L-nucleosides with anti-hbv or anti-ebv activity, the method of inhibiting hbv or ebv infection

 

(57) Abstract:

Describes new connections - L-nucleosides of General formula (1), where R is the residue of uracil, thymine, cytosine or purine, which can be substituted by a halogen atom and alkyl, and R" is a hydrogen atom, acyl, alkyl, monophosphate, diphosphate or triphosphate, or its pharmaceutically acceptable salts, which possess anti-HBV or anti-EBV activity. Describes also a method of inhibiting HBV or EBV infection. 2 S. and 11 C.p. f-crystals, 20 ill., table 2.

The invention relates to methods for treating diseases caused by hepatitis B virus (also known as HBV and Epstein-Barr (also known as EBV, which include the introduction of an effective amount of one or more of the active compounds disclosed here, or farmatsevticheskii acceptable derivatives or prodrugs of one of these compounds.

Background of the invention

Diseases caused by hepatitis B worldwide has reached epidemic levels. After two six-month incubation period, during which the carrier of the infection does not know about the infection, HBV infection can cause acute hepatitis and liver damage, stomach pain, jaundice and povysheniya fatal form of the disease, at which destroyed large portions of the liver. Usually patients recover from acute viral hepatitis. However, in some patients, high levels of viral antigens exist in the blood for a long or indefinite period of time, causing chronic infection. Chronic infection can lead to chronic hepatitis. Patients infected with chronic persistent HBV, most often found in developing countries. By mid-1991, in Asia alone, there were approximately 225 million chronic carriers of HBV, and around the world - almost 300 million speakers. Chronic persistent hepatitis can cause dysfunction, cirrhosis and hepatocellular carcinoma, a primary liver cancer.

In Western industrial countries at high risk of HBV infection is exposed to those who are in contact with HBV carriers or samples of their blood. Epidemiology of HBV in fact very similar to the epidemiology of acquired immunodeficiency syndrome, what can explain why HBV infection is common among patients with AIDS (AIDS) or HIV-related infections. However, HBV is much more contagious than HIV.

HBV is the second after tobacco cause cancer is directly enable tumor development or to cause the development of tumors indirectly due to chronic inflammation, cirrhosis and rebirth of cells associated with infection.

The virus of Epstein-Barr (EBV) is a member of the genus Lymphocryptovirus, which belongs to the subfamily gammaherpesvirinae. It's somewhat of lymphotropic. EBV has the classic structure of herpes viruses, double-stranded genome DNA enclosed in eicosapentaenoic nucleocapsid, which, in turn, is surrounded by a lipid membrane with the viral glycoproteins. Amorphous shell protein occupies the space between the envelope and the nucleocapsid.

All human herpes viruses infect cells and replicate them to some extent, but EBV is doing it more efficiently. Most importantly, the pathogenesis and the host response of EBV infection is much more dependent on infection of lymphocytes than is the case with other human herpes viruses.

Currently, EBV is considered the cause of lymphoproliferative disorders of B-cells and is associated with various other serious and chronic diseases, including rare-like syndrome with progressive mononucleosis and oral hairy leukoplakia in patients with AIDS (AIDS). The assumption that EBV is the main cause of chronic fatigue, not vydergivanii blood. More than 85% of patients with acute phase of infectious mononucleosis secrete EBV.

EBV is associated with cancer. At least two groups of patients at risk of developing associated with EBV lymphomas: those who were transplanted kidney, heart, bone marrow, liver or thymus in combination with immunosuppressive therapy, and patients with AIDS. Cancers associated with EBV include lymphoma Burkitt and nasopharyngeal carcinoma.

In light of the fact that hepatitis b virus and the virus of Epstein-Barr have a strong and often tragic effects on the infected patient, there is an urgent need for an effective pharmacological agents for the treatment of patients infected with these viruses, which would be of low toxicity in relation to the media.

Therefore, the aim of the present invention is to provide compounds, compositions and method of treating diseases caused by hepatitis B.

Another objective of the invention is to provide compounds, compositions and method of treatment of a disease caused by a virus Epstein-Barr.

Summary of the invention

A method of treatment of the media and, in particular, the human is about the number of L-nucleoside of the formula (1)

< / BR>
where R represents a purine or pyrimidine base.

In a preferred embodiment, offered nucleoside in the form specified enantiomer and almost without corresponding enantiomer (i.e., in the form of enriched enantiomer substances, including enantiomerically pure form).

In one preferred embodiment, the active compound is 2'-fluoro-5-methyl-L-arabinofuranosyluracil (also named as L-FMAU). This compound is an effective antiviral agent against HBV and EBV and has low cytotoxicity. Other specific examples of active compounds include N1-(2'-deoxy-2'-fluoro - L-arabinofuranosyl)-5 - ethyluracil, N1-(2'-deoxy-2'-fluoro - L-arabinofuranosyl)-5 - iodotyrosine and N1-(2'-deoxy-2' -fluoro - L - arabinofuranosyl)-5-iodouracil.

Alternatively, the proposed L-nucleoside for the treatment of HBV or EBV, which contains 2'-arabinogalactan group, for example, L-archimidean, L-fludarabin, L-ORGANOTIN and L-ariansen, as shown in Fig. 1.

Disclosed here L-nucleosides and their pharmaceutically acceptable derivatives or pharmaceutically acceptable compositions containing these compounds, are useful for the prevention and treatment of HBV and the technical inflammation of the liver, caused by HBV, cirrhosis, acute hepatitis, fulminant hepatitis, chronic persistent hepatitis, and fatigue. Similarly, these compounds can be used for the treatment of diseases associated with EBV. These compounds or compositions can also be used to prevent or delay the development of clinical disease in patients who are positive in relation to anti-HBV or anti-EBV antibody or HBV - or EBV-antigens, or those who have been in contact with HBV and EBV.

In another embodiment, the active compound or its derivative, or its salt can be introduced in combination or alternation with another anti-HBV agent, or anti-EBV agent, including those listed previously, or anti-HIV agent. Usually, when alternative therapy, an effective dose of each agent is administered serially, whereas combination therapy effective dose of two or more agents are administered together. The doses depend on the speed of absorption, inactivation and excretion (highlight) medications, and other factors known to practitioners. It should be noted that the magnitude of the dose will also depend on the severity of the condition requiring relief. It should also be noted that for each individual patient specific dose regimes and schemes with Vija doctor conducting the treatment.

Limitiruyuschie examples of antiviral agents that can be used in conjunction with open connections here, include (-)-enantiomer of 2-hydroxymethyl-5-(5-ferocity-1-yl)- 1,3-oxathiolane (FTC); (-)-enantiomer of 2-hydroxymethyl-5-(cytosine-1-yl)- 1,3-oxathiolane (ZTS); carbovir, acyclovir, interferon, AZT, DDI (2',3'-dideoxyinosine), DDC(2',3'-dideoxycytidine), L-DDC, L-5-F-DDC and D4T.

Brief description of drawings

Fig. 1 is an illustration of selected L-nucleosides, which are included in the scope of the present invention:

L-FMAU (2'-fluoro-5-methyl-L-arabinofuranosyluracil), L-FIAU(2'-fluoro-5-iodo - L-arabinofuranosyluracil), L-FC (2'-fluoro - L-arabinofuranosylcytosine), L-FIAC (2'-fluoro-5-iodo - a-L-arabinofuranosylcytosine), L-2-C1-2' -F-2'-deoxyactein, L-FEAU (2'- fluoro-5-ethyl - L-arabinofuranosyluracil), L-archimidean L-fludarabine, L-ORGANOTIN and L-urinatin.

Fig. 2 is a graph of the percentage of viable cells on the concentration of the medicine for L-FMAU in H1 cells.

Fig. 3 is a schematic illustration of a 1-O - acetyl-2,3,5-tri-O-benzoyl - L-ribofuranose (compound 10).

Fig. 4 is a diagram of an alternative obtain 1-O - acetyl-2,3,5-tri-O-benzoyl - L-ribofuranose idofuranose (compound 13).

Fig. 6 schematically represents a method for obtaining N9-(3', 5'-di-O-benzoyl-2'-deoxy-2'-fluoro - L-arabinofuranosyl)-2,6 - di-chloropurine (compound 15) and N9-(2'-deoxy-2'-fluoro - L-arabinofuranosyl)- 2,6-di-chloropurine (compound 16).

Fig. 7 represents a method of obtaining a number of 2'-deoxy-2'- fluoro - L-arabinofuranosyl-pyrimidine (compound 17-24).

Fig. 8 represents a method of obtaining the N1-(2'-deoxy-2'- fluoro - L-arabinofuranosyl)-5-iodotyrosine) (compound 22).

Fig. 9 is a method of obtaining 9- -L - arabinofuranosyladenine.

Fig. 10 is an alternative way to obtain 1-O - acetyl-2,3,5-tri-O-benzoyl - L-ribofuranose (compound 10) of 1,2-di-O-isopropylidene - a-L-xylofuranose (compound 3).

Fig. 11 is a graph of the concentration of L- (-)-FMAU in plasma in mice after oral administration in a time-dependent (cross, 10 mg/kg injected twice a day (bid) for 29 days prior to pharmacokinetic studies, and then studies conducted on the thirtieth day with the introduction of the same concentration; shaded circles - administration of a 50 mg/kg twice daily for 29 days before the study, and then on the thirtieth day of the survey in the introduction).

Fig. 12 is a graph of the concentration of L- (-)-FMAU in the liver of mice after oral administration of time (cross, 10 mg/kg injected twice a day for 30 days to pharmacokinetic studies, and then studies conducted on the thirtieth day with the introduction of the same concentration; shaded circles - administration of a 50 mg/kg twice a day for 30 days before the study, and then on the thirtieth day conduct of the study after administration of the same concentration; where there's no shading mugs - introduction for the first time 50 mg/kg on the first day of the study).

Fig. 13a represents the change in weight of animals after 30 days in the control female mice BDF1.

Fig. 13b and 13c represent the change in weight after 30 days of administration to female mice BDF1 doses of 10 mg/kg (13b) and 50 mg/kg (13c) twice a day drug L-(-)-FMAU. Shows the weight of the animals represent the average and standard deviation for 5-7 mice.

Fig. 14-20 present a clinical analysis of the plasma of mice after administration of L-(-)-FMAU at a concentration of 10 mg/kg (three mice) or 50 mg/kg (three mouse) twice a day for 30 days. Fig. 14 is a graph of the concentration of total bilirubin in the plasma of mice in mg/DL. Fig. 15 is a graph of the concentration of mice in mg/DL (deciliter). Fig. 17 is a graph of the concentration of AST (SCOT, serum glutamic axalingua transaminase levels in plasma of mice in U/L. Fig. 18 is a graph of the concentration of ALT (SGPT, serum glutamic pyruvate-transaminase levels in plasma of mice in U/L. Fig. 19 is a graph of the concentration of lactic acid in the plasma of mice in mmol/L. Fig. 20 is a graph of the concentration of lactic dehydrogenase in plasma of mice in U/L.

