β-l-2'-deoxynucleosides used in hepatitis b treatment

FIELD: medicine, pharmacology, bioorganic chemistry, pharmacy.

SUBSTANCE: invention relates to the effective using amount of β-L-2'-deoxynucleoside of the formula (I) or (II) used in manufacturing a medicinal agent used in treatment of hepatitis B, pharmaceutical compositions containing thereof, and methods for treatment of hepatitis B. Proposed agent shows the enhanced effectiveness in treatment of hepatitis B.

EFFECT: enhanced and valuable medicinal properties of agent.

83 cl, 6 tbl, 11 ex

 

This invention relates to methods for treating hepatitis b virus (also referred to as "HBV"), which include the introduction of a master if necessary, either separately or in combination - effective amount of one or more active compounds described in the application, or pharmaceutically acceptable prodrug or salt of one of these compounds.

HBV is second only to tobacco as a cause of human cancer. The mechanism by which HBV causes cancer is not known, although suggest that he can start the development of a tumor or indirectly, to start the development of a tumor through chronic inflammation, cirrhosis and cell regeneration associated with infection.

Hepatitis b virus reaches epidemic levels around the world. After an incubation period of from two to six months, during which the owner knows nothing about infection, HBV infection can lead to acute hepatitis and liver disease that causes pain in the abdomen, jaundice and elevated levels of certain enzymes in the blood. HBV can cause fulminant hepatitis, rapidly progressive and fatal form of the disease, which destroyed a massive portions of the liver.

Usually after acute hepatitis patients recover. However, in some patients, high levels of viral antigen with Granada in the blood for a long or indefinite period of time, causing chronic infectious disease. Chronic infection can lead to chronic persistiruuschem hepatitis. Patients infected with chronic persistent HBV, found mostly in developing countries. By mid-1991 in one of Asia had approximately 225 million chronic carriers of HBV, and worldwide nearly 300 million speakers. Chronic persistent hepatitis can cause fatigue, cirrhosis and hepatocellular carcinoma, a primary liver cancer.

The inhabitants of industrialized Nations high risk of HBV infection include groups of people who have been in contact with HBV carriers or samples of their blood. Epidemiology of HBV is very similar to the epidemiology of acquired immunodeficiency syndrome (AIDS; AIDS), which explains why HBV infection is common among patients with AIDS or AIDS related complex. However, HBV is more contagious than HIV (HIV).

However, later with the help of genetic engineering were received vaccines, which are widely used at the present time. Unfortunately, no vaccine can help patients who are already infected with HBV. Also it is shown that daily treatment α-interferon, genetically engineered protein is encouraging, but this therapy is successful only about one-third of treated patients the century In addition, interferon cannot be assigned for oral administration.

Identified a number of synthetic nucleosides which exhibit activity against HBV. (-)-Enantiomer VSN-189, known as 3TC, declared Liotta, et al. in U.S. Patent 5539116, approved by the U.S. Food and Drug Administration for the treatment of hepatitis C. see Also European Patent application EPA 0494119 A1, filed by BioChem Pharma, Inc.

CIS-2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxa-tolan ("FTC") has activity against HBV. Cm. International Application WO 92/15308; Furman, et al., "The Anti-Hepatitis In Virus Activities, Cytotoxicities, and Anabolic Profiles of the (-) and (+) Enantiomers of cis-5-Fluoro-1-[2-(Hydroxymethyl)-1,3-oxathiolane-5-yl]-Cytosine" Antimicrobial Agents and Chemotherapy, December 1992, page 2686-2692; and Cheng, et al., Journal of Biological Chemistry, Volume 267(20), 13938-13942 (1992).

Von Janta-Lipinski et al. describes the use of L-enantiomers 5'-triphosphates of 3'-fluoro-modified β-2'-deoxyribonucleosides for inhibition of hepatitis b virus polymerase (J. Med. Chem., 1998, 41, 2040-2046). In particular, 5'-triphosphates of 3'-deoxy-3'-fluoro-β-L-thymidine (β-L-FTTP), 2',3'-dideoxy-3'-fluoro-β-L-cytidine (β-L-FdCTP) and 2',3'-dideoxy-3'-fluoro-β-L-5-methylcytidine (β-L-FMethCTP described as effective inhibitors of the DNA polymerase of HBV.

In the International Application WO 96/13512 Genencor International, Inc. and Lipitek, Inc. claim that some L-ribofuranosyl nucleosides can be used for the treatment of cancer and viruses. In particular, describes the use of this class of compounds for the treatment of cancer and HIV.

In U.S. Patent No. 5565438, 5567688 and 5587362 (Chu, et al.) describes the use of 2'-fluoro-5-methyl-β-L-arabinofuranosyluracil (L-FMAU) for the treatment of hepatitis b virus and Epstein-Barr (Epstein-Barr).

In the International Application WO 92/18517 Yale University and University of Georgia Research Foundation, Inc., describes the use of L-FddC (β-L-5-fluoro-2',3'-dideoxycytidine) for the treatment of hepatitis C.

In this area known synthetic nucleosides β-L-2'-deoxycytidine (β-L-2'-dC), β-L-2'-deoxythymidine (β-L-dT) and β-L-2'-deoxyadenosine (β-L-2'-dA). Antonin Holy first described β-L-dC and β-L-dT in 1972, "Nucleic Acid Components and Their Analogs. CLIII. Preparation of 2'-deoxy-L-Ribonucleosides of the Pyrimidine Series," Collect. Czech. Chem. Commun. (1972), 37(12), 4072-87. Morris S. Zedeck et al., offered for the first time β-L-dA for synthesis inhibition induced enzymes in Pseudomonas testosteroni, Mol. Phys. (1967), 3(4), 386-95.

It is known that certain 2'-deoxy-β-L-Erythro-pentofuranose show antineoplastics and selective antiviral activity. Verri et al. describes the use of 2'-deoxy-β-L-Erythro-pentofuranose-nucleosides as antineoplastics agents and antiherpetic agents (Mol. Pharmacol. (1997), 51(1), 132-138 and Biochem. J. (1997), 328(1), 317-20). Saneyoshi et al. describe the use of 2'-deoxy-L-ribonucleosides as reverse transcriptase inhibitors (I) for control of retroviruses and for the treatment of AIDS Jpn. Kokai Tokkyo Koho JP 06293645 (1994).

Giovanni et al. ISS is adaval 2'-deoxy-β -L-Erythro-pentofuranose partly against virus pseudoleskeella (PRV), Biochem. J. (1993), 294(2), 381-5.

Chemotherapy the use of 2'-deoxy-β-L-Erythro-pentofuranose studied Tyrsred et al. (Biochim. Biophys. Acta (1968), 155(2), 619-22) and Bloch, et al. (J. Med. Chem. (1967), 10(5), 908-12).

In this area it is known that β-L-2'-deoxythymidine (β-L-dT) inhibits timedancing (TK) of herpes simplex virus type I (HSV-I). lotti et al. in the International Application WO 92/08727 described that β-L-dT selectively inhibits the phosphorylation of D-thymidine by TK HSV-I, but not by TC man. Spaldari et al. reported that L-thymidine fosfauriliruetsa-encoded thymidine kinase of herpes virus type I and inhibits the growth of viruses, J. Med. Chem. (1992), 35(22), 4214-20.

In light of the fact that hepatitis b virus reaches epidemic levels worldwide, and has severe and often tragic effects on the infected patient, there is a strong need to provide new effective pharmaceutical agents for the treatment of humans infected with a virus, which are characterized by low toxicity to the host.

Therefore, the purpose of this invention is to provide new methods and compositions for the treatment of humans or other hosts infected with hepatitis C.

Brief description of the invention

Described is a method of treatment of a hepatitis b infection in human is a and other animal owners, which includes the introduction of an effective amount of a biologically active 2'-deoxy-β-L-Erythro-pentofuranose (alternatively referred to in the description as β-L-d-nucleoside or β-L-2'-d-nucleoside or its pharmaceutically acceptable salt or prodrug, administered either individually or in combination, possibly in a pharmaceutically acceptable carrier. The term 2'-deoxy, which is used in the description refers to a nucleoside that contains no substitutional group at the 2'-position.

Described 2'-deoxy-β-L-Erythro-pentofuranose or pharmaceutically acceptable prodrug or salt, or pharmaceutically acceptable compositions containing these compounds, are used to prevent and treat infections of hepatitis b and other related conditions such as anti-HBV antibody positive and HBV-positive conditions, chronic liver inflammation caused by HBV, cirrhosis, acute hepatitis, fulminant hepatitis, chronic persistent hepatitis, and fatigue. These compounds or compositions can also be used prophylactically to prevent or retard the progression of clinical illness in individuals who are anti-HBV antibody positive or HBV-antigen positive or who have been exposed to HBV.

In one aspect of this image is to be placed, derivative of 2'-deoxy-β-L-Erythro-pentofuranose is a compound of the formula:

in which R is selected from the group consisting of H, straight, branched or cyclic alkyl, CO-alkyl, CO-aryl, CO-alkoxyalkyl, CO-aryloxyalkyl, CO-substituted aryl, alkylsulfonyl, arylsulfonyl, aralkylamines, balance of amino acids, mono-, di - or triphosphate or a derivative phosphate; and base is a purine or pyrimidine base, which is optionally substituted.

In another aspect derived from 2'-deoxy-β-L-Erythro-pentofuranose is a β-L-2'-deoxyadenosine or its pharmaceutically acceptable salt or prodrug of the formula:

in which R is H, mono-, di - or triphosphate, acyl or alkyl, or a stabilized phosphate derivative (for the formation of a stabilized nucleotide prodrug).

In another aspect, a derivative of 2'-deoxy-β-L-Erythro-pentofuranose is β-L-2'-deoxycytidine or its pharmaceutically acceptable salt or prodrug of the formula:

in which R is H, mono-, di - or triphosphate, acyl or alkyl, or a stabilized phosphate derivative (to the image is of a stabilized nucleotide prodrug).

In the following aspect, derived from 2'-deoxy-β-L-Erythro-pentofuranose is β-L-2'-deoxyuridine or its pharmaceutically acceptable salt or prodrug described by the formula:

in which R is H, mono-, di - or triphosphate, acyl or alkyl, or a stabilized phosphate derivative (for the formation of a stabilized nucleotide prodrug).

In another aspect, a derivative of 2'-deoxy-β-L-Erythro-pentofuranose is β-L-2'-deoxyguanosine or its pharmaceutically acceptable salt or prodrug having the formula:

in which R is H, mono-, di - or triphosphate, acyl or alkyl, or a stabilized phosphate derivative (to obtain a stabilized nucleotide prodrug).

In the following aspect, derived from 2'-deoxy-β-L-Erythro-pentofuranose is β-L-2'-deoxyinosine or its pharmaceutically acceptable salt or prodrug having the formula:

in which R is H, mono-, di - or triphosphate, acyl or alkyl, or a stabilized phosphate derivative (to obtain a stabilized nucleotide prodrug).

In another aspect, a derivative of 2'-deoxy-&x003B2; -L-Erythro-pentofuranose is β-L-thymidine or its pharmaceutically acceptable salt or prodrug of the formula:

in which R is H, mono-, di - triphosphate, acyl or alkyl, or a stabilized phosphate derivative (to obtain a stabilized nucleotide prodrug).

In another aspect 2'-deoxy-β-L-Erythro-pentofuranose introduced in alternation or in combination with one or more 2'-deoxy-β-L-Erythro-pentofuranose or with one or more other compounds which have activity against hepatitis C. in General, during alternating therapy effective dose of each agent is put on the queue, whereas combination therapy effective dose of two or more substances are introduced together. Dosages depend on absorption, inactivation and excretion rate of drug substances, as well as other factors known to specialists in this field. It should be noted that the size of the dose also change depending on the severity of the condition to his relief. In addition, it is clear that for any particular subject should be equipped with special schemes and schedules, medications, depending on time, according to individual needs and professional assessment by a person having appointed the m composition and overseeing their implementation.

The following aspect of the invention provides a method of treatment of humans infected with HBV that includes the introduction of a therapeutic dose of the prodrug described derivatives of 2'-deoxy-β-L-Erythro-pentofuranose. The prodrug, which is used in the application refers to a connection that turns into a nuke when introduced in vivo. He limiting examples include pharmaceutically acceptable salts (alternatively referred to as "physiologically acceptable salts"), 5' and N4(citizen)- or N6(adenosine)is acylated or alkylated derivatives of the active compounds, or 5'-phospholipid or a 5'-ether lipid active connection.

Brief description of drawings

Figure 1-1A illustrates a General method of obtaining β-L-Erythro-pentofuranose (β-L-dN), using L-ribose or L-xylose as a starting material.

Figure 2 is a graph which illustrates the metabolism of L-dA, L-dC and L-dT in the cells of the ner G2 person depending on the accumulation and decay. Cells were incubated in the presence of 10 μm of the compound.

Figure 3 is a graph which illustrates the antiviral effect β-L-dA β-L-dT and β-L-dC model with chronic hepatitis Groundhog.

Used in the description, the term "essentially in the form of ordinary isomer" or "in isolated who Orme" refers to 2'-deoxy-β -L-Erythro-pentofuranose that there are at least about 95% of the installed stereoconfiguration. In a preferred embodiment, the active compound, at least this degree of purity, enter the host when needed therapy.

Used in the description of the term "hepatitis b and related condition" refers to the hepatitis b and related conditions such as anti-HBV antibody positive and HBV-positive conditions, chronic liver inflammation caused by HBV, cirrhosis, acute hepatitis, fulminant hepatitis, chronic persistent hepatitis, and fatigue. The method according to the invention includes the use of derivatives of 2'-deoxy-β-L-Erythro-pentofuranose prophylactically to prevent or retard the progression of clinical illness in individuals who are anti-HBV antibody positive or HBV-antigen positive or who have been exposed to HBV.

Used in the description, the term alkyl, unless otherwise specified, refers to a saturated straight, branched, or cyclic, primary, secondary, or tertiary hydrocarbon, typically C1to C18preferably C1to C6and, in particular, without limitation, includes methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-b is Teal, isopentyl, amyl, tert-pentyl, cyclopentyl and cyclohexyl.

Used in the description of the term "acyl" refers to a fragment of the formula-C(O)R', where R' represents alkyl; aryl, alkaryl, aralkyl, heteroaromatic, alkoxyalkyl, including methoxymethyl; arylalkyl, including benzyl; aryloxyalkyl, such as phenoxymethyl; aryl, including phenyl, optionally substituted with halogen, C1-C4alkyl or C1-C4alkoxy, or amino acid residue. The term acyl, without limitation, in particular, includes acetyl, propionyl, butyryl, pentanoyl, 3-methylbutyryl, acid succinate, 3-chlorobenzoate, benzoyl, acetyl, pivaloyl, mesilate, propionyl, valeryl, the remainder of Caproic, Caprylic, capric, lauric, myristic, palmitic, stearic and oleic acids.

