A method for the treatment of infections caused by hepatitis b virus

 

(57) Abstract:

The invention relates to medicine. A method for treatment of HBV infection, which consists in the introduction of an effective amount of enantiomerically pure D-deoxyadenylate formula 1.

< / BR>
where R is OH, Cl, NH2or H, and X is chosen from the group comprising alkyl, acyl, monophosphate, diphosphate and triphosphate. The method inhibits DNA replication HBV virion. 18 C. and 7 C.p. f-crystals, 3 tab., 6 Il.

The present invention relates to a method for the treatment of infections caused by hepatitis b virus (also denoted "HBV"), which consists in introducing an effective amount of one or more active compounds disclosed in this application, or a pharmaceutically acceptable derivative or procarcinogen predecessor of one of these active compounds.

HBV is the second after Smoking a cause of causing cancer in humans. The mechanism of induction of cancer by the hepatitis b virus is still unknown, although it has been suggested that this virus can stimulate tumor development, both directly and through chronic inflammation, cirrhosis and cell regeneration associated with denhaut level epidemics. After 2-6 months of the incubation period, during which the infected person is not aware of the existence of HBV infection, this infection can induce the phenomenon of acute hepatitis and liver, causing abdominal pain, jaundice, and increased levels of certain enzymes in the blood. HBV can also cause fulminant hepatitis, which progresses rapidly and often turns into a lethal form of the disease with extensive areas of liver damage.

Patients suffering from acute viral hepatitis usually recover. However, in some patients for a long or indefinite period of time maintain high levels of viral antigen in the blood, causing chronic infection. These infections can lead to chronic persistiruuschem hepatitis. Disease chronic persistent hepatitis is most prevalent in developing countries. For example, by mid-1991 only in Asia, there were 225 million carriers of HBV, and worldwide there are nearly 300 million. Chronic persistent hepatitis can cause fatigue, cirrhosis and malignant hepatoma, i.e., primary liver cancer.

In Western industrialwaste blood. In fact, the epidemiology of HBV infections is very similar to the epidemiology of acquired immunodeficiency syndrome, which is quite explains the wide spread of HBV infection among patients with AIDS, or HIV-infected patients. However, HBV is more infectious virus than HIV.

To immunize patients against HBV was developed a vaccine based on human serum. This vaccine was produced through genetic engineering, and although this vaccine has been very effective, however, when it is producing followers faced a number of serious problems associated with the limited supply of human serum from chronic carriers of HBV infection, as well as with the duration and complexity of the cleanup procedure. In addition, each batch of vaccine produced from sera of different sources should be tested on chimpanzees to guarantee its security. Moreover, this vaccine is ineffective for patients already infected with the virus.

Good results were also obtained with a daily injection of interferon, a genetically engineered protein. Currently, however, has not yet found such a pharmaceutical srecu the spread of HBV infection worldwide reaches the level of the epidemic and often leads to tragic consequences for infected patients, getting a new effective pharmaceutical agents to treat hepatitis in humans, which would have low toxicity, remains a critical problem.

Therefore, another aim of the present invention is to develop a method and composition for treatment of humans or other animals infected with HBV.

In a preferred variant of its implementation of the present invention relates to a method for the treatment of humans infected with HBV, which consists in the introduction of an effective amount of enantiomers clean-D - DIOXOLANYL-nucleoside of the formula:

< / BR>
where R represents OH, Cl, NH2or H, or its pharmaceutically acceptable salt or derivative, optionally in combination with a pharmaceutically acceptable carrier or diluent. The compound in which R is chlorine, represents the (-)-(2R, 4R)-2-amino-6-chloro-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl) purine. The compound in which R is hydroxy, a is a (-)-(2R, 4R)-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl) guanine. The compound in which R is amino, represents hydrogen, represents a (-)-(2R, 4R)-2-amino-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl] purine. The absolute configuration of these compounds has not been determined by the methods of crystallography. The above designations of the compounds were given on the basis of comparison of their structure with the configuration source of sugar that is used to obtain the connection. In another embodiment of the present invention, the patient is administered an effective amount of an enantiomer - L-deoxyadenylate, or racemic mixture of L - and D-deoxyuridylate.

It was found that in the case of ACPD ((-)-(2R,4R)-2-amino-6-chloro-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl)purine) and DAPD ((-(2R-4R)-2-amino-9-[(2-hydroxymethyl)-1,3 - dioxolane-4-yl] adenine)) EC50for inhibition of replication intermediates of HBV - DNA for the synthesis of HBV virion is about 0.1 μm. Test compounds DAPD, ACPD or DG - (- (- (2R, 4R)-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl] guanine) in concentrations up to 300 μm in 2.2.15 cells appreciable cytotoxicity was not observed. In the count of clonogenic assays the three peninsulaand turned out to be largely non-toxic to myeloid and erythroid cells (IC50= from 50 to more than 100 compared to AZT, for which IC50is 1 μm), Satie in concentrations up to 1 mm, not inhibit the enzymes involved in the biosynthesis of purine and pyrimidine, for example, as well as adenoidectomies, purine-nucleosidases, hypoxanthineguanine-phosphoribosyltransferase, adenosines, insinking, citizencane, xanthine oxidase, aldehyde oxidase and antiaging.

Disclosed in this application - -DIOXOLANYL, their pharmaceutically acceptable derivatives or salts, or pharmaceutically acceptable compositions containing these compounds, can be used for the prevention and treatment of HBV infections and other related conditions such as status test positive for antibodies against HBV and HBV - positive, chronic liver inflammation caused by a virus HBV, cirrhosis, acute hepatitis, fulminant hepatitis, chronic persistent hepatitis, and fatigue. These compounds or compositions can also be used for preventive purposes to prevent or slow the progression of the disease in individuals with a positive response to HBV antigen or antibody against HBV, or individuals exposed virus HBV.

In one of the variations is W ill result one or more active compounds of the present invention alternately with one or more other anti-HBV. Examples of such agents against HBV, which can be used in alternating therapy include, but are not limited to, enantiomer or racemic mixture of 2-hydroxymethyl 5-(5-ferocity-1-yl)-1-3-oxathiolane (FTC, see WO 92/147443), its physiologically acceptable derivative or physiologically acceptable salt, (-)-enantiomer or radamiejska a mixture of 2 hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane (also referred to as BCH-189 or ZTS, see publication EPA N 0382526 and WO 91/17159 respectively) of its physiologically acceptable derivative, or a physiologically acceptable salt, enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU) enantiomer or racemic mixture of 2'-fluoro-5-ethyl-arabinosyl-uracil (FEAU), carbovir, or interferon.

