Antiviral nucleosides

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to compounds of formula (I) where R1 is chosen from ethyl, n-propyl, isopropyl or isobutyl, and to its pharmaceutically acceptable salts. Besides, the invention refers to a pharmaceutical composition on the basis of said compounds used for treating a hepatitis C virus (HCV) mediated disease, and also to a method of treating the hepatitis C virus (HCV) mediated disease, and to the method of selective O-acylation nucleoside II for producing O-acyl nucleoside I in an alkaline reaction medium including the stages: (i) dissolution of II and DMAP in a heterogeneous mixture of water and a solvent and addition of a water base for pH control between approximately 7.5 to approximately 12; (ii) optional addition of a sufficient amount of saturated aqueous NaCl for preparing a diphase reaction mixture; (iii) addition of an acidating agent and an accessory base sufficient for pH preservation between approximately 7.5 to approximately 12; (iv) reaction monitoring and interruption of adding said acidating agent and said base after sufficient conversion provided; (v) optional contact of O-acylnucleoside with the pharmaceutically acceptable acid to produce a pharmaceutically acceptable salt.

EFFECT: production of the pharmaceutical composition for treating the hepatitis C virus (HCV) mediated disease.

9 cl, 2 tbl, 8 ex

 

The present invention relates to the acylated nucleosides, which are prodrugs precursor inhibitor RNA-dependent viral RNA polymerase of hepatitis C virus (HCV). With the introduction of these compounds oral route they are easily absorbed from the gastrointestinal tract and effectively turn into blood in the original form of the nucleoside. These drugs are inhibitors of replication of RNA-dependent viral RNA and used as inhibitors of HCV NS5B polymerase, as inhibitors of HCV replication and for the treatment of hepatitis C in mammals.

The invention relates to the nucleoside prodrugs, which are inhibitors of HCV replication. In particular, the invention relates to the use of acylated derivatives of pyrimidine nucleosides, which provide improved absorbate drugs in oral introduction of the nucleoside.

Hepatitis C virus worldwide is a significant health problem and is a major cause of chronic liver disease (Voeg N. et al., J. Hepatol. 2000, 32:98-112). For patients infected with HCV, there is a risk of development of cirrhosis and then hepatocellular carcinoma, therefore, HCV is a major factor for liver transplantation.

According to the world Health Organization worldwide have been infected with 200 mill the million people every year becomes infected, at least 3-4 million people. About 20% of infected individuals overcome infection and free from virus, and the rest can remain carriers of the virus HCV rest of your life. In 10-20% of infected individuals eventually develop cirrhosis or cancer that destroys the liver. The infection occurs when parenteral use of infected blood or its components (products), infected mothers or mothers-media to their offspring. Modern treatment of HCV infections, which boils down to immunotherapy with recombinant interferon-α alone or in combination with nucleoside analogue ribavirin, brings limited therapeutic benefit as quickly develops immunity. Thus, there is an urgent need for improved therapeutic tools that effectively overcame wouldn chronic HCV disease.

According to the classification of HCV is a member of the virus family Flaviviridae, which includes childbirth flaviviruses, pestiviruses and hepaciviruses, which include hepatitis C (Rice RAU/02.10.2006 C.M.Flaviviridae: The viruses and their replication, b: Fields Virology, Editors: Fields, B.N., Knipe D.M., Howley P.M., Lippincott-Raven Publishers, Philadelphia, Pa., Chapter 30, 931-959, 1996). HCV is an enveloped virus that contains a gene from a positively oriented single-stranded RNA of approximately 9.4 KB. The viral genome consists of a 5'-untranslated re the ion (UTR), the longest open reading frame (ORF)that encodes a precursor of polyprotein approximately 3011 amino acids, and a short 3' UTR. 5' UTR is the most highly conserved part of the genome of HCV and important for the initiation and control of broadcast polyprotein.

Genetic analysis of HCV was identified six major genotypes, showing>30% difference in DNA sequences. Each genotype consists of a series of several closely related subtypes, which show a 20-25% difference in nucleotide sequences (Simmonds P., 2004, J. Gen. Vilor., 85:3173-88). More than 80 subtypes already described. In the United States, approximately 70% of infected infected with type 1A and 1b. Type 1b is the most common subtype in Asia. (X. Forns, J. Bukh, Clinics in Liver Disease, 1999, 3:6930716; J. Bukh et al., Semin. Liv. Dis. 1995, 15:41-63). Unfortunately, the infection Type 1 is more resistant to therapy than genotypes of Type 2 or 3 (N.N.Zein, Clin. Environ. Rev., 2000, 13:223-235).

Genetic organization and polyprotein processing part ORF related to non-structural protein, pestiviruses and hepatovirus very similar. These viruses with positive-guided RNA chains have one extensive ORF, encoding all viral proteins necessary for viral replication. These proteins are expressed as polyprotein that Ko and excision is created as a cell and encoded by the virus proteases with obtaining the result of Mature viral proteins. These viral proteins, which are responsible for the replication of viral genomic RNA is localized closer to the end. Two-thirds of the ORF are called non-structural (NS) proteins. And pestiviruses and hepatovirus in Mature non-structural (NS) proteins at the site of aminocore region encoding non-structural proteins, to carboxylic ORF, consistently provides P7, NS2, NS3, NS4A, NS4B, NS5A and NS5B.

NS proteins pestiviruses and hepatovirus distributed in the sequence of domains, which are characteristic of certain functional proteins. For example, the NS3 proteins of viruses of both groups include primary amino acid sequences (motifs)that are typical of serine proteases and helices (Gorbalenya et al. Nature 1988, 333:22, Bazan and Fletterick, Virology, 1989, 171:637-639; Gorbalenya et al., Nucleic Acid Res. 1989, 17. 3889-3897). Similarly NS5B proteins pestiviruses and hepatovirus bear sequence, characteristic of RNA-directed RNA polymerase (Koonin E.V., V.V. Dolja, Grit. Rev. Biochem. Molec. Biol. 1993, 28:375-430).

The actual purpose and function of NS proteins pestiviruses and hepatovirus in the life cycle of viruses, in General, similar. In both cases, the NS3 serine proteinase responsible for the proteolytic processing of precursors of polyprotein below its position in the ORF (Wiskerchen, Collet, Virology, 1991, 184:341-350; Bartenschlager et al. J. Virol. 1993 67:3835-3844; Eckart et al. Biochem. Biophys. Res. Comm. 1993 192:399-406; Grakoui et al. J.Virol. 1993 67:2832-2843; Grakoui et al. Proc. Natl.Acad. Sci. USA 1993 90:10583-10587; Ilijikata et al. J.Virol. 1993 67:4665-4675; Tome et al. J. Virol. 1993 67:4017-4026). NS4A protein in both cases, acts as a cofactor NS3 serine proteinase (Bartenschlager et al. J. Virol. 1994 68:5045-5055; Failla et al. J. Virol. 1994 68:3753-3760; Xu et al. J. Virol. 1997 71:53 12-5322). The NS3 protein of both types of viruses also functions as a helicase (Kim et al. Biochem. Biophys. Res. Comm. 1995 215:160-166; Jin, Peterson Arch. Biochem. Biophys. 1995, 323:47-53; Warrener, Collett J. Virol. 1995 69:1720-1726). Finally, NS5B proteins pestiviruses and hepatovirus have predicted the activity of RNA-directed RNA polymerase (Behrens et al. EMBO 1996 15:12-22; Lechmann et al. J. Virol. 1997 71:8416-8428; Yuan et al. Biochem. Biophys. Res. Comm. 1997 232:231-235; Hagedom, PCTWO 97/12033; Zhong et al. J. Virol. 1998 72:9365-9369).

Currently there are a limited number of approved therapeutic methods, which generally would be considered suitable for treatment of HCV infection. New and existing therapeutic approaches for the treatment of HCV and inhibition of HCV NS5B polymerase are described in the following works: R.G.Gish, Sem. Liver. Dis., 1999 19:5; Di Besceglie, A.M. And Bacon, B. R., Scientific American, October: 1999 80-85; G. Lake-Bakaarm Current and Future Therapy for Chronic Hepatitis With Virus Liver Disease, Curr. Drug Targ Infect Dis. 2003 3(3):247-253; P. Hoffmann et al. Recent patents on experimental therapy for hepatitis With vims infection (1999-2002), Exp.Opin. Ther. Patents 2003 13(11):1707-1723; F. Poordad et al. Developments in Hepatitis With drug therapy during 2000-2002, Exp.Opin. Emerging Drugs 2003 8(1):9-25; M.P. Walker et al.. Promising Candidates for the treatment of chronic hepatitis C, Exp.Opin. Investig. Drugs 2003 12(8): 1269-1280; S. - L. Tan et al. Hepatitis With Therapeutics: Current Status and Emerging Strategies, Natire Rev. Dru Discov. 2002 1:867-881; R. De Francesco et al. Approaching a new era for hepatitis With virus therapy: inhibitors of the NS3-4A serine protease and the NS5B RNA-dependent RNA polymerase. Antiviral Res. 2003 58:1-16; Q.M. Wang et al. Hepatitis With virus encoded proteins: targets for antiviral therapy. Drugs of the Future 2000 25(9):933-8-944; J. A. Wu and Z. Hong, Targeting NS5B-Dependent RNA Polymerase forAnti-HCV Chemotherapy Cur. Dmg Targ.-Inf. Dis. 2003 3:207-219.

In these works mentioned connection immediately as they are created. Combined (Raman) therapy on the basis of two or three products aimed at the same or different purpose, has become a standard therapy to suppress or slow the spread of resistant strains of the virus, and compounds disclosed in the above works, can be used in combination therapy with compounds of the present invention.

Ribavirin (1A; amide 1-((2R, 3R, 4S, 5R)-3,4-Dihydroxy-5-hydroxymethyl-tetrahydro-furan-2-yl)-1H-[1,2,4]triazole-3-carboxylic acid; VIRAZOLE,®) is a synthetic pendulous interferon analog of the nucleoside with antiviral activity of broad-spectrum. Ribavirin has in vitro activity against several DNA and RNA viruses, including Flaviviridae (Gary L. Davis, Gastroenterology 2000 118:S104-S114). In monotherapy ribavirin reduces the levels of serum amino-transferase to normal in 40% of patients, but it does not reduces serum levels of HCV-RNA. Ribavirin also shows significant toxicity, and it is known that he calls the characteristic anemia. Ribavirin is not approved for use against HCV as monotherapy agent, but the use of this compound approved for combination therapy with interferon α-2A and interferon α-2b. Viramidine 1b is a prodrug that is transformed into 1A in hepatocytes.

Interferons (IFNs) are already used for the treatment of chronic hepatitis b for about ten years. IFNs are glycoproteins produced by immune cells in response to viral infection. Recognize two different types of interferons: Type 1 includes several interferons alpha and one interferon β, Type 2 includes interferon γ. Interferon Type 1 is produced mainly by infected cells and protects neighboring cells from de novo infection. IFNs inhibit the replication of many viruses, including HCV, and when used as monotherapeutic means for treatment of hepatitis C With IFN suppresses serum HCV-RNA before redetection levels.

In addition, IFN normalizes the levels of aminotransferases in serum. Unfortunately, the effect of IFN temporary. Termination (break) treatment leads to 70% of cases of relapses and only in 10-15% of cases there is a stable long-term virological response with normal levels of serum albinterferon (L. - B. Davis, see above).

