is used immediately or stored at -80 ^o C.

 100 μl of the cell lysate supernatant was added to the wells of an immobilized antibody plate and incubated for 1 hour at room temperature. Washed 3 times in washing buffer. 100 μl of detecting antibodies were added to each well and incubated for another hour at room temperature. Washed 3 times in washing buffer. A streptavidin-peroxidase conjugate was diluted 1: 400 and 100 μl was added to each well. Incubated for 30 minutes at room temperature. The wells were washed with washing buffer. 100 μl of a solution of a chromogenic substrate was added for 30 minutes. 100 μl stop solution (2.5 N sulfuric acid) was added and the absorbance of the solution in each well was measured on a spectrophotometer or immunoassay reader at a wavelength of 450/540 nm. The measurement was performed no later than 30 minutes after adding stop solution. The results were compared with the control samples included in the kit. It was shown that both in patients with hepatitis B and patients with hepatitis C, a large percentage of cells are enriched in Fas receptors (Tables 7, 8).

 The determination of the content of the Fas receptor in the freshly prepared homogenate of a part of biotaps also testified to the accumulation of Fas receptors in the liver tissue after treatment with GSSG∞inosine, as in hepatitis B and especially in hepatitis C (Table 9). So, in the supernatant of the liver cell homogenate lysate from freshly extracted biopsy specimens, the Fas content was less than 2 u / mg when compared with Fas standards.

 The presence of viral infection led to a significant increase in the content of Fas receptors. Upon repeated analysis after 3 (hepatitis B) or 6 (hepatitis C) months against the background of standard management of patients, the content of Fas receptors remained at the same level. In contrast, with GSSG∞inosin therapy, the content of Fas receptors significantly increased.

The implementation of Fas-dependent apoptosis is the main pathway for programmed cell death of hepatocytes. Given the presence of domains enriched in reactive cysteines in the extracellular part of the Fas receptor, it can be assumed that the GSSG∞inosine preparation promotes aggregation and, thus, activation of Fas receptors due to the wave-like change in the ratio of reduced and oxidized SH groups in the pool of extracellular glutathione. Moreover, the intracellular cascade of caspases that realize Fas-dependent signals of cell death, which are cysteine proteases, can also be activated by GSSG∞inosine.
Conclusion: Activation of Fas-dependent apoptosis helps to cleanse tissue from virus-infected cells. However, due to the virus-dependent synthesis of apoptosis inhibitors, Fas-induced processes are incomplete and then virus-infected cells avoid apoptosis. The increase in the content and activity of Fas receptors using the GSSG∞inosin preparation provides a more complete elimination of virus-infected cells. These results are consistent with evidence of a decrease or absence of hepatitis C virus in the bloodstream after treatment with GSSG∞inosine.
Example 4
Investigation of the inhibition of the ATP-azo / helicase activity of hepatitis C virus by GSSG∞inosine, GSSG-IMP and GSSG-UMP in a culture of normal and virus-infected cells
We evaluated the ability of compounds of oxidized glutathione with inosine (GSSG-I), inosine monophosphate (GSSG-IMP) and uridine monophosphate (GSSG-UMP) to influence the ATP-azo / helicase activity of normal and virus-infected (HEP-2) cells. For this, these substances were incubated with a nuclear lysate of normal lymphocytes obtained from the peripheral blood of healthy donors, or with a HEP-2 homogenate of cells infected with hepatitis C.

Venous blood from healthy donors was collected in heparinized tubes tested for endotoxin absence. The mononuclear fraction of blood leukocytes was obtained by centrifugation in a density gradient ficoll-pack (Pharmacia). The cell concentration was adjusted to 2 • 10 ⁶ cells per 1 ml of complete culture medium (RPMI 1640) containing 20 mM HEPES, 2 mM glutamine, 50 μg / ml gentamicin and 10% fetal calf serum. Cell viability was evaluated in a trypan blue test. Then the cell suspension was again centrifuged to remove residues of ficol with the medium. The precipitate was suspended in 1 ml of physiological saline and lysed with Nonidet 40 to obtain cell nuclei as described (Maniatis et al.). Then the nuclei were lysed and the change in ATP-azo / helicase activity in the presence of the studied compounds was analyzed in the lysate.

0.1 ml, annealed with a P ³² -labeled complementary oligonucleotide (42 nucleotides long), single-stranded phage M13 mp10 phage was added to 0.2 ml of the nucleic acid lysate. Concentrations of phage DNA were preliminarily selected, which made it possible to visually, by radio autography, to reveal both the minimum extension and separation of the labeled oligonucleotide from the DIC of the phage, and the almost complete separation of the oligonucleotide from DNA of the single-stranded form of the M13 phage.

0.1 ml Tris-Hcl buffer solution containing ATP and Mg ⁺⁺ was also added to the incubation mixture. Then 0.1 ml of physiological saline solution of NaCl or 0.1 ml of physiological saline with the addition of the test composite to a final concentration of 10, 50 and 100 μg / ml.

After incubation for 30 minutes, 10 μl of the sample was subjected to vertical electrophoresis in 8% polyacrylamide gel. There was a separation into a high molecular weight fraction, which is a hybrid of a P ³² -labeled oligonucleotide bound to phage DNA, and a low molecular weight fraction - an oligonucleotide released from the hybrid by the action of helicase. Densitometry of radio autographs revealed changes in the ratio of bound and released under the action of helicase P ³² -labeled oligonucleotide.

 When assessing helicase activity in a lysate of a culture infected with hepatitis C virus, the same technique was used, but instead of a nuclear lysate, a cell lysate was used to evaluate the contribution of virus-induced cytoplasmic proteins.

 The research results are presented in table. 10 and 11. As can be seen from the table. 1, none of the studied substances at a concentration of 10 μg / ml causes a change in helicase activity in the lysate of donor lymphocyte nuclei, whereas at concentrations of 50 and 100 μg / ml all three tested compounds are significantly suppressed. GSSG-UMP among them causes the greatest inhibition of ATP-azo / helicase activity.

 In the virus-infected cell lysate, the helicase activity is higher since the depletion of 90% of double-stranded DNA hybrids of the M13 single-stranded domain with the oligonucleotide was achieved at a hybrid concentration of 20 pg / ml compared to 5 pg / ml in the lysate of the donor lymphocyte nuclei. The effect of the test compounds also manifests itself at these higher levels of helicase activity. Significant inhibition of helicase activity was observed at a concentration of these substances of both 50 and 100 μg / ml, but at a concentration of 100 μg / ml, inhibition of ATPase / helicase activity was more pronounced. The maximum inhibitory effect on ATPase / helicase activity was obtained using the GSSG-UMP compound.

Discussion and conclusion:
The high helicase activity in cells infected with hepatitis C virus, compared with donor lymphocytes, can be associated not only with proliferative activity, but also with the expression of the 3rd non-structural protein of hepatitis C virus, which is an enzyme with ATPase / helicase activity. Suppression of helicase activity not only in the lysate of donor lymphocyte nuclei, but also in the lysate of virus-infected cells suggests that the compounds of oxidized glutathione with nucleosides (GSSG-I) or nucleotides (GSSG-IMP, GSSG-UMP) suppress the helicase activity of the non-structural NS3 virus protein hepatitis C, which prevents the synthesis of virus-specific RNA in infected cells.

Example 5
Treatment of acute viral hepatitis B with prolonged course of the drug GSSG∞inosin
Surname, name, patronymic of the patient: K.V.V.

Gender: Male
Age 32 years old
Medical history 661
Diagnosis: Acute viral hepatitis B, replication phase (PCR HBV +), prolonged course, moderate activity.

 Complaints during examination: On weakness, heaviness in the right hypochondrium, nausea, sweating.

 Anamnesis of the disease: Sick from 10/01/98, when there were sharp pains in the small joints of the hand, and then pain and hyperemia, swelling in all joints. From 09.10.98 - darkening of urine. Received for treatment at the clinic of viral infections. After the course of treatment (see below), a high cytolytic syndrome remains.

 Previous treatment: A course of detoxification, antispasmodic, antibacterial (kanamycin 0.5 • 2 times a day IM No. 7), anti-inflammatory (indomethacin 1 tablet, 3 times a day) therapy was carried out.

The course of therapy with the drug ≪GSSG∞inosin≫ from 10/29/98 to 11/21/98:
0 day
Intramuscularly "GSSG∞inosine" 1% - 1 ml daily
1-14 day
Intravenously "GSSG∞inosine" 1% - 1 ml
15-17-19 day
Intravenously "GSSG∞inosine" 3% - 1 ml
20-24 day
Intramuscularly "GSSG∞inosine" 1% - 1 ml
The condition of the patient after completion of the course of treatment: A significant improvement in the condition, which is expressed in the absence of weakness, nausea, heaviness in the disposition of the hypochondrium. The results of laboratory tests - see table. 12, 13 and 14.

 Conclusion: Conducting a course of therapy with the drug "GSSG∞inosin" provided positive dynamics of key indicators of the patient's condition, which is expressed in the normalization of biochemical parameters, cessation of HBV replication, persistence of Hbs Ag, and improvement in general condition. The observation in dynamics (1 and 3 months after the end of the course of treatment) showed stabilization of these indicators and allowed us to regard this condition as early convalescence.

Example 6
Treatment of acute viral hepatitis B, severe course, with GSSG∞inosine
Surname, name, patronymic of the patient: A. Ya. V.

Gender Female
Age 18 years old
Medical history 6006
Diagnosis: Acute viral hepatitis B, replication phase (PCR HBV +), severe course.

 Complaints during examination: On weakness, heaviness in the right hypochondrium, nausea, sweating, fatigue.

 Anamnesis of the disease: Sick from 03/12/99, when there were sharp pains in the small joints of the hand, and then pain and hyperemia, swelling in all joints. From 09.16.99 - darkening of the urine. Received for treatment in the 26th department of the hospital. S.P. Botkin. After the course of treatment (see below), a high cytolytic syndrome remains.