For the treatment of patients can be used any method of alternating therapy. Examples of such methods (which, however, do not limit the scope of the present invention are the following scheme of administration of drugs, within 1-6 weeks is administered an effective amount of one drug, and then within 1-6 weeks is administered an effective amount of a second drug. The scheme of the sequential administration of drugs may also be implementation of the present invention the active compound, its derivative or its salt can be introduced in combination with another agent against virus HBV, including the above tools. Basically, when alternating therapy effective dose of each agent against virus HBV is injected periodically, and in adjuvant therapy, low dose of two or more antiviral agents are administered together, in combination with each other. The dose of antiviral agents depend on the speed of their absorption, inactivation and excretion, as well as other factors well known in the art. For example, doses of drugs may also vary depending on the severity of the patient's condition. In addition, it should be noted that for each individual patient must be installed individual regimen medicines, adjusted in accordance with the needs of the patient, and this individual scheme should be carried out under the supervision of a doctor.

A brief description of the drawings.

Fig. 1 illustrates a method of obtaining enantiomerically pure D-deoxyuridylate.

Fig.2 is a graph illustrating the influence of peridiocally and AZT on the formation of erythroid colonies the AI of the test drugs (AZT, 3 ashdodite-thymidine, APD, (-)-(2R, 4R)-2-amino-9-[(2-hydroxymethyl)-1,3 - dioxolane-4-yl] purine, ACPD, (-)-(2R, 4R)-2-amino-6-chloro-9-[(2 - hydroxymethyl)-1,3-dioxolane-4-yl]purine, DG, (-)-(2R, 4R)-9-(2 - hydroxymethyl)-1,3-dioxolane-4-yl]guanine, DAPD, (-)(-(2R,4R)- 2-amino-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl]adenine).

Fig.3 is a graph illustrating the influence of peridiocally and AZ on the formation of colonies of cells precursors of granulocytes and macrophages, measured as % of control cells, depending on the logarithm of the concentration of the test drug. Abbreviations used in this figure are defined above for Fig.2.

Fig. 4 is a graph illustrating the percentage of inhibition of HBV DNA replication in 2.2.15 cells on day 9, depending on the different concentrations of the tested compounds. Abbreviations used in this figure are defined above for Fig.2 ((-)-PTC means (-)-2-hydroxymethyl-5-(5-ferocity-1-yl)-1,3 oxathiolan). Cm. the table for the corresponding data.

Fig.5 is a graph illustrating the absorption of 5 μm tritium-labeled (-)-(2R, 4R)-2-amino-9-(2-hydroxymethyl)-1,3-dioxolane-4-yl)Adelina (DARD) in Hep2G cells. The extract was obtained 4 hours after DAPD exponere absorption 5 μm tritium-labeled (-)-(2R,4R)-2-amino-9-[(2-hydroxymethyl)-1,3 - dioxolane-4-yl] adenine (DAPD) in cells Ners. The extract was obtained 12 hours after DAPD-exhibiting cells. 1000 number of decays/min, injection 145 μl.

Used in the present description, the term "enantiomerically pure" refers to a nucleoside composition that contains at least about 95%, and preferably 97%, one enantiomer of this nucleoside.

The invention disclosed in this application relates to a method and composition for the treatment of HBV infection in humans and other animals, while this method is the introduction of an effective amount of one or more of the above compounds or their physiologically acceptable derivatives, including 5' - and/or N6alkyl or etilirovannym derivatives or their pharmaceutically acceptable salts, optionally in a pharmaceutically acceptable carrier. Compounds of the present invention have activity against virus, HBV, or they are metabolic precursors of these compounds and may be transformed into compounds with antiviral activity.

In another embodiment, its implementation of the present invention relates to a method of treating patients infected with HBV, namely, Thu is isopycnic enantiomerically pure - D-DIOXOLANYL - polynucleotides. Used in the present description, the term "Proletarskiy predecessor" refers to pharmaceutically acceptable derivative of the above nucleoside, i.e. the specified drug precursor can turn into a specific nucleoside of the present invention after administration in vivo or the precursor itself may or may not have antiviral activity. As examples, not limiting the scope of the present invention can serve as pharmaceutically acceptable salts (sometimes referred to as "physiologically acceptable salts"), 5' and N6-acylated or alkylated derivatives of the active compounds (also denoted "physiologically or pharmaceutically acceptable derivatives"). In one embodiment of the present invention the acyl group is an ester of carboxylic acid, where decarbonising part of the ester groups are selected from straight, branched or cyclic C1-C20of alkyl, alkoxyalkyl, such as methoxymethyl, aralkyl, such as benzyl, aryloxyalkyl, such as phenoxymethyl, aryl, for example phenyl, optionally substituted with halogen, C1-C4-alkyl or C1-C4

Used in the present description, the term "alkyl" means, but is not limited to: methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, amyl, tert-pentyl, cyclopentyl and cyclohexyl.

Used in the present description, the term "acyl" means, but is not limited to: acetyl, propionyl, butyryl, pentanoyl, 3-methylbutyryl, bisaccia, 3-chlorobenzoate, benzoyl, acetyl, pivaloyl, mesilate, propionyl, valeryl and groups, Caproic, Caprylic, capric, lauric, Moratinos, palmitic, stearic and oleic acids. These nucleosides can also be obtained in the form of 5'-esterified lipids, is described in the following works: Kucera, L. A., M. Lyer, E. Leake, A. Raben, Modest E. J., D. L. W., H. C. Piantadosi. 1990. Novel membrane - interactive ether lipid analogs thah inhibit infections of HIV-1 production and induce defective virus formation. AIDS Res Hum Retroviruses. 6-491-501; Piantadosi, C.,T. C. Marasco . J., S. L. morres-Natschke, K. L. Meyer, F. Gumus, J. R. Surles, K. S. Ashaq, L. S. Kucera, N. Lyer, C. A. Wallen, S. Piantadosi, and E. J. Modest 1991 - Synthesis and evaluation of novel ether lipid nucleoside conjugates for anti - HIV activity. J. Med. Chim 34:1408 - 1414; Hostetler, K. Y. D. D. Richman, D. A. Carson, L. M. Stuhmiller, G. M. T. van Wijk and H. van den Bosch. 1992. Greatly enhanced inhibition of human immunodeficiency virus type 1 replication in CEM and HT4-6C cells bu 31-deoxythman. 1990. Synthesis and antiretroviral activity of phospholipid analogs of agidothymidine and other antiviral nucleosides. J. Biol. Chem. 265:6112-6117.