Used in the description, the term "purine or pyrimidine base, without limitation, includes 6-alkylboron and N6-alkylphenyl, N6-acylpyrin, N6-benzylurea, 6-Galperin, N6-vinluan, N6-acetylenic purine, N6-acylpyrin, N6-hydroxyalkylated, N6-dialkylphenol, N2-alkylphenyl, N4-alkylpyridine, N4-arylpyrimidine, 4-benzylpiperidine, N4-galerimizin, N4-acetylene pyrimidines, 4-acyl - and N4-arylpyrimidine, 4-hydroxyethylpyrrolidine, 4-ilkelerimizi, thymine, cytosine, 6-etherimide, including 6-azacytosine, 2 - and/or 4-mercaptopyrimidine, uracil, With5-alkylpyridine, With5-benzylpyrimidines, C5-galerimizin, With5-vinylpyridin, C5-acetylenic pyrimidine, C5-arylpyrimidine, With5-hydroxyalkylated, C5-aminopyrimidine, C5-cyanopyrimidine, With5-nitropyrimidin, With5-aminopyrimidine, N2-alkylphenyl, N2-alkyl-6-thiopurine, 5-azacitidine, 5-azauracil, triazolopyridines, imidazopyridines, pyrrolopyrimidine and pyrazolopyrimidines. Functional groups of oxygen and nitrogen on the base can be protected as necessary or as desired. Suitable protecting groups are well known to specialists in this field and include trimethylsilyl, dimethyl-exelsior, tert-butyldimethylsilyl and tert-butylbiphenyl-silyl, trityl, alkyl groups, acyl groups such as acetyl and propionyl, methanesulfonyl and p-toluensulfonyl.

The term biologically active nucleoside used in the description refers to a nucleoside, which is characterized by the value EC50corresponding to 15 micromol or less in the study in 2.2.15 cells, transfected with the hepatitis virion.

Preferred bases include cytosine, 5-fertilizin, 5-bromatosis, 5-modzitzer, uracil, 5-ftgure the sludge, 5-bromouracil, 5-iodouracil, 5-methyluracil, thymine, adenine, guanine, inosine, xanthine, 2,6-diaminopurine, 6-aminopurine, 6-globulin and 2,6-dichloropurine, 6-bromopurine, 2,6-dibromofuran, 6-iturin, 2,6-viturin, 5-bromonicotinic, 5-bravenewworld, 5-biometrician, 5-promaterials, 5-cryptometrics, 5-cryptomaterial.

2'-deoxy-β-L-Erythro-pentofuranose may be provided in the form of 5'-phospholipid or a 5'-ether lipid, as described in the following references: Kucera, L.S., N.Lyer, E.Leake, A.Raben, Modest E.J., D.L.W., and C.Piantadosi. 1990. Novel membrane-interactive ether lipid analogs that inhibit infectious HIV-I production and induce defective virus formation. AIDS Res Hum Retroviruses. 6: 491-501; Piantadosi, C., J.Marasco C.J., S.L.Morris-Natschke, K.L.Meyer, F.Gumus, J.R.Surles, K.S.Ishaq, L.S.Kusera, N.Iyer, 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. Chem. 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 I replication in CEM and HT4-6C cells by 31-deoxythymidine dephosphate dimyristoylglycerol, a lipid prodrug of 31-deoxythymidine. Antimicrob Agents Chemother. 36:2025-2029; Hostetler, K.Y., L.M.Stuhmiller, H.B.Lenting, H. van den Bosch, and D.D.Richman. 1990. Synthesis and antiretroviral activity of phospholipid analogs of azidothymidine and other antiviral nucleosides. J. Biol. Chem. 265: 6112-7.

2'-deoxy-β-L-Erythro-pentofuranose can be converted into a pharmaceutically acceptable ester by reaction with an appropriate esterification agent, such as galogenangidridy or anhydride. The nucleoside or the pharmacist who Cesky acceptable prodrug may be transformed into its pharmaceutically acceptable salt is an accepted way, for example, by processing the appropriate base or acid. Ester or salt can turn into a source nucleoside, for example, by hydrolysis.

Used in the description, the term "pharmaceutically acceptable salts or complexes" refers to salts or complexes of 2'-deoxy-β-L-Erythro-pentofuranose that retain the desired biological activity of the parent compound and exhibit minimal, if any, show, undesired Toxicological effects. Non-limiting examples of such salts are (a) a salt of joining acids, formed with inorganic acids (e.g. hydrochloric acid, Hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palm acid, alginic acid, polyglutamine acid, naphthalenesulfonic acid, naphthalenedisulfonic acid and polygalacturonase acid; (b) salts of joining the Foundation, formed by cations, such as sodium, potassium, zinc, calcium, bismuth, barium, magnesium, aluminum, copper, cobalt, Nickel, cadmium, sodium, potassium and the like, is whether organic cation, obtained from N,N-dibenziletilendiaminom, ammonia or Ethylenediamine; or (C) combinations of (a) and (b); for example, tannat zinc or the like.

Used in the description, the term "prodrug" refers to a connection that turns into a nuke when introduced in vivo. He limiting examples are the pharmaceutically acceptable salts (alternatively referred to as "physiologically acceptable salts"), 5'- and N4or N6-acylated or alkylated derivatives of the active compounds and 5'-phospholipid and 5'-ether lipid derivatives of active connections.

Modification of the active compounds, especially in the N4N6and 5'-O positions, can affect the bioavailability and rate of metabolism of the active substances, thus providing control over the delivery of the active compounds.

A preferred aspect of the present invention is a method of treatment of HBV infections in humans or other animal hosts, which includes the introduction of an effective amount of one or more 2'-deoxy-β-L-Erythro-pentofuranose derivative selected from the group consisting of β-L-2'-deoxyadenosine, β-L-2'-deoxycytidine, β-L-2'-dose irradiation on neurogenesis, β-L-2'-guanosine, β-L-2'-deoxyinosine and β-L-2 deoxythymidine or their physiologically acceptable prodrugs, on the tea phosphate, 5'or N6-alkilirovanie or acylated derivative or its physiologically acceptable salt, optionally in a pharmaceutically suitable carrier. The compounds of this invention possess anti-HBV activity, or converted to a compound or compounds that exhibit anti-HBV activity. In a preferred aspect, 2'-deoxy-β-L-Erythro-pentofuranose is introduced mainly in the form of ordinary isomer, i.e. at least approximately 95% of the installed stereo-configuration.

Nucleotide prodrugs

Any of the methods in the description of the nucleosides can be introduced in the form of stable nucleating prodrug to increase the activity, bioavailability, stability or, in other words, to change the properties of the nucleoside. The number of known ligands nucleotide prodrugs. In General, alkylation, acylation or other lipophilic modification of mono-, di - or triphosphate of the nucleoside will increase the stability of the nucleotide. Examples of substitution groups that can replace one or more hydrogens on the phosphate fragment, are alkyl, aryl, steroids, carbohydrates, including sugars, 1,2-diacylglycerol and alcohols. Most described in the publication together with R. Jones and N.Bischofberger, Antiviral Research, 27 (1995) 1-17. Any of them can be used in combination with described what ucleotide to achieve the desired actions.

In one aspect of the 2'-deoxy-β-L-Erythro-pentofuranose is provided in the form of a 5'-hydroxyl lipophilic prodrugs. Non-limiting examples of U.S. patents that describe suitable lipophilic substituents that can be covalently to include nucleoside, preferably in the 5'-position of the nucleoside, or lipophilic preparations, include U.S. patent No. 5 149 794 (September 22, 1992, Yatvin et al.); 5194654 (March 16, 1993, Hostetler et al.); 5223263 (June 29, 1993, Hostetler et al.); 5256641 (October 26, 1993, Yatvin et al.); 5411947 (may 2, 1995, Hostetler et al.); 5463092 (October 31, 1995, Hostetler et al.); 5543389 (August 6, 1996, Yatvin et al.); 5543390 (August 6, 1996, Yatvin et al.), 5543391 (August 6, 1996, Yatvin et al.); and 5554728 (September 10, 1996, Basava et al.).

Foreign patent applications that describe lipophilic substituents that can be included in the 2'-deoxy-β-L-Erythro-pentofuranose derivative of the present invention, or lipophilic preparations, include WO 89/02733, WO 90/00555, WO 91/16920, WO 91/18914, WO 93/00910, WO 94/26273, WO 96/15132, EP 0350287, EP 93917054,4 and WO 91/19721.

Additional non-limiting examples of 2'-deoxy-β-L-Erythro-pentofuranose are nucleosides that contain the substituents described in the following publications. These converted 2'-deoxy-β-L-Erythro-pentofuranose can be used for the purposes described in the text, or, in other words, as antiviral AG is, for example, including anti-HBV agents. D.H.W. (1973) Distribution of kinase and deaminase of 1 β-D-arabinofuranosylcytosine in tissues of man and mouse. Cancer Res. 33, 2816-2820; Holy, A. (1993) Isopolar phosphorous-modified nucleotide analogues. In: De Clercq (Ed.), Advances in Antiviiral Drug Design, Vol.I, JAI Press, pp.179-231; Hong, C.I., Nechaev, A., and West, C.R. (1979) Synthesis and antitumor activity of 1-(β-D-arabinofuranosylcytosine conjugates of cortisol and cortisone. Biochem. Biophys. Rs. Commun. 88, 1223-1229; Hong, C.I., Nechaev, A., Kirisits, A.J. Buchheit, D.J. and West C.R. (1980) Nucleoside conjugates as potential antitumor agents. 3.Synthsis and antitumor activity of 1-(β-D-arabinofuranosyi)cytosine conjugates of corticosteroids and selected lipophilic alcohols. J. Med. Chem. 28, 171-177; Hostetler, K.Y., Stuhmiller, L.M., Lenting, H.B.M., van den Bosch, H. and Richmann, D.D. (1990) Synthesis and antiretroviral activity of phospholipid analogs of azidothymidine and other antiviral nucleosides. J. Biol. Chem. 265, 6112-6117; Hostetler, K.Y., Carson, D.A. and Richman, D.D. (1991); Phosphatidylazidothymidine: mechanism of antiretroviral action in CEM cells. J. Biol. Chem. 266, 11714-11717; Hostetler, K.Y., Korba, B., Sridhar, C., Gardener, M. (1994a) Antiviral activity of phosphatidyl-dideoxycytidine in hepatitis B-infected cells and enhanced hepatic uptake in mice. Antiviral Res. 24, 59-67; Hostetler, K.Y., Richman, D.D., Sridhar, C.N., Felgner, P.L., Felgner, J., Ricci, J., Gardener, M.F., Selleseth, D.W. and Ellis, M.N. (1994b) Phosphatidylazidothymidine and phosphatidyl-ddC: Assessment of uptake in mouse lymphoid tissues and antiviral activities in human immunodeficiency virus-infected cells and in rauscher leukemia virus-infected mice. Antimicrobial Agents Chemother. 38, 2792-2797; Hunston, R.N., Jones, A.A. McGuigan, C., Walker, R.T., Balzarini, J. and De Clercq, E. (1984) Synthesis and biological properties of some cyclic phosphotriesters derived from 2'deoxy-5-fluorouridine. J. Med. Chem. 27, 440-444; Ji, Y.H., Moog, C., Schmitt, G., Bischoff, P. and Luu, B. (1990); Monophosphoric acid diesters of 7β-hydroxycholesterol and of pyrimidine nucleosides as potential antitumor agents: synthesis and preliminary evaluation of antitumor activity. J. Med. Chem. 33, 2264-2270; Jones, A.S., McGuigan, C.,Walker, R.T., Balzarini, J. and DeClercq, E. (1984) Synthesis, properties, and biological activity of some nucleoside cyclic phosphoramidates. J. Chem. Soc. Perkin Trans. I, 1471-1474; Juodka, B.A. and Smart, J. (1974) Synthesis of ditribonucleoside a(P→ (N) amino acid derivatives. Coll. Czech. Chem. Comm. 39, 363-968; Kataoka, S., Imai, J., Yamaji, N., Kato, M., Saito, M., Kawada, T. And Imai, S. (1989) Alkylated cAMP derivatives; selective synthesis and biological activities. Nucleic Acids Res. Sym. Ser., 21, 1-2; Kataoka, S., Uchida, R. and Yamaji, N. (1991) A convenient synthesis of adenosine 3',5'cyclic phosphate (cAMP) benzyl and methyl triesters. Heterocycles 32, 1351-1356; Kinchington, D., Harvey, J.J., O'connor, T.J., Jones, B.C.N.M., Devine, K.G., Taylor-Robinson, D., Jeffries, D.J. and McGuigan, C. (1992) Comparison of antiviral effects of zidovudine phosphoramidate and phosphorodiamidate derivatives against HIV and MuLV in vitro. Antiviral Chem. Chemother. 3, 107-112; Kodama, K., Morozumi, M., Saitoh, K.I., Kuninaka, H., Yoshino, H. and Saneyoshi, M. (1989) Antitumor activity activity and pharmacology of 1-β-D-arabinofuranosylcytosine-5'-stearylphosphate; an orally active derivative of 1-β-D-arabinofuranosylcytosine. Jpn. J. Cancer Res. 80, 679-685; Korty, M. and Engels, J. (1979) The effects of adenosine - and guanosine 3',5'-phosphoric acid and benzyl esters on guinea-pig ventricular myocardium. Naunyn-Schmiedeberg''s Arch. Pharmacol. 310, 103-111; Kumar, A., Goe, P.L., Jones, A.S. Walker, R.T.Balzarini, J. and De Clercq, E. (1990) Synthesis and biological evaluation of some cyclic phosphoramidate nucleoside derivatives. J. Med. Chem. 33, 2368-2375; LeBec, C., and Huynh-Dinh, T. (1991) Synthesis of lipophilic phosphate triester derivatives of 5-fluouridine and arabinocytidine as anticancer prodrugs. Tetrahedron Lett. 32, 6553-6556; Lichtenstein, J., Barner, H.D. and Cohen, S.S. (1960) The metabolism of exogenously supplied nucleotides by Escherichia coli., J. Biol. Chem. 235, 457-465; Lucthy, J., Von Daeniken, A., Friederich, J. Manthey, B., Zweifel, J., Schlatter, C. And Benn, M.H. (1981) Synthesis and toxicological properties of three naturally occurring cyanoepithioalkanes. Mitt. Geg. Lebensmittelunters. Hyg. 72, 131-133 (Chem. Abstr. 95, 127093); McGuigan, .Tollerfield, S.M. and Riley, P.A. (1989) Synthesis and biological ealuation of some phosphate triester derivatives of the anti-viral drug Ara. Nucleic Acids Res. 17. 6065-6075; McGuigan, C., Devine, K.G., O'connor, T.J., Galpin, S.A., Jeffries, D.J. and Kinchington, D. (1990a) Synthesis and evaluation of some novel phosphoramidate derivatives of 3'-azido-3'-deoxythymidine (AZT) as anti-HIV compounds. Antiviral Chem. Chemother. 1, 107-113; McGuigan, C., O'connor, T.J., Nicholls, S.R., Nickson, C. and Kinchington, D. (1990b) Synthesis and anti-HIV activity of some novel substituted dialkyl phosphate derivatives of AZT and ddCyd. Antiviral. Chem. Chemother. 1, 355 to 360 above; McGuigan, C., Nicholls, S.R., O'connor, T.J., and Kinchington, D. (1990) Synthesis of some novel dialkyl phosphate derivative of 3'-modified nucleosides as potential anti-AIDS drug. Antiviral Chem. Chemother. 1, 25-33; McGuigan, C., Devine, K.G., O'connor, T.J., and Kinchington, D. (1991) Synthesis and anti-HIV activity of some haloalkyl phosphoramidate derivatives of 3'-azido-3'-deoxythymidine (AZT); potent activity of the trichloroethyl methoxyalaninyl compound. Antiviral Res. 15, 255-263; McGuigan, C., Pathirana, R.N., Mahmood, N., Devine, K.G. and Hay, A.J. (1992) Aryl phoaphate derivatives of AZT retain activity against HIV-1 in cell lines which are resistant to the action of AZT. Antiviral Res. 17, 311-321; McGuigan, C., Pathirana, R.N., Choi, S.M., Kinchington, D. and O'connor, T.J. (1993a) Phosphoramidate derivatives of AZT as inhibitors of HIV; studies on the carboxyl terminus. Antiviral Chem. Chemother. 4, 97-101; McGuigan, C., Pathirana, R.N., Balzarini, J. and De Clercq, E. (1993b) Intracellulal delivery of bioactive AZT nucleotides by aryl phosphate derivatives of AZT. J. Med. Chem.36, 1048-1052.