 Previous treatment: A course of massive detoxification, antispasmodic, antibacterial (gentamicin 40 mg • 2 times a day IM No. 7), anti-inflammatory (indomethacin 1 tablet, 3 times a day) therapy was carried out.

The course of therapy with the drug ≪GSSG∞inosin≫ from 04/08/99 to 05/02/99:
0 day
Intramuscularly "GSSG∞inosine" 1% - 1 ml
1-14 day
Intravenously "GSSG∞inosine" 1% - 1 ml
15-17-19 day
Intravenously "GSSG∞inosine" 3% - 1 ml
20-24 day
Intramuscularly "GSSG∞inosine" 1% - 1 ml
The patient’s condition after the end of the course of treatment: A significant improvement in the condition, which is expressed in the absence of weakness, nausea, heaviness in the right hypochondrium. The results of laboratory tests - see table. 15, 16 and 17.

Conclusion: Conducting a course of therapy with the drug "GSSG∞inosin" provided positive dynamics of objective indicators of the patient's condition, namely:
a) the normalization of biochemical parameters;
b) termination of HBV replication and persistence of HbsAg;
c) improvement in general condition.

 Observation in dynamics (1 month after the end of the course of treatment) showed stabilization of these indicators and allowed us to regard this condition as early convalescence.

Example 7
Treatment of chronic viral hepatitis B with GSSG∞inosine
Surname, name, patronymic of the patient: K. M. D.

Gender Female
Age 41 years old
Diagnosis: Chronic viral hepatitis B (HbsAg +) replication phase (PCR HBV +), moderately expressed degree of activity. Concomitant diseases: chronic cholecystitis, chronic pancreatitis. Obesity II degree.

 Liver biopsy (background): The beam and lobular structure of the liver is preserved. Portal tracts are expanded due to the growth of connective tissue with the formation of port-portal septa. In the periportal connective tissue, moderate lympho-macrophage infiltration (3 points). The inner border plate is partially destroyed, step necrosis (2 points). Focal necrosis is determined in lobules (1 point). Intralobular and noncentral lymphoid infiltration. Vacuumization of the cytoplasm of hepatocytes with degeneration and polymorphism of their nuclei. Matte vitreous hepatocytes are determined. Conclusion: chronic hepatitis with mild activity (Knodel IGA = 6) and moderate fibrosis.

 Liver biopsy (12 months after the end of treatment): The beam and lobular structure of the liver is preserved. Portal tracts are slightly expanded due to the proliferation of connective tissue. In the periportal connective tissue, mild lymphatic macrophage infiltration (1 point). The inner border plate is saved. Minor intralobular and non-central lymphoid infiltration. Conclusion: chronic hepatitis with minimal activity (Knodel IGA = 1) and mild fibrosis.

 Complaints during examination: Weakness, inability to perform normal work.

 Anamnesis of the disease: Was in hospital treatment from 9 to 30.04.97 with a diagnosis of chronic viral hepatitis B. The disease proceeded with moderate cytolytic syndrome. A course of detoxification therapy. During treatment, the ALT level decreased from 620 to 365 U / L, the bilirubin level was normal at discharge.

 Previous treatment: In connection with the replicative activity of the virus (PCR HBV +), a course of acyclovir was prescribed from 04/25/97 for 21 days, and then cycloferon from 05/26/97. During treatment, ALT levels ranged from 421 to 168 U / L. The activity of the virus was suspended for 1 month, then again intensified from 15.10.97.

The course of therapy with the drug "GSSG∞inosin" from 10/15/97:
0 day
Intramuscularly "GSSG∞inosine" 1% - 1 ml
1-14 day
Intravenously "GSSG∞inosine" 1% - 1 ml
15-30 day
Intravenously "GSSG∞inosine" 3% - 1 ml
31-90 day
"GSSG∞inosine" 3% - 1 ml three times a week
The patient’s condition after completion of the course of treatment: For the first time in 7 months of illness, the level of blood biochemical parameters returned to normal (ALT - 33 U / L, bilirubin - 9.0 μmol / L). The patient feels well, there is no weakness, normal exercise tolerance appeared. Laboratory indicators - see table. 18, 19, 20 and FIG. thirty.

Conclusion: the course of therapy with the drug "GSSG∞inosin" provided:
- termination of HBV replication;
- normalization of biochemical parameters;
- normalization of immunological parameters and cytokine status;
- marked improvement in morphological parameters (decrease in Knodel index and fibrosis).

 Thus, the drug "GSSG∞inosine" is an effective drug for the treatment of chronic viral hepatitis B.

Example 8
Treatment of chronic viral hepatitis B, cirrhotic stage, with GSSG∞inosine
Surname, name, patronymic of the patient: A. V. I.

Gender: Male
Age 59 years old
Diagnosis: Chronic viral hepatitis B, cirrhotic stage (PCR HBV +), ascites, portal hypertension.

 Complaints at the beginning of the course of treatment: Weakness, persistent ascites, dizziness.

 Anamnesis of the disease: For the first time was in the clinic for HVHV, cirrhotic stage, ascites in 1995. Subsequently, he was re-treated in April 1997. This deterioration since the beginning of October: shortness of breath and pulling pains in the left hypochondrium and epigastrium intensified. Takes furosemide 2 times a week for 1 tablet. On admission: moderate condition, pale skin, tongue bright, moist, pulse 110 beats. min, BP 140/90, BH 20 dv / min. On the left, below the angle of the shoulder blade, blunting of percussion tone, breathing is not performed. The abdomen is enlarged (abdominal volume 113 cm) due to ascites. There is no edema on the legs. Got a course of detoxification, symptomatic, antibiotic therapy. Conducted infusion therapy with diuretics, proteins.

The course of therapy with the drug ≪GSSG∞inosin≫:
1 course from 11/27/97 to 12/20/97 1% IM 3 times a week,
2 course from 01/12/98 to 02/06/98 1% IM 3 times a week,
3 course from 02/11/98 to 05/14/98 1% IM 3 times a week,
4 course from 06/06/98 to 07/10/98 1% IM 3 times a week.

 The condition of the patient after the end of 1 course of treatment: Condition is satisfactory. She notes a significant improvement, notes cheerfulness, walks. Ascites decreased (abdominal volume 89 cm), adequate diuresis, lack of pain in the left hypochondrium. Clinical and laboratory data are presented in table. 21. At the patient's insistence, repeated courses of GSSG∞inosin are prescribed according to the schemes. The courses of this therapy made it possible to maintain stable well-being, as well as a stable picture of clinical and laboratory data (Table 21).

Conclusion: The course of therapy курсаGSSG∞inosin≫ provided:
- positive correction of lymphocyte and thrombocytopenia;
- termination of HCV replication;
- lack of cytolytic syndrome (normalization of bilirubin content and ALT activity);
- decrease in ascites;
- improving the quality of life: improving well-being and its stabilization during the 8 months of observation, which is expressed in increasing energy, vitality, and the desire to actively conduct further treatment.

Example 9
Treatment of chronic viral hepatitis B, cirrhotic stage, with GSSG∞inosine
Last name, first name of the patient: T. I. N.

Gender: Male
Age 48 years old
Diagnosis:
Primary: Chronic viral hepatitis B (HbsAg +), integration phase (PCR HBV-), cirrhotic stage
Concomitant: Condition after cholecystectomy
Complaints at the beginning of the course of treatment: Weakness, insomnia
Medical history: Chronic hepatitis B was first detected in 1999 during an examination for surgical treatment of cholecystitis. After cholecystectomy performed in January 2000, therapy with GSSG∞inosine was started on April 26, 2000.

The course of therapy with the drug GSSG∞inosine.
1% solution 3 times a week for 12 weeks.

 Ultrasound: Background from 04/18/00 1344. The liver is of normal size, the structure is homogeneous, fine-grained, the contours are even, clear vessels are visible but on the periphery, the structure is densified. Portal vein 13 mm, v. mesenterica superior 14 mm, choledoch 7 mm, spleen of normal size, hypoechoic structure, contours are even, clear. The gall bladder is removed. Pancreas: head 22 mm, body 16 mm, tail 20 mm, the contours are even, the structure is homogeneous, fine-grained, echogenicity is equal to the liver. Kidneys of normal size, bean-shaped. there are no calculi. Conclusion: diffuse changes in the liver.

 The condition of the patient after completion of the course of treatment: notes a significant improvement and the ability to fulfill his professional duties in full force. The course of treatment allowed to improve the clinical and laboratory parameters and the quality of life of the patient (tab. 22, 23 and 24).

Conclusion: a course of therapy with GSSG∞inosin provided:
- correction of thrombocytopenia;
- cessation of cytolytic syndrome (normalization of bilirubin content and ALT activity);
- stimulation of cellular immunity (increase in the content of CD3 +, CD4 +, CD16 / 56);
- marked improvement in portal blood circulation according to dopplerography.

Example 10
Treatment of a patient with chronic viral hepatitis C and chronic viral hepatitis B with GSSG∞inosine
Surname, name, patronymic of the patient: Z. V.V.

Gender: Male
Age 18 years old
Medical history: 1043
Diagnosis: Chronic viral hepatitis C, replication phase (PCR HCV +), moderate degree of activity; chronic viral hepatitis B, integration phase (PCR HBV-), drug intoxication, drug addiction.

 Liver biopsy (background): The beam and lobular structure of the liver is preserved. Portal tracts are slightly expanded due to the proliferation of connective tissue. In the periportal connective tissue, moderate lympho-macrophage infiltration (3 points). The inner border plate is saved. In some segments, the bodies of Kaunsilmen are determined (1 point). Intralobular and pericentral lymphoid infiltration. Vacuolization of the cytoplasm and some nuclei of hepatocytes. Conclusion: chronic hepatitis with minimal activity (Knodel IGA = 4) and mild fibrosis.

