Synthesis of β-l-2'-desoxynucleosides

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

 

Cross-reference to related applications

This invention claims the priority in accordance with preliminary applications for the grant of U.S. patent No. 60/483711, filed June 30, 2003, and 60/558616 filed April 1, 2004.

The technical field

This invention relates to methods of producing 2'-deoxy - or 2'-modified nucleosides, in particular, β-L-2'-deoxythymidine. The present invention is an improved method that is easily scalable for industrial production. Compounds obtained by the process according to the present invention are important as antivirals, antineoplastics means and intermediate products for the synthesis of pharmaceutical compounds and compositions.

The level of technology

HBV is the second cause of human malignancies, second only to tobacco. The mechanism by which HBV induces a malignant tumor is not known, although it is assumed that he can start the development of a tumor or indirectly, to start the development of tumors by chronic inflammation, cirrhosis and cell regeneration associated with infection.

The hepatitis B virus has reached epidemic levels in the world. After an incubation period of two to six months, which is about the owner does not know about the infection, HBV infection can cause acute hepatitis and liver damage that causes abdominal pain, jaundice, and elevated blood levels of some enzymes. HBV can cause fulminant hepatitis, rapidly progressive, often in the form of the deadly disease, which destroyed huge areas of the liver.

Patients usually recover from acute hepatitis. However, in some patients, high levels of viral antigen are retained in the blood for a long or indefinite period of time, causing chronic infection. Chronic infection can lead to chronic persistiruuschem hepatitis. Most often patients infected with chronic persistent HBV occur in developing countries. By mid-1991, there were approximately 225 million chronic carriers of HBV only in Asia but around the world nearly 300 million speakers. Chronic persistent hepatitis can cause fatigue, cirrhosis and hepatocellular carcinoma, a primary liver cancer.

In the application WO 96/40164 filed Emory University, UAB Research Foundation and the national center for scientific research (CNRS), described a number of β-L-2',3'-dideoxynucleosides for the treatment of hepatitis B.

In the application WO 95/07287, also filed Emory University, UAB Research Foundation and the national center for scientific research (CNRS), described 2'- or 3'-deoxy - and 2',3'-dideoxy-β-L-menthofuran sinucleanse for the treatment of HIV infection.

In the application WO 96/13512 filed Genencor International, Inc. and Lipitek, Inc., describe how to get L-ribofuranosylpurine as anticancer agents and antiviral agents.

Idenix Pharmaceuticals, Ltd. describe the 2'-deoxy-L-retroinformacion and their use in the treatment of HBV in U.S. patent No. 6395716; 6444652; 6566344 and 6539837. And see also WO 00/09531. Described is a method of treatment of hepatitis B infection in humans and other animal hosts, which is the introduction of an effective amount of a biologically active 2'-deoxy-β-L-retroinformacion (alternative called β-L-dN or β-L-2'-dN) or its pharmaceutically acceptable salt, complex, ester, or prodrug, including β-L-deoxyribozymes (β-L-dT), β-L-deoxyribozymes (β-L-dC, β-L-desoxyribose (β-L-dU)β-L-deoxyribofuranosyl (β-L-dG), β-L-deoxyribofuranosyl (β-L-dA) and β-L-desoxyribose (β-L-dI), administered either individually or in combination, optionally in a pharmaceutically acceptable carrier. Also claimed 5'and N4(citizen) or N6- (adenosine) acylated or alkylated derivatives of the active compounds or 5'-phospholipids or 5'-ether lipid.

Von Janta-Lipinski et al., in J. Med. Chem., 1998, 41 (12), 2040-2046, describe the use of L-enantiomers of 3'-fluoro-modified β-2'-deoxyribonucleoside-5'-triphosphates for inhibition of the polymerase of hepatitis B. In particular, 5'-triphosphates of 3'-deoxy-3-fluoro-β-L-thymidine (β-L-FTTP), 2',3'-dideoxy-3'-fluoro-β-L-cytidine (β-L-FdCTP) and 2',3'-dideoxy-3'-fluoro-β-L-5-methylcytidine (β-L-FMethCTP) claimed as effective inhibitors of the DNA polymerase of HBV. In addition, von Janta-Lipinski et al. describe the biological activity triphosphate β-L-thymidine (but not β-L-2'-dC) as a nucleoside inhibitor of endogenous DNA polymerase of HBV and DHBV. However evaluated only triphosphorylation β-L-thymidine, but not reported nefosfaurilirovanna form, and the article does not comment about fosfauriliruyutza whether the β-L-nucleosides into cells orin vivoor, more importantly, does not comment on the efficiency of phosphorylation of β-L-thymidinein vivo. Therefore, the article does not mention that β-L-thymidine may have some activity against hepatitis B in the cell orin vivo. Cm. also WO 96/1204.