The question of chair-twist equilibria for the phosphate rings of nucleoside cyclic 3',5'-monophosphates.1HNMR and x-ray crystallographic study of the diasteromers of thymidine phenyl cyclic 3',5'-monophosphate. J. Am. Chem. Soc. 109, 4058-4064; Nerbonne, J.M., Richard, S., Nargeot, J. and Lester, H.A. (1984) New photoactivatable cyclic nucleotides produce intracellular jumps in cyclic AMP and GMP concentrations. Nature 301, 74-76; Neumann, J.M., Herve, M., Debouzy, J.C., Guerra, F.I., Gouyette, C., Dupraz, B. and Huynh-Dinh, T. (1989) Synthesis and transmembrane transport studies by NMR of a glucosyl phospholipid of thymidine. J. Am. Chem. Soc. I'll 4270-4277; Ohno, R., Tatsumi, N.. Harada, M., Imai, K, Mizoguchi, H., Nakamura, T., Kosaka, M., Takatuski, K., Yamaya. T., Toyama, K., Yoshida, T., Masaoka, T., Hashimoto, S., Ohshima, T., Kimura, I., Yamada, K. and Kimura, J. (1991) Treatment of myelodysplastic syndromes with orally administered 1-β-D-rabinofuranosylcytosine-5'-srearylphosphate. Oncology 48, 451-455.

Palomino, E., Kessle, D. and Horwitz, J.P. (1989) A dihydropyridine carrier system for sustained delivery of 2',3'-dideoxynuclesides to the brain. J. Med. Chem. 32, 622-625; Perkins, P.M., Barney, S., Wittrock, R., Clark, P.H., Levin, R. Lambert, D.M., Petteway, S.R., Serafinowska, H.T., Bailey, S.M., Jackson, S., Harnden, M.R. Ashton, R., Sutton, D., Harvey, J.J. and Brown, A.G. (1993) Activity of BRL47923 and its oral prodrug, SB203657A against a rauscher murine leukemia virus infection in mice. Antiviral Res. 20 (Suppl. I), 84; Piantadosi, C., Marasco, C.J., Jr. Morris-Natschke, S.L., Meyer, K.L., Gumus, F., Surles, J. R., Ishaq, for K.S., Kucera, L.S., lyer, N., Wallen, C.A., Piantadosi, S. and Modest, E.J. (1991) Synthesis and evaluation of novel ether lipid nucleoside conjugates for anti-HIV-1 activity. J. Med. Chem. 34, 1408-1414; Pompon, A., Lefebvre, I., Imbach, J.L., Kahn, S. and Farquhar, D. (1994) Decomposition pathways of the mono - and bis(pivaloyloxymethyl) esters of azidothymidine-5'-monophosphate in cell extract and in tissue culture medium; an application of the on-line ISRP-cleaning' HPLC technique. Antiviral Chem. Chemother. 5, 91-98; Postemark, T. (1974) Cyclic AMP and GMP. Annu. Rev. Pharmacol. 14, 23-33; Prisbe, E.J., Martin, J.C.M., McGee, D.P.C., Barker, M.F., Smee, D.F., Duke, A.E., Matthews, T.R. and Verheyden, J.P.J. (1986) Synthesis and antiherpes virus activity of phosphate and phosphonate derivatives of 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine. J. Med. Chem. 29, 671-675; Puech, F., Gosselin, G., Lefebvre, I., Pompon, A., Aubertin, A.M., Dirn, A. And Imbach, J.L. (1993) Intracellular delivery of nucleoside monophosphate through a reductase-mediated activation process. Antivral Res. 22, 155-174; Pugaeva, V.P., Klochkeva, S.I., Mashbits, F.D. and Eizengart, R.S. (1969). Robins, R.K. (1984) The potential of nucleotide analogs as inhibitors of retroviruses and tumors. Pharm. Res. 11-18; Posowsky, A., Kim, S.H., Ross and J. Wick, M.M. (1982) Lipophilic 5'-(alkylphosphate) esters of 1-β-D-arabinofuraosylcytosine and its N 4-acyl and 2.2'-anhydro-3'-O-acyl derivatives as potential prodrugs. J. Med. Chem. 25, 171-178; Ross, W. (1961) Increased sensitivity of the walker turnout towards aromatic nitrogen mustards carrying basic side chains following glucose pretreatment. Biochem. Pharm. 8, 235-240; Ryu, E.K., Ross, R.J., Matsushita, T., MacCoss, M., Hong, C.I. and West, C.R. (1982). Phospholipid-nucleoside conjugates. 3. Synthesis and preliminary biological evaluation of 1-β-D-arabinofuranosylcytosine 5'diphosphate{-},2-diacylglycerols. J. Med. Chem. 25, 1322-1329; Saffhill, R. And Hume, W.J. (1986) The degradation of 5-iododeoxyuridine and 5-bromodeoxyuridine by serum from different sources and its consequences for the use of these compounds for incorporation into DNA. Chem. Biol. Interact. 57, 347-355; Saneyoshi, M., Morozumi, M., Kodama, K., Machida, J., Kuninaka, A. and Yoshino, H. (1980) Synthetic nucleosides and nucleotides. XVI. Synthesis and biological evaluations of a series of 1-β-D-arabinofyranosylcytosine 5'-alkyl or arylphosphates. Chem. Pharm. Bull. 28, 2915-2923; Sastry, J.K., Nehete, P.N., Khan, S., Nowak, B.J., Plunkett, W., Arlinghaus, R.B. and Farquhar, D. (1992) Membrane-permeable dideoxyuridine 5'-monophosphate analogue inhibits human immunodeficiency virus infection. Mol. Pharmacol. 41, 441-445; Shaw, J.P., Jones, R.J., Arimilli, M.N., Louie, M.S., Lee, W.A. and Cundy, K.C. (1994) Oral bioavailaability of PMEA from PMEA prodrugs in male Sprague-Dawley rats. 9th Annbal AAPS Meeting. San Diego, CA (Abstract). Shuto, S., Ueda, S., Imamura, S., Fukukawa, K. Matsuda, A. and Ueda, T. (1987) A facile one-step synthesis of 5'-phosphatidyl-nucleosides by an enzymatic two-phase reaction. Tetrahedron Lett. 28, 199-202; Shuto, S., Itoh, H., Ueda, S., Imamura, S., Kukukawa, K., Tsujino, M., Matsuda, A. And Ueda, T. (1988) A facile enzymatic synthesis of 5'-(3-sn-phosphatidyl)nucleosides and antileukemic activities. Chem. Pharm. Bull. 36, 209-217. One preferred group of phosphate prodrug is a group of S-acyl-2-thioethyl, also referred to as "SATE".

Concomitant or alternating therapy

Found that are resistant to the medication form HV can emerge after prolonged treatment with an antiviral agent. The most typical resistance to the drug occurs by mutation of a gene that encodes an enzyme used in the life cycle of the virus, and most often in the case of DNA polymerase of HBV. Recently it was demonstrated that the effectiveness of drugs against HBV infection can be prolonged, increased or restored by introducing the compound in combination or alternation with a second, and possibly third antiviral compound that induces a mutation, other than that caused by the original drug. Alternatively, pharmacokinetics, biological distribution or other parameters of the medicinal substance can be changed by using such combined or alternating therapy. In General, usually combined therapy is preferred in comparison with alternating therapy because it causes multiple simultaneous stresses of the virus.

Activity against HCV compounds In β-L-2'-dA β-L-2'-dC, β-L-2'dU, β-L-2'dG, β-L-2'-dT, β-L-dI or other β-L-2'-nucleoside represented in the description, or prodrugs, phosphates, or salts of these compounds may be increased by the introduction two or more of these nucleosides in combination or in alternation. Alternative for example, one or more of the β-L-2'-dA β-L-2'-dC, β-L-2'-dU, β-L-2'-dG, β-L-2'-dT, β-L-dI and the other β -L-2'-nucleoside represented in the description, you can type in combination or alternation with 3TC, FTC, L-FMAU, DAPD, famciclovir, penciclovir, BMS-200475, bis POM, RMEA (adefovir, dipivoxil); lobucavir, ganciclovir or ribavirin.

In any of the aspects described in the application, if β-L-2'-nucleoside of the present invention is introduced in combination or alternation with a second nucleoside or nucleoside analog reverse transcriptase inhibitor, which fosfauriliruetsa to the active form, it is preferable that the second connection fosforilirovanii would be the enzyme, which is different from the enzyme, fosforiliruyusciye in vivo selected β-L-2'-nucleoside of the present invention. Examples of kinase enzymes are thymidine kinase, tirosinkinaza, guanethidine, adenosines, deoxycytidine, 5'-nucleotidase and deoxyguanosine.

Getting active connections

Derivatives of 2'-deoxy-β-L-Erythro-pentofuranose according to the invention are known in this field and can be obtained in accordance with the method described Holy, Collect. Czech. Chem. Commun. (1972), 37(12), 4072-87 and Mol. Phys. (1967), 3(4), 386-95.

The usual way to get β-L-Erythro-pentofuranose (β-L-dN) using L-ribose or L-xylose as a starting substances are presented in Figure 1.

Mono-, di - or trifosfatnogo active derivatives of n is sidow can be obtained, as described, in accordance with published methods. Monophosphate can be obtained according to the method of Imai et al., J. Org. Chem., 34(6), 1547-1550 (June, 1969). Diphosphate can be obtained according to the method Davisson et al., J. Org. Chem. 52(9), 1794-1801 (1987). Triphosphate can be obtained in accordance with the method Hoard et al., J. Am. Chem. Soc., 87 (8), 1785-1788 (1965).

The protocols of experiments

Melting points were determined in open capillary tubes on a Gallenkamp apparatus MFB-595-010 M and are uncorrected. Absorption spectrum in the UV were recorded on a spectrophotometer Uvikon 931 (KONTRON) in ethanol. Spectra1H NMR removed at room temperature in DMSO-d6spectrometer Bruker AC 250 or 400. Chemical shift are presented in ppm (ppm), DMSO-d5established in 2,49 ppm as recommended. Materialmen, experiments on the breach of an engagement or 2D-COSY conducted in order to establish the location of the protons. A variety of signals are represented by s (singlet), d (doublet), dd (double doublet), t (triplet), q (quadruplet), br (broad), m (multiplet). All J-values are presented in Hz (Hertz). Mass FAB spectra were recorded in positive (FAB>0) or negative (FAB<0) ion mode mass spectrometer JEOL DX-300. The matrix was 3-nitrobenzyl alcohol (NBA) or a mixture (50:50, V/V) glycerol and diglycerin (GT). The specific rotation was measured on spectropol the meter Perkin-Elmer 241 (stroke length 1 cm) and expressed in units of 10 -1C·cm2·g-1. Elemental analysis was performed using the microprobe "Microanalyses du CNRS, Division de Vernaison" (France). The analyses presented by the symbols of the elements and were within ± 0.4% of theoretical values. Thin-layer chromatography was performed on pre-coated aluminium plates silica gel 60 F254(Merck Art 5554), imaging products was carried out by absorption in the UV, followed by charring with 10% alcohol with sulfuric acid and heating. Column chromatography was performed on silica gel 60 (Merck, Art. 9385) under atmospheric pressure.

Example 1

Stereospecific synthesis of 2'-deoxy-β-L-adenosine

9-(3,5-Di-O-benzoyl-β-L-xylofuranosyl)adenine (3)

A solution of 9-(2-O-acetyl-3,5-di-O-benzoyl-β-L-xylofuranosyl) adenine 2 [reference: Gosselin, G., Bergogne, M. - C.; Imbach, J. - L., "Synthesis and Antiviral Evaluation of β-L-Xylofyranosyi Nucleosides of the Five Naturally Occuring Nucleic Acid Bases", Journal of Heterocyclic Chemistry, 1993, 30 (October-November), 1229-1233] (8,30 g of 16.05 mmol) and 98% hydrazine hydrate (234 ml, to 48.5 mmol) in a mixture of pyridine/glacial acetic acid (4/1, V/V, 170 ml) was stirred at room temperature for 22 hours. The reaction was suppressed by the addition of acetone (40 ml) and was stirred continuously in t is the treatment of one additional hour. The reaction mixture was reduced to half its volume, diluted with water (250 ml) and was extracted with chloroform (2×150 ml). The organic layer was washed successively aqueous saturated solution of NaHCO3(3×100 ml) and water (3×100 ml), dried, filtered, concentrated and evaporated with toluene and methanol. The residue was purified by chromatography on a column of silica gel (0-3% Meon in dichloromethane)to obtain 3 (5,2 g, 68%), precipitated from diisopropyl ether:1H NMR (DMSO-d6): δ 4,5-4,9 (m, 4H, H-2', H-4', H-5' and H-5"), 5,64 (t, 1H, H-3', J2',3'=J3',4'=3,5 Hz), and 6.3 (br s, 1H, OH-2'), of 6.45 (d, 1H, H-1', J1',2'=4,6 Hz), and 7.3 (br s, 2H, NH2-6), of 7.4 to 7.9 (m, 10H, 2 Banzarov), 8,07 and a 8.34 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 476 [M+H]+, 136 [NR2]+, (FAB-) m/z 474 [M-H]-, 134 [In]-; UV (95% ethanol): λMaxim257 nm (ε 16400), 230 nm (ε 29300), λmin246 nm (ε 14800); [α]D20=-64 (with 1.07, CHCl3). Analytical calculation for C24H21N5O4(M=475,45:/60,43; N, OF 4.45; N, 14,73. Found: C, 60,41; N, To 4.68; N, 14,27.

9-(3,5-Di-O-benzoyl-2-deoxy-β-L-threo-pentofuranose)-adenine (4)

To a solution of compound 3 (1,00 g, 2,11 mmol) in dry acetonitrile (65 ml) was added 4-(dimethylamino)pyridine (0,77 g, 6,32 mmol) and phenoxythiocarbonyl (of 0.44 ml, and 3.16 mmol). The mixture was stirred at room temperature for 2 hours. After the oxygen is compete the residue was dissolved in dichloromethane (50 ml) and washed successively with water (2× 30 ml), 0.5 N aqueous solution of hydrochloric acid (30 ml) and water (3×30 ml). The organic layer was dried, filtered and concentrated until dry. Untreated thiocarbanilide intermediate connection directly processed by hydride Tris(trimethylsilyl)silane (0,78 ml, 5,23 mmol) and α,α'-azoisobutyronitrile (AIBN, 0,112 g, 0.69 mmol) in dry dioxane (17 ml) by boiling under reflux for 2 hours. The solvent was removed under vacuum, and the residue was purified by chromatography on a column of silica gel (0-5% Meon in dichloromethane)to obtain pure 4 (0,93 g, 96%) as a foamy mass:1H NMR (DMSO-d6): δ 2,9-3,1 (m, 2H, H-2' and H-2"), 4,6-4,7 (m, 3H, H-4', H-5' and H-5"), and 5.8 (br s, 1H, H-3'), to 6.43 (dd, 1H, H-1', J1',2'=3,1 Hz, J1',2"=7,6 Hz), and 7.3 (br s, 2H, NH2-6), of 7.4 to 7.9 (m, 10H, 2 benzoyl), 8,05 and 8,33 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 460 [M+H]+, 325 [S]+, 136 [BH2]+, (FAB-) m/z 458 [M-H]-, 134 [In]-; UV (95% ethanol): λmax261 nm (ε 14400), 231 nm (ε 26300), λmin249 nm (ε 12000); [α]D20=-38 (1.04, DMSO).