 Liver biopsy (12 months after the end of treatment): The beam and lobular structure of the liver is preserved. Portal paths are not changed. In the periportal connective tissue weak lymph-macrophage infiltration (1 point). The inner border plate is saved. In some segments, the bodies of Kaunsilmen are determined (1 point). Intralobular and pericentral lymphoid infiltration. Vacuolization of the cytoplasm and some nuclei of hepatocytes. Conclusion: chronic hepatitis with minimal activity (Knodel IGA = 2).

 Complaints during examination: Weakness, severe pain in the right hypochondrium, knee joints, spine, joints of the hand.

 Anamnesis of the disease: I noticed pains in the joints of the knees, spine in early August 1997. An examination of the blood revealed an increase in the level of bilirubin to 34.0 μmol / L and ALT - 86 U / L. In a stationary examination from 08.15.97, anti-HCV IgG and the replicative activity of hepatitis C virus were determined.

 Anamnesis of life: I started to use drugs from the age of 14. Their intravenous administration was carried out from the age of 17. At the time of inspection, the dose of heroin is about 2 g per day.

 Previous treatment: Not performed.

The course of the drug ≪GSSG∞inosin≫ from 08.15.97:
0 day
Intramuscularly "GSSG∞inosine" 3% - 1 ml
1-14 day
Intravenously "GSSG∞inosine" 3% - 1 ml
15-30 day
Intravenously "GSSG∞inosine" 3% - 1 ml
The course is held for 3 months.

 The condition of the patient after completion of treatment: The condition is satisfactory. Marks a significant decrease in weakness, the absence of pain in the right hypochondrium, in the joints. According to the patient, the way out of the state of drug withdrawal was almost painless and less long. Normalization of biochemical parameters and the absence of replicative activity of the virus are noted (see tab. 25, 26, 27 and Fig. 31).

Conclusion: Conducting a course of therapy with the drug SSGSSG∞inosin≫ provided a positive trend, which is expressed in the normalization of biochemical, serological parameters, and cessation of HCV replication. Immunological parameters and cytokine status data correlate with the arrest of the infection process and the absence of virus replication. When examining the patient’s peripheral blood lymphocytes by flow cytometry using monoclonal antibodies to FasAg (CD95 +) after treatment, an increase in CD95 ⁺ cells was shown, which indicates the activation of the programmed cell death of virus-infected cells. When observed in dynamics one month and three months after the end of the course of treatment, stabilization of this condition is noted. With a second biopsy 12 months after the end of treatment, a decrease in the Knodel index and the absence of fibrosis are noted.

 Concomitant drug intoxication and withdrawal symptoms when using ≪GSSG∞inosin≫ were stopped earlier and passed with less suffering for the patient.

Thus, the treatment with ≪GSSG∞inosin≫ of chronic hepatitis C with replicative activity and concomitant drug addiction allowed us to achieve the following results:
- normalization of biochemical parameters of blood;
- normalization of hematological parameters;
- lack of replicative activity of virus C;
- improvement of morphological indicators of the state of the hepatic parenchyma;
- normalization of immunological parameters and cytokine status;
- induction of apoptosis of virus-infected cells;
- stable therapeutic effect.

Example 11
Treatment of chronic viral hepatitis C in the stage of cirrhosis with GSSG∞inosine
Surname, name, patronymic of the patient: G. N. V.

Gender Female
Age 48 years old
Diagnosis: Cirrhosis (Child C), chronic viral hepatitis C.

 Ascitic syndrome, varicose veins of the esophagus and cardia of the stomach, stage II.

Complaints about the start of treatment:
Weakness, insomnia, arthralgia, skin itching, gum bleeding
Anamnesis of the disease: Chronic hepatitis C in the cirrhotic stage was first detected in 1993 during a random examination. Ascites began to increase in 1999 before the start of GSSG∞inosin therapy.
The course of therapy with the drug GSSG∞inosine.
1% solution 3 times a week for 12 weeks.

 The condition of the patient after completion of treatment: The condition is satisfactory. She notes a significant improvement (see table. B), fulfills her professional duties in full. The course of treatment allowed to improve the clinical and laboratory parameters and the quality of life of the patient (see tab. 28, 29 and 30).

Conclusion: a course of therapy with GSSG∞inosine provided:
- minimal manifestations of ascites syndrome;
- improvement of portal blood circulation according to dopplerography;
- lack of dopplerographic indicators of portal hypertension (absence of spleno-renal anastomoses);
- almost complete normalization of the content of lymphocytes and platelets;
- improving the quality of life.

Example 12
Therapeutic effects of GSSG∞inosine in an AIDS patient
Patient: M.V. M.

Gender: Male
Age 50 years old
The diagnosis is basic: AIDS. According to the classification of the SDS Atlanta: C3.

 Concomitant diagnosis: Kaposi's sarcoma. Tuberculosis of the intrathoracic lymph nodes, cytomegalovirus infection, candidiasis of the oral mucosa, herpes simplex, eczema, neurosyphilis.

 HIV infection was diagnosed in 1994. Immunoblot testing is positive for p24, p17, p25, p18, p55, p40, p68, p58, p34, gp120, gp160.

Initial state: Severe, temperature 38.5 ^o C, persisting for more than a month, Karnovsky index - 50, progressive deterioration of immunological parameters. Sharp leukopenia (1.8 • 10 ¹² ) and lymphopenia (570) excluded the use of drugs of the AZT group and other antiviral drugs.

The course of treatment with the drug "GSSG∞inosine (D-form)":
"GSSG∞inosine" 3% - 1 ml 3 times a week intramuscularly for 12 weeks.

The therapeutic effect after a month of treatment:
- body temperature of 37.5 ^o C;
- improving the quality of life - Karnovsky index 70;
- there was a tendency towards normalization of immunological parameters (the number of lymphocytes increased from 570 to 832, the ratio of CD4 ⁺ / CD8 ⁺ increased from 0.71 to 0.84);
- viral load of 89,000 copies / ml.

The therapeutic effect after 2 months of treatment:
- body temperature 36.9 ^o C;
- improving the quality of life - Karnovsky index 75;
- there remains a tendency to normalize immunological parameters (the number of lymphocytes is 736, the ratio of CD4 ⁺ / CD8 ⁺ is 0.87; CD4 ⁺ is 199; CD8 ⁺ is 228);
- viral load of 56,000 copies / ml.

Therapeutic effect at the end of treatment:
- body temperature 36.6 ^o C;
- improving the quality of life - Karnovsky index 90;
- there remains a tendency to normalize immunological parameters (the number of lymphocytes is 1350, the ratio of CD4 ⁺ / CD8 ⁺ is 0.87; CD4 ⁺ is 527; CD8 ⁺ is 608);
- viral load of 10,000 copies / ml.

 Laboratory indicators - see table. 31 and 32.

Example 13
Therapeutic effects of Zn ₂ -GSSG-IMP ¹ in an AIDS patient
(Zn ₂ -GSSG-IMP - GSSG zinc salt covalently linked to inosine monophosphate)
Patient: Y. O. I.

Gender Female
Age 40 years old
The diagnosis is basic: HIV infection in the AIDS stage. According to the classification of the SDS Atlanta-C2. Chronic recurrent herpes simplex. Encephalopathy Chronic viral hepatitis B (HbsAg +) is being integrated.

 Upon admission to the clinic of infectious diseases 12.03.99 (Department of AIDS) complains of weakness, fatigue, excessive sweating, especially at night, weight loss, memory loss, soreness in the wrist joints, in the popliteal fossa on the left.

 Objectively: The condition is satisfactory. Nutrition lowered. The skin is pale. Cervical and axillary lymph nodes up to 1 cm in diameter. The mucous membranes of the oral cavity are pale. Above the lungs, percussion sound is not muffled. During auscultation - hard breathing, dry scattered rales. The abdomen is soft, sensitive in the epigastric region, a slight tightness is determined in the right ileal region. Acrocyanosis of the bones of the hands is noted. Scars from multiple herpetic eruptions are visible on the lips. Karnovsky index -75.

The course of treatment with the drug "Zn ₂ -GSSG-IMP" from 03/26/99:
"Zn ₂ -GSSG-IMP" 3% - 1 ml 3 times a week intramuscularly - 8 weeks.

After a month of treatment with the drug "Zn ₂ -GSSG-IMP": The patient notes a decrease in weakness, sweating at night has disappeared. I gained 1.8 kg in weight. The level of lymphocytes in the blood is 1276. CD4 ⁺ - 319, CD8 ⁺ - 370. Karnowski index 80.

After the course of treatment with the drug "Zn ₂ -GSSG-IMP": No complaints. Satisfactory condition: sweating disappeared, became more energetic. The skin has become less pale. I gained 2.5 kg in weight. The level of lymphocytes in the blood is 2295, CD4 ⁺ - 574, CD8 ⁺ - 597. Karnowski index 90.

 The specific dynamics of the indicators of the immunogram and hemogram is reflected in the table. 33 and 34.

conclusions
The course of treatment with the drug "Zn ₂ -GSSG-IMP" in monotherapy provided:
- reduction of viral load;
- improving the quality of life of the patient;
- improvement of immunological parameters;
- stabilization of hematological parameters.

Example 14
Therapeutic efficacy of GSSG∞inosine in chlamydial infections
Patient: S.I.M., 29 years old.

 Diagnosis: Genitourinary chlamydia occurring with systemic manifestations. Chronic vesiculoprostatitis. Chronic synovitis of the right knee joint, chronic blepharoconjunctivitis.

