SK8612002A3 - Compositions and methods for L-nucleosides, L-nucleotides, and their analogs - Google Patents

Abstract

Nucleoside and nucleotide compounds and their analogs/prodrugs are provided. Particularly contemplated compounds include 1- beta -L-ribofuranosyl-1,2,4-triazole-3-carboxamide, which may be modified and/or phosphorylated. Contemplated compounds may further be combined with other pharmacological compounds, especially including Ribavirin, antibodies, and cytokines. Preferred uses of contemplated compounds include use as an antiviral compound, anti-inflammatory compound, antineoplastic compound, and as a compound to stimulate cellular growth.

Description

Compositions and methods relating to L-nucleosides, L-nucleotides and analogs thereof

Technical field

The present invention relates to pharmaceutical compositions and their use.

BACKGROUND OF THE INVENTION

There are a large number of different health-inducing effects, many of which result from infections or accumulations of toxic substances in a living body, but this may result in an adverse or contradictory reaction of the immune system against the infected organ. For example, infection with hepatitis C virus (HCV) often leads to a persistent viral infection where an inflamed organ cannot respond immediately to this HCV virus, but rather to an infection induced by an imbalance in the immune response.

Most known methods of treating viral infections can be characterized as either a direct antiviral treatment or an indirect antiviral treatment. In a direct method of antiviral treatment, the virus is selected to act as a suitable direct antiviral drug. For example, HIV-infected patients typically receive a mixture of therapeutic agents to block the spread of viruses, and various groups for direct antiviral treatments are known in the art. For example, some direct antiviral drugs block reverse transcriptase. Reverse transcriptase (RT) inhibitors are typically nucleoside analogs such as AZT, 3TC or ddl. Alternatively, non-nucleotide RT inhibitors, including quercetin, may be used. In vitro, RT inhibitors are typically potent antiviral drugs. However, in vivo, especially during a period of relatively high viral replication rate, the generation of RT inhibitor resistant viral mutants is problematic.

Other direct intiviral drugs block or interfere with protein processing viruses and are commonly known as protease inhibitors. Protease inhibitors are typically highly specific against viral proteolytic enzymes. However, due to their mostly hydrophobic nature, their administration becomes problematic at the desired concentrations. In addition, the development of cross-resistant and often serious side effects of the compounds causes an increase in the difficulty of using protease inhibitors. In order to reduce the development of resistant strains of viruses against many drugs, mixtures of RT inhibitors and protease inhibitors may be overwritten. Although such mixtures have been used relatively successfully for a short period of time, the relatively high incidence of adverse side effects and the theoretical possibility of creating resistant strains of viruses against many drugs still persists.

In an indirect method of antiviral treatment, the immune response to eliciting a viral-induced response may be varied, modulated. For example, immunosuppressive drugs may be used to alleviate the inflammatory condition associated with a particular viral infection; Various immunosuppressive drugs are known in the art. Among others, cyclosporin A is known as an immunosuppressive drug, although it is a very effective immunosuppressive drug, very often used to suppress tissue rejection responses after organ transplantation. However, the use of cyclosporin A may lead to problems due to its overall immunosuppressive action, which makes the patient more susceptible to new infectious diseases. In addition, long-term administration of cyclosporin A is often associated with serious side effects, including excessive hair (hirsutism) and gum enlargement (gingival hyperbath). In addition, the bioavailability of cyclosporin A is at least partially dependent on bile, which may indicate additional problems in hepatitis infection.

Tacrolimus (FK506) can be used as an immunosuppressive drug to overcome at least some of the problems associated with cyclosporin A. For example, Tacrolimus has been found to act in the treatment of atopic dermatitis on the face. Topical application of this immunosuppressive drug resulted in a significant improvement in 95% of treated patients [Alaiti, S., et al .: Tacrolimus (FK506) oitment for atopic dermatitis: A phase I study in adults and children. J. Am.

Acad. Dermatol. 1993 p. 69-76]. In addition, Tacrolimus does not appear to cross the skin barrier, thus eliminating any problems associated with systemic administration.

Although generally well tolerated, treatment with Tacrolimus is, however, limited to topical administration due to the potential for an overall immune risk.

Tacrolimus systematically administered over a long period of time, to develop lymphoproliferative diseases and

If it can often lead to cardiomyopathy.

Many known immunosuppressive drugs provide some alleviation of inflammatory conditions. However, these effects on the organ are not specific when administered systemically. Consequently, the immunity to exogenous and endogenously induced reactions such as bacterial and viral infections, neoplastic or malignant cells, etc. is thus reduced. Thus, a certain window of useful concentrations of immunosuppressive drugs is defined, namely the maximum concentration that is the overall compromise between the immune system condition and the minimum concentration that can provide at least some desired effect.

Although various compounds and methods for treating infectious and inflammatory diseases are known in the art, all or almost all exhibit one or more disadvantages. Therefore, there is a need to provide improved methods and compositions for treating these conditions.

SUMMARY OF THE INVENTION

The present invention is directed to methods and compositions wherein the nucleoside and / or nucleotide drug or analogue thereof is administered to a subject in a concentration or dose effective to achieve the desired pharmaceutical or physiological effect.

In one aspect of the invention, the present invention provides a compound of formula I:

wherein R is H, PO ₃ ² , (PO ₃ ) 2 ^3- or (PO ₃ ) ₃ ^4- group.

Said compounds are optionally further modified by a group which is covalently attached to a carbonyl atom, said compounds of the invention being in the D- or L-configuration.

In another aspect, the present invention provides the use of said compounds for the treatment of a viral infection, wherein they may further be co-administered with a cytokine, preferably IFN-alpha-2b, antibody or Ribavirin (Ι-β-Dribofuranosyl-1,2,4-triazole-3). amide).

In another aspect, the present invention provides the selectivity of said compounds with respect to a pharmaceutical effect on target cells enhanced by modifying said compounds with a modifying group, said modifying group being covalently attached to said drug via a nitrogen atom, and said modified group being enzymatically removed from said drug in a target cell. In particular, said modifying groups include = NH and N (R ¹ ) (R ² ) or = NR ¹ , wherein R ¹ and R ² are independently hydrogen, linear alkyl, branched alkyl, alkenyl, alkynyl, aralkyl, aralkenyl, aralkynyl or aryl and wherein R ¹ and R ² may independently further comprise a nitrogen atom, an oxygen atom, a sulfur atom or a halogen atom.

In another aspect, the present invention provides a method of treating a disease comprising administering to a patient having an inflamed organ in the body said compounds at a dose that causes systemic immunomodulation rather than systemic immunosuppression of the type

I and Type II answers. Effect of Type I and Type I immunosuppression

The response in a given organ of a patient is selective in view of the selectivity of the action of the compounds on the organ.

In yet another aspect, the present invention provides a method of stimulating nerve cell growth, as the compounds disclosed herein have been found to be effective in stimulating nerve cell growth within a given concentration range. In a next step, the compounds are introduced into nerve cells within a given concentration range.

The various objects, features, features, aspects and advantages of the present invention will become more apparent from the following detailed description of various embodiments of the invention.

Detailed description

Claimed compounds

It is generally contemplated that all nucleotides, nucleosides and their corresponding analogs may be suitable for use in conjunction with the theory presented herein, characterized in that all compounds claimed herein may be in either the L-configuration or the D-configuration.

Preferred compounds include, in particular, phosphorylated and non-phosphorylated Levovirin ™ (Ι-β-L-ribofuranosyl-1,2,4-triazole-3-carboxamide of Formula I)

wherein R is hydrogen or a group containing phosphorus or sulfur. When R is a phosphorus-containing group, it is highly preferred that R is a monophosphate, pyrophosphate or triphosphate as shown in Figures IA to 1C.

Depending on the chemical environment (especially the pH), it can be assessed that the phosphate groups may be in their respective mono-, di-, tri- and tetraprotonated forms, and it can also be appreciated that if the phosphate groups are partially or fully deprotonated, salts with one or more monovalent or multivalent cations may be formed. Said cations are in particular alkali metal ions and alkaline earth metal ions such as Mg ²⁺ , Cs ²⁺ , Na ⁺ , etc.

According to alternative aspects of the present invention, R can also be a PO ^{3 2} ', (PO ³ ) 2 ³ ' or (PO ₃ ) ₃ ⁴ 'group wherein one or more oxygen atoms are replaced by a sulfur atom. Although phosphate groups are generally preferred substituents for R, other chemical groups may also be used, and the intended groups include, in particular, mono- or polyanionic groups, preferably with tetragonal geometry. In particular, the compounds include phosphorylated and non-phosphorylated Levovirin ™. In addition, it can be appreciated that these compounds may also have a sugar moiety in the D-configuration, said compound with a sugar D-configuration being, in particular, Ribavirin (1β-D-ribofuranosyl-1,2,4-triazole-3- carboxamide). In addition, the compounds of the effects disclosed herein (i.e., those assessed to be at least some of the antiviral immediately direct which demonstrate the compounds prevent the spread of viruses) should be positive.

Since most organisms have phosphatase moieties, it can be contemplated that the compounds of the subject in the various of this invention may be gradually dephosphorylated, and one or more phosphate groups may be removed over time. For example, a triphosphorylated group can be converted to a diphosphorylated or monophosphorylated compound, or a diphosphorylated compound can be converted to Levovirin ™ in a single reaction.

In terms of assessing the dephosphorylation of phosphorylated Levovirine ™, a significant change in the manner of antiviral action, from direct antiviral action to indirect antiviral action, can be noted. A change from a direct antiviral response to an indirect antiviral response is particularly advantageous as the compounds maintain an antiviral action throughout the extended period of time during metabolism. Therefore, it is highly appreciated that the mode of action of said compounds, in particular phosphorylated Levovirine ™, is carried out at least bimodally - comprising a portion of a direct antiviral action and a portion of an indirect antiviral action.

In relation to the dephosphorylation rate, it can be positively assessed that phosphorylated Levovirine ™ is dephosphorylated much slower than phosphorylated Ribavirin, with a positive effect on the L-configuration of ribose in Levovirine ™. With respect to the part or organ where the dephosphorylation is to be performed, it is appreciated that the dephosphorylation occurs preferably in the liver, but this also applies to other organs and parts, including kidneys, nerve cells and the bloodstream.

In particular, it will be appreciated that all known prodrug forms of said compounds are suitable for use in accordance with the teachings herein, in particular said prodrug forms, including covalent modifications which may be enzymatically (i.e., by aminohydrolase, oxidoreductase or transferase) removed from said compounds.

Examples of particularly suitable prodrugs include those resulting from the addition of a nitrogen-containing group in the carboxyl portion of Levovirine ™, which may be particularly advantageous if the compounds are preferably intended for the liver. For example, Applicants have discovered (unpublished results) that the specificity of Levovirine ™ with respect to the hepatocyte pharmaceutical effect can be enhanced by modifying Levovirine ™ with a nitrogen-containing modification group that is selectively removed from hepatocytes. Structural Formula II below represents Levovirin ™ and Structural Formula III represents Levovirin ™ modified at the carboxyl group to form a carboxamidine group.