6-N-(4-Monoethoxylate)-9-(3,5-di-O-benzoyl-2-deoxy-β-L-threo-pentofuranose)adenine (5)

To a solution of compound 4 (0.88 g, 1.92 mmol) in dry pyridine (40 ml) was added 4-monomethoxypolyethylene (1.18 g, of 3.84 mmol). The mixture was stirred at 60°C for 24 hours. After adding the Oia methanol (5 ml) the solution was concentrated until dry, the residue was dissolved in dichloromethane (50 ml) and washed successively with water (30 ml), aqueous saturated NaHCO3(30 ml) and water (30 ml). The organic layer was dried, filtered, concentrated and evaporated with toluene to obtain 5 (1/01 g, 72%) as a foamy mass:1H NMR (CDCl3): δ of 2.9-3.0 (m, 2H, H-2' and H-2"), 3,62 (s, 3H, och3), 4,6-4,8 (m, 3H, H-4', H-5' and H-5"), 5,85 (pt, 1H, H-3'), 6,44 (dd, 1H, H-1', J1',2'=3,1 Hz, J1',2"=7,3 Hz)and 6.9 (br s, 1H, NH-6), the 6.7 to 6.8 and 7.2 to 7.4 (2m, 24H, 2 benzoyl and MMTr), 7,97 and 8,13 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 732 [M+H]+, (FAB-) m/z 730 [M-H]-; UV (95% ethanol): λmax274 nm (ε 12100), 225 nm (ε 24200), λmin250 nm (ε 5900); [α]D20=-16 (c1, 12, DMSO).

6-N-(4-Monoethoxylate)-9-(2-deoxy-β-L-threo-pentofuranose)-adenine (6)

Compound 5 (0.95 g, of 1.30 mmol) was treated methanolysis solution (saturated at -10° (C) ammonia (40 ml) at room temperature over night. After concentration the residue was dissolved in dichloromethane (60 ml) and washed with water (30 ml). The aqueous layer was extracted twice with dichloromethane (10 ml). The combined organic layer was dried, filtered and concentrated. The residue was purified by chromatography on a column of silica gel (0-5% Meon in dichloromethane)to obtain pure 6 (0,67 g, 98%) as penoobraznaya mass:1H NMR (CDCl3): δ 2,6-2,9 (m, 2H, H-2' and H-2"), and 3.5 (br s, 1H, OH-5'), 3,55 (s, 3H, och3), 3,9-4,0 (m, 3H, H-4', H-5' and H-5"), 4,5-4,6 (m, 1H, H-3'), 6,03 (dd, 1H, H-1', J1',2'=4,0 Hz, J1',2"=8,8 Hz), 7,0 (br s, 1H, NH-6), the 6.7 to 6.8 and 7.1 to 7.4 (2m, 14H, MMTr), 7,40 (d, 1H, OH-3', JH,OH=to 10.6 Hz), 7,80 and 7,99 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 524 [M+H]+, 408 [BH2]+, (FAB-) m/z 1045 [2M-H]-, 522 [M-N]-, 406 [In]-UV (95% ethanol): λmax275 nm (ε 12300), λmin247 nm (ε 3600); [α]D20=+28 (from 0.94, DMSO).

6-N-(4-monoethoxylate)-9-(2-deoxy-5-O-(4-monoethoxylate)-β-L-threo-pentofuranose) adenine (7)

Compound 6 (of 0.62 g of 1.24 mmol) in dry pyridine (25 ml) was treated with 4-monomethoxypolyethylene (0,46 g, 1,49 mmol) at room temperature for 16 hours. After adding methanol (5 ml) and the mixture was concentrated until dry. The residue was dissolved in dichloromethane (60 ml) and washed successively with water (40 ml), saturated aqueous NaHCO3(40 ml) and water (3×40 ml). The organic layer was dried, filtered, concentrated and evaporated with toluene and methanol. The residue was purified by chromatography on a column of silica gel (0-10% Meon in dichloromethane)to obtain 7 (0.71 g, 72%) as a foamy mass:1H NMR (DMSO-d6): δ of 2.21 (d, 1H, H-2' J2',2"=14,3 Hz), 2,6-2,7 (m, 1H, H-2",), 3,1-3,3 (2m, 2H, H-5' and H-5"), 3,64, and the 3.65 (2s, 6H, 2×OCH3), 4,1-4,2 (m, 1H, H-4'), 4,2-4,3 (m, 1H, H-3'), of 5.68 (d, 1H, OH-3', JH,OH=5,2 Hz), 6,24 (d, 1H, H-1', J1'," =7,0 Hz), 6,7-6,8 and 7,1-7,3 (2m, 29H, 2 MMTr and NH-6), 7,83 and 8,21 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 796 [M+H]+, 408 [NR2]+, (FAB-) m/z 794 [M-N]-, 406 [In]-; UV (95% ethanol): λmax275 nm (ε 30900), λmin246 nm (ε 12800); [α]D20=+14 (1,03, DMSO).

6-N-(4-Monoethoxylate)-9-(3-O-benzoyl-2-deoxy-5-O-(4-monoethoxylate)-β-L-Erythro-pentofuranose)adenine (8)

The solution diethylazodicarboxylate (0,38 ml, 2.49 mmol) in dry tetrahydrofuran (20 ml) was added dropwise to a cooled solution (0° (C) nucleoside 7 (0.66 g, 0.83 mmol), triphenylphosphine (0.66 g, 2.49 mmol) and benzoic acid (0,30 g, 2.49 mmol) in dry THF (20 ml). The mixture was stirred at room temperature for 18 hours and was added methanol (1 ml). The solvents were removed under reduced pressure, and the crude substance was purified by chromatography on a column of silica gel (0-5% ethyl acetate in dichloromethane)to obtain compound 8, slightly contaminated with triphenylphosphine oxide.

6-N-(4-Monoethoxylate)-9-(2-deoxy-5-O-(4-monoethoxylate)-β-L-Erythro-pentofuranose)adenine (9)

Compound 8 was treated with a methanol solution (saturated at -10° (C) ammonia (20 ml) at room temperature for 24 hours, then the reaction mixture was concentrated until dry. The residue was dissolved in dichloromethane (30 ml) and washed with water (20 ml). Water is the first layer was extracted with dichloromethane (2× 20 ml)and the combined organic phase was dried, filtered and concentrated. After purification by chromatography on a column of silica gel (0-2% Meon in dichloromethane) was obtained pure compound 9 (0.50 g, 76% from 7) in the form of foams:1H NMR (DMSO-d6): δ about 2.2-2.3 (m, 1H, H-2'), 2,8-2,9 (m, 1H, H-2"), 3,1-3,2 (m, 2H, H-5' and H-5"), 3,64, and the 3.65 (2s, 6H, 2×co3), 3,97 (pq, 1H, H-4'), 4.4 to 4.5 (m, 1H, H-3'), are 5.36 (d, 1H, OH-3', JH, OH=4,5 Hz), 6,34 (t, 1H, H-1', J1',2'=6,4 Hz); 6.8 or 6.9 and 7.1 to 7.4 (2m, 29H, 2 MMTr and NH-6), 7,81 and 8,32 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 796 [M+H]+, 408 [BH2]+, (FAB-) m/z 794 [M-H]-, 406 [In]-; UV (95% ethanol): λmax276 nm (ε 42600), λmin248 nm (ε 23300); [α]D20=+29 (1,05, DMSO).

2'-deoxy-β-L-adenosine β-L-dA)

Compound 9 (0,44 g of 0.56 mmol) was treated with 80% aqueous solution of acetic acid (17 ml) at room temperature for 5 hours. The mixture was concentrated until dry, the residue was dissolved in water (20 ml) and washed with diethyl ether (2×15 ml). The aqueous layer was concentrated and evaporated with toluene and methanol. The desired 2' -deoxy-β-L-adenosine (β-L-dA) (0.12 g, 83%) was obtained after purification by chromatography on a column of silica gel (0-12% Meon in dichloromethane and filtered through a cell Millex HV-4 (0.45 µm, Millipore): melting point 193-194°C (crystallized from water) (Lit. 184-185°C for the L-enantiomer [Cm. Robins, M.J., Khwaja, T.A., Rbins, R.K. J. Org. Chem. 1970, 35, 636-639] and 187-189°for D-enantiomer [Cm. Ness, R.K. in Synthetic Procedures in Nucleic Acid Chemistry; Zorbach, W.W., Tipson, R. S., Eds.; J. Wiley and sons: New York, 1968; Vol 1, pp 183-187];1H NMR (DMSO-d6): δ 2,2-2,3 and 2,6-2,7 (2m, 2H, H-2' and H-2"), 3,4-3,6 (2m, 2H, H-5' and H-5"), 3,86 (pq, 1H, H-4'), a 4.3 and 4.4 (m, 1H, H-3'), of 5.24 (t, 1H, OH-5', JH,OH=5,8 Hz), and 5.30 (d, 1H, OH-3', JH,OH=4,0 Hz), 6,32 (dd, 1H, H-1', J1,2'=6,2 Hz, J1,2"=7,8 Hz), and 7.3 (br s, 2H, NH2-6), 8,11 and 8,32 (2s, 2H, H-2 and H-8); ms: matrix G/T, (FAB+) m/z 252 [N+H]+, 136 [BH2]+, (FAB-) m/z 250 [M-H]-, 134 [In]-; UV (95% ethanol): λmax258 nm (14300 s), λmin226 nm (ε 2100); [α]D20=+25 (1,03 N2O), (Lit. [α]D20=+23 (1,0, N2O) for the L-enantiomer [Cm. Robins, M.J., Khwaja, T.A., Robins, R.K. J. Orq. Chem. 1970, 35, 636-639] and [α]D20=-25 (from 0.47, N2O) for the D-enantiomer [Cm. Ness, R.K. in Synthetic Procedures in Nucleic Acid Chemistry; Zorbach, W.W., Tipson, R. S., Eds.; J. Wiley and sons: New York, 1968; Vol 1, pp 183-187]). Analytical calculation for C10H13N5O3+1.5 N2O (M=278,28): C, 43,16; N, 5,80; N, 25,17. Found: C, 43,63; N, The 5.45; N, 25,33.

Example 2

Stereotypically synthesis of 2'-deoxy-β-L-adenosine (β-L-dA)

Reaction 1:

Starting material: L-ribose (Cultor Food Science, CAS [24259-59-4], party RIB9711013)

Reagents; 95-97% sulfuric acid (Merck; reference number 1,00731,1000); benzoyl chloride (Fluka; reference number is 12930); the sodium sulfate (Prolabo; reference number 28111,365)

Solvents: Methanol, pure for analysis (Prolabo; reference number 20847,295); 99% pyridine (Acros; reference number 131780025); dichloromethane, pure for analysis (Merck; reference number 1,06050,6025); acetic acid, pure for analysis (Carbo Erba; reference number 20104298); acetic anhydride (Fluka; reference number 45830); ethanol 95 (Prolabo; reference number 20823,293)

Links: Recondo, E.F., and Rinderknecht, Eine neue, Einfache Synthese des 1-O-Acetyl-2,3,5-Tri-O-β-D-Ribofuranosides. Helv. Chim. Acta, 1171-1173 (1959).

A solution of L-ribose 140 (150 g, 1 mol) in methanol (2 liters) was treated with sulfuric acid (12 ml) and left at +4°within 12 hours, and then neutralized with pyridine (180 ml). Evaporation gave α,β a mixture of methylribonucleotides 141 in the form of syrup. The solution of this anomeric mixture in pyridine (1,3 liters) was treated with benzoyl chloride (580 ml, 5 mol) with cooling and under mechanical stirring. The solution was left at room temperature for 12 hours and then poured on ice (approximately 10 liters) with continuous stirring. The mixture (oil in water) was filtered through a layer of cellite. The obtained oil layer cellica washed with water (3×3 liters), and then was dissolved with ethyl acetate (3 liters). The organic phase is washed with 5% solution of NaHCO3(2 liters) and water (2 liters), dried over sodium sulfate, filtered and evaporated to obtain 1-O-methyl-,3,5-tri-O-benzoyl-α /β-L-ribofuranose 142 in the form of a thick syrup. The oil was dissolved in acetic anhydride (560 ml) and acetic acid (240 ml). After adding dropwise concentrated acid (80 ml) the solution was kept in the cold (+4° (C) under mechanical stirring for 10 hours. Then the solution was poured on ice (approximately 10 liters) with continuous stirring. The mixture of oily compound in water) was filtered through a layer of cellite. Received kameneobrobnij solid layer cellica washed with water (3×3 liters) and then was dissolved in dichloromethane (2.5 liters). The organic phase is washed with 5% NaHCO3(1 liter) and water (2×2 liters), dried over sodium sulfate, filtered and evaporated to obtain kameneobrobnij solid 143, which crystallized from ethanol 95 (exit 225 g, 44%).

Analysis of 1-O-acetyl-2,3,5-tri-O-benzoyl-β-L-ribofuranose 143:

The melting point of 129-130°C (EtOH 95) (lit. (1) melting point 130-131°)

1H NMR (200 MHz, CDCl3): δ 8,09-7,87 (m, 6H, Harene), a 7.62-7,31 (m, 9H, Harene) to 6.43 (s, 1H, H1), 5,91 (dd, 1H, H3I , J3,46,7 Hz; J3,2a 4.9 Hz), 5,79 (pd, 1H, H2I , J2,34,9 Hz; J1,2<1), 4,78 (m, 2H, H4and H5), 4,51 (dd, 1H, H5I , J5,5'of 13.1 Hz, J5',45,5 Hz), from 2.00 (s, 3H, CH3WITH); (identical to commercial 1-O-acetyl-2,3,5-tri-O-benzoyl-β-D-ribofuranose)

Mass analysis (FAB+, GT m/z 445 (M-OAc)+

Elemental analysis of C28H24O9Designed With 66,66; N 4,79; Detected N

Reaction 2:

Starting material: Adenine (Pharma-Waldhof; reference number 400134,001 party 45276800)

Reagents: Chloride tin fuming (Fluka; reference number 96558); NH3/methanol (methanol, saturated NH3; see page 5); sodium sulfate (Prolabo; reference number 28111, 365)

Solvents: acetonitrile (Riedel-de Hean; reference number 33019; distilled over San2); chloroform, net (Acros; reference number 22706463); ethyl acetate, net (Carlo Erba; reference number 528299)

Links: Saneyoshi, M., and Satoh, E., Synthetic Nucleosides and Nucletides. XIII. Stannic Chloride Catalyzed Ribosylation of Several 6-Substituted Purines. Chem. Pharm. Bull., 27, 2518-2521 (1979); Nakayama, S., and Saneyoshi, M., Synthetic Nucleosides and Nucleotides. XX. Synthesis of Various 1-β-Xylofuranosyl-5-Alkyluracils and Related Nucleosides. Nucleosides, Nucleotides, 1, 139-146 (1982).