Detailed description of the invention

The term "enantiomerically enriched" in the sense used here, refers to a nucleoside composition that includes at least 95%, and preferably, about 97, 98, 99% or 100% of a single enantiomer of that nucleoside.

The term alkyl, as used herein, includes, but is not limited to, C1-C10alkyl groups, including methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, amyl, tert-pentyl, cyclopentyl and cyclohexyl.

The term acyl, in the sense here used, includes, but is not limited to, acetyl, propionyl, butyryl, pentanoyl, 3-methylbutyryl, acid succinate, 3-chlorobenzoate, benzoyl, acetyl, pivaloyl, mesilate, propionyl, is here used, if there are no other definitions, the term aryl refers to phenyl.

The term "protected" in the sense used here, refers to a group if there are other definitions that are attached to the oxygen atom or nitrogen, to prevent their entry into the reaction in the process of obtaining derivatives other fragments of the molecule, which contains these oxygen or nitrogen. Specialists in the field of organic synthesis known to a wide range of oxygen - and asossasiyasi groups.

The term purine or pyrimidine base include, but are not limited to, adenine, N6-alkylphenyl, N6-acylpyrin (where acyl is C(O)alkyl, aryl, alkaryl or aralkyl), N6-benzylurea, N6-halohydrin, N6-vinluan, N6-aceteminphen, N6-acylpyrin, N6-hydroxyalkylated, N6-dialkylphenol, thymine, cytosine, 6-etherimide, 2-mercaptopyrimidine, uracil, N5-alkylpyridine, N5-benzylpyrimidines, N5-kalaidjieva, N5-vinylpyridine, N5-acetylspiramycin, N5-arylpyrimidine, N5-hydroxyalkylated, N5-dialkylphenol, 5 - azacitidine, 5-azauracil, triazolopyridines, imidazopyridines, pyrrolo the TB if necessary or if desired. Suitable protective groups are well known in the art, and include trimethylsilyl, dimethylhexylamine, tert-butyldimethylsilyl and tert-butyldiphenylsilyl, trailer, alkyl groups such acyl groups as acetyl, propionyl, butyl, methylsulphonyl and paratoluenesulfonyl. They specifically include 5-methyluracil (thymine), 5 - iodouracil, cytosine, and 5-ethyluracil.

The present invention also disclosed a method and composition for the treatment of HBV infections, EBV infections and other viral infections in which the virus replicates in humans or other animals-carriers in the same way as HIV, which includes the introduction of an effective amount of one or more of the above L-nucleoside or a physiologically acceptable derivative or physiologically acceptable salts, optionally in a pharmaceutically acceptable carrier. Compounds of the present invention have anti-HBV activity, anti-EBV activity, or into the body in a compound or compounds that exhibit anti-HBV or anti-EBV activity. Disclosed in the present invention compounds can also be used to treat diseases associated with HBV and EBV.

The active compounds can be introduced in the form of any pickup or itself to be active. Limitiruyuschie examples can serve as pharmaceutically acceptable salts (alternatively referred to as "physiologically acceptable salts"), and 5' and the purine - or pyrimidine-acylated or alkylated derivatives of the active compounds (otherwise referred to as "physiologically active derivatives"). In one embodiment, the acyl group is an ester of carboxylic acid (i.e.,- C(O)R'), in which decarbonising fragment (R') of the ester group selected from a branched, unbranched or cyclic C1-C10of alkyl, alkoxyalkyl, including methoxymethyl, aralkyl, including benzyl, such alkoxyalkyl as phenoxymethyl, aryl, including phenyl, optionally substituted by halogen, C1-C4the alkyl or C1-C4alkoxy, sulphonate esters such as alkyl - or aralkylamines, including methanesulfonyl, mono-, di - or tri-phosphate esters, trityl or monomethacrylate, substituted benzyl, trialkylsilyl (for example, dimethyl-tert-Boticelli) or di - phenylmethylene. Optimally, aryl groups in the esters contain a phenyl or benzyl group. The alkyl group may be unbranched, branched or cyclic, and optimally represents the, as described in detail below, or in other ways known to the specialists.

The following synthesis scheme, you can use other standard reagents, including equivalent acids, bases and solvents instead of the above, from among those that are well known in the art. To protect oxygen and nitrogen can be used with a wide range of protective groups, such as that shown in the book: Greene et al., "Protective Groups in Organic Synthesis", John Wiley and Sons, 2nd. Ed., 1991. Suitable protective groups for oxygen and nitrogen include, for example, such tizanidine silyl group as trimethylsilyl, dimethylhexylamine, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, trityl, an alkyl group, such acyl groups as acetyl, propionyl, benzoyl, p-NO2-benzoyl, benzyl or toluyl, methylsulphonyl or p-toluensulfonyl. Functional groups of oxygen and nitrogen in the heterocyclic base must be protected before condensation with sugar.

Suitable reducing agents include NaBH4diisobutylaluminium (DIBAL-H) , lithium borohydride (LiBH4) and sodium bis(2-methoxyethoxy)-aluminiumhydride (Red-Al). Suitable oxidizing agents include aqueous acidic chromium of chislett potassium (KMnO4,), leads to compounds, which lead/pyridine, oxygen over a catalyst of the platinum/carbon, RUO Li4, RUO Li4/NaIO4the sulfoxide/dicyclohexylcarbodiimide (DMSO/DCC) are proton donors, as silver carbonate and carbonate triphenylbismuth, oxidation of Oppenauer (alkoxides of aluminum in acetone) and manganese dioxide (for selective oxidation of allyl or benzyl alcohols in the presence of other alcohols).

The catalysts of the Friedel-(Lewis acid), which can be used in the condensation reaction include SnCl4, ZnCl4, TiCl4, AlCl3, FeCl3BF3-diethyl ether and BCl3. These catalysts require anhydrous conditions, since the presence of water reduces their activity. These catalysts can also be inactivated in the presence of organic solvents with active hydrogens such as alcohols and organic acids. These catalysts are usually used in solvents such as carbon disulfide, methylene chloride, nitromethane, 1,2-dichloroethane, nitrobenzene, tetrachlorethane, chlorobenzene, benzene, toluene, dimethylformamide, tetrahydrofuran, dioxane or acetonitrile. Anhydrous luminiare insoluble in carbon disulfide (Niedballa, et al., J.Org. Chem. 39, 25 (1974)). The preferred Katalizator altoviti in the same conditions, mentioned previously catalysts for Friedel -. The reaction proceeds at a temperature in the range -10 to 200oC. Disilylgermane can be done using a variety of reagents, including acetic acid, triperoxonane acid, hydrogen fluoride, n-tetrabutylammonium, potassium fluoride, pyridine HCl.

As is shown in Fig. 3, from L-xylose (1a), a key intermediate compound 1-O-acetyl-2,3,5-tri-O-benzoyl - L-ribofuranose (10) get a full yield of 20% (L. Vargha, Chem.Ber., 1954, 87, 1351; Holy, A., et al. , Synthetic procedures in Nucleic Acid Chemistry, VI, 163-67). As is shown in Fig. 4, compound 10 can also be obtained from a more expensive source material L-ribose (Holy , A., et al., Synthetic Procedures in Nucleic Acid Chemistry, VI, 163-67). Fig. 3 represents an alternative synthesis of compound 10 (yield 53%), which consistently foryouth C2(J. Org. Chem. , 1985, 50, 3644-47) to obtain 1,3,5-tri-O-benzoyl-2-deoxy - 2-fluoro-L-arabinofuranose (13), which condense with different bases through octabromodiphenyl sugar with obtaining 2'-deoxy-2'-ferramenta.recomendo with different outputs.

1,2-Di-O-isopropylidene - a-L-xylofuranose (3)

To 650 ml of anhydrous acetone are added 4 ml of concentrated sulfuric acid, 5 g molekulyarnihm for 36 hours. The reaction mixture was filtered and thoroughly washed with acetone, and the combined filtrate is neutralized with ammonium hydroxide and then evaporated to dryness. Add 200 ml of ethyl acetate, then filtered and evaporated, obtaining oil, which is dissolved in 250 ml of 0.2% HCl solution and stirred at room temperature for 2.5 hours, the pH set in size 8 with saturated solution of NaHCO3, then is evaporated to dryness, the residue is extracted with ethyl acetate. After removal of the solvent receive a yellow oil, compound 3 (41.7 g, 82.3 per cent).

1H-NMR (CDCl3): 5,979 (d, J = 3,78 Hz, 1H, H-1); 4,519 (d, J = 3.6 Hz, 1H, H-2); 4,308 (Shir.D., 1H, H-3); 4,080 (m, 3H, H-4 and H-5); of 1.321 (s, 3H, CH3); 1,253 (s, 3H, CH3).

1,2-di-O-isopropylidene-3,5-di-O-o-tamilselvan- -L-xylofuranose (4)

Compound 3 (40 g, 210 mmol) is stirred in 500 ml of anhydrous pyridine at 0oC, with TsCl (75 g, 393 mmole) dissolved in 100 ml of CHCl3by adding dropwise. 3 hours later, in the same way add the second portion of TsCl (50 g, 262 mmole) in 50 ml of CHCl3. The resulting mixture was stirred at room temperature for 24 hours, then cooled at 0oC add 10 ml of water, then stirred at room temperature for 30 minutes. The reaction with the x-4), a saturated solution of NaHCO3(200 ml x 2), dried over magnesium sulfate. After removal of the solvent receive a brown syrup, which after crystallization from EtOH gives compound 4 in the form of a solid white color (to 103.8 g, 99%).

1H-NMR (CDCl3): 7,282, 7,836 (m, 8H, OT); 5,849 (d, J = 3,51 Hz, 1H, H-1); 4,661, 4,779 (m, 2H, H-3 and H-4); 4,193 (DD, 1H, H-2); 4,011 (d, 2H, H-5); 2,448, 2,478 (2s, 6H, -OTs); 1,261, 1,320 (2s, 6H, CH3).

1,2-di-O-acetyl-3,5-di-O-p-tamilselvan- , - xylofuranose (5)

Compound 4 (70 g, to 140.5 mmole) is dissolved in 700 ml of glacial AcOH and 100 ml Ac2O, prokopeva at 0oC 50 ml of concentrated sulfuric acid. The resulting solution was stirred at room temperature overnight and then poured into 1 l of a mixture of ice-water, extracted with CHCl3(200 ml x 4), washed with saturated sodium bicarbonate solution, dried over magnesium sulfate. After removal of solvent in vacuo get connection 5 in the form of syrup (84,2 g, crude yield of 110%).