(II)

In particular, modification of Levovirine ™ by a modifying group (preferably containing nitrogen) that can be selectively removed from the target cell (e.g., hepatocyte) helps (1) to increase the selectivity of Levovirine ™ relative to the target cell, thereby reducing the total dose (2). to achieve the desired effective concentration; and (3) reduces the overall toxicity in the non-target cells. Further, the modifying group is covalently bonded to the carboxyl atom of the carboxamide group.

In other alternative aspects, the nitrogen-containing modifying group is not limited to the = NH group, but may also include various primary and secondary amines. In general, suitable modification groups have the structure N (R ¹ ) (R ² ) or = NR ¹ , wherein R ¹ and R ² are independently hydrogen, linear or branched alkyl, alkenyl, alkynyl, aralkyl, aralkenyl or aralkynyl, aryl all of which may further comprise heteroatoms including nitrogen, oxygen, sulfur or halogen. However, it is most preferred that the alternative modifying groups are enzymatically removable from Levovirine ™ and particularly when said enzymes include aminohydrolase such as liver deaminases (e.g. adenosine or cytosine deaminase), liver deamidases (e.g. deamidase) and liver transaminases (glutamate-pyruvate transaminase).

Without being limited to the inventive concept, the modifying groups may inactivate Levovirin ™ or prevent subsequent activation of the once modified Levovirin ™ present in a cell that is not a target cell. However, nitrogen-containing modifying groups can also prevent metabolic activation of the modified Levovirine ™. With respect to the degree of modification of Levovirine ™, it may be noted that the modification may include an organosynthetic modification, an enzymatic modification, or a daily synthesis to produce a modified Levovirin ™.

With respect to enzymatic removal of the modifying group, it may be noted that, depending on the target cell type and the modifying group, enzymatic removal may be of great importance. Enzymatic removal may include enzymes from a variety of groups, including hydrolases, transferases, lyases, and oxidoreductases, particularly adenosine and cytosine z. preferred deaminases, arginase, transaminase, and arylamidase subgroups. It can further be appreciated that said enzymes for enzymatic removal of the modifying group may be exclusively expressed in target cells, but according to alternative aspects of the invention suitable enzymes may also be expressed in cells other than the target cells if all the enzyme is not present subjected to expression, in all cells in a system containing cells. Further, it can be appreciated that said enzymes are preferably subjected to natural expression (i.e., not recombinant) in the eventual target cells under normal and / or pathological conditions. For example, it is known that glutamate-pyruvate transaminase is constitutively subjected to expression with high selectivity in liver cells and therefore may be a suitable enzyme for removal of a modification group. Alternatively, it is known that cytosine deaminase is expressed in relatively large amounts in colon cancer cells, but not or only in small amounts in normal colon cells.

Synthesis of the above compounds

In general, all known methods used in the synthesis of D-nucleotides, D-nucleosides and their possible analogs can be used to synthesize the compounds disclosed herein in the L-configuration (e.g., replacing the sugar moiety in the D-configuration with the corresponding sugar moiety in the L-configuration). An example of a synthetic scheme for the synthesis of Levovirin ™ (Ι-β-L-ribofuranosyl-1,2,4-triazole-3-carboxamide) is shown in Figure 2.

Synthesis of 1,2,3,5-tetra-O-acetyl-β-L-ribofuranose (1)

To a stirred solution of L-ribose (50.0 g, 333.33 mmol) in anhydrous methanol (500 mL) at room temperature was added freshly prepared, anhydrous, methanolic HCl (40 mL, prepared by bubbling anhydrous HCl gas into methanol at room temperature for 15 minutes). at 0 ° C until weight increase to 4 g) by syringe, under argon atmosphere. After addition of methanolic HCl, the reaction mixture was stirred at room temperature for 3-4 hours. Anhydrous pyridine (100 mL) was added and evaporated to dryness at very low pressure (high vacuum) below 40 ° C. This process was repeated twice with anhydrous pyridine (100 mL). The residue was dissolved in anhydrous pyridine (250 mL) and cooled to 0 ° C with an ice bath under an argon atmosphere. To this cold stirred solution was added acetic anhydride (100 mL) via dropwise addition over 15 minutes. After the addition of acetic anhydride, the reaction mixture is stirred again at room temperature for 24 hours while excreting moisture. The reaction mixture was then evaporated to dryness. The residue was partitioned between ethyl acetate (400 mL) and water (400 mL) and extracted into EtOAc. The aqueous layer was then extracted again with EtOAc (100 mL). The combined EtOAc extracts were washed with water (400 mL), saturated NaHCO ₃ (2 x 300 mL), water (300 mL), and brine (200 mL). The organic extract was dried over anhydrous Na ₂ SO ₄ , filtered and the filtrate evaporated to dryness. The residue was co-evaporated with anhydrous toluene (2 x 150 mL) at very low pressure (high vacuum). The dry oily residue (92 g, 95%) was used for the next reaction without further characterization.

The syrup (92 g) from the above reaction was dissolved in glacial acetic acid (300 mL) and treated with acetic anhydride (75 mL) at room temperature. The solution was then cooled to 0-5 ° C in an ice bath and under an argon atmosphere. Concentrated H ₂ SO ₄ (21 mL) was then slowly added over 15 minutes. After H ₂ SO ₄ was added, the reaction mixture was further stirred at room temperature for 14 hours, poured into crushed ice (500 g) and stirred until the ice melted. Water (500 mL) was then added and extracted with CHCl 3 (2 x 300 mL). The chloroform extract was washed with water (3 x 400 mL), saturated NaHCO ₃ (2 x 300 mL), water (200 mL), and brine (200 mL). The washed organic extract was dried over anhydrous MgSO ₄ , filtered and evaporated to dryness to give an oily residue (99 g). This residue was co-evaporated with anhydrous toluene (200 mL) and dissolved in ethyl ether (200 mL) to give colorless crystals over one day after cooling to 10 ° C. The crystalline solid was filtered, washed with hexane / ether (2: 1, 50 mL) and dried to give

60.5 g of product.

Synthesis of methyl-1- (2,3,5-tri-O-acetyl-β-L-ribofuranosyl) -

1,2,4-triazole-3-carboxylate (3) and methyl 1- (2,3,5-tri-acetyl-β-L-ribofuranosyl) -1,2,4-triazole-5-carboxylate (4)

In a RB flask (500 mL) was placed a mixture of methyl 1,2,4-triazole-3-carboxylate (25.4 g, 200 mmol), 1,2,3,5-tetra-acetyl-β-L-ribofuranose (63.66) g, 200 mmol) and bis (p-nitrophenyl) phosphate (1.0 g). This flask was then placed in a preheated oil bath at a temperature of 165 ° C to 175 ° C under an aqueous vacuum aspirator while stirring for 25 minutes. The removed acetic acid accumulates in an ice-cold collection vessel, which is located between the aspirator and the RB flask. The flask is then removed from the oil bath and cooled. After reaching a temperature of 60 ° C to 70 ° C, EtOAc (300 mL) and saturated NaHCO ₃ (150 mL) were introduced and extracted into EtOAc. Subsequently, the aqueous layer was extracted again with EtOAc (200 mL). The combined EtOAc extracts were then washed with saturated NaHCO ₃ (300 mL), water (300 mL), and brine (200 mL). The organic extract is dried over anhydrous

Na 2 SO 4, filtered and the filtrate evaporated to dryness. The residue was dissolved in EtOH (100 mL) and diluted with MeOH (60 mL) then cooled to 0 ° C over 12 h to give colorless crystals. The resulting solid was filtered, washed with a minimum amount of EtOH (20 mL), then dried under high vacuum (high vacuum) over solid NaOH to give 60 g (78%). The filtrate was evaporated to dryness and purified on a silica column using a mixture of saturated CHCl ₃ and EtOAc (9: 1) as eluent. Two products are isolated from the filtrate: the fast-moving product 8.5 g (11%) and the slow-moving product 5 g (6.5%). The slow-moving product corresponds to the crystalline product. The fast-moving product was found to be (4). obtained in the form of a foam. Combined yield (3) was 65 g (84%) ·

Synthesis of Ι-β-ribofuranosyl-1,2,4-triazole-3-carboxamide (5)

Methyl 1- (2,3,5-tri-O-acetyl-β-L-ribofuranosyl) -1,2,4-triazole-3-carboxylate (62 g, 161 mmol) was placed in a steel pressure vessel, bomb, and treated with freshly prepared methanolic ammonia (350 mL, prepared by bubbling anhydrous HCl gas into anhydrous methanol at 0 ° C until saturated) at 0 ° C. The steel bomb was sealed and stirred at room temperature for 18 hours. The steel bomb was then cooled to 0 ° C, opened and the contents evaporated to dryness. The residue was then treated with anhydrous methanol (100 mL) and evaporated to dryness. The residue is then triturated with acetone to give a solid which is filtered and washed with acetone. The solid was then dried overnight at room temperature and dissolved in a mixture of hot EtOH (600 mL) and water (10 mL). Upon cooling the hot solution, colorless crystals formed, which were filtered off, washed with acetone and dried under very low pressure (vacuum). Further concentration of the filtrate affords additional additive. Total yield 35 g (89%).

According to an alternative aspect of the present invention, it is disclosed that the synthesis of Levovirine ™ may also utilize one or more enzymatic conversions. For example, acetylation of Lribose can be carried out with a suitable acetyl transferase (e.g. EC 2.3.1.xx). In another example, formation of the carboxamide group from the corresponding methyl ester may be facilitated by a simple or dual enzyme system comprising an esterase (e.g. EC 3.1.1.xx) and / or an aminotransferase (e.g. EC 2.6.1.XX). According to another example, Levovirin ™ can then be enzymatically converted, to the corresponding mono-, di- or triphosphate (e.g. EC 3.1.3.xx or EC 3.1.4.xx).

It is further contemplated that catalysts other than bis (p-nitrophenyl) phosphate may be used in amounts other than 1 g. The altered amount (i.e. molar proportion) of the catalyst may advantageously increase the selectivity of the reaction towards a higher yield of the desired Nj isomer (L-ribose attached to the Ni atom of the triazole ring) to exceed the N ₂ isomer. For example, suitable amounts of bis (p-nitrophenyl) phosphate include from 3 mmol to 30 mmol or more. Alternatively, amounts less than 3 mmol (from 0.3 mmol to 2.99 mmol) may be included as appropriate. According to further alternative aspects of the present invention, it is not necessary to limit the catalyst to bis (p-nitrophenyl) phosphate only, and alternative catalysts include ptoluenesulfonic acid, trichloroacetic acid and pnitrobenzoic acid.

In particular, from the point of view of the reaction temperature, it is contemplated that lower temperatures may help further increase the selectivity of the reaction to a higher yield of the desired Ni isomer so as to exceed the N ₂ isomer. Therefore, it is contemplated herein that suitable temperatures for the condensation reactions between the triazole moiety and the ribose moiety lie in the range of from 155 ° C to 165 ° C, more preferably in the range of 145 ° C to 165 ° C, most preferably in the range 130 ° C to 165 ° C. 165 [deg.] C.