In the European application for patent No. 0352248 A1 Johansson et al. describe the use of compounds L-ribofuranosyl for the treatment of hepatitis B.

Verri et al. describes the use of 2'-deoxy-β-L-retroinformacion as antineoplastics funds and as antiherpetic funds (Mol. Pharmacol. (1997), 51(1), 132-138 and Biochem. J. (1997), 328(1), 317-20). Saneyoshi et al. demonstrated the use of 2'-deoxy-L-ribonucleosides as reverse transcriptase inhibitors (I) to fight against retroviruses and for the treatment of SLEEP is a, Jpn. Kokai Tokkyo Koho JP06293645 (1994).

Giovanni et al., in particular, it was tested 2'-deoxy-β-L-retroinformacion against virus pseudoleskeella (PRV), Biochem. J. (1993), 294 (2), 381-5.

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

Morris S. Zedeck et al. first described β-L-dA for inhibition of the synthesis of inducible enzymes inPseudomonas testosteroni, Mol. Phys. (1967), 3 (4), 386-95.

In addition, derivatives of cytosine applicable as intermediates for such medicines, as cytidinediphosphocholine, generic name citicoline.

In the publication of U.S. patent No. 20030083306, Idenix Pharmaceuticals, Ltd., described 3'-prodrugs of 2'-deoxy-β-L-nucleosides for the treatment of HBV. Cm. also WO 01/96353.

In U.S. patent No. 4957924 Beauchamp describes the various therapeutic esters of acyclovir.

17-21 April 2002, the conference of the European Association for the study of the liver in Madrid, Spain, Siihnel et al., Gilead Sciences, Inc., presented the poster, which shows that the combination of adefovir with β-L-2'-deoxythymidine gives an additive effect against HBVin vitro.

Synthesis of nucleosides

Ways of getting nukes and intermediate furanosyl compounds are well known in the prior art. In 1952 Pratt et al the market reported the synthesis of L-deoxythymidine (LdT) of arabinose (J. W. Pratt et al., J. Am. Chem. Soc., 1952, 74: 2200-2205). The way of synthesis described Pratt, is the formation of methylglucoside from L-arabinose, followed by transformation into methyldithiocarbamate and restoration to detoxifer. Alternative 2-hydroxy-group was converted to the corresponding group nelfinavir, which was then subjected to reductive cleavage to obtain a final product LdT (J. W. Pratt et al., J. Am. Chem. Soc., 1952, 74:2200-2205; H. Urata et al., Nucleic Acids Res., 1992, 20:3325-3332).

Variations synthesis LdT was carried out by Shull et al., Sznaidman et al., Wang et al. and Stick et al., each of them was converted to L-arabinose in methyl-2'-deoxyribofuranosyl through intermediate glycol (B.K. Shull et al., J. Carbohydr. Chem., 1996, 15:955-64; M. L. Sznaidman et al., Nucleosides, Nucleotides &Nucleic Acids, 2002, 21:155-63; Z.X. Wang et al., Nucleosides, Nucleotides &Nucleic Acids, 2001, 20:11-40; and R.V. Stick et al., Aust. J. Chem. 2002, 55:83-85).

In 1969 Niedballa and Vorbruggen describe the process for obtaining β-nucleosides through a combination of similarvideo N-heterocyclic compounds, in particular pyrimidine, 1-O-alkyl - or preferably 1-acyl-protected sugar, such as 1-acyl-protected ribose, deoxyribose, arabinose or glucose. In the reaction used reagent Friedel-as a catalyst, and the reaction proceeded at ambient temperatures (DE 1919307, Schering Aktiengesellschaft). The inventors have noted that this method is unexpectedly gave almost exclusively the β-anomeric is already installed, and could work in the case of uracil and cytosine, but not in the case of thymidine (DE 1919307, examples 1-10 and 12-15).

In my examples Niedballa and Vorbruggen said only about 1-O-acetyl-, 1-acetyl - 1-O-methyl-derivatives of the compounds of ribose, desoxyribose and arabinofuranose as initial reagents (DE 1191307, examples 1-16). Never used 1-halogenide. Moreover, the authors of the inventions noted that the use of 1-halogenalkane as reagent is undesirable because of its instability (DE 1191307; JP 63026183, Sato et al.). In one example, in which casinowe base was subjected to interaction with the sugar 2'-deoxyribose, the reference compound was 1-O-methyl-2-deoxy-3,5-decolourise (DE 1919307, example 7). It is not unexpected that in this reaction formed β-anomer almost complete elimination of the α-anomer, as it is known that 3'-ester derivative of ribose, it is generally preferable to form the β-anomer in comparison with α-anomeric product.