Methyl-3,5-di-O-p-tamilselvan- , - xylofuranose (6)

The crude product 5 (80 g) is stirred in 500 ml of 1% HCl mixture/CH3OH at room temperature for 30 hours. The solvent is removed under reduced pressure, the residue is dissolved in 300 ml of CHCl3, washed NAPA (60 g, 90% of 4).

Methyl-2-O-benzoyl-3,5-di-O-p-tamilselvan- , - xylofuranose (7)

Syrup of compound 6 (60 g, 127 mmol) was dissolved in 200 ml of pyridine and stirred at 0oC, prokopeva the benzoyl chloride (40 ml, 345 mmol) and the resulting solution was stirred at room temperature for 17 hours. Its concentrated to about 50 ml, then poured into 300 ml of a mixture of ice-water, extracted with CHCl3washed 3 N. H2SO4and saturated NaHCO3, dried over magnesium sulfate. After evaporation of the solvent to obtain compound 7 in the form of syrup (71 g, 97%).

Methyl-2,3,5-tri-O-benzoyl- , -L-ribofuranose (9)

Syrup of compound 7 (70 g) and sodium benzoate (100 g, 694 mmole) are suspended in 1200 ml of DMF and stirred with a mechanical stirrer while boiling under reflux for 16 hours. The mixture is cooled to room temperature and poured into 1 l of a mixture of ice-water, extracted with ether, dried over magnesium sulfate. After evaporation of the solvent receive syrup (50 g, 8a and 8b), which is dissolved in 180 ml of pyridine and benzoylurea (BzCl, 20 ml, 172 mmole) for 17 hours at room temperature. After processing the receive connection 9 in the form of syrup brown (48 g, 83% from 7).

1-O-acetyl-2,3,5-tri-O-benzoyl - L-riboside and 16 ml of concentrated sulfuric acid at a temperature of from 0oC to room temperature for 17 hours. Then poured into 1 l of a mixture of ice-water, extracted with chloroform (200 ml x 4). The combined extract was washed with saturated sodium bicarbonate solution and dried over magnesium sulfate. After removal of the solvent receive a syrup of brown, which is treated with ethanol and receive connection 10 in the form of a solid white color (8.8 g, 32%). So the melting of 124.7oC, the literature dealing with. 129-130oC; D form: 130-131oC []D= -45,613 (from 1.0, CHCl3); D. shape: []D= +44,2.

1H-NMR (CDCl3): 7,317, 8,134 (m, 15H, OBz), 6,437 (s, 1H, H-1), 5,835 (m, H-2 and H-3), 4,649 (m, 3H, H-4 and H-5), 2,003 (s, 3H ).

1-O-acetyl-2,3,5-tri-O-benzoyl - L-ribofuranose (from L-ribose)

L-ribose (5 g, a 33.3 mmole) are suspended in 120 ml of 1% HCl/ MeOH and stirred at room temperature for 3 hours, resulting in a get a clear solution. The reaction is quenched by adding 30 ml of anhydrous pyridine, and then evaporated under reduced pressure. The remaining syrup is evaporated together with pyridine (30 ml x 2), then dissolved in 80 ml of anhydrous pyridine and stirred at 0oC, prokopeva the benzoyl chloride (20 ml, 172 mmole). After stirring at room temperature for 17 hours the reaction is complete. Add 10 ml of water, elewaut in 150 ml of a mixture of ice-water, extracted with chloroform (50 ml x 4), washed successively 3 N. H2SO4, (30 ml x 2), saturated sodium bicarbonate (30 ml x 3) and dried over magnesium sulfate. After removal of the solvent to obtain compound 9 as 13 g of syrup.

The crude compound 9 was dissolved in 80 ml of HBr/AcOH (45%, weight/volume) and stirred at room temperature for 1.5 hours. To this mixture is added glacial acetic acid (50 ml), the resulting solution was stirred at 0oC, slowly adding 34 ml of water to maintain the temperature below 7oC. and Then stirred at room temperature for 1 hour, poured into 200 ml of a mixture of ice-water, extracted with chloroform (50 ml x 5). The combined extracts are washed with saturated sodium bicarbonate solution, dried over magnesium sulfate. After removal of the solvent to obtain 13 g of a syrup which is dissolved in 40 ml of anhydrous pyridine, stirred at 0oC. Then was added dropwise acetic anhydride (14 ml, of 148.4 mmole). After completion of the reaction, the reaction mixture was poured into 150 ml of a mixture of ice-water, extracted with chloroform (50 ml x 4), washed successively 3 N. H2SO4, (30 ml x 2), saturated sodium bicarbonate solution (50 ml x 2) and dried over what society white (9.2 grams, 53,7% of L-ribose).

1,3,5-tri-O-benzoyl - L-ribofuranose (11)

Compound 10 (9 g, 17,84 mmole) is stirred in 100 ml of CH2Cl2at 0oC, while adding 70 ml of CH2Cl2containing HBr (3.2 g, 30.5 mmole), immediately. The resulting mixture is stirred at a temperature of 0oC for 3.5 hours, add 55 ml of water, the resulting mixture was stirred at room temperature for 18 hours. The organic layer emit, washed with saturated sodium bicarbonate solution and dried over magnesium sulfate. After evaporation of the solvent receive the foam, from which after recrystallization from CH2Cl2and n-hexane get the connection 11 in the form of a solid white color (6.2 g, 75,5%). So melting point 137-138oC, from the literature: 140-141oC []D= -81,960 (0,55, CHCl3; D the form: []D= +83,71.

1H-NMR (CDCl3): 7,312, 8,187 (m, 15H, OBz), 6,691 (d, J = 4.59 Hz, H-1); 5,593 (DD, J4-3= 6,66 Hz, J2-3= 1.8 Hz, 1H, H-30); 4,637, 4,796 (m, 4H, 4-2, H-4 and H-5); 2,3 (Shir. OH).

1,3,5-tri-O-benzoyl-2-O-amidosulfuric- -L - ribofuranose (12)

Compound 11 (5,94 g, 12,84 mmole) is stirred in 50 ml of anhydrous CH2Cl2at -15oC (a mixture of dry ice-CCl4). Successively added anhydrous DMF (15 ml) and sulfurylchloride (3.2 m temperature for 4 hours. Three servings add imidazole (9.7 g, 12,78 mmole), cooling in an ice bath. Then it is stirred at room temperature for 17 hours. The resulting mixture was poured into 150 ml of a mixture of ice-water, the aqueous phase extracted with CH2Cl2(50 ml x 3). The combined organic layers washed with water and dried over magnesium sulfate. Purified through column chromatography (hexane: EtOAc/5:1 to 1:1), resulting in a receive connection 12 in the form of a solid white color (3,7, 49%). So melting 124,5 output reached 125.5oC; literature dealing with. 129oC; []D= -68,976, D form: +66,154.

1H-NMR (CDCl3): 6,9, and 8.2 (m, 18H, Ar-H); to 6.67 (d, J = 4.4 Hz, 1H, H-1); 5,59 (DD, 1H, H-3); a total of 5.21 (DD, 1H, H-2); from 4.5 to 4.8 (m, 3H, H-4 and H-5).

1,3,5-tri-O-benzoyl-2-deoxy-2-fluoro - L-arabinofuranose (13)

A suspension of compound 12 (3.33 g, 5,62 mmole), KHF2(1,76 g, 22,56 mmole) in 30 ml of 2,3-butanediol is stirred in an argon atmosphere. The mixture is heated to 150oC by adding 1 ml of HF/H2O (48%, 27.6 mmole), the mixture was stirred at 160oC for 1.5 hours. For damping the reaction add a mixture of brine-water, and then the resulting solution was extracted with methylene chloride (50 ml x 4). The combined extracts washed with brine, water, saturated sodium bicarbonate solution, dried over messagefilename, and then EtOAc receive a syrup, which crystallized from 95% EtOH, receiving the connection 13 (1.3 g, 49,8%). So melting 77-78oC (literature. 82oC).

1H-NMR (CDCl3): 7,314, 8,146 (m, 15H, OBz); 6,757 (d, J = 9.1 Hz, 1H, H-1); 5,38 (d, J = 48,5 Hz, 1H, H-2); 5,630 (DD, J = 22,5 Hz, 1H, H-3); 4,768 (m, 3H, H-4 and H-5).

N9-(3', 5'-di-O-benzoyl-2'-deoxy-2'-fluoro - L-arabinofuranosyl)-2,6-di-globulin (15)

Compound 13 (464 mg, 1 mmol) dissolved in 10 ml of methylene chloride, adding to 1.4 ml of HBr/AcOH (45% weight/volume). The resulting solution was stirred at room temperature for 24 hours and then evaporated to dryness. The residue is dissolved in 20 ml of methylene chloride and washed with water, saturated sodium bicarbonate solution, dried over magnesium sulfate. After filtration and evaporation receive premasagar connection 14 (100%, based on TLC data).

At the same time suspended 2,6-di-globulin (378 mg, 2 mmole) in 15 ml of HMDS (hexamethyldisilazane) and 2 mg of ammonium sulfate, and then refluxed for 2 hours. HMDS is evaporated under high vacuum in a nitrogen atmosphere to obtain similarvideo base white.

Premasagar 14 is dissolved in 25 ml of anhydrous 1,2-dichloroethane. The resulting solution was added to Siciliana about is her. Add chloroform (30 ml) and washed successively with water (20 ml x 2), saturated sodium bicarbonate solution (20 ml x 2), saturated NaCl (20 ml x 2) and dried over magnesium sulfate. From CHCl3crystallized compound 15 (105 mg, 19.7 percent). So melting 158oC; D form: 158oC.

UV (methanol)max= 238,5 nm, 273,0 nm.

1H-NMR (300 MHz, DMSO-d6): 8,82 (d, J = 1.5 Hz, 1H, H-8); 7,49, with 8.33 (m, 10H, OBz); 6,767 (DD, JH-H= 4 Hz, KF-H= to 13.8 Hz, 1H, 1-H'); 5,854 (DQC, J = 67,4 Hz, 1H, H-2'), 5,910 (m, 1H, H-3'); 4,751 (m, 3H, H-4' and H-5').

M9(2'-deoxy-2'-fluoro - L-arabinofuranosyl)- 2,6-di-globulin (16)

Compound 15 (70 mg, of 0.13 mmole) is dissolved in 25 ml of saturated NH3/CH3OH in a sealed steel bomb and heated at 90oC for 6 hours. After removal of the solvent under reduced pressure get semi-liquid substance is yellow, which is purified using preparative TLC (9: 1/CHCl3: CH3OH). After lyophilization from water and 1,4-dioxane get the white powder - connection 16 (30 mg, 75%). UV (H2O) pH2,max= 212,00 nm, 263,5 nm ( 6711); pH7,max= 213,5 nm, 263,00 nm ( 7590); pH11, max= 213,5 nm, 263,00 nm ( 5468).