Adenine (19.6 g, 144 mmol) suspended in acetonitrile (400 ml) with 1-O-acetyl-2,3,5-tri-O-benzoyl-β-L-ribofuranose 143 (60 g, 119 mmol). To this suspension was added a steaming tin chloride (22 ml, 187 mmol). After 12 hours the reaction mixture was concentrated to small volume (about 100 ml), and added acidic sodium carbonate (110 g) and water (120 ml). The obtained white solid (tin salt) was extracted with hot chloroform (5×200 ml). The combined extracts were filtered through a layer of cellite. The organic phase is washed with 5%solution of NaHCO 3and water, dried over sodium sulfate, filtered and evaporated to obtain compound 144 (60 g, colourless foam). The foam was treated with methanol saturated with ammonia (220 ml) in a closed vessel at room temperature under stirring for 4 days. The solvent is evaporated under reduced pressure and the resulting powder was suspended in ethyl acetate (400 ml) by boiling under reflux for 1 hour. After filtration, the powder was recrystallized from water (220 ml) to obtain L-adenosine 145 (24 g, crystals, 75%).

Analysis β-L-adenosine:

The melting point 233-234°With (water) (lit. (4) melting point 235-238°)

1H NMR (200 MHz, DMSO-D6): δ 8.34 per 8,12 and (2s, 2H, H2and H8), 7,37 (1s, 2H, NH2), 5,86 (d, 1H, H1'I , J1',2'6,2 Hz), 5,43 (m, 2H, OH)2' and OH5'), 5,19 (d, 1H, HE3', J and 3.7 Hz), 4,60 (m, H2'), 4,13 (m. 1H, H3'), of 3.94 (m, 1H, H4'), 3,69-to 3.49 (m, 2H, H5 aand H5'b), (identical to commercial D-adenosine)

Mass analysis (FAB+, GT) m/z 268 (M+N)+, 136 (BH2)+

Reaction 3:

Reagents: 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane (Fluka; reference number 36520); sodium sulfate (Prolabo; reference number 28111,365)

Solvents: 99% pyridine (Acros; reference number 131780025); ethyl acetate, net (Carlo Erba; reference number 528299); acetonitrile (Riedel-de Haen; reference number 33019)

References: Robins, M.J., et al., Nucleic Acid Related Compounds. 42. A General Procedure for the Efficient Deoxygenation of Secondary Alcohols. Regiospecific and Stereoselective Conversion of Ribonucleosides to 2'-Deoxynucleosides. J. Am. Chem. Soc. 105, 4059-4065 (1983).

To L-adenosine 145 (47,2 g, 177 mmol)suspended in pyridine (320 ml)was added 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane (63 ml, 201 mmol)and the mixture was stirred at room temperature for 12 hours. The pyridine is evaporated, and the residue was distributed in ethyl acetate (1 liter) and 5% solution of NaHCO3(600 ml). The organic phase was washed 0,5N HCl solution (2×500 ml) and water (500 ml), dried over sodium sulfate, filtered and evaporated to the dry state obtained dry matter was led from acetonitrile to obtain compound 146 (81 g, 90%).

Analysis of 3',5'-O-(1,1,3,3-tetraisopropyl-1,3-disiloxanyl)-β-L-adenosine 146

The melting point 97-98° (acetonitrile) (lit. (5) D enantiomer, melting point 98°C)1H NMR (200 MHz, CDCl3): δ 8,28 and to 7.95 (2s, 2H, H2and H8), 5,96 (d, 1H, J1',2', 1,1 Hz), 5,63 (s, 2H, NH2), 5,10 (dd, 1H, H3', J3',4'7,6 Hz, J3',2'5,5 Hz), of 4.57 (dd, 1H, H2'I , J2',1'1,2 Hz; J2',3'a 7.6 Hz), 4,15-3,99 (m, 3H, H4'H5'aand H5'b), and 3.31 (sl, 1H, OH)2'), was 1.06 (m, 28H, isopropyl protons)

Mass analysis (FAB-, GT) m/z 508 (M-N)-, 134 (C)-, (FAB+, GT) m/z 510 (m+H)+, 136 (BH2)+

Reaction 4:

Reagents; 99% CI is ethylaminomethyl (Acros; reference number 1482702050); 99% familiarisation (Acros; reference number 215490050); Tris(trimethylsilyl)silane "TTMSS" (Fluka; reference number 93411); α,α'-azoisobutyronitrile "AIBN" (Fluka, reference number 11630); sodium sulfate (Prolabo; reference number 28111,365)

Solvents: acetonitrile (Riedel-de Haen; reference number 33019); ethyl acetate, net (Carlo Erba; reference number 528299); dioxane, pure for analysis (Merck; reference number 1,09671,1000); dichloromethane (Merck; reference number 1,06050,6025); methanol (Carbo Erba; reference number 309002);

References: Robins, M.J., Wilson, J.S., and Hansske, F., Nucleic Acid Related Compounds. 42. A General Procedure for the Efficient Deoxygenation of Secondary Alcohols. Regiospecific and Stereoselective Conversion of Ribonuc-leosides to 2'-Deoxynucleosides. J. Am. Chem. Soc. 105, 4059-4065 (1983).

Connection 146 (34 g, 67 mmol) was added acetonitrile (280 ml), DMAP (16.5 g, 135 mmol) and familiarisation (10,2 ml, 73 mmol). The solution was stirred at room temperature for 12 hours. The solvent was evaporated and the residue was distributed between ethyl acetate (400 ml) and 0.5 N HCl solution (400 ml). The organic layer was washed with 0.5 N HCl solution (400 ml) and water (2×400 ml), dried over sodium sulfate, filtered and evaporated to the dry state to obtain the intermediate compound as a pale yellow solid. Untreated 147 was dissolved in dioxane (ml) and was added AIBN (3.3 g, 20 mmol) and TTMSS (33 ml, 107 mmol). The progressive solution was heated to boiling with a back hall is dildocam and was stirred for 2 hours. The reaction mixture was concentrated to a yellow oil, which was chromatographically (eluent - dichloromethane/methanol 95/5) to obtain compound 148 (23 g, colorless foamy mass, 70%). Aliquots were led from ethanol/petroleum ether.

Analysis of 3',5'-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane)-2'-deoxy-β-L-adenosine 148:

The melting point 110-111°C (EtOH/petroleum ether) (Lit.(5) melting point 113-114°C (EtOH))

1H NMR (200 MHz, CDCl3): δ 8,33 and 8,03 (2s, 2H, H2and H8), 6,30 (dd, 1H, H1', J 2,85 Hz, J 7,06 Hz), 5,63 (sl, 2H, NH2), 4,96 (m, 1H, H3'), 4,50 (m, 2H, H5 aand H5'b),2,68 (m, 2H, H2'aand H2'b), a 1.08(m, 28H, isopropyl protons)

Mass analysis (FAB+, GT) m/z 494 (M+H)+, 136 (BH2)+

Reaction 5:

Reagents: ammonium fluoride (Fluka; reference number 09742); silica gel (Merck; reference number 1,07734,2500)

Solvents: Methanol, pure for analysis (Prolabo; reference number 20847,295); dichloromethane, pure for analysis (Merck; reference number 1,06050,6025); ethanol 95 (Prolabo; reference number 20823,293)

References: Zhang, W., and M.J. Robins, Removal of Silyl Protecting Groups from Hydroxyl Functions with Ammonium Fluoride in Methanol. Tetrahedron Lett., 33, 1177-1180 (1992).

A solution of 3',5'-O-(1,1,3,3-tetraisopropyl-1,3-disiloxane)-2'-deoxy-L-adenosine 148 (32 g, 65 mmol) and ammonium fluoride (32 g, mmol) in methanol was stirred while boiling under reflux for 2 hours was Added silica gel, and the mixture was carefully evaporated to obtain a white powder. This powder was applied on a column of silica gel, which was suirable dichloromethane/methanol 9/1. The appropriate fractions were combined and evaporated to obtain a white powder, which was led from ethanol 95 (12.1 g, 75%).

Analysis of 2'-deoxy-β-L-adenosine 149:

The melting point 189-190°C (EtOh 95) (identical to commercial 2'-deoxy-D-adenosine)

1H NMR (200 MHz, DMSO-D6): δ 8,35 8,14 and (2s, 2H, H2and H8), 7,34 (sl, 2H, NH2), 6.35mm (dd, 1H, H1', J 6,1 Hz, J a 7.85 Hz), 5,33 (d, 1H, OH)2', J 4,0 Hz), 5,28 (dd, 1H, H3'J of 4.95 Hz, J and 6.6 Hz), 4,42 (m, 1H, OH)5'), 3,88 (m, 1H, H4'), 3,63-to 3.52 (m, 2H, H5'aand H5'b), a 2.71 (m, 1H, H2'a), 2,28 (m, 1H, H2'b). (identical to commercial 2'-deoxy-D-adenosine

αD+260(0.5 water) (commercial 2'-deoxy-D-adenosine -25° (0.5 water)).

UV λmax260 nm (ε 14100) (H2O).

Mass analysis (FAB+, GT) m/z 252 (M+N)+, 136 (BH2)+

Example 3

Stereospecific synthesis of 2'-deoxy-β-L-cytidine

1-(3,5-Di-O-benzoyl-β-L-xylofuranosyl) uracil (11)

The hydrazine hydrate (1.4 ml, 28.7 mmol) was added to a solution of 1-(2-O-acetyl-3,5-di-O-benzoyl-β-L-xylofuranosyl)-uracil 10 [Cm. Gosselin, G., Bergogne, M. - C., Imabach, J-L., "Synthesis and Antiviral Evaluation of β-L-XylofuranosyI Nucleosides of the Five Naturally Occuring Nucleic Acid Bases", Journal ofHeterocyclic Chemistry, 1993, 30 (Oct.-Nov.), 1229-1233] (4,79 g, 9,68 mmol) in pyridine (60 ml) and acetic acid (15 ml). The solution was stirred over night at room temperature. Acetone was added (35 ml) and the mixture was stirred for 30 minutes. The reaction mixture was evaporated under reduced pressure. The obtained residue was purified by chromatography on a column of silica gel [eluent: stepwise gradient of methanol (0-4%) in dichloromethane to obtain 11 (3.0 g, 68%), which was led from cyclohexane/dichloromethane: melting point=111-114°;1H NMR (DMSO-d6): δ 11,35 (br s, 1H, NH), of 7.9 to 7.4 (m, 11H, 2C6H5Co, H-6), 6,38 (d, 1H, OH-2', JOH-2'=4,2 Hz), 5,77 (d, 1H, H-1', J1'-2'=1,9 Hz), of 5.55 (d, 1H, H-5, J5-6=8 Hz), 5,54 (dd, 1H, H-3', J3'-2'=3,9 Hz and J3'-4'=1,8 Hz), 4.8 (m, 1H, H-4'), 4,7 (m, 2H, H-5' and H-5"), 4,3 (m, 1H, H-2'); MS: FAB > 0 (matrix GT) m/z 453 (M+H)+, 105 (C6H5CO)+; FAB < 0 (matrix GT) m/z 451 (M-H)-, 121 (C6H5CO2)-, 111 (C)-; Analytical calculation for C23H20N2O8.H2O: C, 58,09; N, AMOUNTS TO 4.76; N, 5,96. Found: C, 57,71; H, Was 4.42; N, 5,70.

1-(3,5-Di-O-benzoyl-β-L-arabinofuranosyl) uracil (12)

To a solution of 1-(3,5-di-O-benzoyl-β-L-xylofuranosyl)-uracil 11 (8 g of 17.7 ml) in a mixture of anhydrous benzene-DMSO (265 ml, 6:4, V/V) was added anhydrous pyridine (1.4 ml), dicyclohexylcarbodiimide (up 10.9 g, 53 mmol) and dichloracetic acid (0.75 ml). The resulting mixture was stirred at the room for the Noah temperature for 4 hours, then was diluted with ethyl acetate (400 ml) and the solution was added oxalic acid (4.8 g, 53 mmol) in methanol (14 ml). After stirring for 1 hour the solution was filtered. The filtrate was washed with saturated solution of NaCl (2×500 ml), a 3% solution of NaHCO3(2×500 ml) and water (2×500 ml). The organic phase was dried over Na2SO4, then was evaporated under reduced pressure. The obtained residue was solubilizers in a mixture of absolute EtOH-benzene (140 ml, 2:1, V/V). When 0°to this solution was added NaBH4(0.96 g, of 26.5 mmol). After stirring for 1 hour the solution was diluted with ethyl acetate (400 ml), then filtered. The filtrate was washed with saturated NaCl (400 ml) and water (400 ml). The organic phase was dried over Na2SO4, then was evaporated under reduced pressure. The crude substance was purified by chromatography on a column of silica gel [eluent: stepwise gradient of methanol (0-3%) in dichloromethane to obtain 12 (5,3 g, 66%), which was led from acetonitrile: melting point = 182-183°;1H-NMR (DMSO-d6): δ 11,35 (br s, 1H, NH), 8,0-7,5 (m, 11H, 26H6CO, H-6), 6,23 (br s, 1H, OH-2'), x 6.15 (d, 1H, H-1', J1'-2'=4 Hz), 5,54 (d, 1H, H-5, J5-6=8,1 Hz), lower than the 5.37 (t, 1H, H-3', J3'-2'=J3'-4'=2,6 Hz), 4,7-4,6 (m, 2H, H-5' and H-5"), and 4.5 (m, 1H, H-4'), 4,4 (m, 1H, H-2'); MS: FAB > 0 (matrix GT) m/z 453 (M+H)+, 341 (S)+, 113 (BH2)+, 105 (C6H5CO)+-, 121 (C6H5CO2)-, 111 (C)-; Analytical calculation for C23H20N2O8; WITH, 61,06; N, TO 4.46; N, TO 6.19. Detected, 60,83; N, 4,34; N,6,25.

1-(3,5-Di-O-benzoyl-2-deoxy-β-L-erythropoiesis)uracil (13)

To a solution of 1-(3,5-di-O-benzoyl-β-L-arabinofuranosyl)uracil 12 (5,2 g of 11.4 mmol) in anhydrous 1,2-dichloroethane (120 ml) was added phenoxythiocarbonyl (4,7 ml, 34,3 ml) and 4-(dimethylamino)pyridine (DMAP, 12.5 g, 102,6 mmol). The resulting solution was stirred at room temperature in an argon atmosphere for 1 hour and then evaporated under reduced pressure. The residue was dissolved in dichloromethane (300 ml)and the organic solution was sequentially washed ice of 0.2 N hydrochloric acid (3×200 ml) and water (2×200 ml), dried over Na2SO4, then was evaporated under reduced pressure. The crude substance was evaporated several times with anhydrous dioxane and dissolved in this solvent (110 ml). In an argon atmosphere to the resulting solution was added hydride Tris(trimethylsilyl)silane (4,2 ml, 13.7 mmol) and α,α'-azoisobutyronitrile (AIBN, 0.6 g, 3,76 mmol). The reaction mixture was heated and stirred at 100°C for 1 hour in an argon atmosphere, then cooled to room temperature and evaporated under reduced pressure. The remainder of the imali chromatography on a column of silica gel [eluent: stepwise gradient of methanol (0-5%)], to obtain 13 (2,78 g, 56%), which was led from EtOH: melting point = 223-225°;1H-NMR (DMSO-d6): δ and 11.4 (br s, 1H, NH), 8,0-7,5 (m, 11H,2 C6H5CO, H-6), 6,28 (t, 1H, H-1', J=7 Hz), and 5.5 (m, 2H, H-1' and H-5), 4,6-4,4 (m, 3H, H-4', H-5' and H-5"), and 2.6 (m, 2H, H-2' and H-2"); MS: FAB > 0 (matrix GT) m/z 437 (M+H)+, 3325 (S)+; FAB < 0 (matrix GT) m/z 435 (M-H)-, 111 (B)-; Analytical calculation for C23H20N2O7: C, 63,30; N, TO 4.62; N, 6.42 PER. Found: C, 62,98; N, 4,79; N,6,40.