According to other alternative aspects of the present invention, the selectivity of a given reaction towards a higher yield of the desired Ni isomer to exceed the N ₂ isomer can also be favorably affected by the chemical modification of methyl 1,2,4-triazole-3-carboxylate. Chemical modifications include the formation of a complex structure in which the N ₂ atom, steric hindrance, and direct chemical modifications of the N ₂ atom are involved. For example, the free electron pair at the N ₂ atom of the triazole moiety and the electron donor in the modified carboxylate group can be used for the complex metal ion compound, thereby reducing the availability of the N ₂ atom for interaction with the ribose moiety. Alternatively, the carboxylate group in the methyl-1,2,4-triazole-3-carboxylate may be modified by a relatively built-in group, resulting in preferential and sterically blocking or reducing reactions at the N ₂ atom. Alternatively, the N ₂ atom can be directly modified with a protecting group, with suitable protecting groups including t-Boc and benzyl.

It is further contemplated that a higher yield of the desired Nj isomer that exceeds the N ₂ isomer can be achieved using enzymatic synthesis wherein the ribose moiety (or Lribonucleotide) and the modified or unmodified methyl-

1,2,4-triazole-3-carboxylate serves as a substrate for ribosyltransferase (for example EC 2.4.2.5 or EC 2.4.2.6).

Alternatively, Levovirin ™ can be synthesized via the interaction of the protected L-ribose to 1,2,4-triazole-3-nitrile by converting the nitrile group to the carboxamide as shown in Figure 3. In yet another alternative synthesis, the interaction of the triazole moiety can also be achieved. with a ribose moiety by a reaction in which (for example, benzyl protected) ribose has an -NHNH 2 group attached to a C 1 atom which reacts with N 1. a triazole carboxylate atom wherein the triazole carboxylate is subsequently converted to carboxamidine as shown in Figure 4.

In view of the synthesis of prodrug forms of said L-nucleotides, L-nucleosides and their possible analogs, it would be desirable that, in particular, the synthetic scheme would generally depend on the composition structure of the compound. However, all synthetic methods that may be considered suitable and contemplated synthetic schemes include in vitro syntheses, enzymatic syntheses, in vivo syntheses, and any acceptable combinations thereof. Examples of synthetic schemes for the formation and formation of said prodrugs are given in U.S. Pat. Patent Application Number 09 / 594,410 (supra'i.

Where the L-nucleotides, L-nucleosides and their possible analogs are phosphorylated, it is reported that all methods of incorporating a phosphorous group into the nucleotide, nucleoside, or optional analogs thereof, are suitable. Conversion of contemplated nucleosides to their corresponding phosphoryl forms can be achieved synthetically (Hughes, BG et al., (1983); 2 ', 5'-oligoadenylated and 2', 5'oligonucleotide analogues.) 1. Substrate specificity of the interferon-induced murine 2 ', 5'-oligoadenylate synthetase and enzymatic synthesis of oligomers. Biochemistry, 22: 2116-2126). However, various methods may also be mentioned and enzymatic phosphorylation is included (see, for example, Van Rompay, AR et al., (2000); Phosphorylation of nucleosides and nucleoside analogs by mammalian nucleoside monophosphatecinases; Pharmacol. Ther. 87 (2 to 3); 189) -198) and organochemical phosphorylation in aqueous medium (Schwartz, A. and Ponnamperuma, C. (1968); Phosphorylation of adenosine with linear polyphosphate salts in aqueous solution (Nature 218, 443).

Use of said compounds

In general, the intended compounds can be used for any treatment or therapy of a system that demonstrates a positive response to the administration of the compounds disclosed herein. However, it is highly preferred that the compounds disclosed herein are used for antiviral treatment (as a direct antiviral compound and / or as an indirect antiviral compound) for treatment to modulate the immune system and for treatment related to stimulating cell growth. Particularly suitable uses include administration of the compounds disclosed herein for antineoplastic therapy.

Antiviral treatment

In general, the invention and the medicament and / or the present antiviral medicament may be said to consist of viral infections, a method of step, a composition at a dosage that the compounds of the subject are used as direct as indirect antiviral. which is effective to inhibit the development of a viral infection (ie, a method which relates to a host cell in which one or more viruses act so that the host cell produces one or more copies of the virus, the term producing refers to nucleotide synthesis, protein synthesis) processing and protein folding), wherein such composition comprises at least one of the compounds disclosed herein, preferably at least one compound of Formulas I and III. Preferred doses are in the range of 5 mg / day to 2500 mg / day, more preferably in the range of 50 mg / day to 500 mg / day. However, alternative dosages, routes, methods and forms are also contemplated, and suitable alternative dosages are described below. The use of the compounds disclosed herein is not limited to individual viruses in individual viral infections, and viral infectious diseases include, but are not limited to, HIV infection, HCV infection and HBV infection, RSV infection, influenza viral infectious disease and parachrip viral infectious disease.

Peripheral blood mononuclear cells (PBMCs) are often used to study various serious infections such as hepatitis C, HIV, hepatitis B and various herpes viruses. (Antivir. Chem. Chemother. 2000 July, 11 (4): 291-301; J. Infect. Dis. 1998 Oct., 178 (4): 1189-92; Virology 2000 Mar., 268 (1): 12- 60). PMBCs are infected with a given virus and the cell line is studied in terms of relevant information, what interactions occur between drugs and between PMBCs, and how the infected PMBCs behave during the period of time under different environmental conditions. Based on studies conducted with infected PMBCs, models can be developed that show how the individual pharmaceutical compositions, the environment and / or the condition and conditions act on these PMBCs-infected cells.

The inventors have discovered (unpublished results) that Ribavirin shows a positive response against PMBC-HIV infected cells. Surprisingly, Levovirin ™ also exhibits a similar immunomodulatory profile against PMBC-HIV infected cells through the loss of enzymes in the body necessary for phosphorylation of Levovirin ™ present in the patient. Based on the above observations and other information and tests relating to this issue, it can be stated that Levovirin ™ and the modified Levovirin ™ of structural formula III can be used to treat HIV and similar viruses.

immunomodulation

Figure 5 is a method of immunosuppression of a target organ 500 having a first step 510 in which a drug that reduces both a type 1 and a type 2 response is provided when the drug is administered at a higher concentration than the immunosuppressive concentration, and the type 1 response increases relative to Type 2 when administered at a lower concentration than the immunosuppressive concentration, wherein the drug is preferably accumulated in the target organ. In the next step 520, the patient is administered the drug at a dose that is effective to accumulate the drug in the target organ to an immunosuppressive concentration. Consequently, a method of treating a disease characterized by a patient's inflammation of the liver consists of two steps - in the first step, a compound is provided that contains Levovirine, phosphorylated Levovirine ™ and modified phosphorylated Levovirine ™ or modified

Ribavirin (supra). In a second step, the patient is administered said compound at a dose that (a) causes systemic immunomodulation and not systematic immunosuppression of the Type 1 and Type 2 responses, and (b) causes systemic immunosuppression of the Type 1 and Type 2 responses in the patient's liver.

The term immunosuppression refers to the case where the size of T and / or B cell clones of lymphocytes is reduced or their reactivity, expansion or differentiation is suppressed. Thus, immunosuppression may result from the activation of specific or non-specific T suppressor lymphocytes of either T or B clones, or most or all T or B lymphocytes are affected by the drug. For example, Cyclosporin A and FK506 are relatively specific for T cells, while alkylating agents, e.g. cyclophosphamide are less specific in their action.

As used herein, the term cytokine refers to a group of soluble proteins and peptides that act as humoral regulators at nano- to pico-concentrations and that modulate the functional activities of individual cells and tissues either under normal or pathological conditions. Cytokines also mention interactions between cells directly and regulate placement in the extracellular environment.

As used herein, the term lymphokines refers to a subset of cytokines produced by helper T cells, and can generally be considered to fall into two subclasses of type 1 and type 2. Cells of type 1 produce interleukin 2 (IL-2), a tumor necrotic factor (TNFα) and interferon gamma (IFNγ), and in particular are responsible for cell-mediated immunity, such as delayed type hypersensitivity and antiviral immunity. In contrast, type 2 cells produce interleukins, IL-4, IL-5, IL-6, IL-9, IL-10 and IL-13, and are particularly useful in assisting in cases of humoral immune responses such as those manifested in responses to allergens (e.g., IgE and IgG4 isotopic shift antibodies).

Thus, the terms Type 1 and Type 2 responses are intended to encompass the entire range of effects resulting from induction of type 1 and optionally type 2 lymphocytes. Among other things, these responses include increased production of the corresponding cytokines, increased proliferation of the corresponding lymphocytes, and other other effects associated with increased cytokine production, including the effects on motility. The type 1 response is generally characterized by an increase in IL-2, TNFα and IFNγ, while the type 2 response is generally characterized by an increase in IL-4, IL-5, IL- 6 and IL-10.

As used herein, the term drug preferably accumulates in the target organ refers to a selective target organ mechanism that results in a relative increase in drug uptake or retention in that target organ compared to other tissues or organs. The mechanism may thus include active delivery by transporters, receptors, vesicles, etc., but may also be based on physicochemical principles, including the pH dependency of the dosed drug, differences in solubility of the drug in different ionic strength alternating agents, chemical or enzymatic modifications in target organs or target cells, etc.

Preferably, Ribavirin is administered to patients with HCV (hepatitis C virus) infection; Ribavirin is orally administered to a patient at a single dose of 600 mg / day for 180 days. Generally, a single dose of 600 mg / day is lower than the systemic immunosuppressive concentration, but is effective for the preferential accumulation of Ribavirin in the target organ and will increase significantly in the liver. O in the liver. Concentration (here the liver) will be enriched immunosuppressive Ribavirine is known to cause an increase in type 1 response compared to type 2 response and to decrease type 1 and type 2 response at relatively high concentrations. Examples are summarized in International Patent Application Number PCT US98 / 00634, issued Jan. 13, 1998, which is incorporated herein by reference.

According to alternative aspects of the present invention, it is not necessary to limit said drug to a lan

Ribavirin; alternative drugs include the compounds (supra) disclosed herein, especially modified and unmodified Levovirin ™ and phosphorylated Levovirin ™. Other alternative therapeutics include the compounds provided herein when the alternative compounds reduce both the Type 1 and Type 2 responses at an immunosuppressive concentration and increase the Type 1 response compared to the Type 2 response at a lower concentration than the immunosuppressive concentration.

Various viral infections other than HCV infection, including arbovirus infections, may also be presented to the patient. Consequently, there is no restriction to the liver, brain, but may also include thymus that the other lungs, the spleen, the disease, presented according to a specific structure (i.e. in which the body of the drug). For example, other organs, such as etc. Generally, the highly affected kidneys can be treated, not the invention, the drug accumulation schemes are preferential accumulation. Ribavirin and Levovirin ™ are preferably and both type 1 and type 2 responses are reduced at such immunosuppressive concentrations. Here, those diseases in which suppression of the immune response in the liver is desirable, such as hepatitis C, autoimmune / lupoid hepatitis, liver transplant recipients, etc.