A certain limitation of early IFN therapy is rapid elimination (to irens) protein from the blood. Chemical derivatization IFN using polyethylene glycol (PEG) resulted in significantly improved pharmacokinetic properties of proteins. PEGASIS® is a conjugate of interferon α-2b and 12 kDa mono-methoxy PEG. (VA Luxon et al., Clin. Therap.2002 24(9): 13631383; A. Kozlowski and J.M.Harris, J. Control. Release 2001, 72:217-224).

Interferon α-2A and interferon α-2b has recently been approved as a means for monotherapy in the treatment of HCV, ROFERON-A® is a recombinant form of interferon α-2A. PEGASIS® (Roche) is paglierani (i.e. modified polyethylene glycol) form of interferon α-2A. INTRON-A® (Sobering Corporation) is a recombinant form of interferon α-2b, a PEG-INTRON® (Schering Corporation) is paglierani form of interferon α-2b.

Other forms of interferon-α, as well as interferon β, γ, τ and ω, are currently undergoing clinical trials for treatment of HCV. For example, INFERGEN® (interferon of alfacon-1) from biterMune, OMNIFERON® (natural interferon) from Viragen, ALBUFERON®, Human Genome Sciences, REBIF® (interferon (3-1) from Ares-Serono, Omega Interferon from BioMedicine, Oral Interferon Alpha from Amarillo Bosciences, and interferon γ-1b from InterMune tested,

Combined therapy of HCV ribavirin and interferon-α at the present time has shown itself to be the best therapy. The combination of ribavirin and PEG-IFN (see below) leads to prolonged reaction to the virus in 54-56% of patients. SVR reaches 80% for Type 2 and 3 HCV. (Walker, see above). It is Oleniy, this combination gives also side effects that cause clinical problems. With subcutaneous injection of IFN-α associated symptoms such as depression, flu-like symptoms and skin reactions and hemolytic anemia associated with ribavirin, administered continuously.

Other macromolecular compounds, which are ongoing preclinical and clinical trials, in terms of treatment of viral hepatitis C are: Interleukin-10 by Schering-Plough, IPSO 1 from bitermenron, Merimebodlib (VX-497) from the Vertex, HEPTAZYME® by RPI, IDN-6556 from Idun Pharma, XTL-002 from XTL, HCV/MFS9 from Chiron, CIVACIR® (immune globulin for hepatitis C) from NABI, ZADAXIN® (thymosin α-1) from SciClone, thymosin plus pegylated interferon from SciClone, CEPLENE®; therapeutic vaccine Epimmune/Genencor, a therapeutic vaccine from Merix, therapeutic vaccine, ChronVacC, Tripep.

Other makromolekulare methods include ribozymes, targeted to HCV RNA. Ribozymes are small molecules of natural origin with their endonuclease activity which catalyzes sequence-specific cleavage of RNA. An alternative approach is the use of antisense oligonucleotides, RNA binding and stimulating mediated RNA asain splitting.

Currently already identified a number of potential molecular targets for the development and testing them as anti-HCV therapeutic agents, such as the er, NS2-NS3 autoprotease, N3 protease, N3 helicase and NS5B polymerase. RNA-dependent RNA polymerase is absolutely necessary for replication of single-stranded sense RNA genome, and therefore this enzyme is of considerable interest to the medical and palazetto.

Nucleoside inhibitors of NS5B polymerase can act either as a non-natural substrate, which results in the termination circuit, or as a competitive inhibitor competes with the binding of the nucleotide to the polymerase. In order to function as a chain terminator, nucleoside analogue must be absorbed by the cell and converted in vivo triphosphate to compete for customers nucleotide binding to the polymerase. This conversion to the triphosphate is commonly by cellular kinases, which leads to additional structural requirements for the potential of the nucleoside polymerase inhibitor. Unfortunately, this limits the direct assessment of nucleosides as inhibitors of HCV replication in studies conducted on cells amenable to in situ phosphorylation.

B=adenine, thymidine, uracil, citizen, guanine and gipoksantin.

In WO 0190121, published on November 29, 2001, J.-P.Sommadossi and .Lacolla described and gave examples of activity and the t-HCV polymerase in part 1'-alkyl - and 2'-alkynylamino formulas 2 and 3. In WO 01/92282 published on 6 December 2001, J.-P.Sommadossi and .Lacolla disclosed and described examples of treating Flaviviruses and Pestiviruses 1'-alkyl - and 2'-alkynylamino formulas 2 and 3. In WO 03/026675, opublikovannoi April 3, 2003, G. Gosselin has described the use of 4'-alkynylamino formula 4 for treating Flaviviruses and Pestiviruses.

In WO 2004003000, published January 8, 2004, J.-P.Sommadossi et al. described 2'- and 3'-prodrugs based on 1'-, 2'-, 3'- and 4'-substituted β-D and β-L-nucleosides. In WO 2004/002422, published January 8, 2004, described 2'-C-methyl-3'-O-malinowy ether ribofuranosylthiazole for the treatment of Flaviviridae infection. Idenix reported clinical trials related compounds NM283, which I guess is the valine ester 5 and analogue 2 cytidine (B=cytosine). In WO 2004/002999, published January 8, 2004, J.-P.Sommadossi et al. he described a series of 2' or 3' prodrugs of 1', 2', 3' or 4'-branched nucleosides for the treatment of infections Flaviviridae, including HCV infections.

In WO 2004/046331, published June 3, 2004, J.-P.Sommadossi et al. described 2'-branched nucleosides and mutation Flaviviridae. In WO 03/026589 published 3 April 2003, G.Gosselin et al. described methods of treatment of viral hepatitis C using 4'-modified nucleosides. In WO 2005009418, published February 3, 2005, R.Storer et al. described purine nucleoside analogues for the treatment of diseases caused by, including Flaviviridae.

Other patent applications rusk is to see the use of certain nucleoside analogues for the treatment of viral hepatitis C. In WO 01/32153, published may 10, 2001, R.Storer described derivatives of nucleosides for treatment of viral diseases. In WO 01/60315, published on 23 August 2001, .Ismaili et al. described methods of treatment or prevention of infections caused by Flaviviruses, using derivatives of nucleosides. In WO 02/18404, published March 7, 2002, R.Devos et al. described 4'-substituted nucleosides for treatment of viral HCV. In WO 01/79246, published on 25 October 2001, .A.Watanabe described derivatives of 2' or 3'-hydroxymethyl-nucleosides for treatment of viral diseases. In WO 02/32920, published April 25, 2002, and in WO 02/48165, published on 20 June 2002, L.Stuyver et al. described derivatives of nucleosides for treatment of viral diseases.

In WO 03/105770, published on 24 December 2003, C. Bhar et al. described a number of carbocyclic nucleoside derivatives which inhibit RNA-dependent RNA polymerase of the virus. Nucleosides disclosed in this publication are mainly 2'-methyl-2'-hydroxy-substituted nucleosides. In WO 2002/057425, published on 25 July 2002, S.S.Carroll et al. described derivatives of nucleosides that inhibit RNA-dependent viral polymerases, and methods of treatment of HCV infections.

In WO 02/057287, published on 25 July 2002, S.S.Carroll et al. described related 2α-methyl and 2β-methylribose derivatives, in which the base of neobythites is but substituted 7H-pyrrolo[2,3-(1]pyrimidine radical 6. In the same application shows one example 3β-methyl-nuke. S.S.Carroll et al. (J.Biol.Chem. 2003 278(14):11979-11984), described the inhibition of HCV polymerase 2'-O-methylcytidine (6A). In WO 2004/009020, published on January 29, 2004, D..Olsen et al. described a number dinucleotide derivatives as inhibitors of RNA-dependent RNA polymerase of the virus.

PCT publication number WO 99/43691, in the name of Emory University, entitled "2'-Formulised", describes the use of certain 2'-pornological for treatment of HCV. US patent No. 6348587 issued by Emory University, entitled "2'-Formulised, shows the number of families 2'-pornological, useful in the treatment of hepatitis b, HCV, HIV and abnormal cell proliferation. Both configurations 2'-forsometimes disclosed.

Eldrup et al. (Oral Session V, Hepatitis With Virus, Flaviviridae; 16thInternational Conference on Antiviral Research (Apr. 27, 2003, Savannah, Ga.)) describe the relationship between structure and activity of 2'-modified nucleosides in terms of inhibition of HCV.

Br et al. (Oral Session V, Hepatitis With Virus, Flaviviridae; 16thInternational Conference on Antiviral Research (Apr. 27, 2003, Savannah, Ga.) p A75) described the synthesis and pharmacokinetic properties of nucleoside analogues as potential inhibitors of replication of HCV RNA. The authors reported that 2'-modified nucleosides show high inhibitory activity studies on cell replicons.

Olsen et al. (Oral Session V, Hepatitis C Virus, Flaviviridae; 16thInterntional Conference on Antiviral Research (Apr. 27, 2003, Savannah, Ga.) p A76) also showed the effect of 2'-modified nucleosides on the replication of HCV RNA.

Non-nucleoside allosteric inhibitors of HIV reverse transcriptase have proven to be an efficient therapeutic effect alone or in combination with nucleoside inhibitors and protease inhibitors. Several classes of non-nucleoside HCV NS5B inhibitors have already been described and are currently at the stage of testing, including benzimidazole (.Hashimoto et al. WO 01/47833, .Hashimoto et al. WO 03/000254, P.L.Beaulien et al. WO 03/020240:A2; P.L.Beaulien et al. US 6448281 Bl P.L.Beaulien et al. WO 03/007945 A1); indoles (P.L.Beaulien et al. WO 03/0010141 A2); benzothiadiazine, for example 7 (D.Dhanak et al. WO 01/85172 A1; D. Dhanak et al. WO 03/037262 A2; .J.Duffy et al. WO 03/099801 A1; D.Chai et al. WO 2004052312, D.Chai et al. WO 2004052313, D.Chai et al. WO 02/098424, J.K.Pratt et al. WO 2004/041818 A1; J.K.Pratt et al. WO 2004/087577 A1), tifany, for example 8 (..Chan et al. WO 02/100851)

Benzothiophene (D..Young and T.R.Bailey WO 00/18231); β-ketopropane (S.Attamura et al. US 6492423 B1, A.Attamura et al. WO 00/06529); pyrimidines (.Gardelli et al. WO 02/06246 A1); pyrimidinedione (T.R.Bailey and D.C.Young WO 00/13708); triazine (K.-H. Chung et al. WO 02/079187 A1); derivatives of rhodanine (T.R.Bailey and D.C.Young WO 00/10573, J.C.Jean et al. WO 01/77091 A2); 2,4-dioxopregna (R.A.Love et al. EP 256628 A2); derivatives of phenylalanine (M.Wang et al. J.Biol. Chem. 2003 278:2489-2495).

To develop a new strategy for anti-HCV therapy was purposefully selected NS3 protease. In WO 98/22496 published 28 is flanged 1998, M.R.Attwood et al. described mechanism based on inhibition of the active sites of proteases (M.R.Attwood et al. Antiviral Chemistry and Chemotherapy 1999 10:259-273; M.R.Attwood et al. Preparation and use of amino acid derivatives as anti-viral agents, German Patent Pub. DE 19914474). In WO 98/17679, published on 30 April 1998, R.D.Tung et al. revealed the mechanism of action of peptide inhibitors for NS3 protease.