 Anamnesis: A patient suffers from chronic prostatitis for 8.5 years. It was treated 4 times with the help of modern antibiotics (macrolides, fluoroquinolones, doxycycline). The last 2 courses of treatment were carried out against the background of immunotherapy (Cycloferon, Viferon). In 1999, the last time he was treated in the clinic of urology of St. Petersburg honey. Un-ta, but after 3 months. a relapse of the disease has developed in the form of exacerbation of chronic prostatitis and synovitis of the right knee joint; Also worried were pain in the eyes, photophobia, discharge from the eyes after sleep. On this occasion, he was treated by an ophthalmologist (ointment with tetracycline, erythromycin), but no positive effect was observed. Since March 1992 he received: bicillin, kanamycin, doxycycline, tetracycline, trichopolum, tinidazole, metacyclin, sumamed, tsifran, zanocin, abactal, rulide, cycloferon, viferon.

 In September 2000, on the recommendation of specialists from the military traumatology and orthopedics clinic of the Military Medical Academy, the patient was referred for consultation to the military medical clinic of infectious diseases, where an in-depth examination and complex therapy using the drug GSSG∞inosine + antibiotics were performed. Etiotropic therapy was carried out after determining the antibiotic sensitivity of chlamydia isolated on the cell culture from the ejaculate and synovial fluid of the joint.

 When handling: satisfactory condition, complaints of general weakness, malaise, prolonged (more than 1.5 months) subfebrile temperature. periodic pain in the perineum and in the region of the right knee joint, as well as swelling of soft tissues and hyperemia of the skin in the joint.

 In the secret of the prostate gland are signs of chronic prostatitis. During puncture of the right knee joint, 8 ml of turbid synovial fluid was obtained. After the puncture, the edema somewhat decreased.

 A direct immunofluorescence method (PIF) in the secretion of the prostate gland and synovial fluid detected reticular and elementary bodies of C. Trachomatis in significant quantities. The titer of antibodies in blood serum to C. trachomatis (in the NIFM - indirect immunofluorescence method) is 1: 128, in synovial fluid - 1: 64. The pathogen culture was isolated in pure form on the cell culture from ejaculate and articular fluid, the sensitivity to antibiotics was determined.

 The dynamics of the main laboratory parameters are presented in table. 35.

 Conclusion: the use of the drug “GSSG∞inosin” according to the scheme: 1 ml 3% solution once every 2 days for 10 days (5 injections) before antibiotic therapy and similarly during antibiotic therapy (8 days - maxaquin and 8 days - vilprafen) - provided a persistent clinical and bacteriological effect. Sanitation of the body from chlamydia was established by the main indicators immediately after treatment, and according to PCR - 3 months after the course of therapy. GSSG∞inosine helped to reduce the normalization time of the main immunological gels (T- and B-lymphocytes, IL-1, IL-2, IL-8, IFN-α and γ).

Example 15
Therapeutic efficacy of GSSG∞racil in mixed infections: herpetic and chlamydial
Patient: Mr. M. M.

Diagnosis: Chlamydia. Genital herpes. Chronic pyelonephritis
Anamnesis: For 5 years, suffers from chronic pyelonephritis and chlamydial herpes infection. It was treated with all types of antibiotic therapy (fluoroquinolones, macrolides, tetracyclines), including accompanied by drugs such as roferon, cycloferon. The use of cycloferon caused toxic manifestations in the form of fever (t up to 40 ^o C), alopecia, skin rashes. In 1996, she was treated in the urology department of the Central Health Unit 122 for exacerbation of chronic pyelonephritis.

 Over the 5 years of continuous treatment, the titer of antibodies to chlamydia trachomatis decreased to 64. One month after the end of each treatment course, it increased above 150.

From December 1993 to August 1997 she received the following drugs:
Trichopolum, Tinidazole, Bicillin, Timogen, Summamed, Metacyclin, Dalacin S, Marveron, Doxycycline, Cifran, Zanocin, Macropen, Clotrimazole, Rheoferon, Abactal, Rovamycin, Tarivid, Rulid.

 In April 1999, the patient applied for treatment to the Department of Urology and Andrology of the Medical Academy of Postgraduate Education, where she was treated with the new drug GSSG∞ uracil in combination with antibiotic therapy.

 The treatment condition is satisfactory, complaints of weakness, periodic pain in the knee joints and lumbar region, and discharge in the form of whites.

 In the urine, weak leukocyturia. Autoantibodies to kidney tissues in titer 1: 4 are detected in the blood (complement binding reaction "in the cold" according to V. I. Ioffe and K. M. Rosenthal).

 The dynamics of peripheral blood are presented in table. 36.

Treatment protocol:
- 2 courses of antibiotic therapy (doxycycline 0.1 g 2 times a day for 10 days 3-12 days of treatment; clacid 0.25 2 times a day for 10 days 21-30 days of treatment);
- drug GSSG∞ uracil according to the scheme (1-30 days of treatment).

 After a month of treatment: Condition is good. No complaints. Blood and urine counts are normal.

Conclusion: the use of the drug GSSG∞ uracil provided:
- clinical recovery of the patient;
- the absence of signs of chlamydia and herpes according to laboratory data (bacteriological analysis, PCR, serological studies);
- lack of autoantibodies to the tissues of the kidneys, normalization of urinalysis.

Example 16
Therapeutic effects of Li ₂ -GSSG-IMP) in a patient with respiratory tuberculosis
Patient: N.F. V.

Gender: Male
Age 30 years old
The main diagnosis: Infiltrative tuberculosis of the upper lobe of the left lung in the phase of decay and seeding, CD (+).

 Concomitant diagnosis: Chronic bronchitis in the acute phase.

 Anamnesis: Pulmonary tuberculosis was detected in prisons in May 1998. Treatment in a prison hospital from May to August 1998 had no effect, the process progressed. After his release in August 1998, he continued treatment in a tuberculosis hospital in St. Petersburg. However, no improvement was achieved, and as of October 1998 massive bacterial excretion persisted, and the decay cavity in the lung tissue increased. Mycobacterium tuberculosis resistance to streptomycin and isoniazid has been established.

Initial state: Moderate, evening temperature 37.5 ^o C, night sweats, weakness, cough with purulent sputum discharge, Karnovsky index - 50, progressive deterioration of immunological parameters. Leukocytopenia, lymphocytopenia, monocytosis, stab neutrophil shift. In sputum CD (+) by microscopy up to 100 in the field of view. X-ray in the upper lobe of the left lung - infiltration with a decay cavity of 4x3 cm and foci of screening in the lower lobe of the left lung.

The course of treatment with the drug "Li ₂ -GSSG-IMP" as an accompaniment of anti-TB chemotherapy (isoniazid, rifampicin, pyrazinamide, amikacin).

"Li ₂ -GSSG-IMP" 3% - 1 ml intramuscularly 1 time per day, 5 times a week, course duration 3 months.

The therapeutic effect after a month of treatment:
- normalization of body temperature;
- the disappearance of night sweats;
- improving the quality of life - Karnovsky index 70;
- the patient gained 4 kg in weight;
- improvement of hematological parameters, there is a tendency to increase the level of IL-12, normalization of the ratio of Th ₁ / Th ₂ ;
- decreased bacterial excretion (BK + 2-4 in the field of view by microscopy), sputum acquired a serous character.

The therapeutic effect after 2 months of treatment:
- body temperature 36.7 ^o C;
- improving the quality of life - Karnovsky index 75;
- there is a tendency to normalize immunological parameters;
- cessation of bacterial excretion (BK-method of microscopy).

Therapeutic effect at the end of treatment:
- body temperature 36.6 ^o C;
- improving the quality of life - Karnovsky index 90;
- normalization of immunological parameters;
- lack of bacterial excretion (three times by the BK-flotation method);
- X-ray: closure of the decay cavity in the upper lobe of the left lung with an outcome in a linear scar (see Figs. 31 and 32).

 The change in the main laboratory parameters is given in table. 37.

Conclusion: the use of the drug "Li ₂ -GSSG-IMP" in the complex anti-tuberculosis therapy for 3.5 months ensured:
- cessation of bacterial excretion;
- healing of the decay cavity in the lung tissue;
- Overcoming drug resistance of Mycobacterium tuberculosis.

 These effects were not achieved by prior conventional chemotherapy for 6 months of treatment.

Экспериментальное исследование гепатопротекторной активности препарата GSSG∞инозин
Изучение гепатопротекторной активности препарата GSSG∞инозин проведено на доступных и воспроизводимых моделях токсического гепатита, так как известно, что механизмы гепатотоксичности - воспаление, цитолиз и холестаз - универсальны и неспецифичны вне зависимости от агента, индуцирующего повреждение печени. Для сравнения использовали известные гепатопротекторы "Эссенциале" (Essentiale, Rhone-Poulenk Rorer) и "Легалон" (Legalon, Madaus).Example 17
Hepatoprotective efficacy of the drug

An experimental study of the hepatoprotective activity of the drug GSSG∞inosine
A study of the hepatoprotective activity of the drug GSSG∞inosin was carried out on accessible and reproducible models of toxic hepatitis, since it is known that the mechanisms of hepatotoxicity - inflammation, cytolysis and cholestasis - are universal and non-specific, regardless of the agent that induces liver damage. For comparison, hepatoprotectors Essentiale (Essentiale, Rhone-Poulenk Rorer) and Legalon (Legalon, Madaus) were used.

 When modeling toxic hepatitis, dichloroethane and acetaminophen were used. Both compounds were introduced into the stomach to male rats of standard weight (160-170 g) through a metal atraumatic probe in doses of 500 and 400 mg / kg, respectively, 1 time per day for 4 days in a mixture with olive oil (1: 1). In addition, the combined administration of dichloroethane and acetaminophen at doses of 300 and 250 mg / kg, respectively, was used. After 10 days, the presence of toxic hepatitis was confirmed in all experimental animals by the morphological picture of the liver. Since that time, for 10 days, GSSG∞inosine (5 mg / kg, i / m) and comparison preparations: Essentiale ampoules (1 ml / kg iv in the tail vein) and Legalon (silymarin ) (100 mg / kg w / w).