In particular, the method of the present invention is not intended to provide antiviral treatment, but is intended to at least partially suppress an immune response in a target organ that is infected with a virus. The target organ of the immune response is reported to be particularly useful in hepatitis C, where the HCV virus does not directly contribute to the organ damage, but rather to the imbalance between the type 1 response and the type 2 response that is caused by the infection. Thus, a method of treatment with a drug that both specifically reduces the type 1 response and the type 2 response in the liver of a HCV infected patient to prevent hepatitis damage prophylactically as well as therapeutically may be mentioned. Both Ribavirin and Levovirin ™ are particularly advantageous because they exhibit excellent human tolerance, long-term prophylaxis, long-term action.

As regards administration (route, dosage, schedule, deadline, etc.) of Ribavirin or said alternative compounds, the same conditions as described below can be applied. It can also be further noted that Ribavirin or the alternative compounds disclosed herein can be used in a general overall health situation, and vice versa, in a clinical therapeutic situation. Accordingly, Ribavirin or alternative compounds herein may also be used to improve digestion. For example, one or more of said compounds may be administered to individually promote poor digestion - preferably poor digestion attributed to liver status, as is the case with hepatitis B or C infections, or other conditions characteristic of liver inflammation. In such cases, digestion can be improved by administering to the patient Ribavirin or Ribavirin as the compound below in amounts that are common in systemic immunosuppression, but the amount accumulated in the liver then reaches a concentration that corresponds to immunosuppression in the patient's liver.

As another example, not the therapeutic or clinical use of Ribavirin or the alternative compounds in humans, it may be noted that Ribavirin, or alternatively, the compounds listed below may be used in amounts that are common in systemic immunosuppression but are accumulated in the liver to improve human skin coloring. concentration that corresponds to immunosuppression in the patient's liver.

Ribavirin (1- (5-deoxy-β-D-ribofuranosyl) -

1,2,4-triazole-3-carboxamide) or Levovirin ™ (1- (5-deoxyβ-L-ribofuranosyl) -1,2,4-triazole-3-carboxamide) or any of the mono-, di- or triphosphorylated forms. The amount administered is preferably that sufficient to produce systemic immunomodulation of the Type 1 and Type 2 responses and hepatic suppression of both the Type 1 and Type 2 responses. The most preferred amounts are in the range of 300 mg / day to 800 mg / day, although in some individual cases, this range may be 50 g / day to 100 mg / day, up to as high as the range 2000 mg / day to 2400 mg / day. Effects in relation to other organs, including the brain or other organs known to have significant accumulation of Ribavirin, are also considered.

Stimulation of neuronal growth

As used herein, the term neuronal growth stimulation refers to any method in which cell growth and / or division of either initiated growth occurs.

from resistant cells, or accelerating growth and / or cell division, the term neural refers to all cells that are directly or indirectly included in

propagation of cognitive, sensory or motor signals. For example, nerve cells can be said to be directly involved in signal propagation, while myelin sheath cells or supporting nerve tissue cells are indirectly involved based on their isolation function or structural / metabolic support provided to nerve cells. Similarly, receptors can be considered as nerve cells within the scope of this definition. In contrast, cells that form in the lower and upper layers of the diaper are not considered nerve cells in this context and are not directly or indirectly involved in the propagation of cognitive, sensory or motor signals.

Surprisingly, the inventors have discovered that Ribavirin is effective in stimulating neuronal growth and can now state that a variety of phosphorylated analogs may be effective in a similar manner. In one experiment, Levovirin ™ was found to be effective in stimulating the growth of unipolar nerve cells in vitro at concentrations ranging from 0.5 μΜ to 500 μΜ. Subsequently, Levovirin ™ on 5.0 μΜ culture medium can be used to stimulate unipolar nerve cell growth.

Furthermore, methods of stimulating neuronal growth are not limited to unipolar nerve cells, but that various alternative cells, including bipolar and multipolar nerve cells, may be included in this field. In addition, according to alternative aspects of methods of stimulating neuronal growth, individual cell types in a diverse diverse nerve cell population may be targeted. For example, the target may be unipolar, bipolar and multipolar cells in complex neuronal structures such as the brain, spinal cord, or eye. Thus, neurons may be part of neuronal tissue that comprises at least four of the following cell types: astrocytes, dendrocytes, myelin sheath cells, supporting nerve tissue cells, unipolar nerve cells, bipolar nerve cells, multipolar nerve cells, and receptor cells.

Thus, these methods are not necessarily intended solely to stimulate neuronal growth in cell culture. According to other alternative aspects of the present invention, it may further be noted that cells may be stimulated in tissue culture, but in particular, nerve cells may be stimulated in vivo. In vivo stimulation of neuronal growth can be used as a prophylactic method of treatment or as a therapeutic treatment. For example, the methods of the present invention can be used to prevent demyelinating diseases (diseases related to the loss of nerve fiber pulp) or neurovegetative diseases such as Alzheimer's disease or Parkinson's disease, or as a preventive treatment before surgical intervention in patients. Methods of treatment contemplated herein include reversing or ameliorating the attenuation after asphyxia, traumatic, toxic, infectious, degenerative, metabolic, ischemic, or hypoxic insulins, or irritable states.

Consequently, as shown in Figure 6, the method 60Q improves patient coordination, where in a first step 610 it is examined whether it is phosphorylated or non-phosphorylated

Ribavirin or Levovirin ™ effective to stimulate nerve cell growth in vivo over a given concentration range. In the next step 620, the patient receives an amount of phosphorylated or non-phosphorylated Ribavirin or Levovirin ™ that is effective to stimulate the growth of at least some of the patient's nerve cells. In particular, improvement in eye-hand coordination can be mentioned.

In a preferred way of improving patient coordination, it is investigated whether Ribavirin is effective in stimulating nerve cell growth in vivo in a concentration range of 0.5 μΜ to

500 μΜ, in the case of sciatic nerve, Ribavirin is orally administered at a dose of 1200 mg / day to support the traumatic condition.

However, other conditions other than the traumatic condition of the nerve sciatica, including mechanical and chemical damage to a large number of nerve cells, infections of nerve cells with bacteria and / or viruses, and degenerative diseases are contemplated for the patient. In view of the patient's medical condition, it can be said that according to the present invention it can stimulate nerve cells to a very wide extent, and in particular it is said that the stimulated cells pass mainly between the patient's brain and free muscles or between the patient's brain and skin receptors.

Another class of methods disclosed herein includes improving the condition of a tactile receptor or other patient's sensitivity or sensitivity. Yet another group of methods disclosed herein includes improving macroscopic and microscopic motor control.

Without wishing to be bound by theory or limitations, it may be noted that administration of Ribavirin, Levovirine or any of the mono-, di- or triphosphorylated forms of Ribavirin or Levovirine ™ may affect changes in η

a type 1 response or a type 2 response in a patient, which may, under concomitant circumstances, lead to a condition in which nerve cells are protected or to stimulate neuronal growth. Thus, it is noted that the compounds of the present invention can be administered as part of a treatment for a patient's disease in a dosage range effective to cause the patient to increase the lacquer type response and reduce the type response.

2. With respect to in vivo administration (route, dosages, schedule, deadline, etc.) of Ribavirin or alternative compounds disclosed herein is as described below.

Antineoplastic treatment

Further, the compounds of the present invention may be used as antineoplastic agents for the treatment of solid or lymphatic tumors, which include neoplasms of various cancerous tumors, connective tissue malignancies and lymphomas, but particularly include acute myeloid leukemia and chronic myeloid leukemia in sudden turnovers during disease. Further, administration of the compounds disclosed herein for antiviral treatment is usually carried out according to the method, dosages, schedule and terms used with known D-nucleotides, D-nucleosides and their possible analogs in antineoplastic therapy.

Administration of said compounds

With respect to the administration of the compounds disclosed herein, they may be administered according to any suitable protocol, in any suitable pharmaceutical form. Oral administration of these compounds is preferred. Other alternative routes of administration may be appropriate according to alternative aspects of the present invention, however, it has been found that the individual routes of administration are highly dependent on chemical stability, bioavailability, dosage, formulation and / or the desired pharmacokinetic / pharmacodynamic properties of the compounds disclosed herein. Appropriate administration will therefore depend on local delivery (eg in the form of ointment, spray, cream, etc.), parenteral systemic delivery _| (for example by inhalation) and whether it is direct or indirect delivery to the bloodstream (for example by intravenous or intramuscular injection, etc.).

molds the administration of said compounds may vary. For example if it has a medicine or medicinal product reasonable stability in gastrointestinal transit

Suitable undesirable chemical or enzymatic modifications include syrup, tablets, gel capsules, powder and the like. On the other hand, when there is the crossing _t of the compounds of the gastrointestinal system of the absorption and penetration through the gastrointestinal tract into the blood stream, problems may arise and if the suitable forms such as injectable solutions or suspensions (e.g., physiological saline, with modified buffered value pH 7.2 to 7.5).

In terms of dosages of the compounds, various suitable dosages may be used, with dosages typically ranging from 1 mg to several 100 mg, even higher. For example, if the compounds disclosed herein are secreted or metabolized at a relatively low rate, or if long-term is desired. treatment, then doses are typically in the range of 5 mg / day to 200 mg / day. On the other hand, if these compounds have relatively low bioavailability or if metabolic conversion (e.g., dephosphorylation) is relatively rapid, then dosages are typically in the range of 100 mg / day to 2500 mg / day.

As regards the dosage of L-nucleosides, especially Levovirine ™, it can further be noted that Levovirine ™ does not appear to be phosphorylated in vivo, at least not in hepatocytes and erythrocytes, and since the antiviral effect of Ribavirin addicted to not expected effect. In phosphorylation, an expert that Levovirin ™ should (which attempts would be antiviral Russian published) does not demonstrate that in a given direct area there are not actually a direct antiviral effect. Surprisingly, the antiviral effect of Levovirin ™ is demonstrated at a dose not exceeding 200 mg / day, preferably in the range of 10 mg / day to 200 mg / day, more preferably in the range of 50 mg / day to 200 mg / day, and even more preferably in the range of 50 mg / day to 100 mg / day. Thus, it is experimentally supported that the results obtained when investigating the dosage of Levovirine correspond to five times lower serum concentrations than the equivalent dose of Ribavirin.

Experiments have shown that Ribavirin is removed from serum by adequate phosphorylation in red blood cells (see, for example, Homma et al .: High Performance Liquid Chromatographic Determination of Ribavirin in Whole Blood Asses Disposition in Erythrocytes; Antimicrob. Agents

Chemother. (1999), 43 (11): 2716-2719. Once phosphorylated

Ribavirin can no longer leave the cell. Consequently, red blood cells act as a ribavirin trap and higher doses of Ribavirin are then required to reach a given serum level. Levovirin ™ is not phosphorylated and therefore does not tend to accumulate in red blood cells. As a result, red blood cells do not play the role of Levovirin ™ wells, and therefore lower doses of Levovirin ™ serum are sufficient to achieve the desired serum level.