In WO 99/07734, published on 18 February 1999, and in WO 00/09543, published August 9, 1999, M. Llinas-Brunet et al. described peptide protease inhibitors. In WO 00/59929, published on 12 October 2000, Y.S.Tsantrizos et al. described macrocyclic tripeptides, which are powerful inhibitors of HCV NS3 protease. A series of related patents Boehringer-Ingleheim reveals similar between protease inhibitors and has already led to the identification of tripeptides derived BILN 2061 (M. Llinas-Brunet et al. Bioorg. Med. Chem. Lett. 2000 10(20):2267-70; J.Med.Chem. 2004 47(26): 6584-94; J.Med.Chem. 2004 47(7): 1605-1608; Angew. Chem. Int. Ed. Eng. 2003 42(12): 1356-60).

Other Tripeptide inhibitors discovered by Bristol-Myers Squibb, have been described, inter alia, in WO 03/099274, published on 4 December 2003, in WO 2004/032827, published on 22 April 2004, in WO 03/053349, published July 3, 2003, in WO 2005/046712, published on 26 may 2005, and in WO 2005/051410, published on 9 June 2005 In WO 2004/072234, published on 26 August 2004 and in WO 2004/093798, published on 4 November 2004, Enanta Pharmaceuticals were described and other Tripeptide protease inhibitors. In WO 2005/037214 published 28 Aprile,, L.M.Blatt et al. describe other Tripeptide derivatives inhibiting HCV NS3 protease In WO 2005/030796, published 7 April 2005, S.Venkatraman et al. described macrocyclic inhibitors of the NS3 serine protease of HCV. In WO 2005/058821, published June 30, 2005, F.Velazquer et al. described inhibitors of HCV NS3/NS4a serine protease. In WO 02/48172, published on 20 June 2007, Z. Zhu described varilite as inhibitors of the NS3 protease. In WO 02/08187 and in WO 02/08256, published on 31 January 2002, A.Saksena et al. described peptide inhibitors of HCV NS3 protease. In WO 02/08251, published on 31 January 2002, M.Lim-Wilby et al. described peptide inhibitors of the NS3 protease. In the US 6004933, published on 21 December 1999, L.W.Spruce et al. described heterocyclic derivatives of peptides, which inhibit cysteine proteases, including HCV endopeptidase.

We also study and substratespecific inhibitors of the NS3 protease, such as derivatives of 2,4,6-trihydroxy-3-nitrobenzamide (Sudo K. et al., BBRC 1997 238:643-647; Sudo K. et al. Antiviral Chemistry and Chemotherapy 1998 9:186), including RD3-4082 and RD3-4078, the former of the above-mentioned substituted by amide 14C-chain, and the latter is subjected to the steam treatment-phenoxyphenyl group.

SCH 68631, phenanthridine, an inhibitor of HCV protease (Chu M. et al., Tetrahedron Lett. 1996 37:7229-7232). In another example, the same authors SCH 351633, isolated from the fungus Penicillium griseofulvum, was described as a protease inhibitor (Chu M. et al. Bioorg. Med. Chem. Lett. 1999 9:1949-1952) Nanomolecular effectiveness against HCV NS3 proteases enzyme has already been achieved by the design of selective inhibitors based on the macromolecule eglin .Eglin with, isolated from leeches, is a strong inhibitor of several serine proteases such as protease a and b from S. griseus, a-chymotrypsin, chymase and subtilisin (Qasim M. A. et al., Biochemistry 1997 36:1598-1607).

Thiazolidine derivatives which show relevant inhibition in a study HPLC with reversed phase in relation to the fused protein NS3/4A and the substrate NS5A/5B (Sudo K. et al. Antiviral Research, 1996 32:9-18), especially compound RD-1-6250 carrying merged cinnamonny residue, substituted long-chain-alkyl, represent RD4 6205 and RD4 6193. Thiazolidine and benzanilide identified N. Kakiuchi et al. in FEBS Let. 1998 421:217 to 220 and N. Takeshita et al. Anal. Biochem. 1997 247:242-246.

Imidazolidinone as inhibitors of the NS3 serine protease of HCV are disclosed in WO 02/08198 from Sobering Corporation, published on 31 January 2002, and in WO 02/48157 from Bristol Myers Squibb, published 20 June 2002. In WO 02/48116, published June 20, 2002, P. Glunz et al. described pyrimidinone inhibitors of the NS3 protease.

Other enzyme targets for anti-GCS-therapies include HCVIRES site (Internal Entry Site) and the HCV helicase. About the IRES inhibitors were messages from Immusol, Rigel Pharmaceuticals (R803) and Anadys (ANA 245 and ANA 246). Vertex described inhibitor of helicase HCV.

Concomitant therapy that can be suppressed resistant mutant strains, became obsidionalis approach in antiviral chemotherapy. Described here nucleoside inhibitors can be combined with other nucleotide sequence that is sidname inhibitors of HCV polymerase, nucleoside inhibitors of HCV polymerase inhibitors and HCV protease. As it emerged and developed other classes of drug against HCV, for example, inhibitors of virus entry inhibitors of helicase, IRES inhibitors, ribozymes and antisense oligonucleotides, they can also be excellent candidates for use in combination therapy. Derivatives of interferon has already been successfully combined with ribavirin and interferon or chemically modified interferon will be useful in combination with the described here nucleosides.

Derivatives of nucleosides are often effective anti-virus (such as HIV, HCV, Herpes simplex, CMV) and anti-cancer chemotherapeutic agents. Unfortunately, their practical use is often limited by two factors. First, poor pharmacokinetic properties often limit the absorption of nucleoside from the digestive tract and the intracellular concentration of nucleoside derivatives, and second, suboptimal physical properties limit the choice of the drug compound, which could be used to increase the level of separation of the active ingredient.

Albert introduced the term "prodrug"to describe the connection, which in itself has no biological activity, but capable of metabolically transformirovalsya to become active drug substance (A.Albert, Selective Toxicity, Chapman and Hall, London, 1951). Lately there are many reviews about the prodrugs (.Ettmayer et al. J.Med.Chem. 2004 47(10):2393-2404; .Beaumont et al. Curr. Dmg Metab. 2003 4:461-485; H.Bundgaard, Design of Prodrugs: Bioreversible derivatives for various functional groups and chemical entities in Design of Prodrugs, H.Bundgaard (ed) Elsiver Science Publishers, Amsterdam 1985; G.M.Paulette et al. Adv. Drug Deliv. Rev. 1997 27:235-256; R.J.Jones and N.Bischofberger, Antiviral Res. 1995 27:1-15, and .R.Wagner et al., Med. Res. Rev. 2000 20:417-45).

Despite the fact that metabolic transformation can kataliziruetsa special enzymes, often by hydrolases, the active substance may also be regenerated in the nonspecific chemical processes.

To pharmaceutically acceptable prodrugs refer to a compound that is metabolized, for example hydrolyzed or oxidized, in the host with the formation of compounds of the present invention. If bioconversion should not be education toxicologically burdened fragments. Typical examples of prodrugs include compounds that have biological labile protective groups associated with the functional balance of active compounds. Alkylation, acylation and other lipophilic modification of the hydroxyl group in the sugar residue are used when creating pronucleotides. These pronucleotides can hydrolyze or deaccelerate in vivo to obtain active compounds.

Factors ogranichivaya and oral bioavailability, often absorption from the gastrointestinal tract and secretion during the first passage of the intestinal wall and liver. Optimization transcellular absorption when passing through the digestive tract requires D7,4) is greater than zero. Optimization of the distribution coefficient, however, does not ensure success. Maybe the prodrug should be avoided in the enterocytes of the active factors of migration during infiltration. Intracellular metabolism in enterocytes can lead to passive transport or active transport of the metabolite pumps drip back into the intestinal cavity. The prodrug should also be resistant to unwanted biotransformation in the blood before you reach the target cells or receptor.

Although the alleged prodrugs can sometimes be created by using rational thinking based on the chemical functionality of a given molecule, chemical modification of the active component leads to a completely different, new molecular structure, which can exhibit undesirable physical, chemical, and biological properties that are missing in connection predecessor. The requirements for identification of metabolites can be a challenge, if numerous metabolic pathways of lead to many different metabolites. So identifying acacia prodrugs remains a controversial and generate problems of occupation. Moreover, evaluation of pharmacological properties of potential prodrugs is difficult and expensive. The pharmacokinetic results obtained in animal models may differ, if you extracted them to humans.

The object of the present invention is to provide new compounds, methods and compositions for the treatment of the subject host infected with hepatitis C.

The present invention relates to new dialling derivatives of 4-amino-1-((2R, 3R, 4R, 5R)-3-fluoro-4-hydroxy-5-hydroxymethyl-3-methyl-tetrahydro-furan-2-yl)-1H-pyrimidine-2-it (also known as (2R)2'-deoxy-2-methyl-2-fluoro-citizen).

The compounds of this invention have a structure according to formula I.

where R1selected from the group consisting of C2-5unbranched or branched amine, C2-5unbranched or branched alkenyl, C2-5unbranched or branched quinil, C2-5lower haloalkyl,3-6cycloalkyl and C2-4alkoxy; or represents its corresponding hydrate, solvate or salt the addition of acids.

Compounds of the present invention are useful in the treatment of disorders caused by HCV.

The invention also includes methods of treatment of HCV described in this invention compounds is s and pharmaceutical compositions including these compounds.

In one embodiment, the present invention features a compound according to formula I, where R1has the above values.

In another embodiment, the present invention features a compound according to formula I, where R1represents ethyl, n-propyl, ISO-propyl, n-butyl or isobutyl.

In another embodiment, the present invention features a compound according to formula I, where R1represents ethyl or isopropyl.

In another embodiment, the present invention features a compound according to formula I, where R1represents isopropyl, and the connection is cleaners containing hydrochloride or sulfate salt.

In another embodiment, the present invention features a compound according to formula I, where R1represents isopropyl, and the connection is cleaners containing hydrochloride salt.

In another embodiment, the present invention features a compound according to formula I, where R1represents ethoxy, n-propoxy or isopropoxy.

In another embodiment, the present invention features a compound according to formula I for use in therapy, particularly for use in the treatment of diseases caused by HCV virus.

Another variant of the invention concerns the use of compounds of formula I for the manufacture of Lekarstvo the th means for treatment of the disease, caused by HCV virus.

The compound of the formula I may, in particular, be used to prepare drugs for injection to the needy in this patient in therapeutically effective area, preferably in a dose of from 0.1 to 10 g per day, more preferably in a dose of 0.5 to 7 g / day and most preferably in a dose of from 1.0 g to 6.0 g / day.

The compound of the formula I can also be used for the preparation of a medicinal product, which may optionally include a therapeutically effective amount of at least one immune system modulator, for example, interferon, chemically derivatizing interferon, interleukin, tumor necrosis factor or colony stimulating factor and/or at least one antiviral agent that inhibits replication of HCV, such as HCV protease inhibitor, another nucleoside HCV polymerase, non-nucleoside HCV polymerase, an inhibitor of helicase HCV, an inhibitor of primacy HCV or merged inhibitor of HCV.

Another variant of the present invention provides a method of treating diseases mediated by HCV virus comprising the administration to a patient in need, a therapeutically effective dose of a compound according to formula I, which is described above.

Another variant of the present invention provide the supports method for the treatment of disease, mediated by the HCV virus comprising the administration to a patient in need, a dose of from 0.1 g to 10 g per day of the compounds according to formula I, which is described above.

In another embodiment of the invention, the dose is 0.5-7 g / day, and in another embodiment, the dose is 1.0 to 6.0 grams per day.

In another embodiment, the present invention provides a method of treating diseases mediated by HCV virus comprising co-administration to a patient in need, a therapeutically effective dose of a compound according to formula I, which is described above and a therapeutically effective amount of at least one immune system modulator and/or at least one antiviral agent that inhibits replication of HCV.