In subsequent patents Vorbruggen et al. referred to his earlier method of synthesis (1969) as "very unfavorable"as the separation of salts of Lewis acids or catalysts for Friedel-formed during the reaction, it was necessary in many labour-intensive stages in the final processing, which gave a lower percentage of the yield of the final product (DE 2508312 equivalent to UK application GB 1542442). In GB 1542442 was reported by the Deputy is not in the way that Lewis acids trimethylsilyloxy esters of inorganic acids and about the original reagents, which was a 1-halogen-, 1-O-alkyl - or 1-O-acyl-sugar. As indicated earlier, in all the illustrative examples used source reagent 1-O-acetyl-β-D-ribofuranose, and therefore, was not unexpected obtained β-anomeric product with almost complete absence of α-anomer (GB 1542442, examples 1-13).

Similarly, in U.S. patent 4209613 Vorbruggen described one-step synthesis of nucleosides, which was in collaboration similarvideo the base of the nucleoside with 1-O-acyl-, 1-O-alkyl - or 1-halogen derivative of a protected sugar in the presence of a catalyst of the Friedel-selected from any of the catalyst from the group of Lewis acids (US 4209613). As indicated earlier, in all the illustrative examples used source reagent 1-O-acetyl-β-D-ribofuranose, and again was not unexpected obtained β-anomeric product with almost complete absence of α-anomer (US 4209613, examples 1-16).

In U.S. patent 5750676, Vorbruggen et al. reported method, which consisted in the interaction of free sugars with N-heterocyclic base in the presence cilleruelo agent and an inert solvent containing a Lewis acid, the improvement consisted in perselisihan sugar free. Was not made clarifications on anomeric ratios of the product, and does not say about the advantage of one Lewis acid. However, p is emery showed what was required multiple stages of processing, to obtain the final products, a clear disadvantage for the production on an industrial scale (US 5750676, examples 1-3).

Another way of obtaining nukes, reported by Vorbruggen et al., was the synthesis in a single vessel using complex trialkylsilyl ester of an inorganic or strong organic acids, the main catalyst for Friedel -, the base of the nucleoside, and 1-O-acyl-, 1-O-alkyl - or 1-halogen-substituted derivative, protected derivative of sugar (US 4209613).

Intermediate product in the form of a chloro-sugar

Chlorine-sugar is an important intermediate product in the formation of LdT and there are numerous methods of its synthesis. Non-limiting examples of the synthesis of chlorine-sugars include the following methods.

Isbell, Bock et al. and Lundt et al. reported the synthesis of LdT from D-xylose by the way, which is included in the intermediate 1,4-lactone (H.S. Isbell, Methods in Carbohydrate Research, 1963, 2:13-14; K. Bock et al., Carbohydrate Research, 1981, 90:17-26; K. Bock et al., Carbohydrate Research, 1982, 104:79-85; and I. Lundt and R. Madsen, Topics in Current Chemistry, 2001, 215:177-191).

Bock et al. and Humphlett was using D-galactose as an initial matter, which was subjected to oxidative cleavage and bromisovali, receiving D-lexikalische. Subsequent phases of the electoral hydrolysis and transformations gave the intermediate chloro-sugar, which then can the be used to obtain LdT (K. Bock et al., Carbohydrate Research, 1981, 90:17-26; K. Bock et al., Carbohydrate Research, 1979, 68:313-319; K. Bock et al., Acta Chem. Scand. B, 1984, 38:555-561; and W.J. Humphlett, Carbohydrate Research, 1967, 4:157-164).

Bock et al. also received LdT from D-gluconolactone by treatment of the latter with an aqueous solution of bromine and hydrazine and then with an excess of an aqueous solution of potassium hydroxide with the formation of primary epoxide. Then they were regrouping Payne primary epoxide in the secondary epoxide on the lactone, and subjected to the lactone oxidative cleavage with formation of an intermediate chlorine-sugar, which then can be used to obtain LdT (K. Bock et al., Carbohydrate Research, 1979, 68:313-316; K. Bock et al., Acta Chem. Scand. B, 1984, 38:555-561). Some referred journal articles Bock et al. he described the formation of a chloro-sugar of D-gluconolactone and bromirovanii D-mannono-1,4-lactone.

Liotta and Hager reported the synthesis of chlorine-sugar from commercially available lactone method of synthesis, which involves the step stereoselective cyclization, as well as the synthesis method, in which use intermediate aldehyde and modified Horner-Emmons Wittig reaction (D. C. Liotta et al., Tetrahedron Letters, 1992, 33:7083-7086; and US 5414078).