The schedule of administration may be varied, with the schedules herein being comprised of single single doses throughout the treatment, multiple daily single doses throughout the treatment, multiple daily doses, and permanent dosing (e.g., permanent infusion, implanted osmotic pump, etc.). ) for at least some time throughout the treatment. It is usually preferred to maintain constant delivery of the compounds according to appropriate schedules, but intermittent delivery is also appropriate (i.e., at least once a first dose is administered, followed at least once by a plurality of doses that are lower than the first dose). As to the duration of treatment, it may be noted that the appropriate duration may vary between individual administrations for several days, several weeks, several years, even longer. For example, if the compounds disclosed herein are used in cell culture, single or relatively short-term administration may be appropriate. On the other hand, when the compounds are administered for the treatment of acute hepatitis, a suitable duration of treatment may range from a few days to several weeks. Similarly, in the treatment of chronic hepatitis by administering the compounds herein, it may be appropriate to prolong the administration to one year or to several years.

According to yet other alternative aspects of the present invention, the compounds disclosed herein may be combined with additional excipients, pharmaceutically active agents, to aid in the treatment of various diseases, particularly viral infections. In addition, these additional pharmaceutically active adjuvants may be administered separately or together and, if administered separately, the administration may take place simultaneously or separately, in a predetermined order. Such additional auxiliaries include and / or pharmaceutically active and immunomodulatory components, for example nucleoside substances in particular. antiviral

Antiviral agents include immunomodulatory protease analogs of the inhibitor component, nucleotide (particularly Ribavirin) and include, for example, cytokines (e.g., interferon α and γ,

IL2, IL4, IL6, IL8, IL10, and IL12).

Other additives include, for example, tolnaphthalate, Fungizone ™, and Amphoteracin;

Niclocide such as antiparasitic agents ™, Vermox ™ and Flagyl ™

Immodium ™, Lomotil ™ adjuvant, pharmaceutical antifungal agents Lotrimin ™, Mycelex ™ such as Mii-related agents and Phazyme ™; γ, Adriamycin ™

Mithracin ™, active agents,

Imuran ™, such as interferon and α

Methotrexate ™, infra

Cyclocort ™, and salicylic acid; ergotamine compounds; hitherto not mentioned, dermatological

Denorex ™, Reagents,

Florone ™, intestine, antitumor Cytoxan ™,

Thiazofurin ™, Taxol ™;

Aclovate ™.

TM t

such as

Oxsoralen preparations, tar further hydrocortisone; Valisone ™; metabolic migraine steroids and immunosuppressors, including cyclosporins, Floron ™, Lidex ™ such as Diprosone ™,

Topicort ™ and agents such as insulin and other medicines that do not fall into the above categories.

Preferred combinations of the compounds disclosed herein with interferon

According to a particularly preferred aspect of the present invention there is provided a synergistic combination of Levovirine ™ and at least one interferon, preferably IFN-α-2b. For Levovirin ™ there is less scope taking activity.

typical yet not only that it acts significantly as Ri.bavirin phosphorylated in erythrocytes, it proves antiviral

Consequently, immunomodulatory, it can be stated that the pharmaceutical action of diseases, interferon, can be TM treatment especially in

Levovirine ™ with significant Ribavirin. For example, doses of Levovirine ™ in combination to treat HCV infections may be proposed in the range of 1 mg to 600 mg, more preferably in the range of 10 mg to 400 mg, even more preferably in the range of 50 mg to 300 mg, and most preferably lower hepatitic. synergistic to indicate with the interferon required in the range of 100 mg to 300 mg. Equivalent synergistic doses of Ribavirin are reported in the range of 600 mg to 800 mg.

In another aspect of the present invention, the synergistic combination of Levovirine ™ and interferon results in reduced toxicity, compared to the combination of Ribavirin and interferon, at equivalent effective doses, particularly due to the lack of significant phosphorylation in erythrocytes.

another other aspect can be mentioned,

In addition, the synergistic combination of Levovirine ™ and interferon specifically allows the target to be a liver due to the lack of phosphorylation of Levovirine ™ in parts other than the liver, particularly with regard to erythrocytes.

With respect to the concomitant administration of Levovirin ™ and interferon, all suitable methods and protocols are acceptable, and it is particularly preferred that Levovirin ™ and interferon are administered according to a protocol similar to known protocols for Ribavirin and interferon. For example, Levovirin ™ may be administered orally, while interferon may be administered by subcutaneous injection. In general, schedules and methods that are independent of one another and independent of whether and for how long both drugs are present in measurable concentrations at the same time in the bloodstream can be used for the concomitant administration of Levovirine ™ and interferon. Further, effective doses of Levovirine ™ can be designed according to effective doses of Ribavirin in the liver when Ribavirin was administered.

In particular, Levovirin ™ is disclosed herein, but chemical modifications thereof, including prodrug forms such as modified Levovirin ™ (Ι-β-L-ribofuranosyl-1,2,4-triazole-3-carboxamide), mono-, di- and triphosphorylated Levovirin ™ and stereochemical variants (e.g. enantiomers, isomers, etc.). Examples of such suitable chemical modifications and prodrug forms are described in U.S. Pat. No. 09/594410, (supra). In addition, suitable drugs other than Levovirin ™ and its variants are also suitable drugs, and in particular alternative drugs, including specific liver prodrugs with an amine or amide group that may be enzymatically deaminated. deamidated.

With respect to interferon, it can be stated that co-administration with Levovirin ™ is not limited to IFN-α-2b, but natural and synthetic fragments, isoforms and consensus forms of interferon-alpha form may also be included in this co-administration. In addition, interferons other than interferon-alpha are also suitable, including interferon beta and its natural and synthetic fragments, isoforms and consensus forms.

In particular, interferon has been reported so far, but cytokines other than interferon and chemokines, including IL-2, IL-12 and TFN, may also be suitable. In particular, the pegylated forms of the interferons herein (i.e., said polyethylene glycol-associated interferons) are also suitable for use in conjunction with the teachings herein.

Combination of said compounds with a second compound that binds a viral protein or cytokine

If a combination of the compounds disclosed herein with other pharmaceutically active agents is used in antiviral therapy, it may be particularly noted that such combinations may comprise the compounds disclosed herein with direct and indirect antiviral activity and a second compound that enhances the overall antiviral effect (the total antiviral effect consists of from a direct antiviral effect and an indirect antiviral effect), wherein the second compound specifically binds a viral protein or cytokine.

In this regard, they are more preferred herein.

if preferred by the compounds in this combination, the nucleoside analogue and still the nucleoside analogue is

Ribavirin carboxamide).

antiviral (1- (5-deoxy-β-D-furanosyl) -1,2,4-triazole-30) Ribavirine is known to have direct when inhibiting RNA and DNA effect, viral replication [Huffman et al., Antimicrob.

Agent s

Chemother.

(1973) 3:

235; Sidwell et al., Science

177: 705] the effect of indirect antiviral T mediating and promoting a type 1 response, a patent application of which in the type response described in U.S. Pat.

incorporated herein by reference. However, compounds, especially L-nucleoside alternative compounds, that have an antiviral effect may be used. For example, cells to suppress such as number 09/156 646, which is also mentioned by a variety of analogs, as well as having direct and indirect ones, if a very high concentration of L-nucleoside analogs is desired, Levovirin ™ can be used.

Further, depending on the chemical nature of the first compound, the first compound may have a significantly greater direct antiviral effect or a significantly greater indirect antiviral effect. Said direct antiviral effects include inhibition of viral replication, e.g., inhibition of reverse transcriptase, said direct antiviral effects comprising a shift of type / type 2 equilibrium towards a type 1 or type 2 response as described in U.S. Pat. It should also be noted that an indirect antiviral effect may include suppression of a type 1 or type 2 response, as described in U.S. Pat. Advantageously, Application Provisional Application Number 60/172 097 (supraj. Shift Type / Type2 toward Type 1 or Type 2 Response or Suppress Type / Type 2 Response can be advantageously controlled or controlled by the first compound, wherein the dosage of the first compound is determined by shifting or suppressing Type 1 or Type 2 responses.

For the second compound, it is preferred that the compound comprises an antibody (e.g., a monoclonal or polyclonal antibody). However, from the standpoint of the alternative necessary occurring synthetic forms obtained by some aspects of the subject matter of the present invention, define this antibody on nature, but that antibodies (for example, from molecular development technologies) antibodies. Particularly desirable are fragments of the invention, non-antibody forms include antibodies or antibody fragments produced by recombinant cells or where the molecular weight of the second compound could be relatively low (ie, less than 75 kDa). Said antibody fragments include Fab, any F (ab) _2, and any scFab. In addition, suitable antibodies can be modified by introducing additional properties, for example, a reporter group, a second affinity moiety (e.g., a bispecific antibody), or a pharmaceutically active molecule. For example, the reporter group may include a radioisotope or a metal that is detectable in vivo by imaging devices (e.g., magnetic resonance imaging). Said pharmaceutical active molecules may include reverse transcriptase inhibitors, protease inhibitors or cytotoxic agents. The production of recombinant and non-recombinant antibodies is well known in the art (see, for example, Current protocols in Immunology;

John Wiley & Sons (1999); Edited by: John Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strober), all known methods can be used to produce them in accordance with the teachings herein. Antibodies are typically administered by injection (e.g., by intravenous injection) and the actual doses are in the range

0.01 mg to several 10 mg, but lower doses may be considered if necessary.

Further, the ability of the second compound to bind to a protein or to a cytokine is particularly advantageous if it results in inactivation of the viral protein and / or cytokine; it can also be stated that this inactivation can be carried out by various mechanisms. For example, virus inactivation may be achieved by antibody-mediated precipitation (i.e., by the formation of a molecular network between antibodies and viruses). Alternatively, the ability of the second compound to bind may inactivate the virus by blocking or other obstructive proteins or other viral surface structures that are essential for virus ineffectiveness or propagation. Further, the second compound may then bind to non-structural viral proteins, including viral polymerases and proteases. Such binding targets (i.e., hapten) include, for example, proteins such as gp 120/41 of the HIV virus, but also proteins such as reverse transcriptase of the HIV virus. Other viral proteins also include proteins from HIV virus, hepatitis virus, influenza virus and RSV virus. With respect to cytokines, it can be noted that inactivation can be achieved by sequestering the cytokine from the accumulation of cytokines. For example, if the hapten for the second compound is a type 1 cytokine, then the cytokines contemplated include interleukin-2, interferon-gamma, and tumor necrosis factor-beta, but in cases where the hapten for the second compound is a type 2 cytokine, the cytokines considered include interleukin-4, interleukin-5, and interleukin-10.

In general, inactivation of a virus or cytokine by a second compound may have a large number of desired effects, which effects may or may not be demonstrated as an additional additive or synergistic effect in combination with the first compound. For example, in cases where the first compound has a direct antiviral effect (resulting in a significant reduction in viral titer), the second compound may even further reduce this viral titer by precipitation of the remaining viruses. Alternatively, the second compound may then reduce the number of infectious viral particles by binding to viral components that are essential for contagion. Furthermore, where the second compound binds a cytokine, the second compound shifts the type 1 / type 2 balance towards a type 1 response by sequestering one or more type 2 cytokines from the accumulated cytokines, thereby helping to restore cellular immunity, thereby significantly reducing the viral load. Alternatively, if the first compound has an indirect antiviral effect (even here it results in a significant reduction in viral titer), then the second compound can further reduce the viral titer by precipitation of the remaining viruses. Thus, alternatively, one or more type 1 and / or type 2 cytokines may be sequestered with a second compound or a mixture of second compounds, thereby fine-tuning (i.e. modulating) the type 1 and / or type 2 response induced by the first compound.