In another embodiment, the present invention provides a method of treating diseases mediated by HCV virus comprising co-administration to a patient in need, a therapeutically effective dose of a compound according to formula I, which is described above and a therapeutically effective amount of at least one modulator of the immune system, and the immune system modulator is an interferon, interleukin, tumor necrosis factor or colony stimulating factor.

Another variant of the present invention pre is provides a method of treating diseases, mediated by the HCV virus comprising co-administration to a patient in need, a therapeutically effective dose of a compound according to formula I, which is described above and a therapeutically effective amount of at least one modulator of the immune system, and the immune system modulator is an interferon or chemically derivationally interferon.

Another variant of the present invention provides a method of treating diseases mediated by HCV virus comprising co-administration to a patient in need, a therapeutically effective dose of a compound according to formula I, which is described above and a therapeutically effective amount of at least one other antiviral compounds.

Another variant of the present invention provides a method of treating diseases mediated by HCV virus comprising co-administration to a patient in need, a therapeutically effective dose of a compound according to formula I, which is described above and a therapeutically effective amount of at least one other antiviral compound that is an inhibitor of HCV protease, other nucleoside inhibitor of the HCV polymerase, non-nucleoside inhibitor of the HCV polymerase, an inhibitor of helicase HCV, inhib the torus of primacy HCV or merged inhibitor of HCV.

Another variant of the present invention provides a pharmaceutical composition comprising a compound according to formula I, which is described above, is mixed with at least one pharmaceutically acceptable carrier, diluent or excipient.

Another variant of the present invention provides a method of obtaining a compound according to formula I, which is described above, and the method includes a stage (i)to(v)listed in paragraph 15 of the claims and illustrated in the examples. The method includes processing I in basic aqueous organic medium, which may be homogeneous or two-phase with allermuir agent, as defined here, in the presence of DMAP and a sufficient amount of base to maintain the pH of the solution at pH of at least about 7.5.

This method allows to carry out the acylation without concomitant reaction of the heterocyclic base.

The expression "a" or "an" value, in the sense used here, means one or more of such units, for example, the connection is related to one or more compounds or at least one connection. So the terms "a" (or "an"), "one or more" and "at least one" can be used here vzaimozatmeniya way.

The terms "optional" or "optionally" in the second sense, in which they are used here, means that the subsequently described event or circumstance may occur, but not necessarily, because the description includes instances when the event or circumstance occurs and instances when it doesn't. For example, "optional link means that the link may be present and may not be, and description then includes single, double, or triple bond.

The term "independently" is used here to show that the variable (value) is used in any example without regard to the presence or absence of a variable with the same or a different definition within the same connection. So, in the connection in which R appears twice and is defined as independently carbon or nitrogen, both R can be carbon, both can be ashtami, or one R can be carbon, and the other nitrogen.

The term "alkenyl" used here is meant unsubstituted uglevodorodnyi chain radical of 2-10 carbon atoms, having one or two olefinic double bonds [preferably one olefinic double bond], "C2-10alkenyl" when used here means alkenyl consisting of 2-10 carbon atoms. Examples are vinyl; 1-propenyl, 2-propenyl (allyl) or 2-butenyl (crotyl).

The term "alkyl" means here nerasvetlena the Yu or branched chain saturated monovalent hydrocarbon residue, consisting of 1-10 carbon atoms. The term "lower alkyl" means straight or branched residue of a hydrocarbon chain, comprising from 1 to 6 carbon atoms. "C2-10alkyl" means here an alkyl comprising from 1 to 10 carbon atoms. Examples of alkyl groups include (but without limitation to) the lower alkyl groups include methyl, ethyl, propyl, i-propyl, n-butyl, i-butyl, t-butyl or pentyl, isopentyl, neopentyl, hexyl, heptyl and octyl. The term (ar) alkyl or (heteroaryl) alkyl indicates that the alkyl group is optionally substituted aryl or heteroaryl respectively.

The term "quinil" here means radical, which represents an unbranched or branched hydrocarbon chain of 2 to 10 carbon atoms, preferably from 2 to 5 carbon atoms, and having at least one triple bond. "C2-10quinil" means here quinil consisting of 2-10 carbon atoms. Examples are ethinyl, 1-PROPYNYL, 2-PROPYNYL, 1-butynyl, 2-butynyl or 3-butynyl.

The term "cycloalkyl" here means a saturated carbocyclic ring containing from 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. "C3-7cycloalkyl" means here cycloalkyl containing 3-7 carbon atoms in the carbocyclic ring.

The term "haloalkyl" means here Nera is extensive or branched alkyl chain, as defined above, in which 1, 2, 3 or more hydrogen atoms replaced by halogen. "C1-3haloalkyl" means here haloalkyl with 1-3 carbon atoms and 1 to 8 halogen substituents. Examples are 1-vermeil, 1-chloromethyl, 1-methyl bromide, 1-iodomethyl, trifluoromethyl, trichloromethyl, tribromoethyl, triacetyl, 1-foretel, 1-chloroethyl, 1-bromacil, 1-codetel, 2-foretel, 2-chloroethyl, 2-bromacil, 2-codetel, 2,2-dichloroethyl, 3-bromopropyl or 2,2,2-triptorelin.

The term "alkoxy", as used here, refers to-O-alkyl group in which alkyl is defined above. Examples are methoxy, ethoxy, n-propyloxy, i propyloxy, n-Butylochka, i bucalossi, t-Butylochka. "Lower alkoxy" means here alkoxygroup with "lower alkyl" group as defined above. "C1-10alkoxy" means-O-alkyl, where alkyl represents a C1-10.

The term "dianiline derived, used here, means derivateservlet nucleoside derivative, as described here, in which the 3'- and 5'-hydroxy refer to air-OC(=O)R1and OC(=O)R2where R1and R2such as defined in paragraph 1.

The term "allerease agent" used herein refers to the anhydride, halide acid, chlorocarbonate (for example, ethylchloride) or activated derivative of N-substituted alpha amino acids. The term"anhydride" in the sense used here refers to compounds of the General structure R 1C(O)-O-C(O)R1where R has the meaning given in paragraph 1. The term "haloalkylthio" refers here to the compounds of the General formula R1C(O)X, where X is halogen. The term "allmydata" refers here to the compound of General formula R1C(O)X, where X is an N-imidazolium. The term "activated derivative" connection in the sense used here refers to the transient reactive form of the original compound that makes the connection active in the desired chemical reaction, in which the original connection is only moderately reactive or even preaction-able. Activation is achieved by the formation of a derivative or chemical groups in the molecule with a higher content of free energy than the parent compound, which gives an activated form greater susceptibility to reaction with another reagent. In the context of the present invention, the activation carboxypropyl extremely important and relevant activating agents or groups that activate carboxypropyl are described in detail below.

Of particular interest from the point of view of the present invention cause anhydrides, carboxylic acid chlorides and carboxylic acids.

The phrase "heterogeneous aqueous solvent mixture" means here a mixture of water and organic co-solvent, in the result of which gives a two-phase or heterogeneous mixture. This heterogeneous aqueous solvent mixture may be formed of a co-solvent with limited water solubility or ionic strength of the aqueous component may be adjusted to limit the solubility of the co-solvent in the aqueous phase.

The term "alkali metal hydroxide" refers to the connection MES, where M represents lithium, sodium, potassium or cesium, "bicarbonate of an alkali metal" refers to a group of M2CO3where M is sodium or potassium, and the carbonate of an alkali metal" refers to a group of M2CO3where M is sodium or potassium. The person skilled in the art understand that other bases can be used in the scope of the present invention to maintain the pH at a desired level.

Abbreviations used in this description, include acetyl (AC), acetic acid (SPLA), 1-N-hydroxybenzotriazole (HOBt), atmosphere (Atm), liquid chromatography over high pressure (HPLC), methyl (Me), tert-butoxycarbonyl (Vos), acetonitrile (MeCN), di-tert-butyl-pyrocarbonate or b anhydride (RE2O), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), benzyl (EAP), butyl (BU), methanol (Meon), benzyloxycarbonyl (cbz or Z), melting point (MP), carbonyldiimidazole (CDI), MeSO2(mesyl or Ms), 1,4-diazabicyclo [2.2.2]octane (DABCO), mass spectrum (ms), methyl-t-butyl ether (MTBE), 1,5-diaza icicle [4.3.0]-non-5-ene (DBN), 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU), N-methylmorpholine (NMM), N-organic (NMP), 1,2-dichloroethane (DCE), N,N'-dicyclohexylcarbodiimide (DCC), dichromate pyridinyl (PDC), dichloromethane (DCM), propyl (WG), pounds per square inch (psi), diisopropylethylamine (DIPEA, Hunigs' Base), pyridine (Rog), room temperature, rt, or RT, N,N-dimethylacetamide (DMA), tert-butyldimethylsilyl or t-BuMe2Si, (TBDMS), 4-N,N-dimethylaminopyridine (DMAP), triethylamine (EtsN or TEA), N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), triperoxonane acid (TFA), thin layer chromatography (TLC), ethyl acetate (tO), tetrahydrofuran (THF), diethyl ether (Et2O), trimethylsilyl or Me3Si (TMS), ethyl (Et), monohydrate p-toluenesulfonic acid (TsOH or pTsOH), H-Me-C6H4SO2or tosyl (Ts), isopropyl (i-Pr), N-urethane-N-carboxyanhydride (UNCA), ethanol (tO). Usually used nomenclature, including prefixes "normal" (n) "after" (i), secondary (sec-), "tertiary" (tert-) and "new" (neo), have their ordinary meaning, when used in conjunction with alkyl residues. (J.Rigaudi and D.R.Klesney, Nomenclature in Organic Chemistry, IUPAC 1979 Pergamon Press, Oxford).

Representative examples of compounds covered by this invention and falling under the scope of the present invention, are presented in Table I. These examples and their implementation are intended for specialists in this area could have been more easy is to understand and to reproduce the present invention. This should not be construed as limiting the scope of the invention, but only for the purpose of illustrative purpose and representativeness of the data submitted.

In General, the nomenclature used in the description of this application is based on AUTONOM™ 4.0 computerized system of the Beilstein Institute for recreating IU systematic nomenclature. There is a discrepancy between the depicted structure and name assigned to this structure, it is believed that depicts the structure has more weight and influence. In addition, if stereometry patterns or parts of patterns not shown, by means of, for example, relief or dashed lines, the structure of this structure should be interpreted as covering all isomers of this structure. The numbering system for these ring systems is as follows.

Connect and receive

Inhibiting activity (2R)-2'-deoxy-2'-fluoro-2'-C-methylcytidine against HCV polymerase already described (J.L.Clark et al. J. Med. Chem. 2005 48(17):5504-8; J.Clark, published application US No. 2005/0009737, both publications incorporated herein in its entirety by reference). In clinical practice it is advisable to enter the high-dose I-6 to quickly inhibit HCV polymerase, and then lower viral levels in the conditions under which minimizes ways the ability of the virus to mutate, and the selection of resistant strains. Significantly high levels may impede the achievement of the purposes of the original nuke.