- Deoxyadenylate can be converted into a pharmaceutically acceptable ester by reaction with an appropriate etherification agent, such as galogenangidridy or anhydride.

Nucleoside or its pharmaceutically acceptable derivative can be converted in their pharmaceutically acceptable salt in a standard way, for example by treatment with an appropriate base. Ester or salt can be, in turn, converted into the original nucleoside, for example, by hydrolysis.

The active compound can be obtained in the form of pharmaceutically acceptable salts. Used in the present description, the term "pharmaceutically acceptable salts or complexes" refers to salts or complexes of nucleosides that retains the desired biological activity of the parent compound and do not have or have minimal toxic effects. As examples of such salts is not limiting, however, the scope of the present invention) may be: (a) acid additive salts formed with inorganic acids (e.g. hydrochloric acid, bromododecane key is Tami, such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamula acid, alginic acid, polyglutamine acid, naphthalenesulfonate acid and polygalacturonase acid, (b) primary additive salts formed with cations such as sodium, potassium, zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, Nickel, cadmium, etc., or organic cations such as N,N-dibenzyl-ethylene-diamine, ammonium, or Ethylenediamine; or (C) a combination of salts (a) and (b), for example, cannot zinc, etc.

Modifications of the active compound, and in particular, N6- and 5'-O-positions can affect the bioavailability and rate of metabolism of the active compound and thereby to provide flexible delivery of active ingredient to the desired tissues or organs.

The active compound or its pharmaceutically acceptable derivative or salt can also be mixed with other active materials that do not have an adverse effect on the activity of the compounds or materials that provide additional desirable effect, such as anteprime anti-virus HBV or HIV.

Enantiomerically pure-D-dioxolane-nucleosides can be obtained by the method, described in detail below and are also described in PCT /US 91/09124. This method involves first obtaining (2R,4R) - and (2R,4S)-4-acetoxy-2-(protected oxymethyl)-dioxolane from 1,6-anhydromannose, i.e. sugar, which contains all the necessary stereoisomers to obtain enantiomerically pure final product, including diastereomer with the correct configuration at 1 - position of the sugar (which is 4'-position in the formed then the nuke).

Then (2R, 4R) and (2R,4S)-4-acetoxy-2-(protected oxymethyl)-dioxolane condense with the appropriate heterocyclic base in the presence of SnCl4another Lewis acid, or trimethylsilyltrifluoroacetamide in an organic solvent, such as dichloroethane, acetonitrile or methylenchlorid, resulting in a gain stereochemical net deoxyadenylate.

However, it should be borne in mind that when you obtain enantiomerically pure DIOXOLANYL need to ensure that the reaction conditions were not strong acid as the strong acid can help break dioxolane ring. The above reaction should provoditsya if this is the reaction time should be minimized.

Racemic mixture of deoxyadenylate can be obtained as described in EPA publication 0382526, - L-enantiomer can be isolated from racemic mixtures of known methods, for example by a method involving the use of chiral HPLC columns.

The method of obtaining active compounds is illustrated in Fig.1 and in example 1. Starting material (compound 1) was obtained in the manner described in PCT/US 91/09124 (in this application the compound 8). 2,6-disubstituted purine derivatives were synthesized by condensation of acetate with 1 similarbank 6-chloro - 2-porporino, resulting in the received mixture / = 1/1,3 isomers 2 and 3. Initially, the N7-isomer was again transformed in the N9-isomer, stirring over night at room temperature. An analytical sample was obtained by separation of mixtures into individual isomers 2 and 3 using preparative TCX using CH2Cl2- acetone (19:1) as developing solvent. However, to obtain the final products 10-15 mixture of isomers 2 and 3 were treated with ammonia in DME (Robins M. I; Ugnanski, B. Nucleic acid related - compounds. 34. Non aqueous Diasotisation with tert - butyl nitrite. Introduction of fluorine, chlorine and bromine at C-2 of purine nucleosides - Can. J. Chemistry,

1981, 2608), guanine 8 and 2,6-diamino 9 were obtained by processing the isomer 4. 2-mercaptoethanol NaOme and ammonia in ethanol, respectively. Loose nukes 10-15 were obtained in good yield after processing the corresponding 5'-similarbank nucleoside n-Bu4NF

- isomers 12 and 13 were obtained in the same way as - isomers.

Example 1.

To obtain enantiomerically pure D-deoxyadenylate

(2R, 4R ) and (2R,4S)-9-[(2-(tert-butyldiphenylsilyl)oxymethyl]- 1,3 dioxolane-4-yl]-6-chloro-2-toburen (2 and 3)

A mixture of 2-fluoro-6-chloropurine (of 4.05 g, 23,47 mm) and ammonium sulfate (catalytic amount) in hexamethyldisilazane (940 ml) was heated under reflux for 2 hours. The resulting solution was concentrated in anhydrous conditions, resulting received 2 fluoro-6-globulin in the form of a white solid. Then to a cooled (0oC) and posted the solution similarvideo 2-fluoro-6-chloropurine (5,69 g, 23,69 mm) and compound 1 (7,84 g, 19,57 mm) in dry methylene chloride (175 ml) was added TMS-OTF (to 4.41 ml, 23,44 mm).

The reaction mixture was heated to room temperature and stirred for 16 hours, after which all originally derived condensed N7the product was developed in N9-isomer. Then the reaction was suppressed saturated Suha under reduced pressure. The residue was dissolved in ethyl acetate (200 ml), washed with water and saline, dried with anhydrous sodium sulfate, filtered and evaporated, resulting in the obtained solid residue, after purification with the aid of column chromatography on silica gel (20% E ARS in hexane) was obtained mixture - anomer 8 - anomer 9 (1,3:1, / ) as white solid crystals (6,30 g, 62,8%).

An analytical sample was purified using preparting TCX using CH2Cl2- acetone (19:1) as developing system, and received connection 2 (Rf= 0.50) and 3 (Rf= 0,55) for NMR characterization: UV (Meon)max269,0 nm.