 The effectiveness of treatment was judged by the clinical picture, body weight dynamics, relative liver mass, bilirubin content, transaminase activity, phosphatases, ceruloplasmin level, total protein, blood serum lipids, glycogen, glutathione, SH-groups, liver cytochromes, exercise tests ( hexenal test, bromosulfoalein test) and histological picture of the liver.

 It was established that the dependence on the type of toxicant, the death of experimental animals by the 20th day of the experiment ranged from 40% (in the group etched with dichloroethane) to 80% (in the group of animals exposed to the combined effects of dichloroethane and acetaminophen) (Figs. 34, 35 and 36). The most pronounced liver damage was observed during combined intoxication of the two chemical agents listed above, which was confirmed by a change in the biochemical parameters of the functional state of hepatocytes: an increase in the activity of transaminases, phosphatase, lactate dehydrogenase, a decrease in serum total protein, lipids, a reserve of sulfhydryl blood groups, an increase in the levels of bilirubin, and ceruloplasmin a significant slowdown in the excretion of bromsulfalein.

 The presence of toxic hepatitis was confirmed by morphological studies of liver tissue.

 In animals that died or were euthanized 1 day after oral administration of dichloroethane, signs of acute toxic hepatitis were observed. Microscopy revealed: the hepatocyte cytoplasm is densely filled with small and large drops of fat. Fatty degeneration of the liver was diffuse, vacuolization of the cytoplasm of hepatocytes, swelling of cell bodies was observed.

 A similar pattern was observed with the introduction of acetaminophen. In the case of a combined lesion, the morphological picture of liver biopsy samples was more pronounced and total. In addition to the changes in the hepatic parenchyma described above, foci of lobular collication necrosis were observed.

 When using a combined model of animal intoxication in the liver, the stores of glycogen, reduced glutathione, and levels of detoxifying cytochromes decreased more significantly. The defeat of the liver was characterized by a decrease in its functional activity in the form of a decrease in the rate of decomposition of hexenal and excretion of bromosulfalein. The use of the drug GSSG∞inosin was more effective than the use of legalon and essentials.

 The use of the drug GSSG∞inosin significantly improved the general condition of animals, prevented their death, and had a positive effect on morphometric, biochemical, and functional parameters reflecting the state of metabolism of the liver tissue.

 The use of GSSG∞inosine provided 100% survival of experimental animals with 80% death in the control; 45% mortality in the group receiving legalon, and 30% in the treatment of essentials (Fig. 36).

 In the study of biochemical parameters, obvious reliable signs of liver damage were determined: violation of protein synthesizing and lipid synthesizing functions, activation of the cytolytic syndrome and cholestasis in the conditions of the formation of toxic hepatitis with a combined effect on the body of the above toxicants - dichloroethane and acetaminophen. The therapeutic use of the drug GSSG∞inosin contributed to the restoration of the basic functions of the liver and the normalization of all the main indicators characterizing toxic liver damage (see tab. 38, 39).

 The indicators directly characterizing the state of liver metabolism (glycogen stores, glutathione level, microsomal enzyme activity, and relative liver mass) in animals treated with GSSG∞inosine did not practically differ from the background values (see Table 40).

 Thus, the presented results suggest that the drug GSSG∞inosine is an effective hepatoprotector. With a 10-day course of administration in rats at a dose of 5 mg / kg, the drug exerted a pronounced therapeutic effect in combined liver lesions with dichloroethane and acetaminophen, normalizing negative changes in the biochemical status and morphological parameters of the liver. The therapeutic use of the drug GSSG∞inosine ensured 100% survival of experimental animals with 80% death in the control; 45% death of animals treated with legalon, and 30% mortality on the background of the introduction of essentials (see tables. 38, 39 and 40).

Example 18
Therapeutic efficacy of GSSG∞inosine in experimental toxic liver cirrhosis
In the experiments we used nonlinear white male rats weighing 180-240 g, sourced from the Rappolovo nursery of the Russian Academy of Medical Sciences, 60 animals in total.

 To obtain experimental cirrhosis of the liver in white rats, the technique of Jezequel, A. M. et al. (1989) [27], according to which animals were injected intraperitoneally with a 1% solution of dimethylnitrosoamine (DMNA) in a single dose of 1.0 ml / kg for 3 consecutive days of the week, followed by a 4-day break, after which the injections were resumed. DMNA was diluted with saline. To obtain severe cirrhosis of the liver, it was enough to conduct 4 such injection cycles.

 For histological examination in animals after ephtanasia, anesthesia excised samples of liver tissue from the central and right lateral lobes. Fragments of the liver were fixed with a mixture of formalin, alcohol and acetic acid. Prepared paraffin sections with a thickness of 4 μm, which were stained with hematoxylin-eosin and van Gieson.

 In order to objectify the assessment of the influence of the studied drugs on the development of connective tissue in the liver during experimental toxic damage to DMNA, the area occupied by the connective tissue was measured in the range of 5-7 hepatic lobules on 6 sections of the liver. Morphometry was performed on preparations stained by van Gieson, on which collagen fibers were stained red. Using computer processing, a scanned image of a liver section at a magnification of x200 was subjected to color binarization with the release of red-colored collagen fibers. Based on the number of pixels that form the selected objects, the relative area occupied in the liver tissue by the developed collagen (connective tissue) was calculated.

 The measurement results were subjected to statistical processing. The significance of differences in the average samples was carried out using Student's test at a significance level of p = 0.05.

 Evaluation of morphological changes was carried out on preparations with "blind" labeling, which reduces the risk of aggravation of experimental results.

 The presence of the pathological process and the effectiveness of the drugs was evaluated by clinical signs (animal weight, eating food, appearance, ascites, mobility, death), as well as by pathomorphological signs during autopsy of animals and subsequent histological study of liver preparations.

 After the formation of the pathological process, i.e., after 4 weeks. after the introduction of DMNA by him, all animals were randomly divided into 4 groups of 12 individuals. Group 1 animals did not receive treatment, group 2 animals received saline, 3 - GSSG∞inosine, 4 - comparison drug - Heptral. Group 5 (12 animals) were intact animals of the same population for morphometric studies.

 Each group of animals was divided into two subgroups, one of them the drugs were administered for 3 weeks, after which they evaluated the effectiveness of the morphological method; animals of the second subgroup were treated for 6 weeks. The drugs were injected intramuscularly every other day into the thigh muscle.

 The results obtained indicate the following.

 The first experimental group (12 rats) consisted of individuals who did not receive any treatment after completing the course of DMNA administration. These animals lagged behind in weight (see Table 41), for 3-4 weeks they were inactive, poorly ate food, had dull and fallen off hair. Morphological and histological studies conducted 3 weeks after the cessation of DMNA administration (6 animals) showed ascites and signs of portal hypertension in these individuals. The study of preparations stained by van Gieson allowed us to determine the maturation of connective tissue in septa, as a result of which the latter acquired a regular structure. Gleams of the central veins located in the septal nodes were often occluded and replaced by connective tissue. Collateral circulation was carried out along the extended modified sinusoids enclosed in an ordered regular septic fiber skeleton. Hepatic cells underwent dystrophic changes. On preparations stained with hematoxylin-eosin, a large number of cells with signs of pronounced protein-hydropic dystrophy were detected. Hepatocyte nuclei were partially pyknotized, and in some cases lysed.

 Compared with the changes described above, corresponding to a 3-week period after the administration of a toxic agent, observation of animals (6 individuals) for 6 weeks. allowed us to establish the absence of spontaneous regression of pathological changes. So, when stained according to van Gieson in animals after 6 weeks. after application of DMNA, thickened connective tissue septa with dilated venous collaterals filled with blood were noted. When stained with hematoxylin-eosin, the preservation of the intensity of degenerative changes in the liver in the form of granular and protein-hydropic dystrophy of hepatocytes is shown.

 The second control group (12 rats) consisted of individuals receiving saline. During daily monitoring of animals of this group, it was found that they, like animals of group 1, for a long time (3-4 weeks) after completion of the DMNA administration, did not eat well, lagged behind in weight, had a poor appearance - fallen off dirty hair ; they were lethargic, inactive. Morphological and histological studies revealed that 3-week administration of a 0.9% sodium chloride solution did not affect the formation of pseudo-segments. Despite the appointment of saline, hemorrhages from dilated microvessels developed, which led to a violation of the lobule structure, dystrophy and death of hepatocytes with lysis of cell nuclei. Even with a 6-week administration of a 0.9% sodium chloride solution, the centro-central connective tissue septa clearly limiting the portal lobules were clearly preserved. In septa and in places of destroyed central veins, dilated venous collaterals were visible. When stained with hematoxylin-eosin, pronounced protein-hydropic degeneration of hepatocytes with activation of the process of fibrogenesis was noted.

 Treatment of group 3 animals (12 rats) with GSSG∞inosine led to a rapid improvement in their condition. During daily observation, 7 days after the start of therapy with this drug, an increase in physical activity, an increase in feed intake, an improvement in appearance, and an increase in the weight of almost all experimental rats of this group were noted. On histological preparations of rat liver receiving GSSG∞inosine for 3 weeks, a decrease in the number of connective tissue septa was noted, collagen fibers were distributed mainly perisinusoidally in the form of thin filaments and in the periportal zone of the lobules. When staining preparations with hematoxylin-eosin, a significant decrease in the number of cells with protein-hydropic dystrophy was noted. Macrophage elements were markedly activated. The introduction of GSSG∞inosine for 6 weeks was accompanied by a significant decrease in the amount of connective tissue. There were no signs of intraorgan circulatory circulation. In this course of treatment on preparations stained with hematoxylin-eosin, a statistically significant decrease in the number of dystrophic changes in hepatocytes and cells with lysed nuclei was noted. Dystrophic changes are presented mainly in the form of granular and hyaline droplet degeneration of individual hepatocytes. Resorption of connective tissue septa by activated phagocytes was observed on the preparations. The number of lymphocytes in the connective tissue was increased.