Prodrugs include a strategy to improve the pharmacokinetic and physical properties of the compound and thereby to optimize bioavailability. In US publication No. 2005/0009737 offers General approaches to nucleotide prodrugs based on 2'-deoxy-2'-fluoro-2'-metrocluster. Despite the fact that the candidate prodrug looks deceptively simple, the identification of compounds with appropriate physico-chemical and pharmacodynamic properties, in vivo transformation and security profile - there is a complex variety of tasks that require critical experiments. Difficulties in the identification of prodrugs for oral administration include maintaining significant water solubility, lipophilicity and chemical stability at the same time, it allows for fast and efficient extraction of active rest after injection. Additionally, there must be minimized important nestersky metabolism and indirect conveyor clearance Prodrugs (.Beaumont et al. Curr. Dmg Metab. 2003 4(6):461-485). In addition, inhibition or induction of cytochrome P450-enzymes can cause adverse interactions medication with medication, which is undesirable. It has been unexpectedly OBN is hidden, the diesters of lower Akilov I-6 indicated in Table I, have significantly improved the bioavailability of I-6.

Pharmacokinetic characteristics of potential prodrugs was evaluated in rats and monkeys to try to minimize intraspecific variability and genetic polymorphisms that may lead to intra-specific artifacts. In the sense that the enzymatic transformation responsible for the hydrolysis of ester bonds, the specific affinity of the prodrug may depend on the specific patterns of esterase and/or peptidases, which can catalyze the transformation. It has often been shown that esterase activity in rats is significantly higher than in humans (J.A.Fix et al. Pharm. Res, 1990 7(4):384-387; W.Li et al. Antimicrob. Agents Chemother 1998 42(3):647-653). Another potentially important parameter is the bioconversion of citizenobama Foundation in the uridine using desaminase. Although triphosphates cicadina and uridine are inhibitors of the polymerase in vivo phosphorylation of uridine less efficient than cytidine. So conclude that elevated levels of uridine undesirable. About the lack of efficiency shown derivative of uridine in the Hcl replicon (J.Clark et al. J.Med. Chem., see above), it was reported.

1. Cmaxrepresents a peak concentration 1-7 in the blood.

2. AUC(cyt)- the area above the curve is La citizenobama of nuke.

3. AUC(urd)- the area above the curve oryginalego of nuke.

4. EU90in the replicon is equal to 5.40±2,6 µm (J.Clark et al. J.Med.Chem., see above).

Unexpectedly, it was found that C2-5alkyl diesters I-6 exhibit excellent proletarienne properties. Blood and rats, and monkeys were observed significantly higher levels of fluorinated nucleotides. Moreover, the ratio of cytidine to uridine in fluorinated original basis of the above in both samples, when the nucleoside is administered in the form of diapir.

In addition, befire capable of forming two different monoether and neeterificirovannah nuke. Pharmacokinetic analysis in this situation may be complex, if all samples are contained in the blood in significant quantities. The presence of many metabolites in the blood adds extra time when establishing whether the prodrug safe. Unexpectedly, the hydrolysis of both esters were absolutely easy, and the only important metabolism detected in the blood in addition to the original nucleoside, was 3'-monoether, which quickly turns into I-6.

In order to further assess the potential functioning of the prodrugs in humans, estimated transport imaginary prodrugs through SASO-2 cells. SASO-2 cells are usually used to assess the potential absorption/permeability mo is ekul (G.Gaviraghi et al. in Pharmacokinetic optimization in drug research. Biological, Pharmacokinelic and Computatinal Strategies. B.Testa et al. eds. Wiley Interscience VCH, Zurich 2001 pp.3-14). It was found that the permeability of SASO-2 is acceptable for C2-5alkyl diesters I-6.

In addition to effective in vivo biotransformation of the prodrug for oral administration must also demonstrate appropriate physico-chemical properties for the formation of medicines and to ensure absorption from the intestines to the Department, where necessary and desired biotransformation may occur. Especially relevant are the solubility, partition coefficient and stability in the liquid environment of the stomach and intestinal region. Values of all these parameters is shown in Table 2.

Table 2
Ko/w1lodD2Solubility3SGF4T1/2(h) 37°CSIF5T1/2(L) 37°C
I-13,39ND70,01618,6>8,411
I-22,33 1,670,18625,1866
I-31,270,93,78972
I-43,00ND0,0139>711>1511
I-52,20ND0,13>10,411>1111
I-6-0,94ND76>1211>1211

1. The calculated partition coefficient octanol-water.

2. The experimentally determined distribution coefficient at pH 7.4.

3. Solubility in aqueous medium, water or buffer pH 6.5 (mg/ml).

4. Stability in simulated gastric juice (SGF; pH 1,2).

5. Stability in simulated liquid bowel (SIP; pH 7.4).

6. Solubility in SGF is 13.3 mg/ml

7. not determined

8. Environment one is by SIF, solubility in SGF is 13.6 mg/ml

9. The environment is SIF, solubility in SGF is 0.16 mg/ml

10. Measured in water and buffer at pH 6.

11. A slight degradation was observed during this period of time.

12. Was determined at pH 5.0.

The distribution coefficient oil/water" Po/w(mixed P is calculated as Po/w) is an important characteristic for orally administered drugs. Drug substance must have substantial water solubility to dissolve in the composition and the fluids of the gastrointestinal tract (GIT) in order to interact with endothelial cells in the stomach and intestines, and a significant solubility in fats, in order to overcome the lipid bikinie membrane in these cells and instantly get into the bloodstream. The optimal range of log P of the compound for oral bioavailability is between 1 and 3, which are characteristic of the compounds of the present invention.

The expression of Po/wused here is called the partition coefficient octanol/water. Expression mixed Po/win the sense used here refers to the computed Po/w. A computer program, with which we compute the Po/wavailable to chemists pharmacists and doctors. The term "distribution coefficient" refers to the experimentally determined coefficie is the distribution between octanol and aqueous buffer solution. The partition coefficient and distribution coefficient are similar, but the last one is a function of the pH of an aqueous solution, whereas the Po/windependent of pH.

Solubility in the form of orally taken prodrugs should be higher than at least about 0.1 mg/ml for optimal composition, and the half-life in artificially-induced gastric juice and artificially simulated intestinal fluid should be 1-2 hours, 2-4 hours, respectively, in order to provide sufficient time to pass through the stomach and be absorbed in the intestine.

Compounds of the present invention in a standard way get a single stage, by acylation 1-6 in an aqueous organic solvent. The solvent may be either a homogeneous aqueous solution, or a two-phase solution. pH of the aqueous organic support at 7.5 by addition of base to neutralize the acid produced by the acylation. The base may be a hydroxide of alkali or alkaline earth metal or Quaternary amine. The reaction is carried out in the presence of DMAP, which, as is well known to specialists in this field, is the catalyst for acylation. The advantage claimed of this method is that the target product can be obtained without acylation of heterocyclic base is receiving. Does not require any protective groups that avoids the stage of protection and unprotect. The process is further described in the following examples.

Dosage and introduction

Compounds of the present invention can be part of many used oral dosage forms in combination with the media. For oral administration may be tablets, coated tablets, hard and soft gelatine capsules, solutions, emulsions, syrups or suspensions. Compounds of the present invention is effective when introduced in the form of suppositories, among other types of administration. The most common and standard method of introduction is usually oral using standard daily dosing regimes, which can be adjusted according to severity of disease and patient response to antiviral treatment.

The compound or compounds of the present invention and their pharmaceutically acceptable salts, together with one or more carriers, excipients and diluents can be in the form of pharmaceutical compositions or unit dosage forms. These pharmaceutical compositions and unit dosage forms may include standard ingredients in the adopted usually proportions, with added active compounds or without them, and the unit dosage forms can with erati kind of an effective amount of the respective active ingredient, when used in the established per diem ranges of dosages. The pharmaceutical compositions can be used in solid form, e.g. as tablets or filled capsules, in semi-solid form, in the form of powders, compositions with a continuous selection of the active principle, or in liquid form, for example, in the form of suspensions, emulsions or filled capsules for oral administration; or in the form of suppositories for rectal or vaginal application. Typical product should contain from about 5% to about 95% active compound or compounds (weight/weight). It is believed that the term "drug" or "dosage form" includes liquid and solid composition based on active connections, and the person skilled in the art it will be clear that the active ingredient may be included in the various preparations depending on the preferred dose and pharmaceuticals.

The term "filler" used herein means a compound that is used for the preparation of pharmaceutical compositions and in General is a safe, non-toxic, is not unacceptable in biological or in some other sense, and includes excipients that are acceptable for veterinary use, as well as pharmaceuticals for humans. Compounds of the present invention can introduce themselves, but should enter the ü in combination with one or more suitable pharmaceutical excipients, diluents or carriers selected with regard to the type of administration and standard pharmaceutical practice.

"Pharmaceutically acceptable salt" as a form of the active ingredient may also first to give the desired pharmacokinetic property of the active ingredient that is missing from his mesolevel forms, and may even positively affect the pharmacodynamics of the active ingredient in terms of its therapeutic activity in the body. The term "pharmaceutically acceptable salt" of the compound in the sense used here means a salt that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. Such salts include: (1) salt accession acids, formed with inorganic acids such as hydrochloric acid, vodorodomobilej acid, sulfuric acid, nitric acid, phosphoric acid and the like, or formed with organic acids, such as glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, 3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, succinic acid, methanesulfonate, econsultation, 1,2-ethicalfashion, 2-hydroxyethanesulfonic, benzosulfimide, 4-harbe solargorilla, 2-naphthalenesulfonate, 4-toluensulfonate, camphorsulfacid, louisanna acid, gluconic acid, glutamic acid, salicylic acid, Mukanova acid and tpole to be understood that all references to pharmaceutically acceptable salts include forms accession solvent (solvate) or crystal forms (polymorphs), according to this definition here, or salt attaching the same acid.

Drugs in the solid phase include powders, tablets, pills, capsules, suppositories, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavouring agents, soljubilizatory, lubricants, suspendresume agents, binders, preservatives, dezintegriruetsja tablets agents or encapsulating material. In powders, the carrier is usually a pulverized solid substance which is a mixture of pulverized active ingredient. In tablets, the active ingredient, generally, as a rule, is mixed with carrier having the necessary binding capacity in suitable proportions and compacted to the desired size and shape. Suitable carrier materials include (but without limitation the following: magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragakant, matilal is olosu, carboxymethylcellulose sodium, viscoplastic wax, cocoa butter and other Drugs of solid dosage forms may contain, in addition to the active component, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickening means, solubilizing agents, etc.

Liquid compositions are also suitable for oral administration include liquid compositions, including emulsions, syrups, elixirs, and aqueous suspensions. They include drugs in solid form, which, as implied, turn to drugs in liquid form almost immediately after their use. The emulsion can be prepared in solutions, for example, aqueous propylene glycol, or they may contain emulsifying agents such as lecithin, sorbitan monooleate or (gum) and acacia. Aqueous suspensions can be prepared, dispersive it is the active component in water with viscous material such as natural or synthetic resins, polymers, methylcellulose, sodium carboxymethylcellulose or other well-known suspendresume agents.

Compounds of the present invention can be prepared as compositions for administration in the form of suppositories. Viscoplastic wax such as a mixture of glycerides of fatty acids or cocoa butter, is first melted and the active component is dispersed to homogen the condition, for example, by stirring. Rasslablenaja homogeneous mixture is then poured into billet form of a certain size, she is allowed to cool and harden.

Compounds of the present invention can be prepared as a means for vaginal use. Pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in this field, are suitable and appropriate.

Suitable compounds along with pharmaceutical carriers, diluents and excipients are described in: Remington: The Science and Practice of Pharmacy 1995, published E.W.Martin, Mack Publishing Company, 19th edition, Taston, Pennsylvania. Experienced chemist-pharmacist can modify the structures in the framework of the instructions to the description and to create in the numerous formulations for a particular type of introduction, ensuring the stability of the compositions of the present invention and are not bringing harm to their therapeutic activity.