(-)-(2R, 4R)-2-Amino-9-[[2-[(tert-butyldiphenylsilyl)oxymethyl] - 1,3-dioxolane-4-yl] -6-globulin (4), (-)-(2R, 4R)-9[[2- [(tert-butyldiphenylsilyl)oxy] methyl)-1,3-dioxolane-4-yl] -2 - ferdinan (5), (+)-(2R,4S)-2-amino-9-[[2-[(tert-butylbiphenyl - silyl)oxymethyl]-1,3-dioxolane-4-yl]-6-globulin (6), and (+)- (2R,4S) -9[[2-[(tert-butyldiphenylsilyl)oxymethyl]-1,3-dioxolane-4 - yl]-2-ferdinan (7)

Dry ammonia gas was barbotirovany in stirred solution of coupled 2 and 3 (6.25 g 12,18 mm) in ME (125 ml) over night. The solvent is evaporated under reduced pressure, and the residue was subjected to chromatographic separation on a column with silicagel crystalline solid. UV (Meon)max= 309,5 nm. Analysis (C25H28ClN5O3Si), C, H, Cl, n 5 (Rf= 0,21 1.12 g, 18.6 per cent), colorless needle crystals, UV (Meon)Amax= 261,0 268,0 (DM) nm.

Analysis (C25H26FN5O3Si) C, H, F, N, 6 (Rf=0,43, 1,60 g 25,76%), white crystals, UV (Meon)max= 261,0, 269,0 (DM). nm. Analysis (C25H28FN5O3Si) C, H, F, No. 7 (Rf=0,12, 0.96 g, 16%), microcrystalline solid, UV (methanol)max= 261,0, 269,0 (DM) nm.

Analysis of C25H28FN5O3Si) C, H, F, n

(-)-(2R, 4R)-2-Amino-6-chloro-9-[(2-hydroxymethyl)-1,3 - dioxolane-4-yl] purine (10)

A solution of compound 4 (0,47 g of 0.91 mm) in THF (20 ml) was treated with IM n-Bu4NF/THF (1.1 ml, 1.1 mm), resulting in received 10 (Rf= 0,50 0.21 g, 84%) as a crystalline solid, which was recrystallized from Meon: UV (H2O)=307,0 nm ( 8,370) (pH 7), 307,5 ( 8,590) (pH 2), 307,0 ( of 8.800) (pH 11).

Analysis (C9H10ClN5O3) C, H, Cl, N

(-)-(2R,4R)-2-fluoro-9-[(2-hydroxymethyl)-1,3-dioxolane-4 - yl]-adenine (II)

A solution of compound 5 (0.56 g, 1,12 mm) in THF (20 ml) was treated with IM n-Bu4NF/THF (1.35 ml of 1.35 mm), resulting in the received connection 22 (0.24 g, 85%) as a white crystalline solid, which re is 60,8 ( 16,700), 268,5 (DM) ( 13,200) (pH 11).

Analysis (C9H10FN5O3) C, H, F, n

(-)-(2R,4R)-9-[(2-Hydroxymethyl)-1,3-dioxolane-4-yl]guanine (14)

A mixture of compound 4 (0.29 grams, 0,57 mm), H, CH2CH2OH, (0.51 ml) and 1.0 NaOMe/MeOH (of 11.6 ml) in the Meon (20 ml) was heated under reflux for 3 hours. The reaction mixture was cooled and neutralized glacial acetic acid.

Then the solution is evaporated to dryness, the residue was dissolved with CHCl3was filtered , the filtrate was concentrated to dryness and received the crude compound 8 (0.21 g 75%), then without additional purification was subjected to desilicious to obtain compound 3 (0.07 g, 61%) as a microcrystalline solid, which was precrystallization of the Meon: UV (H2O)max= 252,0 ( 8,730) (pH 7), 254,4 ( 12,130), 277,5 (SD.) ( 8,070) (pH 2), 264,3 ( 10,800) (pH 11).

Analysis (C9H11N5O4G, H, n

(-)-(2R, 4R)-2-Amino-9-[(2 hydroxymethyl)-1,3-dioxolane-4 - yl] -adenine (15).

The steel cylinder was loaded connection 4 (0.28 g 0.55 mm) in anhydrous ethanol (20 ml), saturated with ammonia, and heated at 90oC for 6 hours. After cooling, the solvent is evaporated in vacuum and the obtained compound 9 (0.26 g, 95%) was desirerable in accordance with the x acicular microcrystals, which was recrystallized from MeOH: UV (H2O)max= 279,0 nm ( 8,040) (pH 7) 290,0 ( 7,070) (pH 2), much as 278.8 ( 7,580) (pH 11).

Analysis ( C9H12N6O3C, H, n

(-(-(2-R, 4R, )-2-Amino-9-[(2-hydroxymethyl)-1,3-dioxolane-4 yl] purine can be obtained by restoring the connection 10 with the use of a number of reducing agents, including palladium charcoal and hydrogen gas or a hydride of anti-and azobisisobutyronitrile.

Ability - D-dioxolane-nucleosides to inhibit HBV can be measured using different experimental techniques. Used in this experiment analysis to assess the ability of compounds of the present invention to inhibit the replication of HBV is described in detail in the work of Korba and Gerin, Antiviral Res 19: 55-70 (1992). Below, for purposes of illustration and not limiting of the scope of the present invention, the results of the assessment of the toxicity and anti-HBV virus activity of (-)-(2R,4R,)-2-amino-6-chloro-9-[2 - hydroxymethyl)-1,3-dioxolane-4-yl] -purine, (-)-(2R, 4R)-2-amino-9-] 2 - hydroxymethyl)-1,3-dioxolane-4-yl] -adenine, and (-)-(2R,4R)-9- [(2-hydroxymethyl)-1,3-dioxolane-4-yl]guanine. Other compounds of the present invention were evaluated in the same way.

Evaluation of the antiviral activity of the s). All wells plates were seeded with the same density and at the same time.

Because of the natural differences in the levels of intracellular and extracellular HBV DNA statistically significant estimates of P less than 0.05 was considered only the magnitude of inhibition that 3 times (for DNA HBV virion) or 2.5 times (for intermediates of DNA replication HBV) exceeded the average levels for these forms of HBV-DNA in untreated cells (Korba &Gerin, Antiviral Res. 19: 55-70, 1992). The levels of integrated HBV DNA in each cell DNA preparation (which in these experiments remain constant in relation to cell mass) was used to calculate the levels of intracellular forms of HBV DNA, thus avoiding experimental deviations in the block hybridization assays.