 Observation of the animals of group 4 (12 rats) treated with Heptral showed that for a long time (3 weeks from the start of therapy) their condition did not improve: they ate poor food, did not gain weight, had a poor appearance, were inactive. Morphological and histological studies revealed that after a 3-week course of therapy, the severity of acute phase inflammatory processes in the portal tracts was at the control level (saline treatment). On the preparations, pseudo-segments limited by connective tissue septa containing small collaterals of the central veins, anastomosing between themselves and forming connections with the vessels of the portal tract were clearly visible. Hepatocytes underwent profound dystrophic changes, with the most significant manifestation of protein-hydropic dystrophy. Only the appointment of Heptral for 6 weeks led to some positive effect, although lower than with GSSG∞inosine, the percentage of collagen in the liver decreased, the severity of inflammatory changes decreased, siderophages appeared in the composition of the connective tissue. When analyzing the state of hepatocytes, even after a 6-week course of heptral therapy, it was possible to note the presence of a fairly significant number of cells with hydropic changes. All this testified to the insufficiently effective treatment of experimental cirrhosis with Heptral.

For an objective assessment of the relative content of connective tissue in the liver of rats of experimental and control groups, morphometry of fuchsinophilic (i.e., connective tissue, collagen) fibers was performed on sections of the liver. The data obtained are presented in table. 42. It was found that treatment of toxic cirrhosis with GSSG∞inosine for 3 weeks is accompanied by a decrease in the number of
collagen in the liver 1.6 times compared with the control of saline; at the same time, the use of heptral was accompanied by a decrease in collagen in the liver by only 1.1 times. With a 6-week course of therapy, the difference in effect was even more pronounced: Pyyp∞inosine reduced the amount of connective tissue by 3.27 times, and Heptral - by 1.7 times. All this allows us to conclude that the appointment of GSSG∞inosin promotes involution of connective tissue in cirrhosis. All this allows us to conclude that the appointment of GSSG∞inosin promotes involution of connective tissue in cirrhosis.

 Thus, the high therapeutic effect of GSSG∞inosine on the model of experimental cirrhosis caused by dimethylnitrosoamine was established during daily observation of experimental animals and confirmed by histological studies. On the liver preparations of rats of the control group (DMNA without treatment), hepatocyte death sites, the development of inflammation in the walls of the small central veins, and the formation of connective tissue septa were clearly observed. Hepatocytes underwent dystrophic changes, protein-hydropic dystrophy was most pronounced. The use of saline did not positively affect the development of liver damage: in this case, mature connective tissue septa and dystrophic changes in the hepatocytes of the formed pseudo-segments were visible on the histological preparations. Heptral treatment caused only a slight decrease in the number of connective tissue and the severity of acute phase inflammatory processes in the portal tracts, as well as degenerative changes in liver cells, and the positive effect of heptral therapy was established only with a 6-week course of therapy. On the contrary, in animals treated with GSSG∞inosin, after 3 weeks from the start of treatment, a significant decrease in the amount of connective tissue was observed, even more significant during therapy for 6 weeks. Histological examination of liver preparations from these animals revealed only single small connective tissue septa containing microvessels, and no significant protein-hydropic changes in hepatocytes were revealed, which allows us to judge a rather high hepatoprotective effect of this drug.

 Conclusion: The use of the drug GSSG∞inosine at a dose of 10 mg / kg 3 times a week for 6 weeks with experimental liver cirrhosis in white rats caused by dimethylnitrosoamine directly correlated with involution of connective tissue in this organ, and also contributed to the restoration of damaged hepatocytes. The comparison drug Heptral (S-adenosyl-methionine) had a significantly lower therapeutic effect.

Example 19
Clinical and experimental efficacy of GSSG∞inosine, GSSG∞adenin and GSSG∞5-methylcytosine in the treatment of especially dangerous infections: tularemia and melioidosis
The therapeutic efficacy of GSSG∞inosine, GSSG∞adenine and GSSG∞5-methylcytosine was evaluated on white mice that were immunized with a virulent culture of the causative agent of melioidosis, chemical and live tularemia vaccines.

 The experiments used adult randomly bred male mice weighing 18-21 g, sourced from the Rappolovo nursery RAMS. For immunization, commercial preparations were used. To study the effect of GSSG∞inosine on nonspecific resistance to melioidosis, the drug was administered to mice at different doses (0.5; 2.5; 12.5 and 62.5 mg) once subcutaneously in mice in a volume of 0.2 ml per 1, 6, 12 and 18 hours before infection with the virulent culture of the causative agent of melioidosis strain 59361, after which the mortality rate of animals was determined (percentage of dead animals, average life expectancy - life expectancy) with an observation period of 1 month.

 When studying the effect of GSSG ад adenine on the non-specific resistance of mice to live tularemia vaccine (HFV), GSSG ад adenine was administered intraperitoneally at a dose of 2 mg / mouse (0.5 ml) 6 hours before subcutaneous inoculation of animals with HFV in various doses, followed by determination of LD50 of WTF in the control and experimental groups.

When studying the effect of GSSG∞5-methylcytosine on the protective activity of the "C-complex" isolated from the causative agent of tularemia (conditionally designated by us as the "chemical tularemia vaccine" - HTV), this "vaccine" was administered to mice subcutaneously in different doses without GSSG∞5-methylcytosine and in combination with it. After 21 days, some of the animals were infected with a virulent strain of the causative agent of melioidosis (10 LD50), and others - ZHV in a lethal dose (4 • 10 ⁶ m cells). For urgent prophylaxis of tularemia, GSSG∞5-methylcytosine was administered once 6 hours before vaccination with HTV, and to increase immunity to melioidosis twice: 6 hours before vaccination and 6 hours before infection.

 The results of the experiments were evaluated statistically.

 It was established that on the 18th day after infection of animals 300 LD50 of the virulent strain 59361 of the causative agent of melioidosis GSSG∞inosin protected 34% of mice from death at 100% death in the control. On the 30th day of observation, there was a decrease in the number of surviving animals to 17%, while at the same time, a significant increase in the LSS index (p <0.05) of dead mice treated with GSSG∞inosine was recorded. The most active in protecting against melioidosis was the dose of GSSG∞inosine 2.5 mg, administered to the animal 1, 6, 12 or 18 hours before infection.

 The results of an experiment on the study of the therapeutic properties of GSSG∞adenine in relation to the causative agent of tularemia are presented in table. 56.

 It was shown that the administration of GSSG∞ adenine to animals increased their resistance to ZHTV; at the same time, the LD50 of VHT for mice injected with GSSG∞adenine was 4 times higher than in intact animals.

 The ability of GSSG∞5-methylcytosine to increase the immunogenicity of HTV against pathogens of melioidosis and tularemia was also studied. HTV was used by us simultaneously as a nonspecific and specific defense factor against melioid infection.

 It was noted that vaccination of ZhTV mice in doses of 10, 50, and 250 μg protected from death up to 25% of the animals, while the LSS of the fallen mice only slightly exceeded that in the control group of intact animals (p> 0.05). A two-time administration of GSSG∞5-methylcytosine (6 hours before HTV vaccination and 6 hours before infection) contributed to an increase in animal survival (by 12%) and a significant increase in life expectancy (p <0.05) (Table 57).

 It should be noted that the dose of this series of HTV of 250 μg turned out to be toxic for mice: out of 12 animals vaccinated with this dose, 8 died, while in the group of animals treated with 250 μg of HTV, 2 mg of GSSG∞5-methylcytosine out of 12 mice survived before infection 11.

 When studying the possibility of GSSG∞5-methylcytosine to increase specific immunity to tularemia, it was shown that a single administration of GSSG∞5-methylcytosine 6 hours before vaccination with 25-100 μg of HTB increased the immunogenic properties of this drug, which was expressed in an increase in animal survival (by 14- 17%) and in a significant increase (p <0.05) in the life expectancy of patients with infection with a lethal dose of ZHTV (Table 58).

 Studies have shown the ability of drugs of the GSSG∞inosin group to increase the body's resistance to melioidosis and tularemia. An important feature of the drugs is immunomodulatory, systemic cytoprotective and, especially, hepatoprotective effect. The data obtained on a significant decrease in the toxicity of a dose of HTB 250 mg for mice using GSSG∞5-methylcytosine, introduced 6 hours before vaccination, indicate its antibacterial and cytoprotective effect.

 Thus, new derivatives of oxidized glutathione and nucleic acid elements when used according to a certain technique provide a reliable level of protection for mice from melioidosis and tularemia; reduce the residual toxicity of tularemia "C-complex", which can be used to create comprehensive preventive and therapeutic drugs.

 At the same time, these data suggest that the studied drugs are effective in infections caused by prions.

Sources of information:
1. Consensus Statement. EASL International Consensus Conference on hepatitis CJ Hepatol. 1999, 30: 956-961.

 2. Naoumov N. V., Hepatitis C virus-specific CD4 + T cells: do they help or damage? Gastroenterology, 1999, 117: 1012-1014.

 3. Tran A., Yang G., Doglio A. et al. Phenotyping of intrahepatic and peripheral blood lymphocytes in patients with chronic hepatitis C. Dig. Dis. Sci. 1997, 42: 2495-2500.

 4. Gerlach J. T., Diepolder H. M., Jung M. K. et al. Recurrence of hepatitis C virus after loss of virus-specific CD4 + T cell response in acute hepatitis C. Gastroenterology, 1999, 117: 933-941.

 5. Cramp M. E., Carucci P., Rossol S. et al. Hepatitis C vims (HCV) specific immune response in anti-HCV positive patients without hepatitis C viraemia. Gut, 1999, 44: 424-429.

 6. Eckels D. D., Tabatabail N., Bian T. H. et al. IN vitro human TH-cell response to a recombinant hepatitis C virus antigen: failure in IL-2 production despite proliferation. Hum. Immunol. 1999, 60: 187-199.