Modification of the compounds of the present invention, in order to make them more soluble in water or other means, for example, can be easily performed due to minimal modifications (e.g., due to selection of the composition of salts), which is well known to specialists in this field. Also within the competence of specialists in this field and is able to modify the type of administration and dosage R. the bench in relation to a specific connection with the to manage the pharmacokinetics of the claimed compounds for maximum beneficial effect on patients.

The term "therapeutically effective amount" used herein means the amount required to reduce symptoms in patients. The dosage should individual conditions in each individual case. These dosages can vary widely depending on numerous factors such as the severity of the disease that should be treated, the age and General health of the patient, other medications that a patient receives, the type and form of administration, and the preference and experience of the attending physician. During oral introduction of a daily dosage of between about 0.1 and about 10 g/day is acceptable in monotherapy and/or combination therapy. The preferred daily dosage is between about 0.5 and up to about 7.5 g/day, more preferably 1.5 and about 6.0 g/day. In General, the treatment begins with a large initial "Loading dose"to quickly weaken or eliminate the virus with subsequent reduction of the dose to a level sufficient to prevent the recurrence of infection. Any expert in this field, prepared for the treatment of diseases described here, will be able, without undue experimentation, and on the basis of their own knowledge, experience and description of this application to install a therapeutically effective amount of the compounds of the present invention for the disease and the patient.

Therapeutic efficacy can be established on the basis of experiments on the functioning of the liver, but without limiting the levels of proteins that are serum proteins (e.g., albumin, clotting factors, alkaline phosphatase, aminotransferases (e.g., alaninetransaminase, aspartate transaminase levels), 5'-nucleosidase, γ-glutamyltranspeptidase and others), synthesis of bilirubin, synthesis of cholesterol, and synthesis Belevich acids; metabolic function of the liver, including (but not limited to, carbohydrate metabolism, amino acid metabolism and ammonia.

Alternative therapeutic efficacy may be recorded by measurement of HCV-RNA. The results of these tests will allow to optimize the dose.

In embodiments of the invention the active compound or salt can be administered in combination with another antiviral agent such as ribavirin, another nucleoside HCV polymerase, non-nucleoside HCV polymerase inhibitor of HCV protease, HCV inhibitor of helicase or merged inhibitor of HCV. With the introduction of active compound or its derivative or salt in combination with another antiviral agent activity of the parent compound may be increased. When the treatment is a combination therapy, such introduction may be joint or sequential with respect to nucleoside derivative. "The joint is doing" in the sense used here includes the introduction of agents at the same time or at different times. The introduction of two or more agents at the same time can be achieved using a single composition containing two or more active ingredients or almost simultaneous introduction of two or more dosage forms of an active agent.

It should be understood that the data here are links to the treatment applied to the prevention and treatment of already existing conditions. In addition, the term "treatment" HCV infection, in the sense used here, also includes the treatment and prevention of a disease or condition associated with HCV infection or mediated 4CV infection, or clinical symptoms.

Example 1

(2R, 3R, 4R, 5R)-5-(4-amino-2-oxo-2H-pyrimidine-1-yl)-4-fluoro-4-methyl-3-propionyloxy-tetrahydrofuran-2-ymetray ether propionic acid (I-3)

To a suspension of I-6 (30 g, 0,116 mmol), DMAP (1.4 g, 11,57 mmol) in THF (300 ml) and water (150 ml) is added TEA (35.1 g, 0,347 mol) to obtain a clear solution (pH about 11). The reaction mixture was cooled to 5-10°C. and dropwise began to add propionic aldehyde (30.1 g, 0,23/mol), stirring two-phase reaction mixture. The pH of the register and support at around 11-12, at the same time Podkopaeva solution of KOH. The course is followed by means of HPLC dialysis and after was added propionate, the reaction mixture contained the ü about 52% of diapir, 30% of monoamino and 15% of the original substance. Under the conditions defined above, again added dropwise addition of propionic anhydride (45,2 g, 0,347 mol). The reaction mixture is left overnight without stirring. HPLC of the organic phase showed ...96% diapir and 2% of monoether. This phase is collected and the aqueous phase extracted with THF (100 ml). The combined organic phases are washed with Brin. The organic phase is filtered, evaporated and the residue dissolved in water (CA. 500 ml) and then diluted BRA (CA. 100 ml). The resulting mixture was slowly cooled to room temperature under stirring. The resulting precipitate is filtered off, washed with water and heptane, and dried at about 60°C under vacuum for CA. 60 hours to get to 3.45 g (80.3 per cent) 1-3 purity 90,7% according to HPLC.

Example 2

(2R, 3R, 4R, 5R)-5-(4-amino-2-oxo-2H-pyrimidine-1-yl)-4-fluoro-3-isobutyryloxy-4-methyl-tetrahydrofuran-2-ymetray ether somaclonal acid (I-2)

To a cooled on ice suspension I-6 (970, 3,74 mol) and DMAP (50 g, 0,412 mol) in THF (10 l) is added TEA (2.3 kg, 16.5 mol) and water (7 l), the result is a clear solution. To the mixture while mixing, slowly add isobutyramide (3 equivalent), maintaining the temperature at CA. 0°C. a further 1.2 and then to 0.7 equivalent of isobutyramide add up until HPLC shows that the reaction is virtually the ski is moving towards completion (only CA. 1,95 kg). The reaction mixture is acidified with conc. HCl to pH CA. 6.4 and the organic phase washed with EtOAc (2×10 l). The combined extracts washed with water (1×15 l). The organic phase is filtered and concentrated in vacuo. The residue is dissolved in IPA (CA. 20 kg) and add heptane (14,2 kg). The solution is heated to CA. 74-75°C with getting a clean solution, then CA. 5 l is removed by distillation. The resulting solution is slowly cooled to room temperature. The precipitate formed at CA. 42-43°C. Cooling to 5°C continues slowly, then during the night carry out the mixing. The obtained solid residue is filtered off and the filtrate is washed with a mixture of IPA/heptane (1:8) (13,4 kg), dried under vacuum at about 60-70°C with getting 1,295 kg (86,65%) 1-2 purity 99,45% according to HPLC.

Example 3

Isobutyl ester (2R, 3R, 4R, 5R)-5-(4-amino-2-oxo-2H-pyrimidine-1-yl)-4-fluoro-2-msobuttoniconandcaption-4-methyl-tetrahydrofuran-3-yl ester carboxylic acid (I-4).

Suspension 1-6 (700 mg), DMAP (33 mg) in THF (7 ml) diluted with Brin (7 ml). Add dilute NaOH until pH becomes CA. 11. The reaction mixture was cooled in an ice bath and added dropwise isobutyl chloroformate (1,11 g) in a mixed two-phase reaction mixture, maintaining the pH to about 11 by adding NaOH, if necessary. HPLC analysis shows mainly dicarbonate, zagryazneny the sa. 15% of monocarbonate. To a cooled on ice solution add additional 1 EQ. isobutylphthalate. HPLC shows almost completion of the reaction. The resulting mixture is left overnight at room temperature. Add tO (CA. 50 ml) and the pH of the aqueous phase adjust to 7.5 conc. HCl. The phases are separated, the organic phase is washed with water (3x) and evaporated to dryness to obtain a colorless precipitate (CA. 1.22 g). The precipitate is dissolved in hot acetone, the resulting transparent solution, which is slowly cooled to room temperature, resulting in deposition of sediment. The residue was dissolved in IPA (CA. 20 ml), which then leads to the formation of a precipitate, which is filtered and sequentially washed with IPA-heptane, then dried with receipt of 0.85 g (68.5 per cent) 1-4 with a purity of 97.5% according to HPLC.

Example 4

Propyl ester (2R, 3R, 4R, 5R)-5-(4-amino-2-oxo-2H-pyrimidine-1-yl)-4-fluoro-4-methyl-2-propoxycarbonyl-tetrahydro-furan-3-yl ester carboxylic acid; hydrochloric acid salt (I-5)

The suspension I-6 (700 mg, 2,70 mmol), DMAP (33 mg, 0.27 mmol) in THF (7 ml) and diluted with Brin (7 ml) is added a dilute STAKE to maintain a pH of CA. 11. The reaction mixture is cooled to CA 5°C and a two-phase reaction mixture with stirring, added dropwise n-propylchloride (1.0 g). HPLC showed formation of the desired product along with sa. 20% of monocarbonate. Two additional the s portion of propylphosphonate add (2×1 equivalent) to a cold solution until until HPLC analysis shows that the reaction has come to completion. The reaction mixture was diluted with tO (50 ml) and the pH of the aqueous solution was adjusted to CA. 6.5 conc. HCl. The phases are separated, and the organic phase washed with water (3x), is evaporated to dryness to obtain a colorless precipitate (CA. 1.1 g). Hot IPA solution acidified with 4N Hcl (CA. 1 ml) and evaporated to dryness. The resulting precipitate pererastayut hot tO (CA. 115 ml) and stirred over night at room temperature. The resulting solid mass is filtered and the residue is washed with a mixture of Meon/heptane (1:1). The remaining residue is dried in vacuum at about 60°C to obtain 0,325 g (25,8%) 1-5 with a purity of 97.5% according to HPLC.

Example 5

Determination of pharmacokinetic parameters in rats

Used intact rats male mouse IGS Wistar Han Rats Crl:WI (GLxx/BRL/Han) IGS BR (Hanover Wistar)weighing 200-250 g groups of three animals were used for each level doses of the compounds. The animal was organized by the normal access to food and drink during the experiment. The investigated substance was prepared as a suspension in water containing Captex355EP, Capmul MCM, EtOH and propylene glycol (30:20:20:30) at a dose equivalent to 10 mg/kg I-6 and was administered orally via a stomach tube. Blood samples (0.3 ml) was collected from the thus treated rats at 0.25, 0.5, and 1, 3, 5, and 8 hours from the cervical cannula and 24 hours after cardiac Pontiac samples was added potassium oxalate/NaF and kept them on ice during the entire sampling procedure. The samples were subjected to rotation in a refrigerated centrifuge at 4°C with the speed with which it was possible, and plasma samples were stored at -80°C in the freezer prior to analysis. Aliquots of plasma (0.05 ml) was mixed with 0.1 ml of acetonitrile. Added internal standard (0.05 ml in water) and 0.02 ml of pure solvent. A series of calibration standards were prepared by mixing 0.05 ml aliquots of plasma from neobrabotannim rats with 0.1 ml of acetonitrile, 0,02 ml aliquot of a standard solution in methanol:water (1:1) and 0.05 ml aliquot of the internal standard in the water. Each plasma sample and calibration standard was carefully which on the vortex and then centrifuged at 3000 rpm for 5 minutes to precipitate the protein. Supernatant (100 l each) after centrifugation was transferred into a 96-well plate containing 200 ml aqueous mobile phase for LC/MS/MS analysis.