Typical values for the levels of DNA HBV virion in untreated cells is in the range from 50 to 150 PG per milliliter of culture medium (average is about 76 PG/ml). The intermediate products of the replication of intracellular HBV DNA in untreated cells is in the range from 50 to 100 PG/μg of cellular DNA (on average, about 74 PG/µg).

Basically, the inhibition of intracellular levels of HBV-DNA in the processing of protiven HBV.

For example, the hybridization results of the analyses carried out for these experiments revealed the equivalence of approximately 1.0 PG of intracellular HBV DNA/μg of cellular DNA for 23 gapimny copies per cell and 1.0 PG of extracellular HBV DNA/ml culture medium for 3 to 105viral particles/ml

Were carried out tests on toxicity to determine whether the observed antiviral effects the overall effect on the viability of the cells. Used for this purpose the method was based on the absorption of the dye (neutral red) and is a standard and widely used analysis of cell viability in different systems virus-host, including HBV viruses (herpes simplex virus) and HIV. The procedure of this analysis is illustrated in more detail in the table below.

Compound used in the form of 40 mm mother solutions in DMSO (frozen on dry ice). For this purpose were made daily aliquot samples and frozen at -20oC so that each aliquot was subjected to only one cycle of freezing and thawing. Then daily subjects aliquots were thawed, suspended ativanyou activity the compounds were tested at concentrations of 0.0 and 1 μm. In tests on the toxicity of the compounds were tested in five koncentracija comprising up to 300 microns.

The following tables have used the following abbreviations: ACPD: (-)-(2R, 4R)-2-amino-6-chloro-9-[(2-hydroxymethyl)-1,3 - dioxolane-4-yl] purine, DAPD: (-)-(2R,4R)-2-amino-9-[(2 - hydroxymethyl)-1,3-dioxolane-4-yl]adenine, and Dioxolane-C: (2R,4R)-9-[(2-hydroxymethyl)-1,3-dioxolane-4-yl]guanine.

Example 2.

The toxicity of compounds

Evaluated the ability of the enantiomers ACPD, DAPD and dioxolane-C to inhibit the growth of the virus in cultures of 2.2.15 cells (cells HepC2 transformed by the virion hepatitis). As shown in table 1, for the tested compounds, taken at the concentrations used to evaluate the antiviral activity, significant toxicity was observed (for untreated cells was observed more than 50% inhibition of the absorption of the coloring). Compound was nontoxic to the 2.2.15 cells at a concentration of 100 μm. At a concentration of 300 μm of the compounds were found to moderate toxicity, however, all three compounds were less toxic at this concentration than ddC.

Analysis of toxicity were carried out in 96-well flat-bottomed tablets for the cultivation of tissues. Clickie with the scheme, used for the analysis of antiviral activity. Each compound was tested at 4 concentrations and culture for each concentration used in three copies. To determine the relative level of toxicity was assessed by the degree of absorption of neutral red. For the quantitative analysis used the optical density of internalized painting at 510 nm (A510). The values obtained were expressed as a percentage of the average values of A50(+ standard deviation) obtained for 9 separate cultures of untreated cells, cultured in the same 96 - well tablets that compound. The percentage absorption of colouring for 9 of the control cultures in 40 tablet was 100 + 3. In these assays for compounds 2', 3'- ddC taken at a concentration of 150-190 μm, was observed 2-fold decrease in the absorption of color (compared to the levels observed for untreated cultures (Korba &Gerin, Antiviral Res. 19:55-70, 1992) (see tab. 1).

Example 3.

Activity against hepatitis b virus

As shown in table 2, the levels of DNA HBV virion and the levels of intermediates of replication of intracellular HBV (HBV R1) within the normal variance remain constant for detoxication 2',3 '- ddC) induced a significant suppression of HBV replication - DNA concentration. Preliminary studies have shown that this analytical system (Korba &Gerin, Antiviral, Res 19:55 70, 1992), at a concentration of 9-12 µm 2', 3' - ddC usually observed 90% suppression of HBV - R1 (relative to the average levels in untreated cells).

All three compound are strong inhibitors of HBV replication, and the degree of suppression of their DNA virion HBV and HBV - R1 comparable to or greater than the degree of suppression observed when using compounds 2', 3' - ddC.

Example III

Toxicity for erythroid progenitor cells (BF U-E) person

Fig. 2 is a graph illustrating the influence of the number of selected peridiocally and AZT on the formation of colonies of erythroid progenitor cells (BF U-E) of a person, measured as % of control cells, depending on the concentration (μm). As can be seen from the figure, the four investigated deoxyadenylate, namely APD, (-)-(2R,4R)-2-amino-9-[(2-hydroxymethyl)-1,3 - dioxolane-4-yl]purine, ACPD,

(-)-(2R, 4R)-2-amino-6-chloro-9-[2-hydroxymethyl)-1,3 - dioxolane-4-yl]purine, DG (-)-(2R,4R,)-9-[(2-hydroxymethyl - 1,3-dioxolane-yl]guanine, and DAPD, (-)-(2R, 4R)-2-amino-9-[(2 - hydroxymethyl)-1,3-dioxolane-4-yl]adenine, was found significantly MNI precursor cells of granulocytes and macrophages man

Fig. 3 is a graph illustrating the effect of ACPD, DG, DAPD, APD and AZT on the formation of colonies of precursor cells of granulocytes and macrophages person, measured as % of control cells, depending on the logarithm of the concentration of the test drug. As can be seen from the figure, unindictability was found significantly less toxicity, i.e., had higher IC50than AZT for this cell line.

Example 5.

Inhibition of replication of HBV - DNA

Fig. 4 is a graph illustrating the percentage of inhibition of HBV DNA replication in 2.2.15 cells on the 9th day, depending on different concentrations of the tested compounds using a narrower range of concentrations than was used in example 1. Table 3 summarizes EC50and EC90for virion HBV and HBV - RI, the values of cytotoxicity and selectivity index of for DG, DAPG, ACPD, FTC and DDC.

Example 6

Fig. 5 is a graph illustrating the absorption of 5 μm tritium-labeled (-)-(2R,4R)-2-amino-9-[(2-hydroxymethyl)- 1,3-dioxolane-yl] adenine (DAPD) in cells Hep G2. The extract was obtained after 4 hours after treatment of cells DAPD (1000 number of disintegrations min-PM, injection of 80 ál.)ate way in trifosfatnogo form.