1H-NMR (300 MHz, BMSO-d6): 8,279 (d, J = 1.5 Hz, 1H, H-8); 7,908 (Shir. s, 2H, NH2); 6,321 (DD, JH19 Hz, 1H, H-3'); 3,841 (m, 1H, H-4'); 3,636 (d, 2H, H-5').

N1-(2'-deoxy-2'-fluoro-3', 5'-di-O-benzyl-L - arabinofuranosyl)-thymine (17)

To a solution of compound 13 (400 mg, 0,86 mmole) in anhydrous CH2Cl2(10 ml) is added hydrogen bromide in acetic acid (45% W/V, 1.5 ml), the resulting solution was stirred at room temperature for 17 hours. After removal of solvent and co-evaporation with toluene receive connection 14.

At the same time thymine (215 mg, 1,72 mmole) is refluxed in NM (25 ml) in a nitrogen atmosphere for 17 hours to obtain a homogeneous solution. After evaporation of the solvent receive sililirovany thymine.

A solution of compound 14 in 50 ml of dichloroethane is added to Siciliana Cimino and the resulting solution was boiled under nitrogen atmosphere for 3 days. Add water, and then extracted with CHCl3. The organic layer is washed with water, brine and dried over magnesium sulfate. After evaporation of the solvent to obtain the crude product, which was purified using preparative TLC using 2% MeOH/CHCl3to obtain compound 17 (235 mg, 58%). So melting point 99-101oC. UV (methanol): 230, 264 nm []D= +22,397..

1H-NMR (CDCl3): 7,343-8,389 (the d, 1H, H-3'); 4,812 (d, 2H, H-5'); 4,466 (m, 1H, H-4'); 1,775 (s, 3H, CH3).

Elemental analysis (CH24H21N2O7F), C: 61,01; H: 4,57, N: 5,73, F: 3,92.

N1-(2'-deoxy-2'-fluoro - L-arabinofuranosyl)- thymine (18)

Compound 17 (145 mg, 0,309 mmole) is treated with NH3/CH3OH at room temperature for 18 hours. After evaporation of the solvent and purification by using preparative TLC (15% MeOH/CHCl3) get the connection 18 (70 mg, 87,5%). So melting point 174-175oC. UV: 264 nm, []D= -104,36.

1H-NMR (DMSO-d6): 11,401 (s, 1H, NH); 7,575 (s, 1H, H-6); 6,093 (DD, JH-H= to 4.41 Hz, JF-H= 15.6 Hz, H-1'); 5,844 (d, 1H, 3'-OH); 5,019 (dt, JF-H= 53,3 Hz, 1H, H-2'); 5,087 (t, 1H, 5'-OH); 4,194 (DQC, 1H, H-3'); 3,647 (m, 3H, H-4' and H-5'); 1,781 (s, 3H, CH3).

Elemental analysis: (C10H13N2FO5), C: 44,80; H: EQUAL TO 4.97; N: 10,04; F: 7,03.

N1-(2'-deoxy-2'-fluoro-3', 5'-di-O-benzoyl - L-arabinofuranosyl)-5-ethyluracil (19)

To a solution of compound 13 in anhydrous dichloromethane (10 ml) is added hydrogen bromide in acetic acid (45% weight/volume of 0.97 ml, 5,385 mmole). This solution was stirred at room temperature for 13 hours, after solvent evaporation and co-evaporation with toluene receive connection 14.

At the same time 5-ataecina 5 hours in nitrogen atmosphere to obtain a homogeneous solution.

The solution similarvideo Foundation is evaporated to dryness, avoiding contact with moisture. To the resulting syrup add a solution of compound 14 in anhydrous 1,2-dichloroethane (10 ml). The reaction mixture was stirred at 95oC in nitrogen atmosphere for 20 hours, then evaporated in vacuum to dryness to obtain a solid yellow color, which is mixed with CH3OH/CHCl3(1:1) and filtered. The resulting filtrate is evaporated to obtain the residue, which is treated through column chromatography (CH3OH/CHCl3, 0-1%) to obtain a solid white color, compound 19 (0,557 g, 100%).

1H-NMR (DMSO-d6): for 11.55 (s, 1H, NH); 7,51, 8,08 (m, 10H, Ar-H); to 7.32 (s, 1H, H-6); 6,29, 6,37 (DD, JH-H= 3,7 Hz, JF-H= 20 Hz, 1H, H-3'); 5,47-the 5.65 (DD, JF-H= 54 Hz, 1H, H-2'); 4,62-4,82 (m, 3H, H-4' and H-5'); 2,01, 2,09 (q, 2H, ); of 0.85 (t, 3H, CH3).

N1-(2'-deoxy-2'-fluoro - L-arabinofuranosyl)-5 - ethyluracil (20)

Compound 19 (500 mg) was dissolved in methanolic ammonia (50 ml) and stirred at room temperature for 44 hours. The resulting solution is evaporated to dryness to obtain a solid white color (0.4 g), which is treated on a chromatographic column with silica gel (CH3OH/CHCl3, 0-5%) to obtain a solid substance be'er>1H-NMR (DMSO-d6): 11,42 (s, 1H, NH); EUR 7.57 (s, 1H, H-6'); 6,10, 6,17 (DD, JH-H= 5.0 Hz, JF-H= 14 Hz, 1H, H-1'), 5,88 (Shir.s, 1H, 3'-OH); 5,14, 5,19 (m, 2H, H-2' and 5'-OH); to 4.98 (t, 1H, H-3'); 4,22, to 4.28 (m, 1H, H-4'); 3,55, of 3.78 (m, 2H, H-5').

Elemental analysis (C11H15N2O5F: C: 7,93; H: 5.56MM; N: 10,06; F: 6,68.

N1-(2-deoxy-2'-fluoro-3', 5'-di-O-benzoyl - L-arabinofuranosyl - N4-benzoyl-5-iodotyrosine (21)

To a solution of compound 13 (150 mg, 0,323 mmole) in anhydrous methylene chloride (5 ml) is added hydrogen bromide in acetic acid (45% weight/volume of 0.29 ml, near 1.615 mmole). The reaction mixture was stirred at room temperature for 9.5 hours. After solvent evaporation and co-evaporation with toluene receive the connection 15 in the form of a yellow syrup.

At the same time, N4-benzoyl-5-iodotyrosine (550 mg, near 1.615 mmole) is suspended in HMDS (8 ml) with ammonium sulfate (3 mg) and refluxed for 5 hours in nitrogen atmosphere to obtain a homogeneous solution.

The solution similarvideo Foundation is evaporated to dryness, avoiding contact with moisture. To the resulting syrup add a solution of compound 14 in anhydrous 1,2-dichloroethane (10 ml). The reaction mixture was refluxed in nitrogen atmosphere during which forma (30 ml). The precipitate is filtered and washed with chloroform. The obtained filtrate and washings are combined and evaporated to obtain a syrup brown. The resulting mixture was separated using a chromatographic column with silica gel (CH3OH)CHCl3, 0-1%) to obtain a solid white color, compound 21 (100 mg, 45%).

1H-NMR (CDCl3); 11,40 (Shir.s, 1H, NH); 7,26, to 8.20 (m, 17H, Ar-H, H-6 and NH); 6,36, 6,44 (DD, JH-H= 2,8 Hz, JF-H= 21 Hz, 1H, H-1'); 5,62, of 5.68 (DD, 1H, H-3'); 5,39, to 5.56 (DD, 1H, H-2'); 4,58, is 4.85 (m, 3H, H-4' and H-5').

N1-(2'-deoxy-fluoro - L-arabinofuranosyl)-5 - iodotyrosine (22)

Compound 21 (100 mg, of 0.27 mmole) is treated with saturated NH3/MeOH (60 ml) at room temperature for 24 hours. In the chromatographic processing on silica gel (0-10% CH3OH/CHCl3) get the connection 22 (35 mg, 71%) as a solid white color. []D= -65,4 (from 0.34, CH3OH). UV (MeOH)max= 293 nm.

1H-NMR (DMSO-d6): of 8.04 (s, 1H, H-6); 6,74, 7,94 (s, 1H, NH); 6,01, between 6.08 (DD,H-H= 3,9 Hz, JF-H= 16.6 Hz, 1H, H-1'), to 5.85 (d, 1H, 3-OH), 5,17 (t, 1-H, 5'-OH); to 5.08 (t, 1H, H-2'); 4,89 (t, 1H, H-3'); 4,15-to 4.23 (m, 1H, H-4'), 3,63-with 3.79 (m, 2H, H-5').

N1-(2'-deoxy-2'-fluoro-3',5'-di-benzoyl - L-arabinofuranosyl) -5-iodouracil (23)

To a solution of compound 13 (260 peremeshivayte at room temperature for 36 hours, and then evaporated to dryness. The residue is dissolved in 20 ml of CH2Cl2, washed with water (10 ml), saturated sodium bicarbonate solution (10 ml) and dried over magnesium sulfate. After filtration and evaporation receive premasagar connection 14 in the form of syrup.

At the same time 5-iodouracil (270 mg, 1.12 mmole) is suspended in 10 ml of HMDS and refluxed for 36 hours to obtain a homogeneous solution. It is evaporated

in vacuum to dryness. To this add a solution of compound 14 in anhydrous 1,2-dichloroethane, and the resulting solution was refluxed under nitrogen atmosphere for 1.5 days. Add CHCl3(20 ml) and the resulting solution washed successively with water (10 ml), brine (10 ml) and saturated sodium bicarbonate solution (10 ml) and dried over magnesium sulfate. After removal of the solvent receive a syrup, which crystallized from CH2Cl2to obtain compound 23 in the form of a solid yellow color (237 mg, 73%). Part of it (70 mg) is recrystallized from 2-isopropanol to obtain solid white (67 mg). UV (methanol): max= 230,0 nm 276 nm.

1H-NMR (CDCl3): 8,4, and 7.3 (m, 12H, Ar-H); of 6.29 (DD, JH-H= 2,43 Hz, JF-H= 21,6 G>N1-(2'-deoxy-2'-fluoro - L-arabinofuranosyl)-5 - iodouracil (24)

Compound 23 (40 mg, 0,069 mmole) is dissolved in 25 ml of saturated NH3/MeOH and stirred at room temperature for 24 hours and then evaporated to dryness. The resulting syrup is purified using preparative TLC (5:1/CHCl3:MeOH) to obtain compound 24 as a solid (19 mg, 74%). UV (MeOH):max= for 280.5 nm.