Analysis of samples:

Prodrugs were analyzed using high performance liquid chromatography tandem mass spectrometry (HPLC/MS/MS). To separate the used Thermo Aquasil C18 of 4.6×50 ml column (5 µm). For the ionization process used Electrospray lonization (ESI). Mobile-dose And contained 5 mm ammonium acetate in water with 0.1% formic acid and mobile phase b contained Meon with 0.1% formic acid. The elution was performed with the following gradient with a flow rate of 1 ml/min:

Time%%
0 min955
0.5 min955
0,8 min8515
1.5 min8515
2,8 min1090
3,8 min1090
3.9 minutes955
5 min955

Example 6

Determination of pharmacokinetic parameters in monkeys

There were three male Cynomolgus monkeys weighing 8-10 kg during the experiment the animals were allowed free access to feed and water. The weight of the animals during the entire period of drug administration, and adverse reactions in animals was recorded. The investigated substance was prepared as an aqueous suspension, containing hypromellose (2910, 50 SDR), USP, Polysorbate 80, N, benzyl alcohol, NF (5,0, 4.0 and 9.0 mg/ml) and sterile water (quantity sufficient to obtain 1.0 ml), at a dose equivalent to 10 mg/kg 1-6 and 0.5 mg/kg was administered orally via a stomach tube. Blood samples (0.5-1.0 ml) were collected through 0,083, 0,25, 0,5, 1, 3, 5, 8 and 24 hours. Sample treatment and analysis were performed as described in experiment on rats. A sample of 5 ml of urine was taken from each monkey before the introduction of the dose for 0-8 hours. Urine samples were stored at -80°C and analyzed using LC/MS/MS. Standard curves were constructed for pure animal plasma containing NaF and potassium oxalate.

Example 7

SASO Protocol

Materials:

Dry Krebs-Henseleit buffer, the dihydrate of calcium chloride and sodium bicarbonate were obtained from Sigma (st. Louis, MO). SASO-2 cells (Passage ≈ 100) were obtained from Roche Basel. DMEM (enriched environment, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and serum bull were obtained from JRH Bioscience (Lenexa, KS). MEM antamanide amino acids, L-glutamine, penicillin and streptomycin were obtained from GIBCO, habs, Life Tech. LLC (Grand Island, NY). Liners in wells for cell culture (6.5 mm diameter, 1,12 cm2, pore size of 0.4 μm)were obtained from Costar (Cambridge, MA).

Cell culture:

Cells were grown in 75 cm2the flask while maintaining the temperature at 37°C in an atmosphere of 5% CO2and 95% air. The culture medium containing DMEM/enriched environment with new the m 5% serum bull, 25 mm HEPES, 1% MEM nineteenth amino acids, 1% L-glutamine, 100 U/ml penicillin and 100 WG/ml streptomycin. Culture was massirovala every week in the ratio of breeding 1-3. To study the permeability of SASO-2 cells in the passages of the room 110-120 were seeded at a density of 400,000 cells/cm2on portable polycarbonate filters inserted in the ear and allowed to grow for 7 days before use.

Krebs-Henseleit Buffer:

Krebs-Henseleit bicarbonate buffer containing 10 mm glucose and 25 m l2with specified pH 6.5 and 7.4 are prepared according to the instructions on the package. Dry salt quantitatively dissolved in about 90% of the desired volume of Millipore water. The dihydrate of calcium chloride and sodium bicarbonate added before setting the pH using 1N. HCl or 1N. NaOH. An additional portion of Millipore water was added when the solution needs to be brought to final volume. The solution was sterilized by filtration using a membrane with a porosity of 0.22 micron and kept in the refrigerator (~ 20°C) before use.

Preparation of cell cultures:

Differentiated cells were obtained from Cell Culture Core Facility in a stable at 37°C in the atmosphere of 5% CO2and 95% air. Inserts containing SASO-2 monolayers were washed at 37°C balanced Krebs-Henseleit buffer, pH 7.4.

Method:

Cell inserts (inserts) used as d is fusionio camera. pH Krebs-Henseleit buffer in the apical and basolateral cameras 6.5 and 7.4, respectively, and the initial substrate concentration on the apical side was 100 µM. Cells were preincubated with a test connection to the apical chamber for approximately 30 minutes at 37°C in an atmosphere of 5% CO2and 95% air. Experiments were started when the cell paste with 100 μm of compounds in Krebs-Henseleit buffer pH 665 were transferred to a new tablet with a pre-balanced buffer in basolateral the camera. Samples from the donor side at 0 minute, as well as in the donor and receiver sides after 30 minutes was collected for analysis.

Control after the experiment:

To assess the transformations in the diffusion system used Lucifer Yellow. After the last sampling for determination of the studied compounds Lucifer Yellow was added to the apical chamber to detect the initial concentration of 100 µm. After 60 minutes incubation, 250 l were removed from the basal chamber and examined.

Calculation of permeability coefficient (Parr):

Parrwas calculated by the following formula:

Papp=(V×dC)/(A×Cabout×dt)=(cm/sec)

where V is the volume (cm3) accept the solution, And the surface area (cm2) inserts, Withabout- initial concentration (nm), a dC/dt is the change of concentration in the host (the Republic of the faithful) the camera over time, that is, the slope (nm/min) (tangent angle) concentrations in the receiving chamber with respect to time. Concentration at each sampling point corrected to calculate the aliquots, deleted or moved donor inserts in the new tablets, depending on the experiment.

Example 8

The pharmaceutical compositions of the claimed compounds for the introduction of different methods were prepared as described in example 8.

Composition for containing moisture pelletized composition for oral administration of (A):

Ingredient% weight./weight
Intragranular Active ingredient (I-2)50,0
Mannitol (Partech M200) USP39,0
Povidone, CSO USP5,0
Sodium crosscarmellose NF4,0
Cleansing water USPfor granulation
Extramedullary Crosscarmelose USP2.0

The ingredients are mixed, granularit and distribute in hard gelatin capsules containing 500 mg of active with the unity. The composition for oral administration (In):

Ingredient% weight./weight
Active ingredient (1-2)50,0
Mannitol (Partech M200) USP30,0
Povidone, CSO USP5,0
Sodium crosscarmellose NF9,5
Microcrystalline cellulose5,0
Magnesium stearate0,5

Ingredients harvested granularit, using as solvent water. The composition is then dried to form tablets containing 500 mg of active compound, in an appropriate tabletirujut device.

Composition for oral use (S):

td align="center"> Propylparaben
Ingredient% weight./weight
Active connection6 g
Carboxymethylcellulose1 g
Methylparaben0.15 g
0.05 g
Flavora 0.035 ml
Dyes0.5 mg
Distilled waterto 100 ml
Hydrochloric acidto pH ~4

The ingredients are mixed to obtain a suspension for oral administration Composition for suppositories (D):

Ingredient% weight./weight
The active ingredient1%
Polyethylene glycol 100074,5%
Polyethylene glycol 400024,5%

The ingredients are melted together and stirred on the steam bath. Then pour into molds 2.5 g total weight.

Features (characteristics) of the invention disclosed in the foregoing description, or the following claims, expressed in their specific forms or in the special terms, in order to give an idea of their function or method or process for achieving results, respectively, m the gut, either by itself or in combination with each other, be used to reproduce the invention in its most varied forms.

The above invention is described in detail using illustrative examples, in order to achieve clarity and understanding. The person skilled in the art it will be obvious that all changes and modifications that can be made, fall under the scope of the claims. Therefore, it is clear that the above description be regarded as illustrative and not restrictive. The scope of this invention should therefore be determined not with reference to the above description, but should instead conform to the following claims along with the full scope of equivalents that are covered by the claims.

All patents, patent applications and publications cited here included as references in their entirety in all respects to the same extent as if each individual patent, patent application or publication would be presented similar to a single mark.

1. The compound of the formula I

where R1selected from the group consisting of ethyl, n-propyl, isopropyl or isobutyl, and
its pharmaceutically acceptable salt.

2. The compound according to claim 1, in which R1is ethyl or isopropyl.

3. The compound according to claim 1, in which R1is isopropyl, and the connection is a hydrochloric acid or sulfate salt.

4. The compound according to claims 1-3, intended for the treatment of diseases mediated by the hepatitis C virus (HCV).

5. The pharmaceutical composition intended for the treatment of diseases mediated by the hepatitis C virus (HCV), comprising a therapeutically effective amount of a compound according to claims 1 to 4 in combination with one pharmaceutically acceptable carrier, diluent or excipient.

6. A method of treating diseases mediated by the hepatitis C virus (HCV), comprising the administration to a patient in need, a therapeutically effective dose of a compound according to claims 1-4.

7. The method according to claim 6, characterized in that the patient is given a dose of from 0.1 to 10 grams per day.

8. The method according to claim 6 or 7, further comprising introducing at least one immune system modulator and/or at least one antiviral agent that inhibits replication of HCV.

9. How selective O-acylation of nucleoside II for receiving the O-acyl nucleoside I in alkaline reaction conditions

where R1selected from the group consisting of ethyl, n-propyl, isopropyl or isobutyl, which includes stages:
(i) dissolving II and DMAP in a heterogeneous mixture of water and solvent and added the water a basis for regulating the pH from about 7.5 to about 12;
(ii) the optional addition of a sufficient quantity of saturated aqueous NaCl to obtain a two-phase reaction mixture;
(iii) adding Alliluyeva agent and additional grounds, sufficient to maintain the pH from about 7.5 to about 12;
(iv) reaction monitoring and interruption of adding the specified Alliluyeva agent and specified grounds upon reaching the conversion of a sufficient level;
(v) optional contacting O-alloclasite with a pharmaceutically acceptable acid for the formation of pharmaceutically acceptable salts.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to a pyrimidine nucleoside compound of general formula (1) , in which one of X and Y is a cyano group and the other is hydrogen; R1 is hydrogen, (R3)(R4)(R5)Si- or a carbonyl group which includes an alkyl monosubstituted with an amino group; R2 is hydrogen or (R6)(R7)(R8)Si-, provided that at least one of R1 and R2 is not hydrogen; or R1 and R2 together form a 6-member cyclic group -Si(R9)(R10)-, where each of R9 and R10 is a straight or branched alkyl; R3, R4 and R5 denote a straight or branched alkyl optionally substituted alkoxy, or cycloalkyl; R6, R7 and R8 denote a straight or branched alkyl optionally substituted alkoxy, cycloalkyl or phenyl, or to pharmacologically acceptable salts thereof. The invention also relates to a range of specific compounds of formula (1) or to their pharmacologically acceptable salts: 5'-O-triisopropylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-diethylisopropylsilyl-2'-cyano-2,-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-dimethylthexylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-(dimethyl-n-octylsilyl)-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-dimethylthexylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-diethylisopropylsilyl -2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-(tert-butyldimethylsily)-2'-cyano-2'-desoxy-1-β-O-arabinofuranosylcytosine; 3'-O-triisopropylsilyl-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 3'-O-dimethylthexylsilyl-5'-O-(L-valyl)-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; 5'-O-(L-valyl)-3'-O-(tert-butyldimethylsilyl)-2'-cyano-2'-desoxy-1-β-D-arabinofuranosylcytosine; and 3'-O-cyclopropyl-diisopropylsilyl-2'-cyano-2'-desoxy-1-β-D- arabinofuranosylcytosine.

EFFECT: obtaining formula (1) compounds or their pharmacologically acceptable salts for preparing a medicinal agent for treating tumours.