Example 7.

Fig. 6 is a graph illustrating the absorption of 5 μm tritium-labeled (-)-(2R,4R)-2-amino-9[(2-hydroxymethyl)- 1,3-dioxolane-4-yl] adenine (DAPD) in cells Hep G2. The extract was obtained after 12 hours after treatment of cells with compound DAPD (1000 number of disintegrations min-PM, injection 145 μl. ). The data obtained showed that after 4-hour incubation with the compound, labeled with tritium, there is a high intracellular levels of triphosphate.

Compounds of the present invention and their pharmaceutically acceptable salts, proletarienne precursors and derivatives can be used for the prevention and treatment of HBV infections and other related conditions such as condition, characterized by a positive reaction for antibodies against HBV and positive for HBV, chronic liver inflammation caused by a virus HBV, cirrhosis, acute hepatitis, fulminant hepatitis, chronic persistent hepatitis, and fatigue. These compounds or compositions can also be used for preventive purposes to prevent or slow the progression of the disease in individuals with a positive reacts the minds suffering from any of the above conditions, can be introduced effective amount of (-)-(2R,4R)-2-amino-6-chloro-9-[(2 - hydroxymethyl)-1,3-dioxolane-4-yl] purine (-)-(2R, 4R)-9-[(2 - hydroxymethyl)-1,3-dioxolane-4-yl] -guanine, (-)-(2R,4R)-2-amino - 9-[(2 hydroxymethyl)-1,3-dioxolane-4-yl] -adenine or (-)- (2R,2R)-2-amino-9-[2-hydroxymethyl)-1,3-dioxolane-4-yl] purine or their pharmaceutically acceptable salts or a derivative thereof, optionally in a pharmaceutically acceptable carrier or diluent. These active materials can be introduced in any suitable way, for example, pills, parenterally, intravenously, intradermally, subcutaneously, or by local application in liquid or solid form.

The active compound can be introduced in a pharmaceutically acceptable carrier or diluent in an amount sufficient to provide the desired therapeutic effect and at the same time do not have severe toxic effects on the patient.

The preferred dose of the active compound for all of the above conditions or diseases varies from about 1 to 60 mg/kg, more preferably 1-20 mg/kg, and in a wider range of 0.1 to about 100 mg per kilogram body weight of the patient per day. The range of effective doses of pharmaceutically acceptable which it is derived has its own activity, the effective dose can be determined, as described above, based on the weight derived either by other methods known to experts.

The compound of the present invention can be put into the form of any suitable standardized dosage forms, for example, contains 7-3000 mg, and preferably 70-1400 mg of active ingredient per dosage form. Oral medication usually contains 50-1000 mg of the active ingredient.

In the ideal case, if after the injection the maximum concentration of active compound in the blood plasma ranges from about 0.2 to 70 μm, and preferably from about 1.0 to 10 microns. Such concentrations can be achieved, for example, by intravenous injection of 0.1-5%-aqueous solution of the active ingredient, optionally in saline, or by introducing the active ingredient in the form of a bolus. The concentration of the active compound in the pharmaceutical composition depends on the speed of absorption, inactivation and excretion of drugs, as well as other factors well known in the art. In addition, the dose of the active compounds can also vary depending on the severity of the patient's condition. It should be noted, means which is appointed in accordance with the needs of the patient and is performed under the supervision of the attending physician, and the above concentration ranges are presented only for illustrative purposes and should not be construed as a limitation of the scope or application of the claimed composition. The active ingredient may be entered as a single dose, or it can be divided into several doses fewer and put through various time intervals.

Preferably, if the active compound is administered orally. Basically, the oral compositions contain an inert diluent or edible carrier. These compositions can be included in Milutinovi capsules or compressed into tablets. For oral administration for therapeutic purposes, the active compound may be mixed with excipients and used in the form of pills, tablets, or capsules.

A pharmaceutical composition may also include pharmaceutically acceptable binding agents and/or adjuvants.

These tablets, pills, capsules, lozenges, etc. can contain the following ingredients, or compounds of a similar nature: a binder compound, such as microcrystalline whole is such as alginic acid. Primogel or corn starch, a lubricant such as magnesium stearate or Sterotes, power sliding, such as colloidal silicon dioxide, sweetening agent such as sucrose, or flavoring agent such as peppermint, methyl salicylate, or orange flavoring. If the dosage form is a capsule, in addition to the above ingredients, the capsule may contain a liquid carrier such as fatty oil. In addition, standardized dosage forms may contain other materials which modify the physical form of a drug, for example, as coatings made of sugar, shellac or other intersolubility agents.

The active compound or its pharmaceutically acceptable salt, or pharmaceutically acceptable derivative can be introduced as a component part of an elixir, suspension, syrup, wafer, chewing gum, etc. Syrup, in addition to the active compounds, may also contain sucrose as a sweetening agent, preservatives, dyes and fragrances.

The active compound or its pharmaceutically acceptable salt or pharmaceutically acceptable what about the impact on the activity of the compounds of the present invention or materials, providing additional desired action, such as antibiotics, antifungal agents, anti-inflammatory agents or other antiviral agents, such as anti-virus, HBV, CMV or HIV.

Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components: a sterile diluent such as water for injection, saline solution, fatty oil, polyethylene glycol, glycerine, propylene glycol or other synthetic solvents, antibacterially agents, as well as benzyl alcohol or metalcarbon, antioxidants, as well as ascorbic acid or sodium bisulfite, hepatoblastoma agents, as well as ethylenediaminetetraacetic acid; buffers, as well as acetates, citrates or phosphates, and agents, corrective toychest, also, as sodium chloride or dextrose. Parenteral composition can be placed in ampoules, disposable syringes or glass or plastic bottles for mnogokratnogo use.