1H-NMR (DMSO-d6): 11,82 (Shir.s, CONH); 8,24 (s, 1H, H-6); 6,082 (DD, JH-H= 4,45 Hz, JF-H= to 13.7 Hz, 1H, H-1'); 5,947 (d, 1H, 3'-OH); at 5,296 (t, 1H, 5'-OH); 5,07 (dt, JF-H= 53 Hz, 1H, H-2); 4,24 (Shir.d, JF-H= 21 Hz, 1H, H-3'); 3,81, 3,55 (m, 3H, H-4', H-5').

2,3,5-tri-O-benzyl-L-arabinose

As is illustrated in Fig. 9, 30 g (0,2 mol) of powder of L-arabinose (5), and then 0.4 ml of concentrated sulfuric acid is added to 600 ml (14.8 moles) of anhydrous methanol. This suspension is stirred and heated at a low boil under reflux to obtain a clear solution within 2 hours. Then the reaction mixture was neutralized with 1.5 g of sodium bicarbonate, dried over magnesium sulfate, and then evaporated in a vacuum to obtain a thick syrup (compound 6), which is diluted with 50 ml sizeofimage of tetrahydrofuran, and again concentrate (35-40oC bath) for deletion and 30 g of Drierite, 156 g (2,78 mole) of potassium hydroxide and 200 ml (1,74 mole) of benzylchloride. The resulting mixture is heated at a low boil under reflux overnight, cooled, filtered through a thin layer of celite and concentrated in vacuo and then under high vacuum and 100oC (bath). The crude syrupy methyl-2, 3,5-tri-O-benzyl-L-arabinoside (7) is dissolved in 400 ml of glacial acetic acid. Hydrolysis mixture is heated to 65-70oC for 2 hours, concentrated in vacuo to one third volume and poured into 2.5 l of a mixture of ice and water. After the introduction of the seed (the seed crystals were originally received chromatography with elution with dichloromethane), the mixture was incubated in 5oC during the night. The aqueous layer was decanted from the semi-crystalline mass, and the latter is dissolved in 200 ml of dichloromethane. The resulting solution was washed with cold aqueous sodium bicarbonate, dried over magnesium sulfate, filtered through a thin layer of decolorizing charcoal and concentrated in vacuo to obtain a syrup which is dissolved in 200 ml of cyclohexane. After the introduction of the seed solution stand at room temperature for 1 hour and then at 5oC during the night to obtain 27.7 g (33,2%) of 2,3,5-tri-O-benzyl - L-amean)max= 220;

1H-NMR (300 MHz, CDCl3): 3,48-3,62 (m, 2H, H-5); 3,94-4,18 (m, 3H, H-2, 3,4); 5,33 (d, 1H, J = 4,07, H-1); from 5.29 (s, H-1); 7,25-7,33 (m, 15, H-Aromat.).

2,3,5-tri-O-benzyl-1-O-(p-nitrobenzoyl)-L-arabinose (9)

Compound 8 (Fig. 9) (2 g, was 4.76 mmole) dissolved in 7.5 ml of dichloromethane, and to this solution add a solution of 0.95 g (5.1 mmole) of p-nitrobenzaldehyde in a mixture of 5 ml of dichloromethane and 1.5 ml of pyridine. The reaction mixture was kept at room temperature overnight, and then washed sequentially 1 N. hydrochloric acid (10 ml x 2), aqueous sodium bicarbonate (10 ml x 2) and water (30 ml x 3). Moisture is removed by means of Na2SO4obtained solution was concentrated in vacuo to obtain 2.67 g (98,4%) mixture of anomers of compound (9). So melting 68-82oC. Additional purification on a chromatographic column with silica gel (hexane:acetone 8:1) ensure the melting temperature 89-93oC; []2D4= 13,04 (C 6,8, CH2Cl2)

UV (MeOH)max= 215,5 and 258,5,

1H-NMR (250 MHz, CDCl3): 3,54 at 3.69 (m, 2H, H-5); 4,06 (m, 1H, H-4'); 4,22 (m, 1H, H-3'); to 4.33 (m, 1H, H-2'); to 4.38-4,78 (m, 6H, benzyl CH2); 6,50 (d, 1H, J = 2,35, H-1); 7.23 percent and 7.36 (m, 15H, H-Aromat.benzyl group); 8,01-of 8.25 (m, 4H, H-Aromat. nitrobenzyloxy group).

10(2,3,5-tri-O-benzyl-L-arabinosyl)thymine (12)oC) during the night in an atmosphere of nitrogen to obtain a clear solution. Excess hexamethyldisilazane removed in vacuum, avoiding contact with moisture, resulting in a gain syrup connection 11.

Compound 9 (1 g, of 1.76 mmole) are added to 17 ml of dichloromethane, pre-saturated anhydrous hydrogen chloride at 0oC. After 2 hours at 0oC besieged p-nitrobenzoic acid (0.25 g) is removed by filtration and the resulting filtrate was concentrated in vacuo to obtain a syrup, and then kept in a high vacuum at room temperature for 2 hours, resulting in a gain 2,3,5-tri-O-benzyl - L-arabinopyranoside (compound 10).

Thus obtained compound 10 was dissolved in 15 ml of anhydrous dichloromethane, the resulting solution is added to the mixture similarvideo thymine (11) and 3 g of molecular sieves. The reaction mixture was stirred at room temperature in an argon atmosphere for 18 hours. The reaction mixture is diluted with 50 ml dichloromethane and poured into 2 ml of saturated aqueous NaHCO3under vigorous stirring. Appears white precipitate (tin hydroxide), which is filtered off on a layer of celite. The organic layer is separated from water and industrial the blockhead sodium, and then evaporated in vacuo to obtain a syrup, which was purified through column chromatography with silica gel (chloroform:methanol (100:1)) to obtain compound 12 in the form of syrup (0.68 g, 73%). []2D5= -56,71 (C 0,6, CH2Cl2).

UV (MeOH)max= 218, 265;

1H-NMR (300 MHz, CDCl3): to 1.67 (d, 3H, J = 1,11, CH3); 3,66-3,70 (m, 2H, H-5'); 4,06 (m, 1H, H-4'); 4,13 (t, 1H, J = 4,7, H-3'); 4,41, 4,54, 4,56 (m, 6H, benzyl CH2); 6,28 (d, 1H, J = 5,24, H-1'); 7,15-7,33 (m, 15H, H-Aromat.); the 7.43 (d, 1H, J = 1,31, H-6).

1- -L-arabinofuranoside (13)

Pallidiflora (680 mg, 3,835 mmole) is suspended in 100 ml of methanol and restore, shaking at room temperature in a hydrogen atmosphere. Then a solution of 450 mg of compound 12 in 25 ml of methanol is added to the acidified suspension of palladium mobiles. The reaction mixture was shaken at room temperature in a hydrogen atmosphere for 38 hours. After removal of the catalyst the solution is neutralized Dowex (HCO3) to pH 7 and concentrated in vacuo to obtain a solid white color, which after recrystallization from ethanol gives compound 13 (105 mg, 47.7 percent). So Plav. 244-249oC. []2D5= -91,48 (0,25, H2O).

IR (KBr): 1750, 1600 cm-1(CO); UV (MeOH): max= 268 nm;

1 (Shir. s, 1H, C'5-OH, currency); 5,54 (d, 1H, J = 5,23, C'2-OH or C'3-OH, currency); 5,97 (d, 1H, J = with 4.64, H-1'); 7,51 (d, 1H, J = 0,97, H-6); of 11.26 (s, 1H, NH, exchange). Elemental analysis for C10H14N2O6;

Calculated: C 46,51 H 5,46 N 10,85

Found: C 46,67 H 5,63 N 10,56

1-(2,3,5-tri-O-benzyl-L-arabinosyl)cytosine (15)

Cytosine (and 0.61 g, 6 mmol) and ammonium sulfate (2 mg) are suspended in hexamethyldisilazane (15 ml), and everything is refluxed (140oC) in nitrogen atmosphere for 2 hours to obtain a transparent solution. The mixture similarvideo cytosine is evaporated to dryness in a vacuum, avoiding contact with moisture, to obtain compound 14 in the form of syrup.

Compound 9 (2,82 g, 5 mmol) are added to 47 ml of dry dichloromethane, pre-saturated anhydrous hydrogen chloride at 0oC. After 2 hours at 0oC fell in sediment p-nitrobenzoic acid (0,807 g) is removed by filtration, the obtained filtrate was concentrated in vacuo to obtain compound 10, - 2,3,5-tri-O-benzyl-L-arabinopyranoside in syrupy form.

Thus obtained compound 10 was dissolved 28.5 ml of anhydrous dichloromethane, the resulting solution is added to the mixture similarvideo cytosine (14) and 8.3 g of molecular sieves . The reaction is ablaut 50 ml of dichloromethane and 20 ml of water and poured into 2 ml of saturated aqueous NaHCO3under vigorous stirring. Appears white precipitate (tin hydroxide), which is filtered off on a layer of celite. The organic layer is separated from the water and washed with water (30 ml x 3). The aqueous layer was extracted with dichloromethane, the combined dichloromethane layers are dried over sodium sulfate, and then evaporated in vacuo to obtain a syrup, which was purified through column chromatography with silica gel (chloroform:methanol 96:4) to obtain compound 15 in the form of a solid white color, which after recrystallization from 2-propanol is 1,53 g (60%). So melting point 146-148oC []25= -105,2 (C1, CH2Cl2); UV (MeOH):max= of 211.5,min= 272,5, at pH 7;max= of 211.5,min= 284 at pH 9.

1H-NMR (300 MHz, CDCl3): the 3.65 (d, 2H, J = 4,85, H-5'); 4,00 (t, 1H, J = 3,72, H-3'); 4,11 (m, 1H, H-4'); to 4.28 (m, 1H, H-2'); to 4.38-4,55 (m, 6H, benzyl CH2); to 5.56 (d, 1H, J = 7,5, H-5); to 6.39 (d, 1H, J = 4,59, H-1'); 7,12-7,31 (m, 15H, H-Aromat.); 7,63 (d, 1H, J = 7,5, H-6).

Cleaners containing hydrochloride salt (16) 1- -L-arabinofuranosylcytosine obtained by the catalytic hydrogenation of compound 15

Pallidiflora (315 mg, of 1.78 mmole) are suspended in 160 ml of methanol and restore, shaking at room temperature in a hydrogen atmosphere. To this acidic suspension of palladium is atoi temperature for 3 hours. After removal of the catalyst the resulting solution was neutralized Dowex (HCO3), concentrated in vacuo, and then purified using preparative TLC (MeOH:CHCl3, 3:5) to obtain a syrup which is dissolved in 3 ml of methanol, to which was added 1% HCl solution in MeOH to pH 1, concentrated to dryness and triturated with 2-propanol, resulting in a gain of 36 mg (22,1%) of compound 16. So melting 190-194oC []2D5= -115,47 (C 0,07, H2O); UV (H2O)max= 275 nm at pH 7;max= of 209.5 273 at pH 11;max= 280 nm at pH 1.