9 cl, 20 tbl, 1 dwg, 73 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the method of producing 2'-desoxy-β-L-thymidine, which involves reacting 5'-O-trityl- or 5'-O-dimethoxytrityl- substituted 2,2' -anhydro-1 -β-L- arabinofuranosylthymine with a reducing agent RedAl and a complexing agent 15-crown-5-ether in a polar solvent 1,2-dimethoxyethane (DME) or tetrahydrofuran, obtaining 5'-O-trityl- or 5'-O-dimethoxytrityl- substituted 2,2'-desoxy-β-L-thymidine, subjected to protection removal if necessary. The invention also relates to the method of producing 2'-desoxy-β-L-thymidine, which involves reacting L-arabinose with cyanamide with subsequent reaction of the intermediate product - L-arabinofuranosylaminooxazoline - with a cycling or condensing agent, forming 2,2' -anhydro-1-β-L-arabinofuranosylthymine; reaction of the latter with a reducing agent RedAl and a complexing agent 15-crown-5-ether in a polar solvent 1,2-dimethoxyethane (DME) or tetrahydrofuran, obtaining 2'-desoxy-β-L-thymidine, where L-arabinofuranosylaminooxazoline can be protected by trityl or dimethoxytrityl in position 5' before or after reaction with the cycling or condensing agent; and protection removal of optionally protected 2'-desoxy-β-L-thymidine, if this is necessary or desired. Use in the given methods of such a reducing agent as Red-Al, and such a complexing agent as 15-crown -5-ether, causes a reaction of intramolecular protection and production of the required nucleoside product with good output.

EFFECT: compound is of great importance as an antiviral or antineoplastic preparation.

13 cl, 29 dwg, 28 ex

FIELD: chemistry.

SUBSTANCE: invention relates to method of obtaining enriched with β-anomer 2'-desoxy-2',2'-difluorocytidine of formula (I)

, which includes stages: (i) interaction of enriched with α-anomer compound of 1-halogenribofuranose of formula (III) with nucleic base of formula (IV) in solvent obtaining enriched with β-anomer nucleoside of formula (II) , with constant removal of formed in reaction process silylhalogenide of formula R3SiX (V) by distillation using carrier or running inert gas through reaction mixture; and (ii) removal of protective group from enriched with β-anomer nucleoside of formula (II). Invention also relates to method of obtaining hydrate of enriched with β-anomer 2'-desoxy-2',2'-difluorocytidine of formula (I), which at stage (ii) after removal of protective group additionally includes stages of dissolving formula (I) nucleoside in water; heating of obtained solution to temperature from 40 to 60°C; cooling of solution to temperature ranging from 10 to 25°C with or without mixing and without changing pH; and filtering of deposited solid substances.

EFFECT: method improvement.

17 cl, 2 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to (2'R)-2'-dezoxy-2'-fluoro-2'-C-methylnucleoside (β-D or (β-L) , where X represents O; R1 and R7 independently represent H; R3 represents hydrogen and R4 represents NH2; or its pharmaceutically acceptable salt. The invention also pertains to the method of producing the said compounds, which involves glycosylation of N4-benzoylcytosine with a compound of formula 1-4, where R represents methyl, Pg is chosen from C(O)Ph, CH2Ph or both Pg groups can be included in 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene); with further removal of protection of 3'-OPg and 5'-OPg and N-benzoyl of the obtained product.

EFFECT: invented compounds or their pharmaceutically acceptable salts are used as active ingredients against Flaviviridae family viruses in pharmaceutical compositions and liposomal pharmaceutical compositions.

4 cl, 9 tbl, 5 ex, 4 dwg

FIELD: chemistry.

SUBSTANCE: claimed invention relates to method of gemcitabine hydrochloride purification, which includes enriching gemcitabine hydrochloride with its p-anomer, according to which solution of gemcitabine hydrochloride in water is taken with ratio of water to gemcitabine hydrochloride from 3:1 to 12:1 (wt/vol); solution is processed with activated coal, activated coal being taken in amount from 0.1 to 10 wt % of gemcitabine hydrochloride amount in solution; activated coal is removed from solution with formation of filtered solution; concentration of gemcitabine hydrochloride in filtered solution is increased until ratio of filtered solution to gemcitabine hydrochloride equals from 1:1 to 1:5 (wt/vol), efficient for gemcitabine hydrochloride sedimentation; deposited gemcitabine hydrochloride is isolated; and in case admixture content in deposited gemcitabine hydrochloride is not reduced to required level, stages (a)-(e) are repeated. Claimed invention also relates to method of obtaining gemcitabine hydrochloride using claimed purification method.

EFFECT: creation of efficient method of gemcitabine hydrochloride purification.

5 cl, 1 tbl, 5 dwg, 8 ex

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

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

The invention relates to the chemistry of nucleosides, in particular to an improved method for the preparation of 3'-azido-2',3'-dideoxythymidine (azidothymidine, AZT), used in medicine as an antiviral drug for the treatment of patients suffering from acquired immunodeficiency syndrome (AIDS)

The invention relates to a method for obtaining enriched beta-anomer nucleoside of the formula I, where T is fluorine and R is the corresponding nucleoside described in paragraph 1 of the formula

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula (I) where values of the substitutes are disclosed in the patent claim.

EFFECT: compounds can be applied for treating the infections caused by Pneumovirinae subfamily viruses (RSV, PCB).

53 cl, 502 ex, 11 tbl

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine and deals with method of obtaining lice culture influenza vaccine. Essence of the invention includes method of obtaining virus-containing substance by cultivation of one of cold-adapted influenza virus reassortants with enoculation dose, with multiplicity of infection not lower than 0.0001 EID50/cell in MDCK cell culture on micro-carriers, which have concentration not less than 1 g/l, with application as micro-carrier material of porous polypropelene, in supporting serum-free nutritional medium, containing proteolytic enzyme in amount 0.25-50.0 mcg/ml, and stabilising additive, which includes sorbitol, or sucrose, or peptone from soya in concentration 0.5-4.0 wt%, collection of virus-containing liquid after cultivation is carried out at least 2 times when specific influenza virus activity before each collection of virus-containing liquid reaches at least 7.0 Ig EID50/ml, concentration and purification of virus substance from ballast admixtures, introduction into purified substance before drying of stabilising additives, with application as such of either proline, glycene, lactose, glutamine-acidic sodium, sucrose, gelatins in final concentration (1.5-5), (1.5-5), (1.5-10), (1.5-5), (5-30) and (1-10) wt % respectively, or sucrose, gelatose and soya peptone in final concentration (1-8), (1-8) and (1-8) wt % respectively, or sorbitol and gelatose in final concentration (3-8) and (3-8) wt % respectively.

EFFECT: obtaining more thermostable vaccine with high output.

2 cl, 6 ex, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of chemical-pharmaceutical and food industry, namely to field of creating liquid forms of medications and biologically active food additives, with wide spectrum of pharmacological action, namely general health-improving, anti-inflammatory, antimicrobial, antiviral and immunomodelling and antioxidant action. Claimed medication for treatment and prevention of upper airways diseases is made in form of syrup of thallus of Island moss Cetraria islandica (L.) Ach., enriched with complex of biologically active substances, including polysaccharides and lichen acids. Also claimed is method of obtaining medicinal syrup of Island moss. Vegetable raw material before extraction is preliminarily crushed and extraction is carried out with syrup of various concentrations with its further filtering.

EFFECT: method ensures obtaining of final product - syrup of Island moss, maximal substance extraction, as well as preservation of biologically active substances (BAS) in syrup (polysaccharides, lichen acids) during long term in native state, thus, ensuring high medication efficiency.

8 cl, 9 ex, 3 tbl, 3 dwg

FIELD: medicine.

SUBSTANCE: invention relates to compounds of formula

and their pharmaceutically acceptable salts, inhibiting activity of serine protease, in particular, activity of protease of hepatitis C virus HC3-HC4A. Claimed invention also relates to pharmaceutical compositions, which contain said compounds, method of inhibiting serine protease activity.

EFFECT: elimination or reduction of infection with hepatitis C virus.

27 cl, 1 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a compound of formula [I-D1] or pharmaceutically acceptable salt thereof,

,

where each symbol is defined in the claim. The invention also relates to pharmaceutical compositions containing said compound and having HCV polymerase inhibiting activity.

EFFECT: disclosed compound exhibits anti-HCV activity, based on HCV polymerase inhibiting activity and is useful as an agent for preventing and treating hepatitis C.

32 cl, 497 tbl, 1129 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, and concerns compositions and methods for treating severe acute respiratory syndrome (SARS). Substance of the invention includes a pharmaceutical composition for treating severe acute respiratory syndrome SARS, containing a therapeutically effective amount of recombinant TNF-α receptor or thalidomide in a pharmaceutically acceptable carrier. Also, presented is a method of treating SARS with using this composition, as well as a method of preparing the composition.

EFFECT: drug for treating SARS is developed.

43 cl, 2 ex

FIELD: veterinary science.

SUBSTANCE: agent to treat and prevent disbacterial conditions of intestine in birds contains inulin prepared on milk whey, ferment of live microorganisms Lactobacillus acidophilus, Enterococcus faecium, prepared on skimmed milk, potassium iodide, monobasic calcium phosphate, vitamin E (alpha-tocopherol acetate), vitamin D3 (cholecalciferol), vitamin A (retinol acetate) at the following ratio of components, wt %: inulin 0.5-0.6; milk whey 55.281 -51.979; ferment of live microorganisms Lactobacillus acidophilus, Enterococcus faecium, prepared on skimmed milk 5.5-6.5; potassium iodide 1.25-1.35; monobasic calcium phosphate 37-39; vitamin E 0.45-0.55; vitamin D3 0.0125-0.0135; vitamin A 0.0065-0.0075.

EFFECT: agent makes it possible to increase daily average growth of chickens and to reduce cases of disbacterial intestine conditions.

1 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: strain can be used for prevention and diagnostic purposes in medicine. Strain No. VBA-07 is created for producing vaccines and diagnostic preparations. Strain No. VBA-07 surpasses a prototype strain both in efficiency (the amount of infective virus and virus-specific antigen), and in kinetics of HAV antigen accumulation, and in cultivation time. The strain is deposited in the Microorganism Cultures Collections of JSC 'Vector-BiAlgam' and is registered under No.VB-17.

EFFECT: creation of a producer strain with higher efficiency of HAV antigen and with shorter time of HAV cultivation.

7 dwg, 6 tbl

FIELD: medicine.

SUBSTANCE: invention relates to novel peptide compounds, which possess ability to inhibit protease of hepatitis C virus (HCV), their pharmaceutical compositions and application of compounds for obtaining medication for diseases associated with HCV.

EFFECT: improvement of compound properties.

48 cl, 6 tbl, 65 ex

FIELD: chemistry.

SUBSTANCE: agent is an N- and C-substituted peptide selected from n-decyl ether (1-tetradecyl-1,4-diazoniabicyclo[2.2,2.]octan-4-yl)-acetyl-glutamyl-glycyl lysyl-glycine (1), n-decyl ether (1-tetradecyl-1,4-diazoniabicyclo[2.2,2.]octan-4-yl)-acetyl-glutamyl-β-alanyl-arginyl-glycine (2) and n-decyl ether glutamyl-β-alanyl-lysyl-glycine (3).

EFFECT: high antiviral activity of the agent.

3 dwg, 2 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology and can be used for treatment of locally spread resectable esophageal cancer. Method includes radical surgery, polychemotherapy and radiotherapy. Before operation 2 courses of polychemotherapy are carried out in accordance with the scheme: cysplatin 50 mg/m2 1,8 day, gemcitabine -800 mg/m2 intravenously drop-by-drop during half-hour infusion - 2, 9 days with 3-week long interval between courses. After that, surgical intervention in radical volume with two-zonal 2F lymphodissection, with carrying out IORT in single dose 10 Gy onto the bed of removed tumour and ways of regional metastasis. In post-operational period additionally course of DGT is carried out on cervical, supraclavicular zones in TFD -40 Gy.

EFFECT: application of invention makes it possible to increase efficiency of combined treatment due to prevention of tumour process progression and distant metastasis and reduction of number of post-operational complications.

1 ex

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