In the case of intravenous administration, the preferred carriers are physiological saline or sabotere the invention, avoid rapid removal of active compounds from the body, get the composition with controlled release formulation containing the active compound together with a carrier, for example, such as implants, and microencapsulated delivery systems drugs. For these purposes, can be used biodegradable and biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, complex poliorcetes, and polylactic acid. Methods of obtaining such compositions are well known in the art. These materials can also be obtained from the Alga Corporation and Nova Pharmaceutical Inc. As pharmaceutically acceptable carriers, preferably, are also liposomal suspensions (including liposomes targeted to infected cells and conjugated with monoclonal antibodies against viral antigens). Such liposomal suspensions can be obtained by methods well known in the art and described, for example, in U.S. patent N 4522811. So, for example, liposomal compositions can be obtained by dissolving appropriate lipid (such as stearoylethanolamine, stearoylethanolamine, Arago on the surface of the container there is a thin film of dried lipid. Then in the container is injected aqueous solution of the active compound or its monophosphate, diphosphate and/or trifosfatnogo derivatives. After that, the contents of the container is stirred manually for separation of lipid material from the walls of the container and dispersion of lipid aggregates, resulting in receive liposomal suspension.

Receiving phosphate derivatives - D-DIOXOLANYL

Mono-, di-, and trifosfatnogo derivative - D-DIOXOLANYL can be obtained in accordance with the procedure described above.

Monophosphate can be obtained by the method described by Imai and others J. Org.Chem. 34 (6), 1547-1550 (June 1868). For example, about 100 mg - D-dioxolane-nuke and about 280 μl of phosphorylchloride, stirring, subjected to reaction at approximately 8 ml of anhydrous ethyl acetate at about 0oC for about 4 hours. Then the reaction was suppressed using ice. Prominent phase was purified on a column of activated carbon, elwira 5% ammonium hydroxide in a mixture of ethanol and water (1:1). After evaporation of the eluate was received ammonium( - D-dioxolan-nuke)-5'-monophosphate.

Diphosphate can be obtained in accordance with the procedure described Davisson and other J. Org Chem., 52(9), 1794-1801 (1987). - D-Dioxolane-nucleoside nucleoside with mozillateam in perednia at room temperature for about 24 hours followed by treatment of the resulting product by standard methods (for example, by washing, drying and crystallization).

Triphosphate can be obtained in accordance with the procedure described Hoard and other J. Am.hem. Soc., 87 (8), 1785-1788 (1965). For example, - D-DIOXOLANYL activate (by processing imidazolidin in accordance with known procedures), and then treated with pyrophosphate of tributylamine in DMF. As a result of this reaction is mainly triphosphate with some unreacted monophosphate and some diphosphate. After purification using anion-exchange chromatography on DEAE-column receive triphosphate, for example, in the form of Terentieva salt.

The present invention has been described in the preferred examples of its implementation. However, experts in this field it is clear that various changes and modifications of the invention that can be made on the basis of the above detailed description of the invention. It should be noted that these modifications and changes should not go beyond the scope of the following claims.

1. A method of treating an HBV infection in a human or other animal, characterized in that it includes the introduction of an effective treatment for HBV infection if the group, includes hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate,

or its pharmaceutically acceptable salts, with the specified connection, at least 95% does not contain the corresponding L-enantiomer.

2. A method of treating an HBV infection in a human or other animal, characterized in that it includes the introduction of an effective treatment for HBV infection the number of enantiomerically pure D - deoxyadenylate patterns

< / BR>
where R represents NH2X is chosen from the group consisting of hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate,

or its pharmaceutically acceptable salts, with the specified connection, at least 95% free of the corresponding L-enantiomer.

3. A method of treating an HBV infection in a human or other animal, characterized in that it includes the introduction of an effective treatment for HBV infection the number of enantiomerically pure D - deoxyadenylate patterns

< / BR>
where R represents hydrogen or chlorine and X is chosen from the group consisting of hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate,

or its pharmaceutically acceptable salts, with the specified connection, at least 95% free of the corresponding the same time, that includes the introduction of an effective treatment for HBV infection the number of deoxyadenylate patterns

< / BR>
where R is OH, and X is chosen from the group consisting of hydrogen, alkyl, acyl, monophosphate, diphosphate and triphosphate;

or its pharmaceutically acceptable salt.

5. A method of treating an HBV infection in a human or other animal, characterized in that it includes the introduction of an effective treatment for HBV infection the number of deoxyadenylate patterns

< / BR>
where R represents NH2and X is chosen from the group consisting of hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate, or its pharmaceutically acceptable salt.

6. A method of treating an HBV infection in a human or other animal, characterized in that it includes the introduction of an effective treatment for HBV infection the number of deoxyadenylate patterns

< / BR>
where R represents H or Cl, and X is chosen from the group consisting of hydrogen, alkyl, acyl, monophosphate, diphosphate, and triphosphate,

or its pharmaceutically acceptable salt.

7. The method according to PP.1 - 5 or 6, wherein the enantiomerically pure D - deoxyadenylate administered in pharmaceutically acceptable noorallah introduction.

9. The method according to p. 7, characterized in that the carrier part of the capsule.

10. The method according to p. 7, characterized in that the medium is presented in the form of tablets.

11. The method according to PP.1 - 5 or 6, characterized in that the introduction is parenteral.

12. The method according to PP. 1 - 5 or 6, characterized in that the alkyl group is chosen from the group consisting of methyl, ethyl, propyl, butyl, penttila, hexyl, isopropyl, isobutyl, sec-butyl, t-butyl, isopentyl, amyl, t-pentile, cyclopentyl and cyclohexyl.

13. The method according to PP.1 - 5 or 6, characterized in that the acyl group is chosen from the group consisting of acetyl, propionyl, butyryl, pentanoyl, 3-methylbutyryl, besylate, 3-chlorobenzoate, benzoyl, acetyl, pivaloyl, nelfinavir, propionyl, valeryl, kapronata, kaprilat, caprinate, laureate, myristate, palmitate, stearate and oleate.

14. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside under item 1, is injected in doses, alternating doses of the compounds selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or recarbonization (FIAU); enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FEAU), carbovir, or interferon.

15. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside under item 2, is injected in doses, alternating doses of the compounds selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU); enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

16. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside on p. 3, is injected in doses, alternating doses of the compounds selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU); enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU)audica fact, the effective amount of the nucleoside on p. 4, is injected in doses, alternating doses of the compounds selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU); enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

18. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside on p. 5, is injected in doses, alternating doses of the compounds selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU); enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

19. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside on p. 6, administered in doses cerebus ethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane (FIAU); enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

20. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside under item 1, is administered in combination with compound selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU), enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

21. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside under item 2, is administered in combination with compound selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU), enantiomer iene HBV infection in a human or other animal, characterized in that an effective amount of the nucleoside on p. 3, is administered in combination with compound selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU), enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

23. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside on p. 4, is administered in combination with compound selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU), enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

24. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside on p. 5, is administered in combination with the connection, SEL is atilano; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU), enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

25. A method of treating an HBV infection in a human or other animal, wherein the effective amount of the nucleoside on p. 6, is administered in combination with compound selected from the group consisting of (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane; (-)-enantiomer or racemic mixture of 2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane; enantiomer or racemic mixture of 2'-fluoro-5-iodine-arabinopyranosyl (FIAU), enantiomer or racemic mixture of 2'-fluoro-5-etelarinteella (FEAU), carbovir, or interferon.