1H-NMR (300 MHz, DMSO-d6): 3,61 (d, 2H, H-5'), 3,82 (m, 1H, H-4'); 3,93 (m, 1H, H-2' or H-3'); 4,04 (Shir.s, 1H, H-2' or H-3'); 5,18 (Shir.s, 1H, C5'-OH, currency); 5,61 (Shir.s, 1H, C2'-OH or C3'-OH, in the exchange), 5,67 (Shir.s, 1H, C2'-OH or C3'-OH, currency); 6,00 (d, 1H, J = was 4.02, H-1'); 6,01 (d, 1H, J5,6= 7,8, H-5), 7,92 (d, 1H, J5,6= 7,8, H-6), 8,49 (Shir.s, 1H, NH, capable of currency), 9,38 (Shir.s, 1H, NH, capable of currency).

Cleaners containing hydrochloride salt of 1-L-arabinofuranosylcytosine (16), obtained by treating compound 15 trichloride boron

5 ml of 1M boron trichloride in dichloromethane is cooled to -72oC (dry ice-acetone). To a solution of boron trichloride is added slowly a solution of compound 15 (180 mg, 0,351 mmole) in 3 ml dichloromethane. After 2.75 hours (full dichlormethane (10 ml) and the resulting solution is evaporated to dryness (three times, until you get the balance of a white solid substance), add cold saturated sodium bicarbonate solution to establish a pH of 6-7. The resulting mixture was diluted with ethanol, heated to boiling, treated with charcoal and filtered. The resulting filtrate is evaporated to dryness, obtaining a syrup which is dissolved in 3 ml of methanol, add 1% solution of HCl in methanol to pH 1, concentrated to dryness and triturated with 2-propanol to obtain compound 16 (66 mg, 78.4 per cent).

9-(2,3,5-tri-O-benzyl-L-arabinosyl/adenine (18)

Carefully dried compound 9 (5 g, 8.8 mmole) are added to 82 ml of dichloromethane, pre-saturated anhydrous hydrogen chloride at 0oC. After 2 hours of reaction at 0oC fell in sediment p-nitrobenzoic acid (1,53 g) is removed by filtration and the resulting filtrate was concentrated in vacuo to obtain a syrup, which is kept in a full vacuum at room temperature for 2 hours. Thus obtained 2,3,5-tri-O-benzyl-L-arabinopyranoside (compound 10) was dissolved in 50 ml of dichloromethane, and this solution is added to a mixture of 4.5 g (18,8 mmole) of dried N-benzyladenine5(17) and 14.5 g of molecular sieves . The reaction mixture was stirred at room temperature is see through column chromatography with silica gel, using a mixture of hexane-acetone (3: 1, Rf= 0,22). The obtained product is isolated and concentrated in vacuo to obtain a syrup which is dissolved and mixed with methyl ammonia (20 ml) in a bomb stainless steel, and then heated overnight at 50-55oC. the resulting solution was then concentrated under reduced pressure to obtain a semi-solid substance, which after recrystallization from warm isopropyl alcohol gives compound 18 (2.4 g, 50.7 per cent. So melting point 128-129oC. []27D/= -20,04 (1,04 CH2Cl2);

UV (CH2Cl2):max= 213, 258,5.;

1H-NMR (250 MHz, CDCl3): 1,95 (Shir.s, 14, NH, capable of currency); of 3.69 (d, 2H, J = 4,82, H-5'); 4,18-4,30 (m, 6H, benzyl CH2); 4,51 with 4.64 (m, 3H, H-2', 3', 4'); 5,73 (Shir. s, 1H, H capable of currency); of 6.52 (d, 1H, J = 4,00, N-1'); 6,89-6,93, 7,17-7,37 (m, 15 H, H-Aromat.benzyl group); 8,17 (C, 1H, H-2 or H-8); 8,32 (s, 1H, H-2 or H-8).

9- -L-arabinofuranoside (19)

Trichloride boron (5 ml, 1M) in dichloromethane cooled to -72oC (dry ice-acetone). A solution of compound 18 (150 mg, 0,279 mmole) in 5 ml of dichloromethane is added slowly to a solution of boron trichloride. After 3.5 hours of reaction time the cooling bath is removed and the solvent and the gas is removed in vacuum. The residue is dissolved in cold on the Cold saturated solution of sodium bicarbonate is added to establish a pH of 7-8. The resulting mixture was diluted with ethanol, heated to boiling, the suspension is filtered through celite, and the obtained filtrate was concentrated in vacuo to a syrup, which crystallized from water. The connection output was 55 mg (74%). So melting 256-258oC. UV (H2O):max= 259.

1H-NMR (300 MHz, DMSO-d6): 3,62-3,66 (m, 2H, H-5'); of 3.77 (Shir.s, 1H, H-4'); 4,13 (Shir.s, 2H, N-2', 3'); 5,12 (t, 1H, J = 5,4, C'5-OH, is able to exchange) 5,54 (d, 1H, J = 3,78, C'2-OH or C'3-OH, is able to exchange); 5,63 (d, 1H, J = 4,32, C'2-OH or C'3-OH, is able to exchange); and 6.25 (d, 1H, J = was 4.02, H-1'), 7,25 (Shir.s, 2H, NH2capable of currency); 8,13 (s, 1H, H-2 or H-8); 8,18 (s, 1H, H-2 or H-8).

Fig. 10 is an illustration of an alternative way to obtain 1-O-acetyl-2,3,5-tri-O-benzoyl - L-ribofuranose (compound 10) of 1,2-di-O-isopropylidene - a-L-xylofuranose (compound 3).

1,2-di-O-isopropylidene - a-L-xylofuranose (3)

To 650 ml of anhydrous acetone are added 4 ml of concentrated sulfuric acid, 5 g of molecular sieves 80 g of anhydrous copper sulfate (2) and 40 g L-xylose. The resulting mixture was stirred at room temperature for 36 hours. The reaction mixture was filtered and thoroughly washed with acetone, and the combined filtrate is neutralized with ammonium hydroxide and then evaporated to dryness. Add 200 ml of the PRS was stirred at room temperature for 2.5 hours. Due to the saturated NaHCO3set pH 8, and then evaporated to dryness. The residue is extracted with ethyl acetate. After removal of the solvent receive oil yellow connection 3 (41.7 g, 82.3 per cent).

1H-NMR (CDCl3): 5,979 (d, J = to 3.73 Hz, 1H, H-1); 4,519 (d, J = 3.6 Hz, 1H, H-2); 4,308 (Shir. d, 1H, H-3); 4,08 (and, 3H, H-4 and H-5); of 1.321 (s, 3H, CH3); 1,253 (s, 3H, CH3).

5-O-benzoyl-1,2-di-O-isopropylidene - a-L - xylofuranose (25)

Connection 3 (41 g, 215,6 mmole) is stirred in pyridine (150 ml) and CH2Cl2(150 ml) at 0oC. BzCl (27.5 ml, 237 mmole), dissolved in 30 ml of pyridine, are added to this mixture dropwise. After 30 minutes, add 5 ml of water, the mixture is evaporated to dryness, dissolved in EtOAc, washed with saturated sodium bicarbonate solution and dried over anhydrous sodium sulfate. After evaporation of the solvent receive a syrup of orange color, which is crystallized from Et2O, when receiving the connection 20 (36 g). The mother liquor is evaporated to dryness and the residue is purified on a chromatographic column with silica gel (1% CH3OH/CHCl3) to receive another portion of the connection 25 (12 g, total yield 76%). So melting 82-83oC (literature dealing with.1Dform83,5-84,5oC).

1H-NMR (CDCl3): 7,43, 8,07 (m, 5H, Ar-H); 5,97 (d,51, 1,32 (2s, 6H, 2CH3).

5-O-benzoyl-1,2-di-O-isopropylidene - a-L-erythropoietins-3-ulose (26)

Compound 25 (40 g, 136 mmol) is stirred in 450 ml of CH2Cl2. To this mixture add pyridinediamine (PDC, a 30.7 g, 81,6 mmole) and Ac2O (42,3 ml, 448,8 mmole). The resulting mixture was concentrated to 1/5 of its original volume, and then it was added EtOAc (50 ml). The resulting solution is filtered, the resulting filtrate was poured on a layer of silica gel (10 cm x 5 cm), elute EtOAc, the combined eluate concentrate and co evaporated with toluene (50 ml x 2). After crystallization from hexane and EtOAc get a connection 26 in the form of a solid white (38 g, 96%). So melting 91-93oC []D: -132(C, 1,0, CHCl3); the literature dealing with.2Dform: so melting 93-94oC []D: +135 (C, 1,0, CHCl3); IR (KBr): 1773 (ArCO), 1730 cm-1(CO).

1H-NMR (CDCl3): 7,97, 7,42 (m, 5H, Ar-H); 6,14 (d, 1H, J = 4.4 Hz, H-1); 4,74, and 4.68 (m, 24, H-4, H-2); 4,50, of 4.44 (m, 2H, H-5, H-5'); 1,52, the 1.44 (2s, 6H, 2CH3). Elemental analysis for (C15H16O6): Calculated: C 61,64 H 5,52;

Found: C 61,42 H Of 5.53.

1,2-di-O-isopropylidene - a-L-ribofuranose (27)

Compound 26 (37 g, 127 mmol) was dissolved in EtOH/H2O (400 ml/100 ml) at 0oC, adding portions NaBH4(23.3 g, any the filtrate is evaporated to dryness and evaporated together with methanol. After chromatographic processing on silica gel (0-15%, CH3OH/CH2Cl2) and crystallization from EtOAc/hexane get a connection 27 in the form of white needles (19 g, 79%). So melting 86-87oC []D= -31,5 (s, 0,62, CHCl3), the literature dealing with.3Dform: so melting 86-87oC []D= +37(C, 0,59, CHCl3).

IR (KBr) 3356 cm-1(OH).

1H-NMR (CDCl3): of 5.83 (d, 1H, J = 3.98 Hz, H-1); 4,595 (t, 1H, H-2), 4,055, and 3.72 (m, 4H, H-3, H-4, H-5, H-5'); of 2.38 (d, 1H, D2O capable of currency, 3-OH); to 1.83 (t, 1H, D2O capable of currency, 5-OH); 1,58, to 1.38 (2s, 6H, 2CH3); Elemental analysis (C8H14O5): Calculated: C 50,50 H 7,42;

Found: C 50,62 H 7,46.