It could also be noted that the combination of a first compound having a direct and indirect antiviral effect with a second compound where specific binding of the virus and / or cytokine occurs will reduce the viral titer not only mechanically (ie, enzyme inhibition) but also systemically (ie. stimulation / modulation of immunity) by action. In particular, it may be appreciated that antiviral medicaments comprising first and second compounds having synergistic action are preferred, which have the advantage of helping to reduce the effective dose of the first and second compounds. Suitable antiviral medicaments may also be used in prophylactic treatment.

Combination of the above compounds with Ribavirin

In particular, co-administration of Levovirine ™ with Ribavirin reduces adverse side effects and improves tolerability of Ribavirin and / or Levovirine ™. As for the ratio of Ribavirin to Levovirin ™, if Levovirin ™ is Ribavirin. However, when it is a ratio of Ribavirin to Levovirine when co-administered, it is preferred that it is present in at least an equimolar amount that is, respectively.

anemia desirable, may also be used depending on the desired

For example, in a simultaneous administration, a molar to 80 mole% tolerability of the particular route of administration is present.

In an amount ranging from 51, the Levovirin ™ may be alternative ratios, effects and dosages of the hemolytic event or more. However, if limited, at only about 49% mole% of Levovirine ™ administration can be up to 20% mole or less.

Further, Levovirine ™ and Ribavirin are not the same route of administration. Here, the concurrent administration relates

Ribavirin, administration

Levovirine ™ and at a given time the concentration.

present in

Thus, it may be present molar at the same time it is necessary to use the meaning of whatever form is given

Levovirin ™ administration using the concept of administration of the Ribavirin system in that simultaneous and Levovirin ™ in another manner, can be accomplished simultaneously or measurable by administration of one wherein simultaneous in two different can be administered can at times. For example, Ribavirin, while Levovirin ™ may be another case and Levovirin ™ include, includes a protocol according to which Ribavirin is a method of administering temporally orally by intravenous injection. Ribavirin administered orally administered orally QID.

In particular, it should be noted that:

BIDs for various molar fraction variations, especially due to the potentially tailored to the specific needs of modulating cytokine balance, may be co-administered.

Ribavirin and Levovirine be

TM t

desirable biological effects of the patient, including type 1 / type 2, direct antiviral effect, reduced hematotoxic properties, etc.

According to another aspect of the present invention, it is further contemplated that the simultaneous administration of Ribavirin and Levovirin ™ may involve continuous release and / or dose reduction at more frequent intervals. In particular, it should be noted that continuous release and / or dose reduction at more frequent intervals may reduce unwanted side effects and may help increase the direct and / or indirect antiviral effect. In general, the compounds of the present invention may be administered by any system, and it is preferred that the compounds herein be administered to a mammal, preferably a human, into cells or into tissue cultures.

Metabolites of said compounds

Generally, Levovirin ™ administered to the system is metabolically inert, however, the inventors state that Levovirin ™ may also have metabolites, which are illustrated by the following structural formulas IV to VIII:

\ H

(IV) (VI) (VI)

OCOCH;

Structural formula IV shows triazolecarboxamide, structural formula V shows triazolecarboxylic acid, structural formula VI shows L-ribofuranosyltriazolecarboxamide, and structural formula VII shows 5'-acetyl-Libibofuranosyltriazolecarboxamide and structural formula VIII shows 5'-acetylcarbamyl-a-L-triazolecarboxamide.

Although generally speaking, the metabolic products of Levovirine ™ are produced as products of an enzymatic reaction, it has been found that under certain suitable intracellular or extracellular conditions, metabolites may be formed in the cellular system without an enzymatic reaction. The formation of metabolites from Levovirin ™ may include redox reactions (especially oxidation), enzymatically catalyzed reactions (e.g. hydrolysis), and photochemical reactions.

The reaction products described herein are typically degradation products of Levovirine ™, however, it has been found that these metabolites contain products that result from the addition of chemical groups (e.g., glycosylation or acetylation) and that the modified compounds may undergo subsequent degradation in the same or different compartments or portions. While it is generally assumed that these metabolites have a significantly reduced pharmaceutical effect compared to Levovirin ™, it can be positively assessed here that these metabolites have a similar pharmaceutical effect

Levovirin ™. For example, a triazole moiety or a ribose moiety may serve as an effector (e.g., an allosteric inhibitor).

Further, a single particular dose of Levovirine ™ administered to the system is within the range of 20% to 50%, preferably 51% to 75%, more preferably 76% to 99%, and then most preferably 100% excreted in non-metabolized form.

The present invention is further illustrated by exemplary embodiments which illustrate the invention in more detail and in no way limit the scope and scope of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1A-IC show examples of compounds of the present invention.

Figure 2 shows an example of a synthetic scheme for the synthesis of β-β-L-ribofuranosyl-1,2,4-triazole-3-carboxamide.

Figure 3 shows an alternative example of a synthetic scheme for the synthesis of Ι-β-L-ribofuranosyl-1,2,4-triazole-3-carboxamide.

In Figure 4 is another synthetic example shown schemes for carboxamide synthesis. Ι-β-L-ribofuranosyl-l, 2,4-triazol-3- Figure 5 shows a block diagram of an example is shown

a method of immunosuppression of a target organ according to the present invention.

Figure 6 shows a block diagram of an example is shown

of a method of stimulating cell growth according to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

Targeted hepatitis immunosuppression using Ribavirin

Three placebo-control studies were conducted with Ribavirin in the treatment of chronic hepatitis C. These studies included 134 patients treated with Ribavirin and 97 patients received placebo. Two uncontrolled phase II studies were also included, involving 23 patients treated with Ribavirin. The primary response parameter was normalized or reduced by serum ALT. The response was also assessed in terms of eliminating or reducing serum HCV RNA levels and improving liver histology, judging by changes in Knodell scoring.

In all control and non-control studies, using the definitions of response specified in the protocols and the analysis plan, Ribavirin was statistically significantly superior to placebo at normalized and reducing ALT levels during treatment. In integrated analyzes based on all patients in control studies using a uniform definition of response, including normalization of ALT at the end of treatment or a clinically significant level of partial response, it was found that 46% of Ribavirin patients responded to treatment, but in placebo patients were only 4% (p <0.001). In general, patients responded after two to three months of treatment and the response lasted until treatment continued. Thus, no loss of ALT response was found with improvement during treatment. Subsequent withdrawal of Ribavirin at the end of the active phase of treatment responded to 11.5% of patients who had a sustained response throughout the next follow-up period.

No significant trend in favor of Ribavirin was observed in the improvement of liver histology in each of the control studies by assessment of variations in Knodell scoring and variations in scoring for the individual components. Analysis of the data combination by covariance analysis using the Knodell scoring baseline as a covariant yielded a result according to which statistically significant differences were obtained for Ribavirin for overall scoring and for scoring each component. Thus, in control studies, compared to all patients treated with Ribavirin and patients with placebo, Ribavirin had a modest but real effect on improving liver histology. A comparison of the Ribavirin group with ALT respondents and non-responders shows that experience with significantly greater improvements in liver histology has been confirmed in ALT responders than in ALT respondents. The mean value in Knodell scoring was two points for ALT respondents, compared to one point for all Ribavirin-treated patients. A two point decrease in overall Knodell scoring is considered clinically significant from a hepatological point of view. Thus, there is a statistically significant positive correlation between the ALT response and a clinically significant degree of improvement in liver histology.

In all studies, the primary endpoint is defined as a decrease in ALT levels. In all studies, the complete total ALT response is defined as normalization of ALT. The partial ALT response is the end-of-treatment level.

defined as a 50% or greater reduction at the end of treatment by the patient's baseline value, or a 50% or greater reduction to a level not more than

1.5 times the upper limit of normal.

In studies 92-001 and 91-178, treatment groups were compared for the effect of study medication or a symptom relevant to hepatitis. This is not done in study CT00 / 002, as this case takes the form of a report where systematic collection of symptomatic data is not used.

In study 92-001, there is a statistically significant difference in favor of Ribavirin, due to a decrease in fatigue, a reduction in exhaustion. At the end of the treatment and at the end of the study period, patients with a higher proportion of Ribavirin showed some improvement compared to baseline in patient fatigue compared to placebo patients (p = 0.04 at the end of treatment and p - 0.006 at the end of the study period). ). In this study, there were no significant differences between the treatment groups in terms of any of the symptoms.

In study 91-DK-178, significant differences between treatment groups were isolated in individual symptoms at each visit, some in favor of the placebo group and some in favor of the Ribavirin group, but there were no overall trends in favor of one of the treatment groups.

In Table 1 below, response rates for improving liver histology are shown. None of these studies showed any statistically significant differences between treatment groups according to changes in Knodell scoring, although a numerical trend can be observed in favor of Ribavirin. In study CT00 / 002, there is a difference in favor of Ribavirin in one of the secondary parameters (lymphoid clumps, p - 0.05). .

Table 1

Comparison of control study results - liver histology

protocol Definition of the answer v protokoie Analysis of the planned defined response The result 92-001 Comparison of treatment groups equal to pre-to-post-treatment changes according to Knodell scoring for each patient The same No significant difference between treatment groups 91-DK-178 Long-term response: improvement of liver histopathology by "blindly classifying all liver biopsies for the degree of concomitant hepatitis damage using the Wilcox series summation test" Comparison of treatment groups to appearance to change pre- up to Knodell scoring treatment for each patient There was no significant difference between the treated groups CT00 / 002 Improvement in the level of anti-inflammatory activity in pre- to post-treatment Knodell scores for each patient Comparison of treatment groups with respect to pre- to post-treatment changes according to Knodell scoring for each patient No significant difference between treatment groups Pre- to post-treatment, changes in other histological parameters that may be relevant to hepatitis c reduction lymphoid clusters in the Ribavirin group

Further analyzes of various studies were performed. However, to make the results of these analyzes more meaningful, more significant, the control data were combined so that the sample obtained was larger and larger in size.

Results of integrated analyzes

Analysis of Ribavirin response by relevant efficacy criteria - ALT response during Ribavirin therapy

For integrated efficacy analyzes, the following ALT response definitions shall be used:

Complete response: at the end of the treatment, the normal state returns within the given range.

Partial Response: A 50% or greater reduction in normal patient baseline levels at 1.5 times the upper limit of normal at the end of treatment.

Respondent: Meets the definition of either a full or partial response. The respondent definition is determined as a chart of ALT values over time for a group of patients according to the different type of response definition as used in the study. (Data is presented as a graph dependence, where data is plotted as points by which the third-degree equation is translated using

that a smooth curve is created, Reinsch 1967). Three shall be used definition of answer: a. Full Answer = ALT v normal condition within the given range at the end of treatment b. Partial Answer (A) = 50% or greater reduction

in terms of the usual baseline of the patient at 1.5 times the upper limit of normal at the end of treatment

c. Partial response (B) = 50% or greater reduction in terms of patient's baseline at end of treatment.