 7. Ferrari C., Penna A., Bertoletti A. et al. Antiviral cell-mediated immune response during hepatitis B and hepatitis C virus infections. Recent Results Cancer Res. 1998, 154: 330-336.

 8. Fan X. G., Li C. Z. et al. Circulating Th1 and Th2 cytokines in patients with hepatitis C virus infections. Mediators Innamm. 1998, 7: 295-297.

 9. Rehermann B. et al. Cell mediated immune response to the hepatitis C virus. Cur. Top Microbiol. Immunol. 2000, 242: 299-325.

 10. Schlaak J. F., Pitz T., Lohr H. F. et al. Interleukin 12 enhances deficient HCV-antigen-induced Th1-type immune response of peripheral blood mononuclear cells, J. Med. Virol. 1998, 56: 112-117.

 11. Semenenko T. A. Cellular immunity in hepatitis C. Viral hepatitis: achievements and prospects. Inf. Bull. 1 (8), 2000.

 12. Bartholomew M. M., Jansen R. W., Jeffers L. J. et al. Lancet, 1997, 349, No. 9044, 20-22.

 13. Di Bisceglie A. M., Conjeevaram M. S., Eried M. W. et al. Ann. Intern. Med. 1995, 123, 897-903.

 14. Schiff E. R., J. Med. . Virol. , 2000 Jul; 61 (3): 386-91.

 15. Dienstag J. L. et al. , N. Endl. J. med. 1999 Oct 21; 341 (17): 1256-63.

 16. Yao G. B. et al. Long-term efficacy of lamivudine in the treatment of patients with chronic hepatitis B virus infection - a multicenter randomized, double-blind, placebo-controlled trial, DDW 1999.

 17. Pol S., Nalpas B., Bourlierem et al. Combination of ribavirin and interferon-alpha surpasses high doses of interferon-alpha alone in patients with genotype-1b-related chronic hepatitis C. Hepatology, 2000 Jun; 31 (6): 1338-44.

18. The Medical Management of AIDS, 4 ^th edition. Sande M. And Volberding PWB Saunders, Company, 1995.

19. Robert S. Bohinski, "Modern Concepts in Biochemistry", 4 ^th edition, 1987.

20. Harrison's Principles of Internal Medicine, 14 ^th edition, p. 511, 1998.

 21. Apoptosis: a role in neoplasia, C. D. Gregory, 1996.

 22. The Molecular Biology of Apoptosis, D. L. Vaux, A. Strasser; Proc. Natl. Acad Sci. USA 93 (1996).

 23. Cytokines in oncohematology, L. Gracheva, Moscow, 1996.

 24. Paola Galinari et al. Multiple enzymatic activities associated with recombinant NS3 protein of hepatitis C virus. J. Virol. August 1998, p. 6758-6769, Vol. 72, No. 8.

 25. RF patent 2089179, MKI A 61 K 38/00, 1997.

 26. RF patent 2153350, MKI A 61 K 38/00, 38/06, 33/00, 1999.

 27. Jezequel, A. M. et al. Dimethylnitrosamine-induced cirrhosis. Evidence for an immunological mechanism. J. Hepatol. 8, 42-52 (1989).

 28. Guidance materials on the experimental and clinical study of new drugs. Parts 1 and 3 (Official Edition). Pharmacological Committee, M., 1975, 1981.

 29. Requirements for the preclinical study of the general toxic effects of new pharmacological substances (Temporary guidelines). Pharmacological Committee. M., 1985 and the Rules of preclinical safety assessment of pharmacological agents (GLP). RD 64-126-91. M., 1992.

Claims (84)