3,5-di-O-benzoyl-1,2-di-O-isopropylidene - a-L - ribofuranose (28)

Compound 27 (19 g, 100 mmol) is stirred in 300 ml of pyridine at 0oC, prokopeva this BzCl (40 ml, 348 mmole), and then stirred at room temperature for 3 hours. The solvent is evaporated to dryness. The residue is extracted with EtOAc, washed with saturated solution of NaHCO3, dried over sodium sulfate, the solvent is evaporated and crystallized from ether, receiving the connection 28 in the form of a solid white (39 g, 98%). So melting point 83-85oC.

1H-NMR (CDCl3): 8,07, of 7.36 /SUB>H22O7). Calculated: C 66,50 H 5,64;

Found: C 66,32 H 5,57.

1-O-acetyl-2,3,5-tri-O-benzoyl - L - ribofuranose (31)

Compound 23 (38 g, 95 mmol) is stirred in 300 ml of 1% HCl/CH3OH at room temperature for 30 hours, add 20 ml of pyridine, and then the solution is evaporated to dryness. The residue is evaporated together with pyridine (30 ml x 2) and then dissolved in 100 ml of anhydrous pyridine, prokopeva BzCl (17 ml, 146 mmol) at 0oC, then stirred at room temperature for 3 hours. The solvent is evaporated and the residue dissolved in EtOAc, washed with 0.5 N. HCl, then with saturated NaHCO3, dried over sodium sulfate. After evaporation of the solvent to obtain crude compound 30 in the form of syrup. This crude compound was stirred in glacial acetic acid (400 ml), and then Ac2O (100 ml) at 0oC, prokopeva concentrated sulfuric acid (10 ml). This solution was stirred at room temperature overnight. The resulting mixture was poured into a mixture of ice-water, extracted with CHCl3, neutralized with saturated sodium bicarbonate solution, and then dried over magnesium sulfate. After evaporation of the solvent receive light yellow syrup, which crystallized from methanol to obtain UB> (C, 1,0, CHCl3); the literature dealing with.4So melting 129-130oC []D= -43,6 (C, 1,0, CHCl3).

1H-NMR (CDCl3): 7,317, 8,134 (m, 15H, OBz); 6,437 (s, 1H, H-1); 5,835 (m, 2H, H-2 and H-3); 4,649 (m, 3H, H-4 and H-5); 2,003 (s, 3H ).

II. Biological activity

Disclosed here, the compounds can be assessed in terms of their activity against HBV and EBV, as described hereinafter, or by other methods known to experts.

Example 1. Biological activity against HBV

Use hepatoma cells of a person with HBV (2.2.15 cells) in this analysis. These cells are grown to confluence in culture medium with the minimum necessary set with 10% serum, fetal calf. Wednesday replaced with a three-day intervals. Merging start processing the drugs, and then continue it for 3 days. Again add the same concentration of drug after removal of the medium from the cultures and incubated for these cultures for a further 3 days. Wednesday after the six-day processing harvested, and viral particles are precipitated using polyethylene glycol. Then the particles digest, and performed Southern analysis. The resulting blots hybridizing with HBV-specific probe and estimate the number of viral DNA in comparison with the levels of DNA from cultures of individual DNA defined relative to the integrated HBV DNA. Calculate the concentration of drug that causes 50% inhibition of DNA (ID50compared with the control samples. The results obtained are summarized in table 1.

Example 2. Biological activity against EBV

H1 cells support long-term growth phase for two days before primary treatment. Cells are centrifuged at 600 g for 10 minutes at room temperature in order to separate them from the environment containing pre-existing viral particles. H1 cells sown in plates with 24 cells at a density of 1105cells to a cell in 2 ml of fresh medium with medication or without him and incubated at 37oC for 5 days. The medium containing the virion, preserved and used for evaluation of the inhibitory effect of drugs on productivity and gain popularity virus using bioanalysis. The virion is then precipitated from not containing cell medium by centrifugation at 45000 rpm for 90 minutes in the rotor SW-50 Ti (Beckman). Virions resuspended in 1 ml growth medium, and then used to infect 1106of Raji cells within 48 hours. Since the level of EBV DP activity in Raji cells after superantisypware proportional to the amount added of Larionov, hiraniwa EBV DP activity repeatedly diluted with controls. If the cells treated with 1 mm L-FMAU for 6 days, with no evidence of any inhibition of the content of mitochondrial DNA in H1 cells.

Slot-blotting - Number of mitochondrial DNA is determined using the method slot-blotting (2). Treated and untreated cells H1 in the number 2105subjected to lysis in 200 μl 10 mm Tris.HCl (pH 7.5) solution by the method of freeze/thaw. Cell lysate treated with 10 μg/ml RNase A at 37oC for 30 min, and then with proteinase K (100 µg/ml) at 55oC for 2 hours. Equal amounts of 20 x SSC buffer is added to each cell lysate. After boiling for 10 minutes, the samples applied to nylon membranes (Hyhond-N, Amersham Corp.). Radiolucency fragment of the mitochondrial DNA of a person is used as a probe for DNA hybridization. The same membrane probe again Alu human DNA after removal of the probe mitochondrial DNA. The number of mitochondrial DNA in treated and untreated H1 cells counted using a densitometer (LKB Ultroscan XL).

The results obtained are presented in table 2.

The effect of inhibition of the compounds on EBV: Value CD50receive, exposing cells H1 to the influence of different concentration is univelt the values obtained with the control. H1 cells treated for 5 days and determine ID90in the bioassays.

Example 3. The excretion of L-(-)-FMAU from plasma and liver

Evaluate the cleaning plasma and liver of mice from L-(-)-FMAU after oral administration. Mice injected with L-(-)-FMAU, labeled with tritium (regiospecificity 9.3 µmol/mccoury).

Fig. 11 is a graph of the concentration of L- (-)-FMAU in the plasma of mice after oral administration of from time to time, and Fig. 12 is a graph of the concentration of L-(-)- FMAU in the liver of mice after oral administration of from time to time (cross, 10 mg/kg administered twice daily for 30 days prior to pharmacokinetic studies, and then conduct research on the thirty-first day after injection of the same concentration; shaded circles - administration of a 50 mg/kg twice a day for 30 days before the studies, and then on the thirty-first day doing research after the introduction of the same concentration; where there's no shading mugs - introduction for the first time 50 mg/kg on the first day of the study).

In these moments of time (see Fig. 10 and 11) in mice deprived blood from the retro-orbital sinus using processed heparinized capillaries. Plasma is extracted with trichloroacetic acid and neutralise the>As shown in Fig. 10 and 11, the peak concentration of L-(-)-FMAU in plasma and liver was observed after about one hour. The connection is virtually eliminated from plasma and liver after four hours.

Example 4. The toxicity of L-(-)-FMAU for female mice BDF1

Fig. 13a illustrates the change in body weight after thirty days, the control BDF1 female mice. Fig. 13b and 13c illustrate the changes in body weight after 30 days in female BDF1 mice, which is administered 10 mg/kg (13b) and 50 mg/kg (13c) twice a day L-(-)-FMAU. The weight of the representative body are presented as mean and standard deviation for 5-7 mice. As can be seen from Fig. 13 and 13c, L-(-)-FMAU, apparently, has no serious effect on the weight of the mice within thirty days, indicating good tolerance of the connection.

Example 5. Clinical analysis of plasma of mice after treatment with L-(-)-FMAU

In Fig. 14-20 presents the results of clinical analyses of the chemical composition of the plasma of mice after administration of L-(-)-FMAU in doses of 10 mg/kg (three mice) or 50 mg/kg (three mouse) twice a day for thirty days.

Fig. 14 is a graph of the concentration of total bilirubin in the plasma of mice in mg/DL. Fig. 15 is a graph of the concentration of alkaline phosphatase in mouse plasma in U/L. Obeaune in mice fall into a normal range of values for humans (less than 1.2 mg/DL), but the values for alkaline phosphatase slightly above normal levels for humans (30-114 U/l).

Fig. 16 is a graph of the concentration of creatinine in mouse plasma in mg/DL. Creatinine is an indicator of liver function. Except mouse 50-2 levels of creatinine in treated mice did not differ from levels in control mice.

Fig. 17 is a graph of the concentration of AST (SGOT, serum glutamic oxalic transaminase levels in mouse plasma in U/L. Fig. 18 is a graph of concentrations of AST (SGPT, serum glutamic pyruvic transaminase levels in mouse plasma in U/l SGOT and SGPT are indicators of liver function. Except mouse 50-2 (SGOT and SGPT) and mouse 10-2 (only for SGOT) levels of the enzyme-treated mice did not differ from the levels observed in control animals.

Fig. 19 is a graph of the concentration of lactic acid in rat plasma in mmol/L. Fig. 20 represents a graph of concentrations of lactic dehydrogenase in mouse plasma in U/l Lactic acid produced in the muscles during glycolysis. Lactic dehydrogenase (LDH) is present in the form of various isoenzymes in various tissues. The allocation of LDH in plasma may serve as an indicator of powerdrome for control animals.

III. Oligonucleotides

The oligonucleotides of the desired sequences can be modified by replacing one or more of L-nucleosides disclosed here, the nucleotide in the oligonucleotide. In a preferred embodiment, L-nucleoside is placed on one end of the oligonucleotide. The modified oligonucleotide can be used, for example, antisense technology.

Antisense technology relates generally to the modulation of gene expression through a process in which sinteticheski nucleotide hybridized with complementary sequence of nucleic acid to inhibit the transcription or replication (if mistaway sequence is DNA), to inhibit translation (if mistaway sequence is RNA) or to inhibit the processing (if mistaway sequence is pre-RNA). Using this technique, it is possible to modulate a wide range of cellular activities. A simple example is the inhibition of protein biosynthesis at the expense of antisense oligonucleotide associated with the mRNA. In another embodiment, a synthetic oligonucleotide hybridized with specific gene sequence in dvuhtsepochnyj DNA, forming tranceboy complex is but also be used to activate gene expression indirectly, inhibiting the biosynthesis of natural repressor. AOT can be used to suppress the expression of pathogenic genes, such as those that facilitate the replication of viruses, including human immunodeficiency virus (HIV), hepatitis B virus (HBV) and herpes virus, as well as cancers, particularly solid tumors, such as glioma, breast cancer and melanoma.

The stability of the selected oligonucleotide against nucleases is an important factor for applications in vivo. It is known that the 3'-and Exo-nuclease activity responsible for the destruction of the unmodified antisense oligonucleotides in serum (Vlassov centuries, Yakubov, L. A., Prospects for Antisense Nucleic Acid Therapy of Cancer and AIDS, 1991, 243-266, Wiley-Liss, Inc. , New York; Nucleic Acids Res., 1993, 21, 145). In one embodiment, the disclosed here L-nucleotides can be used to minimize the 3'-exonuclease antisense oligonucleotides.