All other patients were considered non-respondents. Graphic dependencies were also constructed for non-responders treated with Ribavirin in each study and for placebo patients (both respondents and non-responders) in each study.

The three complete response curves show that the response was achieved after one-third of the treatment period and has continued since that time. The three partial response curves (A) show a similar response representation. The three partial response curves (B) show a markedly greater variability in ALT levels during treatment. The graph dependence for Ribavirin-treated respondents and the graph dependence for placebo patients show, as expected, a variance in the data that does not show changes that would have any definable pattern or profile during treatment and over the following time periods. Accordingly, it can be concluded that the definition of partial response (B) is not appropriate for an integrated efficacy analysis. Given the consistent nature of the graph dependence of the full response and partial response definitions (A) and the fact that these definitions are meaningful clinically, it was decided that for integrated efficacy analysis the response would be defined as either a full response or a partial response ( A).

The following Table 2 shows the results for each of the studies and for the combined database using the ALT response defined above. The differences between the respondents in the two groups of treated patients are compared using either the Chi-square test (quadrilateral method) or the Fischer exact test.

Table 2

Percent ALT Response Rate

study Ribavirin n / N (%) Placebo n / n (%) Value p 92-001 15/28 (54, 6) 1/30 (3.3) <0.001 91-DK-178 11/29 (37.9) 1/29 (3.4) <0.001 CT00-002 32/70 (45.7) 2/36 (5.6) <0.001 Combined database 15/28 (53.6) 1/30 (3.3) <0.001

n = number of patients with ALT response (integrated definition) N = all patients treated (applicable to the context herein) without those patients whose study was not considered valid.

To identify the ALT response corresponding to the end of treatment, this last valid observation returns to a maximum of two visits that successfully continue for those patients who lack such a correct value. This procedure is used in two of the three control studies (91-DK-178 and CT00 / 002). In nine patients in

Ribavirin group and two patients in the placebo group did not find valid observation. ALT response rates were consistent throughout the three control studies and ranged from 37.9% to 53.6%.

%. After combining the data obtained, ALT was used

In all cases, the rates of ALT by Ribavirin were statistically treated with ALT responses of 5.6%;

45.7%.

in patients excellent, v

In placebo response ‘response significantly placebo. consistently low, and the treatments treated with the group were in the range of 3.3 patients at a rate% to 4.2% after combining the data obtained.

ALT response in the reporting period

Table 3 summarizes the steady-state response rates in studies 92-001 and 91-DK-178 and in the two combination studies. Individual study, analytical, ALT definitions of persistent steady-state response proposed in accordance with the projected intended diagram are used. A sustained steady-state responder is basically a patient with either a normalized ALT response or a partial response at the end of the treatment that meets both of the criteria listed here during the reporting period. It is not possible to provide the same analysis for study CT00 / 002, since only some patients can obtain complete ALT data for the entire course of the study period.

Table 3

Percentage rate of ALT persistent steady-state response - according to logged definitions

study Ribavirin n / N Placebo n / N% 92-001 1/15 (6.7) 0/1 (0.0) 91-DK-178 2/11 (18.2) 0/1 (0.0) Combined database 3/26 (11.5) 0/2 (0.0)

n = number of patients with ALT with a sustained steady-state response

N = all patients with full or partial response at end of treatment (study, analytical, proposed definitions in accordance with projected intended diagram)

According to the analyzes in the two studies, the sustained steady-state response rates were 6.7% and 18.2% for Ribavirin-treated patients, compared to 0% for placebo-treated patients. Due to the small sample size and very low number of placebo responders, statistical analysis was not used.

Improving Knodell scoring

For each of the three controls. Studies show a clear trend in favor of Ribavirin, with changes in overall Knodell scoring and many other scoring components. The same trend is evident when combining data from the three studies reported. This trend applies not only to the scoring improvement but also to the deterioration, which is indicated even if no improvement is observed in any of the two treatment groups, with worsening observed in a few patients in the Ribavirin group. This is a very important observation given that one of the goals in the treatment is to prevent the worsening of the chronic and progressively developing condition. Analysis of the combined data by CMC by the χ-square test (Chi-square test) showed no statistically significant differences. Analysis of the combined data by analysis of variance (as used in studies 92-001 and 91-DK-178) shows a significant difference in favor of Ribavirin for overall Knodell scoring, but not for scoring any of the individual components.

The liver histology data was further examined by covariance analysis, using the Knodell scoring baseline as a covariant. Baseline regression analysis of Knodell scoring versus end treatment scoring for all Ribavirin and placebo treated patients showed that the slope, the slope, is less than 1.0 but greater than zero. This indicates that the baseline of Knodell scoring influences the presumption (expectation) of the outcome of the treatment, regardless of any differences between Ribavirin and placebo. Where noticeably different from

0.1, there is a more suitable test than variance analysis (Fisher 1951). The result of the covariance analysis is shown in Table 4 below. Mean scoring changes for Ribavirin-treated patients are small, but due to the small variance, the placebo differences are statistically significant. It is interesting to note that the improvement was not only noticeable in Knodell's sub-scoring regarding fibrosis and that there was less deterioration in the Ribavirin group compared to the placebo group.

Table 4

Comparison of Ribavirin and placebo according to changes in Knodell scoring from baseline to end of treatment covariance analysis using Knodell scoring as covariant, taking into account all phases of the three combined studies

Mean change in scoring from baseline to end of treatment Knodell component Ribavirin N = 107 Placebo N = 78 Value p Periportal activity and necrosis - 0,40 - 0,01 0,000 .4 Portal inflammation - 0,30. - 0,10 0.020 6 Lobular necrosis - 0, 33 -0.13 0,001 9 fibrosis + 0,07 + 0.25 0,007 1 Overall scoring - Mi - 0.05 0,009 9

Investigation of correlation between ALT response and improvement of Knodell scoring

In each of the three control studies mentioned there is

Ribavirin significantly more effective than placebo, at normalized and decreased levels of ALT response (elevated serum ALT level is a biochemical indicator of hepatitis inflammation). Here, too, there is a correlation between response to Ribavirin therapy in terms of normalization or reduction of ALT levels among patients, and according to improvement in liver histology as determined by Knodell scoring. It has been found that there is a truly consistent trend towards a positive relationship between the ALT response and the improvement of Knodell scoring and that both of these parameters are valid as a decisive method, i. the method that is critical to the assessment. In the combined database, this trend is statistically significant for Knodell scoring (p = 0.008), fibrosis (p = 0.014) and portal inflammation (p = 0.022) using the Cochran-Mantel-Haenszel (CMH) Chi-square test. (Table 5,6,7)

Table 5

Knodell's response status - overall stage

ALT answers

respondent Nerespondent protocol The answer n O, o n O, Ό P-value ' 92-001 improved 7 46.7 4 33, 3 0,099 unchanged 7 46, 7 3 25.0 impaired 1 6.7 5 41.7 91-DK-178 improved 10 90.9 7 38.9 0,021 * unchanged 1 9.1 5 27.8 impaired 0 0, 00 6 33, 3 CTOO-O02 improved 11 42, 3 12 48.0 0, 171 unchanged 11 42, 3 5 20.0 impaired 4 15.4 8 32.0 integrated improved 28 53.8 23 41.8 0.008 * unchanged 19 36, 5 13 23, 6 impaired 5 9.6 19 34, 5

N = n

number of patients in the study number of patients responding in the category% = (n / N) * 100 ² ΟΜΗ statistics * P <Ο, 05

Table 6

Response status according to Knodell evaluation - fibrosis according to

ALT answers

respondent Nerespondent protocol The answer n about 0 n Q. o P-value ' 92-001 improved 2 13.3 1 0.00 0, 434 unchanged 12 80.0 11 91.7 impaired 1 6.7 1 8.3 91-DK-178 improved 5 45, 5 2 11.1 0, 191 unchanged 4 36.4 11  61.1 impaired 2 18.2 5 27.8 CT00-002 improved 6 23, 1 1 4.0 0, 120 unchanged 18 69.2 20 80.0 impaired 2 7.7 4 16, 0 integrated improved 13 25.0 3 5, 5 0.014 * unchanged 34 65, 4 42 76, 4 impaired 5 9.6 10 18.2

N = number of patients in study n = number of patients responding in% = (n / N) * 100

YMH statistics * P <0.05

Table 7

Knodell's Response Status - Portal Inflammation by ALT Response

respondent Nerespondent protocol The answer n O o n O. o P-value ' 92-001 improved 5 33, 3 3 25, 0 0,220 unchanged 9 60, 0 5 41.7 impaired 1 6, 7 1 33.0 91-DK-178 improved 5 45, 5 4 22.2 0,227 unchanged 6 54.5 11 61, 1 impaired 0 0.00 3 16, 7 CT00-002 improved 5 19.2 6 24.0 0, 159 unchanged 21 80, 8 16 64.0 impaired 0 0, 00 3 12.0 integrated improved 15 28.8 13 23, 6 0,022 * unchanged 36 69.2 32 58.2 impaired '1 1, 9 10 18.2 and

N = number of patients in the study n = number of patients responding in% = (n / N) * 100 ^Χ ΟΜΗ statistics * P <0.05

The association between ALT response and improvement in liver histology is further studied,. to quantify the improvement in liver histology in patients responding to Ribavirin and to determine if there is a subset of patients from which a more substantial clinical benefit can be derived from Ribavirin treatment. For each of the three observed groups, ALT responses in Ribavirin-treated respondents and non-responders are compared for the mean change in overall Knodell scoring throughout the treatment. Thus, the analysis shown in Table 8 below quantifies the direct changes according to the data in Table 5.

It can be seen that in each study, there is a correlation between ALT response and improvement in liver histology, with responder ALT responses associated with a greater improvement in liver histology compared to ALT response of non-responders. This effect is surprisingly noticeable especially in study 91-DK-178, where the mean reduction in Knodell scoring is 4.7 for ALT respondents compared to 0.17 for ALT non-responders. A regression model is used to investigate the relationship between the ALT response and changes in Knodell scoring. This showed that in all three studies

ALT responses showed an improvement in liver histology, the reduction may be a significant prediction with respect to

So ALT normalization or correlate with patients in patients in a similar way possible clinical hepatitis in which terms.

by improving the histology of liver C treated with Ribavirin, an ALT response was also obtained to derive substantial benefit

Ribavirin, did not show a clinically relevant response compared to the patients treated with ALT, and there was no benefit in patients receiving placebo.