 

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5 cl, 4 tbl, 4 ex

FIELD: medicine, hepatology, virology.

SUBSTANCE: invention relates to a method for treatment of chronic viral hepatitis C of genotype-2 with moderate activity. Method is carried out by every day parenteral administration of solutions containing a mixture of amino acids, nucleosides and enzyme. In the first day 0.2 ml of 0.01% solution of a solution containing a mixture of amino acids: lysine, arginine, asparagines, glutamic acid taken in the equal amounts is administrated; in the next day 0.2 ml of 0.02% solution containing a mixture of nucleosides: adenosine, inosine, amino acids, guanosine, and enzyme lysozyme also taken in equal amounts. Administration of solution is alternated every day during carrying out the treatment course and each 2 months break in administration of amino acids is carried out for 28-32 days. After 9-11 days of administration of proposed solutions method involves additional course of parenteral administration of interferon-α 2b in the dose 3 ml IU, 3 times per a week for 6 months. The total duration of treatment course is 6 months, 9 days - 6 months, 11 days. Method provides clinical remission and elimination of hepatitis virus and normalization of biochemical indices for 6 months after termination of treatment.

EFFECT: improved method of treatment.

2 tbl, 4 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to geranyl compounds represented by the following formulas (I-1) , (I-2) or (I-3) wherein R1 means compounds of the following formulas: or R2 means a group remaining after removing all carboxyl groups presenting in carboxylic acid chosen from group consisting of malic acid, citric acid, succinic acid, fumaric acid and others; m = 1, 2 or 3; n = 0, 1 or 2, and m + n represent a number of carboxylic groups presenting in indicated carboxylic acid; R3 means p-hydroxyphenyl or mercapto-group. Also, invention relates to derivatives of mevalonic acid represented by the following formula (I-4): wherein R means -CH2OH or CH3. Also, invention to an antitumor agent comprising as an active component geranyl compound of formulas (I-1), (I-2) or (I-3) or derivative of mevalonic acid of the formula (I-4), and optionally a pharmaceutically acceptable carrier or solvent. Also, invention relates to a method for treatment of liver cancer based on using geranyl compound of formulas (I-1), (I-2) or (I-3), or derivative of mevalonic acid of the formula (I-4) and using proposed compounds in manufacturing an antitumor agent. Invention provides using geranyl compounds or derivatives of mevalonic acid as antitumor agents.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

7 cl, 3 tbl, 17 ex

FIELD: biology.

SUBSTANCE: invention relates to compositions containing immunostimulating RNA olygomers and uses thereof. Claimed molecules of immunostimulating RNA having chain length of 5-40 nucleotides and including at least one guanine and at least one uracyl have immunostimulating properties and represent exclusive ligands of one or more Toll-like receptors, including Toll-like receptor 7 (TLR7) and Toll-like receptor 8 (TLR8).

EFFECT: composition and methods for stimulation of immune response activation.

120 cl, 23 ex, 20 dwg

FIELD: medicine, pharmacy.

SUBSTANCE: invention proposes an agent comprising the following components, mg: 2-ethyl-6-methyl-oxypyridine succinate, 25-1500; nicotinamide adenine dinucleotide, 0.5-100, and inositol, 100-1200. Agent can comprise additionally L-carnitine in the amount 10-100 mg. Also, agent can comprise additionally choline alphoscerate in the amount 50-1000 mg. Agent can be prepared in injection or tabletted medicinal formulation or as suppository. Agent provides effective delaying apoptosis and arresting transformation of apoptotic alterations to necrotic alterations in pathological processes of different etiology.

EFFECT: valuable properties and enhanced effectiveness of agent.

4 cl, 2 dwg, 2 tbl, 9 ex

FIELD: medicine, pharmacy.

SUBSTANCE: invention proposes an agent comprising the following components, mg: 2-ethyl-6-methyl-oxypyridine succinate, 25-1500; nicotinamide adenine dinucleotide, 0.5-100, and inositol, 100-1200. Agent can comprise additionally L-carnitine in the amount 10-100 mg. Also, agent can comprise additionally choline alphoscerate in the amount 50-1000 mg. Agent can be prepared in injection or tabletted medicinal formulation or as suppository. Agent provides effective delaying apoptosis and arresting transformation of apoptotic alterations to necrotic alterations in pathological processes of different etiology.

EFFECT: valuable properties and enhanced effectiveness of agent.

4 cl, 2 dwg, 2 tbl, 9 ex

FIELD: medicine.

SUBSTANCE: method involves adding 52-523 mg/l guanosine disodium triphosphate and 3.5-4.7 g/l of sodium chloride to radiopaque preparation before introducing it.

EFFECT: prevented cardiac function disorder; high safety of coronography without worsening radiopaque preparation quality.

1 tbl

FIELD: medicine, oncology, hematology.

SUBSTANCE: method involves the complex using symptomatic, antibacterial, general tonic agents and nonspecific immunomodulating therapy. For this aim, lysozyme hydrochloride powder is given orally in the daily dose 500-1800 mg, 2 times per a day, every day for 20-30 days in combination with dosing sodium nucleinate in the daily dose 100-1500 mg, 2 times per a day, for 20-30 days by oral or sublingual route, and lactulose given orally in the dose 2.5-5 ml, 1-2 times per a day, every day for 10-30 days. Lysozyme hydrochloride is given 0.5-1 h before eating, sodium nucleinate is given after intake of lysozyme hydrochloride directly and lactulose is given 20-30 min before eating, or before eating immediately, or in 3-4 h after intake of lysozyme hydrochloride and sodium nucleinate. Method provides the complex correction of nonspecific resistance of body in patients suffering from leukosis and involving maintenance of immune homeostasis, plasma proteolysis, intestine microecology and reparative processes and improved tolerance of scheduled polychemotherapy.

EFFECT: improved method of treatment.

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