2'-deoxy-β-L-citizen (β-L-dC)

In an argon atmosphere to a solution of 1-(3,5-di-O-benzoyl-2-deoxy-β-L-Erythro-pentofuranose)uracil 13 (2.66 g, 6.1 mmol) in anhydrous 1,2-dichloroethane (120 ml) was added a reagent Lawesson (Lawesson) (1,72 g, 4.26 deaths mmol)and the reaction mixture was stirred while boiling under reflux for 2 hours. The solvent is then evaporated under reduced pressure and the residue was purified by chromatography on a column of silica gel [eluent: stepwise gradient of ethyl acetate (0-8%) in dichloromethane]to obtain 4-thio-intermediate as a yellow foamy mass. The solution of this thio-intermediate (1.5 g, of 3.31 mmol) in methanolic ammonia (previously saturated at -10°and hermetically closed) (50 ml) was heated at 100°tank stainless steel for 3 hours and then was cooled to 0°C. the Solution was evaporated under reduced pressure. The crude substance was purified is by chromatography on a column of silica gel [eluent: stepwise gradient of methanol (0-20%) in dichloromethane]. Finally, the appropriate fractions were combined, filtered through a cell Millex HV-4 (0.45 μm, Millipore)and evaporated under reduced pressure to obtain the desired 2'-deoxy-β-L-citizen (β-L-dC) as a foamy mass (0.6 g, 80%), which crystallized from absolute EtOH: melting point=198-199°;1H-NMR (DMSO-d6): δ to 7.77 (d, 1H, H-6, J6-5=7,4 Hz), 7,10 (br d, 2H, NH2), 6,13 (t, 1H, H-1', J=6,7 Hz), 5,69 (d, 1H, H-5, J5-6=7,4 Hz), 5,19 (d, 1H, OH-3', JOH-3'=4,1 Hz), 4,96 (t, 1H, OH-5', JOH-5'=JOH-5"=5,2 Hz), 4,1 (m, 1H, H-3', of 3.75 (m, 1H, H-4'), and 3.5 (m, 2H, H-5' and H-5"), 2,0 (m, 1H, H-2'), and 1.9 (m, 1H<H-2"); MS: FAB > 0 (matrix GT) m/z 228 (M+H)+, 112 (BH2)+; FAB < 0 (matrix GT) m/z 226 (M-H)-; [α]20D=-69 (0,52, DMSO) [α]20D=+76 (0,55, DMSO) for commercially available hydrochloride D-enantiomer]. Analytical calculation for C9H13N3O4: C, 47,57; N, 5,77; N, BE 18.49. Found: C, 47,35; N, Of 5.68; N, 18, 29.

Example 4

Stereotypically synthesis of 2'-deoxy-β-L-cytidine (β-L-dC)

2-Amino-β-L-arabinofuranose[1',2':4,5]oxazoline (1)

A mixture of L-arabinose (170 g, 1.1 mol)of cyanamide (100 g of 2.38 mol), methanol (300 ml) and 6 M NH4OH (50 ml) was stirred at room temperature for 3 days and then kept at -10°With during the night. The product was collected by suction, washed successively with methanol and ether and drying is whether under vacuum. The output 130 g (66%) of analytically pure compound 1, the melting point 170-172°;1H NMR (DMSO-d6) δ ppm of 6.35 (br s, 2H, NH2), of 5.15 (d, 1H, H-1, J=5,6 Hz), the 5.45 (br s, 1H, OH-3), 4,70 (br s, 1H, OH-5), 4,55 (d, 1H, H-2, J=5,6 Hz), 4,00 (br s, 1H, H-3), the 3.65 (m, 1H, H-4), of 3.25 (m, 2H, H-5, H-5').

Reagents:

L-arabinose: Fluka, >99.5%pure, reference number 10839

The cyanamide: Fluka, >98%, reference number 28330

O2,2'-anhydrous-β-L-uridine (2)

A solution of compound 1 (98,8 g of 0.57 mol) and methylpropionate (98 ml) in 50% aqueous ethanol (740 ml) was boiled under reflux for 5 hours, then cooled and concentrated under reduced pressure to half the original volume. After precipitation with acetone (600 ml) the product was collected by suction, washed with ethanol and ether, and dried. The mother solution was partially concentrated, the concentrate is precipitated with acetone (1000 ml), the solid is collected by suction and washed with acetone and ether to obtain another product weight. Total yield, 80 g (62%) of compound 2, the melting point 236-240°;!H NMR (DMSO-d6) δ ppm 7,87 (d, 1H, H-6, J=7,4 Hz), 6.35mm (d, 1H, H-1', J=5,7 Hz), 5,95 (d, 1H, H-5, J=7,4 Hz), 5,90 (d, 1H, OH-3'), 5,20 (d, 1H, H-2', J=5,7 Hz)to 5.00 (m, 1H, OH-3'), of 4.44 (br s, 1H, H-3'), of 4.05 (m, 1H, H-4'), of 3.25 (m, 2H, H-5, H-5').

The reagent;

Methylpropionate: Fluka, >97%, reference number 81863

3',5'-di-O-benzoyl-O2,2'-anhydrous-β-L-uridine. (3)

To a solution of compound 2 (71,1 g, 0.31 mol) in anhydrous pyridine (1200 ml) dobavlyalost benzoyl (80.4 ml) at 0° And in argon atmosphere. The reaction mixture was stirred at room temperature for 5 hours with the exclusion of atmospheric moisture, and stopped by adding ethanol. The solvent was evaporated under reduced pressure and the obtained residue was evaporated with toluene and absolute ethanol. The crude mixture was then diluted with ethanol, and the precipitate was collected by suction, washed successively with ethanol and ether, and dried. Exit 129 g (95,8%) of compound 3, melting point 254°;1H NMR (DMSO-d6) δ parts per thousand of 8.1 to 7.4 (m, 11H, C6H5CO, H-6), of 6.50 (d, 1H, H-1', J=5,7 Hz), 5,90 (d, 1H, H-5, J=7,5 Hz), 5,80 (d, 1H, H-2', J=5,8 Hz), 5,70 (d, 1H, H-3'), the 4.90 (m, 1H, H-4'), 4,35 (m, 2H, H-5, H-5').

Reagent:

The benzoyl chloride: Fluka, pure for analysis, reference number 12930

3',5'-Di-O-benzoyl-2'-chloro-2'-deoxy-β,L-uridine (4)

When 0°to a solution of compound 3 (60,3 g, 0,139 mol) in dimethylformamide (460) solution was added 3.2 N-HCl/DMF (208 ml, prepared in situ by adding to 47.2 ml acetylchloride at 0°to a solution of 27.3 ml of methanol and 133,5 ml of dimethylformamide). The reaction mixture was stirred at 100°C for 1 hour while removing atmospheric moisture, cooled and poured into water (4000 ml). The precipitate of compound 4 was collected by suction, washed with water and recrystallized from ethanol. The crystals were collected, washed with cold ethanol and ether and dried at on igenom pressure. Exit to 60.6 g (92,6%) of compound 4, the melting point of 164-165°;1H NMR (DMSO-d6) δ to 8.7 ppm (br s, 1H, NH), 8,1-to 7.3 (m, 11H, C6H5CO, H-6), x 6.15 (d, 1H, H-1', J=4,8 Hz), and 5.5 (m, 2H, H-5, H-2')and 4.65 (m, 4H, H-3', H-4', H-5', H-5").

Reagents:

Acetylcholine: Fluka, pure for analysis, reference number 00990

3',5'-Di-O-benzoyl-2'-deoxy-β,L-uridine (5)

A mixture of compound 4 (60,28 g, 0,128 mol)hydride tri-n-butyanova (95 ml) and azobisisobutyronitrile (0,568 g) in dry toluene (720 ml) was boiled under reflux with stirring for 5 hours and cooled. The solid is collected by suction and washed with cold toluene and petroleum ether. The filtrate was concentrated under reduced pressure and diluted with petroleum ether to precipitate additional mass of compound 5. Output 54,28 g (97,2%) of compound 5; melting point 220-221°;1H NMR (CDCl3δ ppm 8,91 (br s, 1H, NH), 8,1-7,5 (m, 11N,6H5CO and H-6), to 6.43 (q, 1H, H-1', J1',2'=5,7 Hz and J1',2"=8,3 Hz), 5,7-5,6 (m, 2H, H-3' and H-5), 4,8-4,6 (m, 3H, H-5', H-5 and H-4'), of 2.8 and 2.7 (m, 1H, H-2'), 2,4-2,3 (m, 1H, H-2").

Reagents:

The hydride is tri-n-butyanova: Fluka, >98%, reference number 90915

Azobisisobutyronitrile: Fluka, >98%, reference number 11630

3',5'-Di-O-benzoyl-2'-deoxy-β-L-4-thio-uridine (6)

A solution of compound 5 (69 g, 0,158 mol) and the reagent Lawesson (74 g) in anhydrous methylene chloride (3900 ml) was heated in an argon atmosphere overnight. After the Ariane solvent the crude residue was purified by chromatography on a column of silica gel [eluent: gradient of methanol (0-2%) in methylene chloride], to obtain a pure compound 6 (73 g) in quantitative yield;1H NMR (CDCl3) δ to 9.5 ppm (br s, 1H, NH), of 8.1 to 7.4 (m, 10H, C6H5CO), 7,32 (d, 1H, H-6, J=7,7 Hz), 6.30-in (dd, 1H, H-1', J=5,6 Hz and J=8,2 Hz), to 6.22 (d, 1H, H-5, J=7,7 Hz), 5,6 (m, 1H, H-3'), 4,7 (m, 2H, H-5', H-5"), and 4.5 (m, 1H, H-4'), 2,8 (m, 1H, H-2'), 2,3 (m, 1H, H-2").

Reagents:

The reagent Lawesson (Lawesson): Fluka, >98%, reference number 61750

2'-Deoxy-β-L-cytosine.

A solution of compound 6 (7,3 g to 0.016 mol) in methanol saturated with ammonia (73 ml)was heated at 100°tank stainless steel for 3 hours. After careful cooling, the solvent is evaporated under reduced pressure. An aqueous solution of the residue was washed with ethyl acetate and evaporated until dry. This procedure was performed on 9 other samples (each of 7.3 g) of compound 6 (total number of 6=73 g). 10 residues were combined, diluted with absolute ethanol and cooled to obtain 7 in the form of crystals. Traces benzamide was removed from the crystals 6 extraction system solid-liquid (boiling under reflux in ethyl acetate for 1 hour). Output 28,75 g (78,6%) of compound 6; melting point 141-145°;1H NMR (DMSO) δ by 8.22 ppm and 8.00 (2 br s, 2H, NH2), 7,98 (d, 1H, H-6, J=to 7.59 Hz), 6,12 (t, 1H, H-1; J=6,5 Hz and J=7,6 Hz), of 5.89 (d, 1H, H-5, J=to 7.59 Hz), 5,3 (br s, 1H, OH-3'), 5,1 (br s, 1H, OH-5'), 4,2 (m, 1H, H-3'), 3,80 (q, 1H, H-4', J=3,6 Hz and J=6,9 Hz), 3,6-3,5 (m, 2H, H-5', H-5"), 2,2-2,0 (m, 2H, H-2', H-2"); FAB < 0, (GT) m/e 226 (M-H)-, 110 (C)-; FAB > 0 (GT) 228 (M+H)+, 112 (B+2H)+; [α]D20- 56,48 (=1,08 in DMSO); UV (pH 7) λmax=270 nm (ε 10000).

Reagents:

A methanolic ammonia solution: pre-saturated with -5°C, tightly closed and kept in the refrigerator.

Example 5

Stereotypically synthesis of 2'-deoxy-β-L-thymidine (β-L-dT)

3',5'-Di-O-benzoyl-2'-deoxy-5-iodine-β-L-uridine (7)

A mixture of compound 5 (105,8 g, 0,242 mol), iodine (76,8 g), CAN (66,4 g) and acetonitrile (2550 ml) was stirred at 80°C for 3 hours, then the reaction mixture was cooled at room temperature, leading to crystallization of compound 7 (86,6 g, 63.5 per cent); melting point 192-194°;1H NMR (DMSO) δ ppm to 8.34 (s, 1H, NH), and 8.2 to 7.2 (m, 11H, 2 C6H5CO, H-6), of 6.31 (q, 1H, H-1', J=5,5 Hz and J=8,7 Hz), and 5.5 (m, 1H, H-3'), 4,7 (m, 2H, H-5', H-5"), and 4.5 (m, 1H, H-4'), and 2.7 (m, 1H, H-2'), 2,3 (m, 1H, H-2"); FAB<0, (GT) m/e 561 (M-H)-, 237 (C)-; FAB > 0 (GT) 563 (M+H)+; [α]D20+39,05 (s=1,05 in DMSO); UV (EtOH 95) νmax=281 nm (ε 9000), νmin=254 nm (ε 4000), νmax=229 nm (ε=31000); Analytical calculation for C23H19IN2O7: C, 49,13; H, 3,41; N, 4,98; I, 22,57; Found: C, 49,31; N, Of 3.53; N,Of 5.05; I, 22,36.

Reagents:

Iodine: Fluka, 99.8 per cent, the reference number 57650

Nitrate of cerium ammonium (CAN): Aldrich, >98.5%with the reference number 21547-3

3',5'-Di-O-benzoyl-2'-deoxy-3-N-toluoyl-β-L-thymidine (9)

It is astory compound 7 (86,6 g, 0,154 mol) in anhydrous pyridine (1530 ml)containing N-ethyldiethanolamine (53,6 ml)was added by portions at 0°p-trouillard (40,6 ml). The reaction mixture was stirred for 2 hours at room temperature, then added water to stop the reaction, and the reaction mixture was extracted with methylene chloride. The organic phase is washed with water, dried over ammonium sulfate and evaporated to the dry state to obtain crude 3',5'-di-O-benzoyl-2'-deoxy-3-N-toluoyl-5-iodine-β-L-uridine (8), which can be used for the next stage without further purification.

A solution of the crude mixture 8, palladium acetate (3,44 g), triphenylphosphine (8.0 g) in N-methylpyrrolidinone (1375 ml) with triethylamine (4.3 ml) was stirred at room temperature for 45 minutes. Then add tetramethylsilane (42,4 ml) dropwise at 0°C in argon atmosphere. After stirring at 100-110°during the night the reaction mixture was poured into water and was extracted with diethyl ether. The organic solution was dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by chromatography on a column of silica gel [eluent: stepwise gradient of ethyl acetate (0-10%) in toluene]to obtain compound 9 in the form of a foamy substance (42,3 g, 48.3%) in 2 stages).1H NMR (DMSO) δ ppm of 8.3 to 7.2 (m, 15H, 2 C6H5CO, 1 sub> 3With6H4CO, H-6), of 6.29 (t, 1H, H-1', J=7,0 Hz), 5,7 (m, 1H, H-3'), a 4.7 to 4.5 (m, 3H, H-5', H-5', H-4'), 2,7-2,6 (m, 2H, H-2', H-2"); FAB < 0, (GT) m/e 567 (M-H)', 449 (M-CH3With6H4CO)-, 243 (C)-, 121 (C6H5Soo)-; FAB > 0 (GT) 1137 (2M+H)+, 569 (M+H)+, 325 (M-B), 245 (B+2H)+, 119 (CH3With6H5CO)+.