 1. Organic salt of oxidized glutathione (GSSG), characterized in that the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form and have a nitrogenous base or nucleoside, or a purine and pyrimidine nucleotide.  2. The organic salt of claim 1, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and a nitrogenous base of purine or pyrimidine nature.  3. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and adenine.  4. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and guanine.  5. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and thymine.  6. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and cytosine.  7. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and uracil.  8. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and 5-methylcytosine.  9. The organic salt of claim 2, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and 4-thiouracil.  10. The organic salt according to claim 2, in which the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and dihydrouracil.  11. The organic salt of claim 1, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and a nucleoside of purine or pyrimidine nature.  12. The organic salt of claim 11, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and adenosine.  13. The organic salt of claim 11, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and guanosine.  14. The organic salt of claim 11, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and inosine.  15. The organic salt of claim 11, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and cytidine.  16. The organic salt of claim 11, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and uridine.  17. The organic salt of claim 11, wherein the counterions are oxidized glutathione, in the formula of which all amino acids are in L-form, and thymidine.  18. Organic salt of oxidized glutathione (GSSG), characterized in that the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and a nitrogenous base, or nucleoside, or a nucleotide of purine or pyrimidine nature.  19. The organic salt of claim 18, wherein the counterions are oxidized glutathione, in the formula of which two chemically unique amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and the nitrogenous base is purine or pyrimidine in nature.  20. The organic salt according to claim 19, in which the counterions are oxidized glutathione, in the formula of which two chemically unique amino acids are represented in D-form, and the remaining amino acids are presented in L-form and adenine.  21. The organic salt of claim 19, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and guanine.  22. The organic salt according to claim 19, in which the counterions are oxidized glutathione, in the formula of which two chemically unique amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and thymine.  23. The organic salt of claim 19, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and cytosine.  24. Organic salts according to claim 19, in which the counterions are oxidized glutathione, in the formula of which two chemically unique amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and uracil.  25. The organic salt of claim 19, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and 5-methylcytosine.  26. The organic salt of claim 19, wherein the counterions are oxidized glutathione, in the formula of which two chemically unique amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and 4-thiouracil.  27. The organic salt of claim 19, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and dihydrouracil.  28. The organic salt of claim 18, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and a purine or pyrimidine nucleoside.  29. The organic salt of claim 28, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and adenosine.  30. The organic salt of claim 28, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and guanosine.  31. The organic salt of claim 28, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and inosine.  32. The organic salt of claim 28, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and cytidine.  33. The organic salt of claim 28, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and uridine.  34. The organic salt of claim 28, wherein the counterions are oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in L-form, and thymidine.  35. An organic salt of oxidized glutathione, characterized in that in it the counterions are oxidized glutathione, in the formula of which all amino acids are represented in L-form, or oxidized glutathione, in the formula of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids are represented in the L-form, and nucleosides of purine or pyrimidine nature, in the formula of which the ribose is presented in the form of β-D-ribose, or in the form of β-D-deoxyribose.  36. The organic salt of claim 35, wherein the counterions are oxidized glutathione in the L- or D-form, and simultaneously two nucleosides, one of which is purine and the other pyrimidine in nature.  37. Organic salt of oxidized glutathione, characterized in that in it the counterions are oxidized glutathione in the L- or D-form and at the same time two nitrogenous bases: one of which is purine, and the second is pyrimidine in nature.  38. An individual substance based on phosphamide derivatives of oxidized glutathione, characterized in that it contains organic cations represented by a number of nucleosides or nucleotides and a phosphamide derivative of oxidized glutathione, the amino acids of which exist in L-form.  39. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with adenosine monophosphate (AMP) is used.  40. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with guanosine monophosphate (GMF) is used.  41. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with inosine monophosphate (IMP) is used.  42. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with cytidine monophosphate (CMP) is used.  43. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with uridine monophosphate (UMF) is used.  44. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with thymidine monophosphate (TMF) is used.  45. The substance according to p. 38, characterized in that as a phosphamide derivative of oxidized glutathione, nucleic acid derivatives of purine or pyrimidine nature are used, in which the ribose is in the form of β-D-ribose or in the form of β-D-deoxyribose.  46. An individual substance based on phosphamide derivatives of oxidized glutathione, characterized in that it contains organic cations represented by a series of nucleosides or nucleotides and a phosphamide derivative of oxidized glutathione, in the structure of which two chemically unambiguous amino acids are represented in D-form, and the remaining amino acids in L- form.  47. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with adenosine monophosphate (AMP) is used.  48. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with guanosine monophosphate (GMF) is used.  49. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with inosine monophosphate (IMP) is used.  50. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with cytidine monophosphate (CMF) is used.  51. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with uridine monophosphate (UMF) is used.  52. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with thymidine monophosphate (TMF) is used.  53. The substance according to p. 46, characterized in that as a phosphamide derivative of oxidized glutathione use a derivative of nucleotides of purine or pyrimidine nature, in which the ribose is in the form of β-D-ribose or in the form of β-D-deoxyribose.  54. An individual substance based on phosphamide derivatives of oxidized glutathione, characterized in that it contains inorganic cations of alkali, alkaline earth and / or transition metals and a phosphamide derivative of oxidized glutathione in L-form or D-form, with a number of nucleosides or nucleotides of purine or pyrimidine nature wherein the ribose is in the form of β-D-ribose or in the form of β-D-deoxyribose.  55. The substance according to p. 54, characterized in that as a phosphamide derivative of oxidized glutathione, a derivative with two nucleotides is used, one of which is purine, and the second is pyrimidine in nature.  56. A pharmaceutical composition exhibiting antiviral, antibacterial, immunorehabilitation and hepatoprotective activity, containing as an active substance at least one of the compounds of claims. 1-55 in an effective amount and a pharmaceutically acceptable carrier or excipient.  57. A pharmaceutical preparation having antiviral, antibacterial, immunorehabilitation and hepatoprotective effects, containing as an active substance at least one of the compounds of claims. 1-55 in an effective amount and a pharmaceutically acceptable carrier or excipient.  58. A method of producing an organic salt of oxidized glutathione and inosine, in which the molecule of oxidized glutathione acts as the anion, and the molecule of inosine (9-β-D-ribofuranosyl-hypoxanthine) acts as the cation, which consists in the fact that the solution of the disodium salt of oxidized glutathione with stirring and room temperature, 1 equivalent of inosine powder is added and left to dissolve completely, after which the clear solution is filtered and freeze-dried.  59. The method of producing inosyl-5-phosphoryl-N-glutathione, which consists in the fact that a phosphamide bond between the amino group of glutamic acid in the molecule of oxidized glutathione and the residue of phosphoric acid in the molecule of inosyl-5-monophosphate is obtained in the reaction of its N-oxysuccinimide activated ester and oxidized glutathione in dimethylformamide at pH 8.0-8.5.  60. The method of exposure to infectious agents of the patient’s body, including localized intracellularly, and / or stimulation of T-cell and humoral anti-infectious immunity, along with providing cytoprotective effects, which consists in introducing into the mammalian body an effective amount of a pharmaceutical product obtained on the basis of chemical interaction disulfide-containing peptides with nitrogen bases or nucleosides or nucleotides of purine or pyrimidine nature. 61. The method according to p. 60, which consists in the induction of apoptosis mechanisms, including by expression of the apoptosis inducer, FAS / APO-1 antigen (CD95 ⁺ ) on the membranes of virus-infected cells, on the membranes of macrophages containing tuberculosis mycobacteria, on the cell membranes infected with mycoplasmas, chlamydia, malaria plasmodium and other infectious agents by administering to the mammalian organism a pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine, GSSG∞urac l, GSSG∞thymine, GSSG∞adenine, GSSG∞guanine, GSSG-inosine monophosphate, GSSG-uracyl monophosphate, GSSG-thymidine monophosphate, GSSG-cytosine monophosphate and other compounds based on the interaction of L- or D-forms of oxidized glutathione and its salts bases or nucleosides or nucleotides of purine or pyrimidine nature.  62. The method of claim 60, comprising administering to the body of a hepatitis C virus-infected mammal, a pharmaceutical preparation containing an effective amount of an active ingredient selected from the group consisting of GSSG∞inosine, GSSG-inosine monophosphate, GSSG-uracil monophosphate and derivatives of L- or D-forms of oxidized glutathione and nucleic acid components in order to inhibit the ATP-azo / helicase activity of hepatitis C virus NS3 to stop replicative activity and eliminate hepatitis C virus, therefore, achieve therapeutic about the effect.  63. The method according to p. 60, which consists in predominantly inhibiting the replicative activity of RNA-containing viruses, in the case of using as the active substance, compounds obtained on the basis of the chemical interaction of the L- or D-forms of oxidized glutathione and its salts with nitrogenous bases or nucleosides or nucleotides of pyrimidine nature.  64. The method according to p. 60, which consists in predominantly inhibiting the replicative activity of DNA-containing viruses, if the active substance is used, compounds obtained on the basis of the chemical interaction of L- or D-forms of oxidized glutathione and its salts with nitrogenous bases or nucleosides or nucleotides of purine nature.  65. The method of claim 60, comprising administering to the mammalian organism a pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine and other oxidized glutathione compounds with nucleic acid components in order to preferentially activate the functional activity of the Th1 group of T- cells, or Th2-group of T-cells, or their regulated ratio, which differentially determines the effectiveness of the immune system of a mammalian organism in relation to the etiotropic factor in infectious diseases. 66. The method according to p. 60, which consists in introducing into the patient's body a viral hepatitis C pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine and / or GSSG-inosine monophosphate to restore the normal ratio of Th1 / Th2 cells and achieve the optimal content of cytokines produced by Th1 and Th2 groups of cells, as well as to increase the endogenous production of IL-2, IL-12, IFN-α and γ.
67. The method according to p. 60, which consists in introducing into the patient's body a viral hepatitis A, B and / or C pharmaceutical preparation containing an effective amount of an active substance selected from the group consisting of GSSG∞inosine and other compounds of oxidized glutathione, its salts, s components of nucleic acids to achieve hepatoprotective effects, including the prevention and / or treatment of complications of chronic viral hepatitis, namely the prevention and treatment of liver cirrhosis and hepatocellular carcinomas.  68. The method according to p. 60, in which the disease is selected from all types of infectious diseases, including viral and bacterial, including anaerobic infections; chlamydial and mycoplasma infections; mycoses (candidiasis); protozoal infections.  69. The method of claim 60, wherein the pharmaceutical preparation of claim 57 is administered parenterally, orally, in the form of inhalation solutions, solutions for local instillations, ophthalmic or intranasal drops, ointments, creams or gels for cutaneous use, and also in the form of suppositories, until a therapeutic effect and / or prevention of complications caused by infectious agents in the process of developing an infectious disease are achieved.  70. A method of treating a subject in need of terminating the replicative activity of viruses, bacteria, or other infect agents, inducing apoptosis mechanisms of infected cells, restoring and stimulating anti-infective capable immunity: T-cell, and / or humoral, systemic cytoprotective effects, which consists in administering to the subject a pharmaceutical a preparation containing an active substance selected from the group comprising biologically active compounds obtained based on the chemical interaction of L- or D-forms of oxidized glutathione, its salts with nitrogen bases, or nucleosides or nucleotides of purine or pyrimidine nature in an amount sufficient to achieve a therapeutic effect.  71. The method according to p. 70, in which the disease is acute with a prolonged course of viral hepatitis B, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine and GSSG-inosine monophosphate.  72. The method of claim 70, wherein the disease is acute viral hepatitis C and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞uracil and GSSG-inosine monophosphate.  73. The method of claim 70, wherein the disease is chronic viral hepatitis B, and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞guanosine, GSSG∞adenosine, GSSG-inosine monophosphate and GSSG-thymidine monophosphate.  74. The method according to p. 70, in which the disease is chronic viral hepatitis C, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞uratsil, GSSG∞cytosine, GSSG∞dihydroracil, GSSG-uracilyl monophosphate, GSSG-cytosine monophosphate, ura -GSSG-inosine. 75. The method of claim 70, wherein the disease is cirrhotic chronic hepatitis and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞uridine, GSSG∞thymidine, Li ₂ -GSSG-inosine monophosphate and Na ₂ -GSSG thymidine monophosphate. 76. The method of claim 70, wherein the disease is pulmonary tuberculosis, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞cytosine, GSSG-5-methylcytosine, Li ₂ -GSSG-inosine monophosphate. 77. The method of claim 70, wherein the disease is genitourinary tuberculosis, and the pharmaceutical is selected from the group consisting of GSSH∞ thymine, Na ₂ -GSSG-guanosine monophosphate and uracil-Li ₂ -GSSG-guanosine monophosphate. 78. The method according to p. 70, in which the disease is AIDS, as well as a cytomegalovirus infection, an infection caused by the Epstein-Barr virus and / or pneumocystis, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞dihydrouracil, GSSG∞4 -thiouracil, as well as Zn ₂ -GSSG-thymidine monophosphate and Ag ₂ -GSSG-uracil monophosphate and uridine∞GSSG∞inosine.
79. The method according to p. 70, in which the disease is an infection caused by herpes viruses, and the pharmaceutical preparation is selected from the group comprising GSSG∞inosine, Li ₂ -GSSG-guanosine monophosphate, and also the D-form Na ₂ -GSSG-cytosine monophosphate and D -form of GSSG∞ uracil.
80. The method according to p. 70, in which the disease is mycosis (candidiasis), and the pharmaceutical preparation is selected from the group consisting of GSSG∞ uridine, GSSG∞ 4-thiouracil and Ag ₂ -GSSG-uracil monophosphate. 81. The method of claim 70, wherein the disease is a mycoplasma infection, and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞adenosine, and Na ₂ -GSSG-adenosine monophosphate. 82. The method of claim 70, wherein the disease is a chlamydial infection, and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞thymine, GSSG∞uridine, GSSG∞guanosine, and Na ₂ -GSSG-guanosine monophosphate. 83. The method of claim 70, wherein the disease is malaria, leishmaniasis, and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞cigosin, and GSSG∞5-methylcytosine.
84. The method of claim 70, wherein the disease is an anaerobic infection and the drug is selected from the group consisting of D-form GSSG∞inosine, (D-cysteine) and D-form

(D-glutamic acid). 85. The method of claim 70, wherein the disease is selected from the group consisting of viral hepatitis A, dysentery and cholera, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, Li ₂ -GSSG-inosine monophosphate and Li ₂ -GSSG-guanosine monophosphate as well as the D form of GSSG∞ uracil (D-glutamic acid). 86. The method of claim 70, wherein the disease is infectious meningitis, and the pharmaceutical is selected from the group consisting of GSSG∞inosine, Li ₂ -GSSG-inosine monophosphate, D-form GSSG∞uracil (D-glutamic acid), GSSG∞5 -methylcytosine, Ag ₂ -GSSG-uracilyl monophosphate. 87. The method according to p. 70, in which the disease is selected from the group of especially dangerous infections, consisting of plague, tularemia and anthrax, and the pharmaceutical preparation is selected from the group comprising GSSG∞inosine, GSSG∞adenine, GSSG∞thymine, GSSG∞5 -methylcytosine, GSSG∞4-thiouracil, D-form GSSG∞uracil (D-glutamic acid), D-form GSSG∞inosine, (D-cysteine), adenine∞GSSG∞thymine.
88. The method of claim 70, wherein the disease is a prion infection (“mad cow disease”), and the pharmaceutical is selected from the group consisting of GSSG∞inosine, GSSG∞uridine, GSSG∞dihydrouracil, Ag ₂ -GSSG-yrapacylmonophosphate, Ag ₂ -GSSG-thymidine monophosphate, as well as uracil monophosphate-Li ₂ -GSSG-inosine monophosphate.  89. The method of claim 70, wherein the disease is selected from the group consisting of influenza and acute respiratory viral infections, and the pharmaceutical preparation is selected from the group consisting of GSSG∞inosine, GSSG∞adenosine, GSSG∞uracil and GSSG∞thymin.

Images