The oligonucleotides of the present invention, which is able to contact polyribonucleotide acid or polydeoxyribonucleotide acid, can be used as antisense agents in such a way as conventional antisense agents. See, basically: Antisense Molecular Biology and S-oligos, Synthesis, 1 (Oct. 1988) (published Synthecell Corp., Rockville, Md. ); 2 J. DiscoverieMartin C. , J. Med.Chem., 1993, 36, 1923-1937. Antisense agents of the present invention can be used, designing antisense agent that is capable of selectively contact with a predetermined sequence polydeoxyribonucleotide acid or sequence polyribonucleotide acid of a cell containing such a sequence (for example, adding antisense agent in the culture medium containing the same cell), so that the antisense agent was captured by the cell, is contacted with a predetermined sequence and would block its transcription, translation or replication. Requirements for the selective binding of the antisense agent is known (for example, length 17 basis for selective binding to the human genome).

IV. Obtaining pharmaceutical compositions

Disclosed here compounds and their pharmaceutically acceptable salts, prodrugs and derivatives can be used for the prevention and treatment of infections with HBV and EBV, as well as other related conditions such as anti-HBV or anti-EBV positive antibodies and HBV - or EBV-positive conditions, chronic liver inflammation caused by HBV, cirrhosis, acute hepatitis, fulminant (transient) hepatitis, hronologicheskij order to prevent or delay the development of clinical disease in individuals with anti-HBV and anti-EBV antibody or HBV - or EBV - positive antigens or those who have been in contact with HBV or EBV.

Patients with any of these conditions can be treated by introducing effective HBV - or EBV-healing amount of one of these active compounds or mixtures thereof, or pharmaceutically acceptable derivative or its salt, optionally in a pharmaceutically acceptable carrier or diluent. The active substance can be entered in any suitable way, for example, parenteral, oral, intravenous, subcutaneous, dermal or topically, in liquid or solid form.

Active connection include pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to the patient a therapeutically effective amount, which does not cause serious toxic effects in the treated patient.

The preferred dose of the active compound for all of these conditions will dose in the range of from about 1 to 60 mg/kg, preferably from 1 to 20 mg/kg of body weight per day, usually from 0.1 to about 100 mg/kg of body weight per day. Effective doses can be calculated based on the weight of the original nucleoside, which is to be paid. If the derivative itself is active, the effective dose m In one embodiment, the active compound is administered in accordance with the instructions on the leaflet for a medicinal product or in Physician''s Desk Reference (table reference of the doctor) at the 3'-azido-3'-deoxythymidine (AZT), 2',3'-dideoxyinosine (DDI), 2'3'-dideoxycytidine (DDC) and 2',3'-dideoxy-2',3'-didehydrothymidine (D4T) for HIV indication.

The compound is usually administered in the form of a separate suitable dosage form, including but not limited to these, a dose containing from 7 to 3000 mg, preferably from 70 to 1400 mg of active ingredient in a unit dose form. Usually convenient oral dose of 50-1000 mg

Ideally the active ingredient should be introduced to achieve peak concentrations of the active compounds in the plasma from about 0.2 to 70 μm, preferably from about 1.0 to 10 microns. This can be achieved, for example, by intravenous injection of a 0.1 to 5% solution of the active ingredient, optionally in saline solution or in the form of a pill with the active ingredient.

The active compound can be prepared in the form of pharmaceutically acceptable salts. In the sense, as used herein, the term pharmaceutically acceptable salts or complexes refers to salts or complexes of nucleosides, which retain the right Bioline toxic effects. Limitiruyuschie examples of such salts can serve as (a) salt accession inorganic acids (e.g. hydrochloric acid, Hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc. and salts of such organic acids as acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, ascorbic acid, benzoic acid, tannic acid, Paveway acid, alginic acid, polyglutamic acid, naphthalenesulfonate, naphthalenedisulfonic acid and polygalacturonic acid; (b) salt accession bases formed with cations such as sodium, potassium, zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, Nickel, cadmium, etc., or with organic cations as educated N,N-dibenziletilendiaminom, ammonia or Ethylenediamine; or (c) a combination of (a) and (b), for example, salt tannate zinc, etc.

Modification of the active compounds, specifically, N6or N4and 5'-O provisions may affect biouswoyaemosti and metabolic rate active compounds, providing control over the flow of active connections.

The concentration of the active compounds in medicinal whom the data specialists. It should be noted that the magnitude of the dose depends on the severity of the patient's condition, which should be facilitated. You should also understand that for a specific subject-specific dose schedule over time should vary according to individual needs and professional experience of the treating physician, and the concentration ranges are only examples and in no way should limit the scope of the invention or the practical application of the claimed compositions. The active ingredient can be taken immediately or it can be divided into several smaller doses and taken with different intervals between meals.

The preferred method of introduction of the active compounds is oral. Oral compositions generally include an inert diluent or edible carrier. They can be enclosed in gelatin capsules or can be pressed into tablets. For the purpose of oral therapeutic treatment of active compound may be incorporated in excipients and used in the form of tablets, pellets or capsules.

Part of the composition can be pharmaceutically compatible binding agents and/or adjuvants.

Tablets, sawing the s: such a binder, as microcrystalline cellulose, resin tragakant or gelatin; excipients, such as starch or lactose, leavening agents such as Aleynikova acid, Primogel or corn starch, such sliding substances as magnesium stearate or Sterotes or colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate or orange concentrate. If the unit dose form is a capsule, it may contain, in addition to the above types also such a carrier liquid, such as body oil. In addition, dosage unit forms can contain various other materials which modify the form of a unit dose, for example, sugar, shellac, or other coating for oral administration.

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

The active compound or pharmaceutically acceptable derivative, or its salt which can Supplement the desired action, for example, antibiotics, antifungal drugs, anti-inflammatory or other antiviral drugs, including anti-HBV, anti-EBV, anticytomegalovirus or anti-HIV or anti-EBV agents.

Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application can include the following components; sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol and other synthetic solvents; antibacterial agents as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfate; chelating agents like ethylenediaminetetraacetic acid; buffers as acetates, citrates or phosphates and such agents for regulating tone as sodium chloride or dextrose. Preparations for parenteral injection can be enclosed in ampoules, disposable syringes or vials with multiple doses, made of glass or plastic.

If the drug is administered intravenously, preferred carriers are physiological saline or superstarcase saline solution with phosphate s connection from rapid excretion from the body, for example, in the form of compositions with variable selection, including implants and microencapsulated delivery systems. You can use biodegradable biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyarteritis and polylactic acid. Methods of obtaining such compositions known in the art. These materials can also be purchased from Alza Corporation and Nova Pharmaceuticals, Inc.

Liposomal suspensions (including liposomes targeted to them are infected cells with monoclonal antibodies to viral antigens) are also preferred as pharmaceutically acceptable carriers. They can be obtained in accordance with methods known in the art, for example as described in U.S. patent N 4522811 (which is included as a reference). So, for example, liposomal compositions can be obtained by dissolving appropriate lipid (lipids) (such as stearoylethanolamine, stearoylethanolamine, arachidonylglycerol and cholesterol) in an inorganic solvent that is then evaporated, and there is a thin film of dried lipid on the surface of the vessel. Then injected into the vessel an aqueous solution of active compound or its monopoply material from the walls of the vessel and dispersing the lipid fragments, resulting receive liposomal suspension.

The present invention has been disclosed with reference to its preferred options. Other variations and modifications of the invention should be apparent from the preceding detailed description of the invention. All these variations and modifications are included in the scope of the attached claims.

1. L-Nucleoside of the General formulas (1)

< / BR>
where R represents the balance of uracil, thymine, cytosine or purine, which can be substituted by a halogen atom and alkyl;

R" represents a hydrogen atom, acyl, alkyl, monophosphate, diphosphate or triphosphate, or its pharmaceutically acceptable salt.

2. L-Nucleoside of the formula (1) under item 1 or its pharmaceutically acceptable salt as an active ingredient of a pharmaceutical composition having anti-HBV or anti-EBV activity.

3. L-Nucleoside of the formula (1) under item 1 or its pharmaceutically acceptable salt with anti-HBV or anti-EBV activity.

4. L-Nucleoside under item 1, which is 2'-fluoro-5-methyl--L-arabinofuranosyluracil.

5. L-Nucleoside under item 1 or 3, selected from the group including N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-ethyluracil,N1-(2'-ASS="ptx2">

6. L-Nucleoside under item 1 or 3, where the base is selected from the group consisting of 5-methyluracil(thymine), 5-iodouracil, cytosine, and 5-ethyluracil.

7. L-Nucleoside under item 2, which is a 2'-fluoro-5-methyl--L-arabinofuranosyluracil.

8. L-Nucleoside under item 2, which is selected from the group including N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-ethyluracil,N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-iodotyrosine and N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-iodouracil.

9. L-Nucleoside under item 2, wherein the base is selected from the group consisting of 5-methyluracil(thymine), 5-iodouracil, cytosine, and 5-ethyluracil.

10. Method of inhibiting HBV or EBV infection, characterized in that the injected L-nucleosil formula (1) under item 1 at a dose of from 0.1 to 100 mg/kg of body weight per day.

11. The method according to p. 10, characterized in that the L-nucleoside is a 2'-fluoro-5-methyl--L-arabinofuranosyluracil.

12. The method according to p. 10, characterized in that the L-nucleoside is selected from the group including N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-ethyluracil, N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-iodotyrosine and N1-(2'-deoxy-2'-fluoro--L-arabinofuranosyl)-5-iodouracil.

13. The method according to p. 10, otlichayuschij.

 

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where

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or

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The invention relates to new compounds of formula I Nu-O-Fa, where O is oxygen, Nu is a nucleoside or nucleoside analogue, including such nitrogen base, as adenine, Esenin, cytosine, uracil, thymine; Fa - acyl monounsaturated C18YPD C20-9-fatty acids, which fatty acid etherification hydroxyl group in 5-position of the sugar portion of the nucleoside or nucleoside analog, or a hydroxyl group, an acyclic chain of an analogue of the nucleoside

The invention relates to medicine

The invention relates to new nukleotidfosfatazu derived from the remnants of lipid esters of General formula I, in which R1, R2represent a linear or branched saturated alkyl chain containing 1-20 carbon atoms; R3, R5represent hydrogen, hydroxyl group; R4represents a hydroxyl group; X represents a sulfur atom, sulfinyl or sulfonyloxy group; Y represents an oxygen atom; b is a purine and/or pyrimidine base, provided that at least one of the residues R3or R5represents hydrogen; their tautomers, their optically active forms and racemic mixtures, or their physiologically acceptable salts with inorganic and organic acids and/or bases, and also to processes for their preparation and medicines containing the above-mentioned connection
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