Reagents:

p-Trouillard, Aldrich, 98%, reference number 10,663-1

Diisopropylethylamine: Aldrich, >99.5%pure, reference number 38,764-9

N-methylpyrrolidinone: Aldrich, >99%, reference number 44,377-8

The palladium acetate: Aldrich, >of 99.98%, reference number 37,987-5

Triphenylphosphine: Fluka, >97%, reference number 93092

Tetramethylol: Aldrich, >99%, reference number 14,647-1

2'-Deoxy-β-L-thymidine.

A solution of compound 9 (42,3 g 0,074 mol) in methanol saturated with ammonia (1850 ml)was stirred at room temperature for two days. After evaporation of the solvent the residue was diluted with water and washed several times with ethyl acetate. The aqueous layer was separated, evaporated under reduced pressure and the residue was purified by chromatography on a column of silica gel [eluent: stepwise gradient of methanol (0-10%) in methylene chloride]to obtain pure 2'-deoxy-β-L-thymidine (are 11.62 g, 64,8%), which crystallized from ethanol; melting point 185-188°;1H NMR (DMSO) δ to 11.3 ppm (s, 1H, NH), of 7.70 (s, 1H, H-6), 6,2 (pt, 1H, H-1'), of 5.24 (d, 1H, OH-3', J=4,2 Hz), to 5.08 (t, 1H, OH-5', J=5,1 Hz), 4,2 (m, 1H, H-3"), and 3.7 (m, 1H, H-4') 3,5 of 3.6 (m, 2H, H-5', H-5"), from 2.1 to 2.0 (m, 2H, H-2', H-2"); FAB < 0, (GT) m/e 483 (2M-H)-, 349 (M+T-H)-, 241 (M-H)-, 125 (C)-; FAB > 0 (GT) 243 (M+H)+, 127 (B+2H)+; [α]D20is 13.0 (C=1.0 in DMSO); UV (pH 1) νmax=267 nm (ε 9700), νmin=234 nm (ε 2000).

Reagent:

A methanolic ammonia solution: pre-saturated with -5°C, tightly closed and kept in the refrigerator.

Example 6

Stereotypically synthesis of 2'-deoxy-β-L-inosine (β-L-dI)

β-L-dI synthesized by diaminononane 2'-deoxy-β-L-adenosine (β-L-dA) according to the method previously described in section 9-D-glucopyranosyl (Cm. I.Iwai, T.Nishimura and .Shimizu, Synthetic Procedures in Nucleic Acid Chemistry, W.W.Aorbach and R.S.Tipson, eds., John Wiley & Sons, Inc. New York, vol.1, pp 135-138 (1968)).

Thus, the solution β-L-dA (200 mg) in a mixture of acetic acid (0,61 ml) and water (19 ml) was heated with sodium nitrite (495 mg)and the mixture was stirred at room temperature overnight. The solution is then evaporated until dry under reduced pressure. An aqueous solution of the residue was applied onto a column of ion exchange resin IR-120 (H+), and the column was suirable water. The appropriate fractions were collected and evaporated until dry, to get a clean β-L-dI, which crystallized from methanol (106 mg, 53%, the output is not optimal): melting point = 209°of 211°C; UV (N2O), &x003BB; max=247 nm;1H-NMR (DMSO-d6)=8,32 and 8,07 (2s, 1H each, H-2 and H-8), 6,32 (pt, 1H, H-1; J=6,7 Hz), 4,4 (m, 1H, H-3'), a 3.9 (m, 1H, H-4'), which is 3.7 and 3.4 (m, 2H partially closed HOD, H-5', 5"), and 2,6 2,3 (2m, 1H each, H-2' and H-2"); mass spectrum (matrix, glycerol-thioglycerol, 1:1, V/V), FAB>0: 253 (m+H)+, 137 (base + 2N)+; FAB<0: 251 (m-H)', 135 (base)'; [α]D20=+19,3 (0,88, N2O).

Anti-HBV activity of active connections

The ability of the active compounds to inhibit the growth of the virus in cultures of 2.2.15 cells (HepG2 cells transformed by the virion hepatitis) can be estimated according to the method, described in detail below.

Conclusions and summary of the study the antiviral effect of these cultural systems and analysis of HBV DNA described Korba and Milman (1991, Antiviral Res., 15:217). Evaluation of the antiviral action was carried out on two separate passages of the cells. All wells in all the tablets were sowed with the same density and at the same time.

Due to genetic variation in the levels of both intracellular and extracellular HBV DNA, only reduction of more than 3.5 times (for DNA HBV virion) or 3.0 times (for intermediates of DNA replication HBV) below average levels of these forms of HBV DNA in untreated cells was considered as statistically significant (P<0,05). The levels of integrated HBV DNA in each cell DNA preparation (which remained constant for the cell e is their experiments) was used to calculate the levels of intracellular forms of HBV DNA, thus, ensuring that the same amount of cellular DNA are compared between separate samples.

Typical values for extracellular DNA of HBV virion in untreated cells ranged from 50 to 150 PG/ml of culture medium (average approximately 76 PG/ml). Intracellular intermediates of replication of HBV DNA in untreated cells ranged from 50 to 100 ug/PG cellular DNA (on average approximately 74 PG/μg of cellular DNA). Generally lower levels of intracellular HBV DNA due to treatment with the antiviral compounds was less pronounced and was slower than the decrease in levels of DNA HBV virion (Korba and Milman, 1991, Antiviral Res., 15:217).

The way in which in these experiments, the analysis of hybridization results in an equivalence of approximately 1.0 PG of intracellular HBV DNA to 2-3 genomic copies per cell and 1.0 PG/ml of extracellular HBV DNA at the 3×105viral particles/ml

Example 7

Investigated the ability trifosfatnogo derivatives β-L-dA β-L-dC, β-L-dU, β-L-2'-dG, β-L-dI β-L-dT to inhibit hepatitis C. Table 1 describes the comparative inhibitory activity of triphosphates β-L-dT (β-L-dT-TP), β-L-dC (β-L-dC-TP), β-L-dU (β-L-dU-TP) and β-L-dA (β-L-dA-TP) in relation to the DNA polymerase of hepatitis b virus marmot (WHV)DNA-α-, β- and γ-olymers person.

Table 1
InhibitorDNA-Polym. WHV IC50DNA-Polym. α Kib(µm)DNA-Polym. βib(µm)DNA-Polym. γib(µm)
β-L-dT-TP0,34>100>100>100
β-L-dA-TP2,3>100>100>100
β-L-dC-TP2,0>100>100>100
β-L-dU-TP8>100>100>100
aIC50: the concentration that causes 50% inhibition

bKiwas determined using activated DNA calf thymus as a matrix for seed and dATP (d) as substrate. Inhibitors were analyzed using the graphical method of Dixon (Dixon). Under these conditions, the average KmDNAαpolymerase person for dATP calculated approximately as of 2.6 μm. DNAβpolymerase man was characterized by a Km of the stationary state of 3.33 μm for dATP. DNAγpolymerase person characterized Km steady state of 5.2 μm.

Use the 8

Investigated the activity against hepatitis b virus -β-L-dA β-L-dC, β-L-dU, β-L-2'-dG β-L-dT - transfected cells ner G-2 (2.2.15). Table 2 illustrates the action β-L-dA β-L-dC, β-L-dU β-L-dT on replication of hepatitis b virus in transfected cells ner G-2 (2.2.15).

Table 2
ConnectionVirionsaHBV EU50(µm)HBV RibEU50(µm)Cytotoxicity IC50(µm)The selectivity index of the IC50/EC50
β-L-dT0,050,05>200>4000
β-L-dC0,050,05>200>4000
β-L-dA0,100,10>200>2000
β-L-dI1,01,0>200>200
β-L-dU5,05,0>200>40
aExtracellular DNA

bIntermediate replication (intracellular DNA)

Example 9

Determined the effect of combinations β-L-dA β-L-dC and β-L-dT on the growth of hepatitis b virus in 2.2.15 cells. Rezultaty.prepodavateli in Table 3.

Table 3
CombinationRatioEU50
L-dC+L-dT1:3.023
L-dC+L-dT1:1.053
L-dC+L-dT3:1.039
L-dC+L-dA1:30.022
L-dC+L-dA1:10.041
L-dC+L-dA1:3.075
L-dT+L-dA1:30.054
L-dT+L-dA1:10.077
L-dT+L-dA1:3.035

Each combination showed anti-HBV activity, which was synergistic. In addition, the combination of L-dA+L-dC+LdT were also synergistic in this model.

Example 10

Determined the inhibition of the replication of hepatitis b virus in 2.2.15 cells by β-L-dA β-L-dC, separately and in combinations. The results are presented in Table 4.

Table 4
aβ-L-2'-deoxyadenosine (µm)bβ-L-2'-deoxycytidine (µm)% inhibitioncC.I.
0,5
0,0524
0,0051
0,595
0,0540
0,00510
0,050,05800,34
0,050,005560,20
0,050,0005500,56
0,0050,05720,35
0,0050,005540,35
0,0050,0005300,16
0,00050,05500,83
0,00050,005150,28
0,00050,00050no analysis
aβ-L-2'-deoxyadenosine: IC50=0,09 ám
bβ-L-2'-deoxycytidine: IC50=0.06 micron
cValues of indicators combinations indicate a synergistic action of the (< 1), additive effect (=1) and the antagonistic action 01)

Example 11

Determined the effectiveness of L-dA, L-dT and L-dC in relation hepadnaviruses (hepadnavirus) infection in woodchucks (Marmota monax), chronically infected with hepatitis marmots (WHV). This animal model of HBV infection is common and is used for the evaluation of antiviral agents against HBV.

Protocol:

The experimental group (n=3 animals/group, were administered the medication, n=4 animals/control)

Group 1control filler
Group 2lamivudine (3TC) (10 mg/kg/day)
Group 3-6L-dA (0,01; 0,1; 1,0; 10 mg/kg/day)
Group 7-10L-dt (0,01; 0,1; 1,0; 10 mg/kg/day)
Group 11-14L-dC (0,01; 0,1; 1,0; 10 mg/kg/day)

Drugs were administered orally via a stomach tube once daily and took blood samples on 0, 1, 3, 7, 14, 21, 28 day and after treatment+1, +3, +7, +14, +28, and +56 days. Evaluate the activity and toxicity was based on the reduction of WHV DNA in serum: dot-blotting, quantitative PCR (polymerase chain reaction, PCR). The results are presented in Figure 3 and in Table 5.

Table 5

protivovirusnaya activity LdA, Ldt, LdC model of chronic HBV infection Groundhog
DaysControlLdALdTLdC
ng DNA WHV per ml serum1,2
0381436423426
139836945123
34121401462
7446102646
1439274120

1LdA, LdT, LdC was administered 10 mg/kg orally once a day
2The limit of detection is 1 ng/ml DNA WHV per ml serum

The data show that L-dA, L-dT and L-dC are highly active on this model in vivo. First, the viral load decreases to undetectable (L-dT) or almost undetectable (L-dA, L-dC) levels. Second it is shown that L-dA, L-dT and L-dC are more active than 3TC (lamivudine) on this model. Thirdly, the re-emergence of the virus was not determined at least within two weeks after discontinuation of L-dT. The fourth curve "dose-response" suggests, is a way of increasing doses of L-dA and L-dC will be manifested in antiviral activity similar to the activity of L-dT. Fifth in all animals treated with medicines, increased weight, and there was no toxicity associated with the drug.

The toxicity of compounds

To determine whether any of the observed antiviral effects due to shared effects on cell viability, conducted a toxicity study. Used method is the determination of the effect β-L-dA β-L-dC and β-L-dT on the growth of cells in Karaganda analysis of human bone marrow when compared with lamuvidine. The results are presented in Table 6.

Table 6
ConnectionCFU-GM (µm)BFU-E (µm)
β-L-dA>10>10
β-L-dC>10>10
β-L-dT>10>10
β-L-dU>10>10
lamuvidine>10>10

Obtaining pharmaceutical compositions

A person suffering from any described in the application by the violation, including hepatitis b, can be treated by administration to the patient an effective amount of β-2'-deoxy-β-L-Erythro-pentofuranose-nucleoside, for example, β-L-2'-deoxyadenosine, β-L-2'-deoxycytidine, β-L-2'-dose irradiation on neurogenesis, β-L-2'-deoxy-guanosine or β-L-2'-deoxythymidine or their pharmaceutically acceptable prodrugs or salts in the presence of a pharmaceutically acceptable carrier or diluent. The active substance can be entered using any appropriate method, for example, oral, parenteral, intravenous, intradermal, subcutaneous, or topically, in liquid or solid form.

The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to provide the patient a therapeutically effective amount of compounds for inhibition of viral replication in vivo, without causing serious toxic effects on the patient treated. The term "inhibitory amount" means the amount of active ingredient sufficient to cause inhibition, which is determined by, for example, such a method as the method described in the application.

The preferred dose of the compound for all of the above conditions is located in the area approximately from 1 to 50 mg/kg, preferably from 1 to 20 mg/kg of body weight per day, mostly from approximately 0.1 to 100 mg per kilogram body weight of recipient per day. The range of effective dosages farmacevtichesky acceptable Proletarsk is and can be calculated on the basis of weight related nucleoside, which must be delivered. If the prodrug is active by itself, an effective dose can be established, as mentioned above, by using the weight of prodrug, or other methods known to experts in this field.

The connection is conveniently introduced into any acceptable standard dosage form, including a form containing 7 to 3000 mg, preferably from 70 to 1400 mg of active ingredient on a standard dosage form, but is not limited to this. Usually conventional 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 μm. This can be achieved, for example, by intravenous injection of 0.1 to 5% solution of the active ingredient, optionally in saline, or introduced in the form of a bolus of the active ingredient.

The concentration of the active compound in the drug composition depends on absorptio, inactivation and excretion rate of drug substances, as well as other factors known to specialists in this field. It should be noted that the magnitude of the doses also vary depending on the severity of the condition, which should be facilitated. In addition, it is clear that for any particular subject should have is tanovich a special regimen of medicines depending on time in accordance with individual need and the professional judgment of an individual, appointing a song or watching their introduction, and that the concentration ranges are examples only and are not intended to limit the scope or application of the claimed composition. The active ingredient can be entered directly or can be divided into several small doses for administration at different intervals of time.

The preferred method of introduction of the active compounds is oral. Oral compositions generally include an inert solvent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic injection of the active compound may be injected with fillers and applied in the form of tablets, lozenges or capsules. Pharmaceutically compatible binding agents, and/or adjuvant substances may be included as part of the composition.

Tablets, pills, capsules, lozenges and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, tragacanth gum or gelatin; an excipient such as starch or lactose, a disintegrator, such as alginic acid, primogel or corn starch; a lubricant such as magnesium stearate or sterate; substance which provides colleenie, such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or a flavoring agent, such as peppermint, methyl salicylate or orange flavor. If the standard dosage form is a capsule, it may contain, in addition to the substances of the above type, a liquid carrier such as fatty oil. In addition, standard forms can contain various other materials which modify the physical form of the drug, for example, coatings of sugar, shellac or other intersolubility substances.

The connection can be entered as a component of an elixir, suspension, syrup, water, chewing gum and the like. In addition to the active compounds, the syrup may contain sucrose as a sweetener and certain preservatives, dyes or colorants or flavorings.