Table 8

study CT00-002 92-001 91-DK-178 Combined database Duration treatments (week) 24 36 48 Mean Knodell change (range) for ALT respondents -1.23 (- 7.4) n = 26 -1.47 (-8.2) n = 15 -4.09 (-9.0) n = 11 -1.90 (-9.4) n = 52 Mean Knode11a change (range) for ALT non-responders -0.96 (-9.4) n = 25 . +0.47 (-3.6) n = 12 -0, 17 (6,7) n = 18 -0.36 (-9.7) n = 55 P-value 0,004 0.000 1 0,002 0.000 1 Mean change value Knodell (range) in all aviribavirin patients -1.10 (-9.4) n = 51 -0, 56 (-8.6) n = 27 -1.65 (-9.7) n = 29 -1.11 (-9.7) n = 107 Mean Knodell change (range) in all placebo patients -0, 09 (-4.4) n = 23 + 0.44 (-3.5) n = 27 -0, 52 (-8.4) n = 27 -0, 51 (-8.5) n = 77 P-value NS NS NS 0.01

* ALT response was defined as normalization at end of treatment or reduction of 50% or more relative to baseline up to 1.5 times above normal at end of treatment ** Regression analysis

In summary, the Phase III program for Ribavirin in chronic hepatitis C consists of three randomized, two blind, placebo-controlled, parallel group studies. A total of 134 patients receiving Ribavirin and 97 patients receiving placebo were randomly selected. The response to the treatment is determined using the following parameters:

1. Reduction or normalization of ALT level (ALT elavation is a biochemical indicator of hepatitis inflammation)

2. Improved liver histology as reported by reduced Knodell scoring (Knodell scoring system quantifies the degree of liver damage by assigning points to different relevant microscopic characteristics and then summarizing these subset of points to obtain a total score)

3. Eliminate hepatitis C virus from the blood or reduce the amount of virus present.

Of the 134 patients in the Phase III studies, 101 were complete with data on ALT level, Knodell scoring, and virus level. Of these 101 patients, 24 patients met the criteria for optimal clinical response to Ribavirin therapy. These criteria are normalized or clinically significantly reduced according to the ALT level and Knodell scoring is also reduced for two or more points. For these 24 patients, a clinical response was obtained without concomitant reduction in blood virus levels. Table 9 below summarizes data on ALT level and Knodell score for 24 patients compared to the remaining 77 patients with lower response rates.

Table 9

respondents N = 24 Nerespondenúí N = 77 Mean baseline ALT (range) for U / L 142.5 (52-269) 176.9 (35-629) Mean end of ALT treatment (range) 'U / L 36.8 (21-62) 91.7 (13-286) Mean Knodell scoring reduction (range) 4.1 (2-9) 0.15 (-7 to +9)

* The upper limit or normal state is 40 U / L

Specific embodiments and applications of nucleosides, nucleotides and analogs thereof have been discovered and disclosed herein. However, it is believed that those skilled in the art are aware of many possible modifications in addition to those described herein, without thereby affecting the inventive method herein. The subject matter of the present invention is therefore not so limited and is exemplified by the following claims. However, both the interpretation and the claim may be further interpreted in the widest possible manner consistent with the context herein. In particular, the terms consisting and comprising may be interpreted as referring to elements, components or steps and steps in a non-exclusive manner, indicating that they refer to elements, components or steps and steps that may be present or used or combined with other elements , components or steps and steps, and may not be mentioned or referenced directly.

Industrial usability

The compounds of the present invention are intended pharmaceutical compositions and are therefore suitable in the pharmaceutical industry. ¹

Claims (54)

PATENT CLAIMS 1. A compound of formula I: wherein R is H, PO ₃ ^2- , (PO ₃ ) 2 ³ or (PO ₃ ) group. 2. A compound of formula IX: 3. Compound of formula (X): (X) wherein W is -N (R ¹⁾ (R ^2), or = NR ^1, wherein R ¹ and R ² are independently hydrogen, linear alkyl, branched alkyl, alkenyl, alkynyl, aralkyl, aralkenyl, aryl or arylkynyl. A compound of formula X according to claim 3, wherein R ¹ and R ² are nitrogen, oxygen, sulfur or halogen. A method of treating a viral infection in a patient, comprising administering a composition comprising the compound of claim 1, claim 2, claim 3 or claim 4 in a dose effective to inhibit the development of the viral infection. The method of treatment of the viral claim 5, wherein the mg / day to 500 mg / day. infection that the patient's dose is in the range of 50 A method of treating a viral claim 5, wherein: 500 mg / day. mg / day to 2 The method of claim 5, wherein the dose is a patient according to a range of 500 viral treatments characterized in that the viral moiety consisting of H'IV HBV infection, RSV infection, an infection selected from infection, infection, and para-influenza virus infection. The patient's infection is an HCV influenza viral infection The method of treating a viral infection of claim 5, wherein the cytokine is co-administered. in a patient to a patient according to below The method of treating a viral infection of claim 9, wherein the patient's cytokine is interferon. 11. A method of treating a viral claim 10, wherein: 12. A method of treating viral claim 5, wherein Ribavirin. A compound according to claim 1, 14. A method of treatment comprising a compound effective to attenuate the viral concurrently according to claim 13 of developing a viral infection in a patient according to said interferon being alpha-2b. infection in a patient according to the invention, wherein R is PO3 ² '. infection in a patient, administers the composition at a dose that is an infection. A compound according to claim 1, wherein R is (PO ³ ) ₂ ³ · 16. A method of treating a viral infection in a patient, comprising administering a composition comprising a compound of claim 15 at a dose effective to inhibit the development of the viral infection. The compound of claim 1, wherein R is (PO 3) 3 ^3- . A method of treating a viral infection in a patient, comprising administering a composition comprising a compound of claim 17, at a dose effective to inhibit the development of the viral infection. 19. A method for increasing the selectivity of a pharmaceutical active molecule with respect to a pharmaceutical effect on target cells, characterized in that a drug substance is provided, wherein the drug substance is a Ι-β-L-ribofuranosyl-1,2,4-triazole-3-carboxamide; the drug substance is modified by a modifying group, wherein the modifying group is covalently attached to the drug substance by a nitrogen atom; wherein the modified moiety is enzymatically removed from said drug in a target cell. 20. A method of increasing the selectivity of a pharmaceutical active molecule with respect to a pharmaceutical effect on target cells, wherein the modifying group is = NH, wherein the modifying group is covalently attached to a carbonyl atom in the drug substance. A method of increasing the selectivity of a pharmaceutically active molecule with respect to a pharmaceutical effect on target cells, wherein the modifying group is N (R ¹ ) (R ² ) or = NR ¹ , wherein R ¹ and R ² are independently hydrogen, linear alkyl, branched alkyl, alkenyl, alkynyl, aralkyl, aralkenyl, aralkynyl or aryl. A method of increasing the selectivity of a pharmaceutically active molecule with respect to a pharmaceutical effect on target cells, wherein R ¹ and R ² independently contain a nitrogen atom, an oxygen atom, a sulfur atom or a halogen atom. 23. A method of treating a disease characterized by inflammation of an organ of a patient, the method comprising: provides a compound according to claim 1, claim 2 or claim 3; and the compound is administered to a patient at a dose that (a) causes systemic immunomodulation and not systemic immunosuppression of the type 1 and type 2 response, and (b) causes systemic immunosuppression of the type 1 and type 2 response in the patient's organ. 24. A method of treating a disease according to claim 23, wherein the compound is a compound according to claim 1. 25. A method of treating a disease according to claim 23, wherein the compound is a compound according to claim 2. 26. A method of treating a disease according to claim 23, wherein the compound is a compound according to claim 3. 27. A method of treating a disease according to claim 23, wherein the organ is a liver infected with a virus. 28. The method of treating a disease according to claim 23, wherein the virus is HCV virus. 29. A method of stimulating neuronal growth, characterized in that it has been found that a compound of formula XI Y wherein Y is β-β-L-ribofuranosyl, β-β-L-ribofuranosyl-5-phosphate, β-β-L-ribofuranosyl-5-diphosphate, β-β-L-ribofuranosyl-5-triphosphate, β-β- D-ribofuranosyl, β-β-D-ribofuranosyl-5-phosphate, 1βD-ribofuranosyl-5-diphosphate, or β-β-Dribofuranosyl-5-triphosphate, is effective to stimulate neuronal growth within a given concentration range; and exclusively neurons with the compound within a given concentration range. 30. The method of stimulating neuronal growth of claim 29, wherein the compound is Ribavirin. 31. The method of stimulating neuronal growth of claim 29, wherein the compound is phosphorylated Ribavirin. 32. The method of stimulating neuronal growth of claim 29, wherein the compound is Levovinn. 33. The method of stimulating neuronal growth of claim 29, wherein the compound is phosphorylated by Levovirin ™. 34. The method of stimulating neuronal growth of claim 29, wherein the compound is administered to a patient as part of the treatment of a disease at a dosage range effective to increase the Type I response and reduce the Type II response in the patient. 35. A method of stimulating neuronal growth according to claim 29, wherein said method is designed to target unipolar nerve cells in neurons. 36. The method of stimulating neuronal growth of claim 29, wherein said method is directed to target bipolar nerve cells in neurons. 37. The method of stimulating neuronal growth of claim 29, wherein the neurons are part of a neuronal tissue comprising at least four of the following cell types: astrocytes, dendrocytes, myelin sheath cells, supporting nerve tissue cells, unipolar nerve cells, bipolar nerve cells , multipolar nerve cells and receptor cells. 38. An antiviral medicine, comprising: a first compound having a direct and indirect antiviral effect; and a second compound that enhances the overall antiviral effect, wherein the total antiviral effect comprises a direct and indirect antiviral effect, and wherein the second compound specifically binds a hapten selected from the group consisting of a viral protein and a cytokine. . The antiviral medicament of claim 38, wherein the first compound comprises a nuclezide analog. The antiviral medicament of claim 38, wherein the nucleoside analog is Ribavirin. The antiviral medicament of claim 38, wherein the nucleoside analog is Levovirin ™. The antiviral medicament of claim 38 or claim 39, wherein the nucleoside analog is in the form of a prodrug. 43. The antiviral medicament of claim 38, wherein the direct antiviral effect comprises inhibiting viral replication. 44. The antiviral medicament of claim 38, wherein the direct antiviral effect comprises a type 1 / type 2 equilibrium shift toward the type 1 response. 45. The antiviral medicament of claim 38, wherein suppression of the type 1 and type 2 responses is included in the direct antiviral effect. 46. The antiviral medicament of claim 38, wherein the second compound comprises an antibody. 47. The antiviral medicament of claim 38, wherein the antibody is selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a synthetic antibody, and an antibody fragment. 48. The antiviral medicament of claim 38, wherein the viral protein is a virus protein selected from the group consisting of HIV virus, hepatitis virus, influenza virus, parachrip virus, and RSV virus. 49. An antiviral characterized transcriptase. medicament in that the viral composition according to the protein of claim 38 is reverse) 50. An antiviral characterized in that the composition according to the invention is a cytokine of the type of claim 38 First 51. An antiviral characterized by a synergistic effect. claim 38, the compound has a medicament according to that the first compound and the second 52. The antiviral medicament of claim 38, wherein at least one of the first compound and the second compound is selectively accumulated in the organ. 53. The antiviral medicament of claim 38, wherein the organ is selected from the group consisting of liver and brain. 54. The antiviral medicament of claim 38, wherein the first compound is Ribavirin and the second compound is an antibody that specifically binds a type 2 cytokine.