The compound or its pharmaceutically acceptable derivative or salt can also be mixed with other active materials that do not impair the desired action, or with substances that Supplement the desired action, such as antibiotics, antifungal agents, anti-inflammatory agents, protease inhibitors, or other nucleoside or non-nucleoside antiviral agents. Solutions or suspensions used for parenteral, intradermal, subcutaneous or local applications to the tion, may include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates, and agents for the maintenance of tone, such as sodium chloride or dextrose. Parenteral preparations can be enclosed in ampoules, disposable syringes or multiple-dose vials made of glass or plastic.

When intravenously, preferred carriers are physiological saline or phosphate buffered saline (PBS).

In a preferred aspect, the active compounds are prepared with carriers that protect the compound against rapid elimination from the body, as, for example, compositions with controlled release, including implants and microencapsulation delivery system. You can use biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyarteritis and polixena acid. Methods of obtaining the same, the compositions, obviously, well-known experts in this field. Substances can be obtained commercially from Alza Corporation.

Liposomal suspensions (including liposomes, targeting infected cells with monoclonal antibodies to viral antigens) are also preferred as pharmaceutically acceptable carriers. They can be obtained in accordance with methods known to experts in this field, for example as described in U.S. Patent No. 4522811. For example, liposomal compositions can prigotovit by dissolving appropriate lipid(s) (such as stearoylethanolamine, stearoylethanolamine, arachidonyl-phosphatidylcholine and cholesterol) in an inorganic solvent that is then pariveda, leaving a thin layer of dried lipid on the surface of the tank. Then an aqueous solution of active compound or its monophosphate, diphosphate and/or trifosfatnogo derived contribute in the tank. The tank is then rotated by hand to separate the lipid substance from the walls of the tank and dispersing the lipid aggregates, creating, thus, the liposomal suspension.

This invention is described with reference to its preferred implementations. Changes and modifications of the invention are obvious to experts in this field from the previous detailed description of the invention. Suppose h is about all these changes and modifications are within the scope of this invention.

1. Applying an effective amount of β-L-2'-deoxynucleoside formula

in the production of drugs for the treatment of infection of hepatitis b virus by host.

2. Applying an effective amount of β-L-2'-deoxynucleoside formula

in the production of drugs for the treatment of infection of hepatitis b virus by host.

3. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salts in the manufacture of drugs for the treatment of infection of hepatitis b virus by host.

4. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salts in the manufacture of drugs for the treatment of infection of hepatitis b virus by host.

5. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable ester in the manufacture of drugs for the treatment of infection of hepatitis b virus by host.

6. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable ester in the manufacture of drugs for the treatment of infection of hepatitis b virus by host.

7. The use according to any one of claims 1 to 6, where β-L-2'-deoxynucleoside at least 95% of the specified enantiomeric form.

8. The use according to any one of claims 1 to 6, where β-L-2'-deoxynucleoside is represented with a pharmaceutically acceptable carrier.

9. The use of claim 8, where the pharmaceutically acceptable carrier is suitable for oral administration.

10. The use of claim 8, where the pharmaceutically acceptable carrier is suitable for intravenous administration.

11. The use of claim 8, where the pharmaceutically acceptable carrier is suitable for parenteral administration.

12. The use of claim 8, where the pharmaceutically acceptable carrier is suitable for intradermal injection.

13. The use of claim 8, where the pharmaceutically acceptable carrier is suitable for subcutaneous injection.

14. The use of claim 8, where the pharmaceutically acceptable carrier is suitable for topical administration.

15. The use of claim 8, where β-L-2'-deoxynucleoside represented as a single dose.

16. The application indicated in paragraph 15, where the unit dose contains from 10 to 1500 mg β-L-2'-deoxynucleoside.

17. The application of clause 15 or 16, where unit is the usual dose is a tablet or capsule.

18. The use according to any one of claims 1 to 6, where the owner is the man.

19. Pharmaceutical composition comprising an effective amount of a combination of the following β-L-2'-deoxynucleosides

and

for treatment of infections of hepatitis b virus by host.

20. Pharmaceutical composition comprising an effective amount of a combination of the following β-L-2'-deoxynucleosides:

and

or their pharmaceutically acceptable salts for the treatment of infection of hepatitis b virus by host.

21. Pharmaceutical composition comprising an effective amount of a combination of the following β-L-2'-deoxynucleosides:

and

or their pharmaceutically acceptable salts or esters for the treatment of infection of hepatitis b virus by host.

22. The pharmaceutical composition according to any one of p-21, where β-L-2'-deoxynucleoside at least 95% of its synthetic enantiomeric form.

23. The pharmaceutical composition according to any one of p-21, where β-L-2'-deoxynucleoside is represented with a pharmaceutically acceptable carrier.

24. The pharmaceutical composition according to item 23, where the pharmaceutically when memy carrier is suitable for oral administration.

25. The pharmaceutical composition according to item 23, where the pharmaceutically acceptable carrier is suitable for intravenous administration.

26. The pharmaceutical composition according to item 23, where the pharmaceutically acceptable carrier is suitable for parenteral administration.

27. The pharmaceutical composition according to item 23, where the pharmaceutically acceptable carrier is suitable for intradermal injection.

28. The pharmaceutical composition according to item 23, where the pharmaceutically acceptable carrier is suitable for subcutaneous injection.

29. The pharmaceutical composition according to item 23, where the pharmaceutically acceptable carrier is suitable for topical administration.

30. The pharmaceutical composition according to item 23, where β-L-2'-deoxynucleoside represented as a single dose.

31. The pharmaceutical composition according to item 30, where the unit dose contains from 10 to 1500 mg β-L-2'-deoxynucleoside.

32. The pharmaceutical composition according to item 30 or 31, where a single dose is a tablet or capsule.

33. The pharmaceutical composition according to any one of p-32, where the owner is the man.

34. Applying an effective amount of β-L-2'-deoxynucleoside formula

as a drug for the treatment of infection of hepatitis b virus by host.

35. Use effectively the sector number β -L-2'-deoxynucleoside formula

as a drug for the treatment of infection of hepatitis b virus by host.

36. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salts as a drug for the treatment of infection of hepatitis b virus by host.

37. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salts as a drug for the treatment of infection of hepatitis b virus by host.

38. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable ether complex as a drug for the treatment of virus infection hepatitis B y the owner.

39. Applying an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable ether complex as a drug for the treatment of infection of hepatitis b virus by host.

40. The use according to any one of p-39, where β-L-2'-deoxynucleoside at least 95% presented the Yong specified in enantiomeric form.

41. The use according to any one of p-39, where β-L-2'-deoxynucleoside is represented with a pharmaceutically acceptable carrier.

42. The application of paragraph 41, where the pharmaceutically acceptable carrier is suitable for oral administration.

43. The application of paragraph 41, where the pharmaceutically acceptable carrier is suitable for intravenous administration.

44. The application of paragraph 41 where the pharmaceutically acceptable carrier is suitable for parenteral administration.

45. The application of paragraph 41, where the pharmaceutically acceptable carrier is suitable for intradermal injection.

46. The application of paragraph 41, where the pharmaceutically acceptable carrier is suitable for subcutaneous injection.

47. The application of paragraph 41, where the pharmaceutically acceptable carrier is suitable for topical administration.

48. The application of paragraph 41, where β-L-2'-deoxynucleoside represented as a single dose.

49. Use p, where the unit dose contains from 10 to 1500 mg β-L-2'-deoxynucleoside.

50. Use p or 49, where a single dose is a tablet or capsule.

51. The use according to any one of p-50, where the owner is the man.

52. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-desoxyn is leased formula

53. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

54. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salt.

55. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salt.

56. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable ether complex.

57. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutical is viable complex ether.

58. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of a combination of the following β-L-2'-deoxynucleosides:

and

or their pharmaceutically acceptable salt, or a complex ester.

59. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salt, or a complex ester, where R is selected from the group consisting of H, -CO-alkyl, CO-aryl, -CO-alkoxyalkyl-WITH-aryloxyalkyl, alkylsulfonyl, arylsulfonyl, Arakishvili, acyl, amino acid residue, mono-, di - or triphosphate or a stabilized phosphate.

60. The method of treatment or prophylaxis of a host infected with hepatitis b virus, comprising an introduction to the host an effective amount of β-L-2'-deoxynucleoside formula

or its pharmaceutically acceptable salt, or a complex ester, where R is selected from the group consisting of H, -CO-alkyl, CO-aryl, -CO-alkoxyalkyl-WITH-aryloxyalkyl, alkylsulfonyl, arylsulfonyl, Arakishvili, acyl, amino acid is these residue, mono-, di - or triphosphate or a stabilized phosphate.

61. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is amino acid residue.

62. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is amino acid residue.

63. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is acyl.

64. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is acyl.

65. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is a mono-, di - or triphosphate or a stabilized phosphate.

66. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutical is Eski acceptable salt, where R is a mono-, di - or triphosphate or a stabilized phosphate.

67. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable ester, where R is selected from the group consisting of H, -CO-alkyl, CO-aryl, -CO-alkoxyalkyl-WITH-aryloxyalkyl, alkylsulfonyl, arylsulfonyl, Arakishvili, acyl, amino acid residue, mono-, di - or triphosphate or a stabilized phosphate.

68. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable ester, where R is selected from the group consisting of H, -CO-alkyl, CO-aryl, -CO-alkoxyalkyl-WITH-aryloxyalkyl, alkylsulfonyl, arylsulfonyl, Arakishvili, acyl, amino acid residue, mono-, di - or triphosphate or a stabilized phosphate.

69. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is selected from the group consisting of H, -CO-alkyl, CO-aryl, -CO-alkoxyalkyl-WITH-aryloxyalkyl, alkylsulfonyl, arylsulfonyl, Arakishvili, acyl, amino acid residue, mono-, di - or trifo the veil, or a stabilized phosphate.

70. The method according to p, where β-L-2'-deoxynucleoside is a compound of the formula

or its pharmaceutically acceptable salt, where R is selected from the group consisting of H, -CO-alkyl, CO-aryl, -CO-alkoxyalkyl-WITH-aryloxyalkyl, alkylsulfonyl, arylsulfonyl, Arakishvili, acyl, amino acid residue, mono-, di - or triphosphate or a stabilized phosphate.

71. The method according to p-70, where the nucleoside is used alternately or in combination with an effective amount of a compound selected from the group consisting of β-L-2-hydroxymethyl-5-(cytosine-1-yl)-1,3-oxathiolane (ZTS), CIS-2-hydroxymethyl-5-(5-fertilizin-1-yl)-1,3-oxathiolane (FTC), β-L-2'-fluoro-5-methyl-arabinofuranosyl-uridine (L-FMAU), β-D-2,6-diaminopyridine (DAPD), famciclovir, penciclovir, 2-amino-1,9-dihydro-9-[4-hydroxy-3-(hydroxymethyl)-2-methylenecycloartanol]-6H-purine-6-she (entecavir, BMS-200475), 9-[2-(phosphono-methoxy)ethyl]adenine (RMEA, adefovir, dipivoxil), lobucavir, ganciclovir and ribavirin.

72. The method according to any of PP-70 β-L-2'-deoxynucleoside at least 95% of the specified enantiomeric form.

73. The method according to any of PP-70 β-L-2'-deoxynucleoside is represented with a pharmaceutically acceptable carrier.

74. The method according to p, where the pharmaceutically acceptable the th carrier is suitable for oral administration.

75. The method according to p, where the pharmaceutically acceptable carrier is suitable for intravenous administration.

76. The method according to p, where the pharmaceutically acceptable carrier is suitable for parenteral administration.

77. The method according to p, where the pharmaceutically acceptable carrier is suitable for intradermal injection.

78. The method according to p, where the pharmaceutically acceptable carrier is suitable for subcutaneous injection.

79. The method according to p, where the pharmaceutically acceptable carrier is suitable for topical administration.

80. The method according to p, where β-L-2'-deoxynucleoside represented as a single dose.

81. The method according to item 80, where the unit dose contains from 10 to 1500 mg β-L-2'-deoxynucleoside.

82. The method according to item 80, or 81, where a single dose is a tablet or capsule.

83. The method according to any of PP-78, where the owner is the man.

All claims priority set from 10.08.1998 according to the application 60/096110; US.



 

Same patents:

FIELD: organic chemistry, chemical technology, medicine.

SUBSTANCE: invention relates to acyclic nucleoside phosphonate derivatives of the formula (1): wherein means a simple or double bond; R1 means hydrogen atom; R2 and R3 mean hydrogen atom or (C1-C7)-alkyl; R7 and R8 mean hydrogen atom or (C1-C4)-alkyl; R4 and R5 mean hydrogen atom or (C1-C4)-alkyl possibly substituted with one or more halogen atoms, or -(CH2)m-OC(=O)-R6 wherein m means a whole number from 1 to 5; R6 means (C1-C7)-alkyl or 3-6-membered heterocycle comprising 1 or 2 heteroatoms taken among the group consisting of nitrogen (N) and oxygen (O) atoms; Y means -O-, -CH(Z)-, =C(Z)-, -N(Z)- wherein Z means hydrogen atom, hydroxy-group or halogen atom, or (C1-C7)-alkyl; Q (see the claim invention); its pharmaceutically acceptable salts or stereoisomers. Also, invention proposes methods for preparing compounds of the formula (1) and their using in treatment of hepatitis B or preparing a medicinal agent designated for this aim.

EFFECT: improved preparing method, valuable medicinal properties of compounds and agent.

16 cl, 10 tbl, 87 ex

FIELD: organic chemistry, medicine, virology.

SUBSTANCE: invention relates to technology of organic compounds, namely, to 5'-aminocarbonylphosphonates d4T that are inhibitors of the human immunodeficiency virus reproduction. Invention describes 5'-aminocarbonylphosphonates d4T of the general formula: wherein R' means hydrogen atom (H), alkyl, aryl; R'' means hydrogen atom (H), alkyl, aryl; R', R'' mean cyclic alkyl; R means alkyl. These compounds are inhibitors of the human immunodeficiency virus reproduction. Invention provides preparing new compounds eliciting valuable biological properties.

EFFECT: valuable medicinal properties of compounds.

2 dwg, 1 tbl, 5 ex

The invention relates to nucleoside analogs of formula (1) in which R1represents H or a group protecting the hydroxyl, R2represents H, a group protecting the hydroxyl group of phosphoric acid, a protected group, phosphoric acid or a group of the formula P(R3R4in which R3and R4are the same or different and represent a hydroxyl group, a protected hydroxyl group, alkoxygroup, allylthiourea, cyanoacetylurea, amino group, substituted alkyl group; And represents alkylenes group containing from 1 to 4 carbon atoms, and a represents a substituted purine-9-ilen group or substituted 2-oxopyrimidine-1-ilen group containing at least one Deputy, selected from hydroxyl groups, protected hydroxyl groups, amino groups, protected amino groups, alkyl groups

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< / BR>
where

< / BR>
or

< / BR>
Cat+is a cation of an alkali metal,

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FIELD: organic chemistry, biochemistry, medicine, virology.

SUBSTANCE: invention relates to derivatives of 2'=amino-2'-deoxynucleosides of the formula:

wherein R means hydrogen atom (H), alkyl, aminoalkyl; R1 means -(R2NR3) wherein R2 and/or R3 means H, -OH, -NH2, alkyl, benzyl under condition that R doesn't represent H or methyl when R2 and R3 mean H. Compounds elicit an antiviral activity with respect to measles and Marburg viruses exceeding that of ribavirin.

EFFECT: valuable properties of compounds.

4 tbl, 2 dwg, 18 ex

The invention relates to a derivative of gemcitabine formula (I), where R1, R2, R3independently selected from hydrogen and C18and C20saturated and monounsaturated acyl groups, provided that R1, R2, R3can't all be hydrogen

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