Schinazi et al., Ravid et al. and Taniguchi et al. describe the methods of obtaining the intermediate chloro-sugars from hydroxylamino acid, which is subjected to cyclization to the derived ribonolactone, which can then be converted into a chlorine-Sakha is (US 6348587 B1, R. F. Schinazi et al.; U. Ravid et al., Tetrahedron, 1978, 34:1449-1452; and M. Taniguchi et al., Tetrahedron, 1974, 30:3547-3552).

Jung et al. reported the use of epoxidation on he sang sharpless on commercially available alcohol to get epoxide, which is then treated with alcohol, getting diol, which is then converted to the acetonide. Acetonide acidified, getting the desired ribofuranose, which was then turned into a chloro-sugar. Alternative epoxies were subjected to hydroborating using oxidation on the Turn, and chlorine-sugar formed from getaway-derivative (M. E. Jung et al., Tetrahedron Letters, 1998, 39:4615-4618).

Yadav et al. and Harada et al. described the synthesis, when used allylbromide and ozonolysis or 2-methyl bromide[1,3]dioxolane without ozonolysis to obtain chlorine-sugar (J. S. Yadav et al., Tetrahedron Letters, 2002, 43:3837-3839; T. Harada et al., Chem. Lett., 1981, 1109-1110), whereas Ohuri et al., Cheng et al. and Abramski et al. reported processing glikas acidic methanol to obtain 2-deoxyribofuranosyl, which is then transformed into the desired chlorine-sugar.

In JP 09059292 Takeya Mori described the synthesis in a single vessel 4-aminopyrimidines of nucleoside 4-hydroxypyrimidinone nucleoside by protecting the hydroxyl groups of the reagent trimethylsilyl groups, followed by interaction with phosphorus oxychloride or 4-chlorophenyltrichlorosilane and aminating aqueous solution of ammonia.

Chu reported a method of obtaining 2'-deso is dinucleotides, which lies in the interaction of nucleoside having a 2'- and 3'-hydroxyl group, with a mixture of Allbreed or acylchlorides and Hydrobromic or hydrochloric acid at moderate temperatures by obtaining a derived halogenosilanes, whose protection was removed with the formation of the desired nucleoside product (US 5200514).

In Nucleosides and Nucleotides, 1996, 15 (1-3):749-769 Kamaike et al. describe the formation of 2'-deoxyribonucleosides by nucleophilic substitution of 4-azolyl-1-β-D-ribofuranosylpurine-2(1H)it is received by the conversion of uridine with [15N]phthalimide in the presence of triethylamine or DBU, obtaining N4-phthaloyl[4-15N]cytidine with high yields.

In JP 71021872 Sankyo Co. Ltd. presented reaction similarvideo bases cytosine, uracil, thymine or azauracil with sugar halide, such as halogenated ribose or glucose, in the presence of a solvent and halide mercury.

D-xylose

Using D-xylose as an initial matter, it is possible to synthesize 2'-deoxynucleoside according to the methods described in the prior art.

Okabe et al. described the synthesis of 2-deoxy-3,5-di-O-para-toluoyl-α-L-iitrobotrepubblica, which can then be subjected to interaction with obtaining β-L-2'-deoxythymidine (LdT) (Okabe et al., J. Org Chem., 1991, 56(14): 492; Bock et al., Carbohydr. Res., 1981, 90:17-26; Bock et al., Carbohydr. Res., 1982, 104:79-85).

Next follows a non-limiting list of methods used to obtain intermediate products of the synthesis of 2'-deoxynucleosides, and in particular, 2'-deoxythymidine from D-xylose.

Takahata et al. and Graf et al. reported the formation of 2,5-dibromo-2,5-dideoxy-D-lyxo-1,4-lactone as a result of interaction of lyxo-1,4-lactone with potassium iodide in acetone (Takahata et al., J. Org. Chem., 1994, 59:7201-7208; Graf et al., Liebigs Ann. Chem., 1993, 1091-1098).

Lundt et al., Bock et al. and Choi et al. describe the inversion of the 5-bromo-2,5-dideoxy-D-thiopentone-1,4-lactone with the formation of 2-deoxy-L-ribono-1,4-lactone (Lundt et al., Topics in Current Chemistry, 2001, 215:177-191; Bock et al., Carbohydr. Res., 1981, 90:17-26; WO 01/72698, Y-R. Choi et al.).

Urata et al. and Zhang et al. reported the conversion of 2-deoxy-3,5-di-O-toluoyl-α,β-L-ribose, 2-deoxy-3,5-di-O-para-toluoyl-α-L-retroinformation either directly from lactol as a result of interaction with hydrochloric and acetic acids, or indirectly through an intermediate 2-deoxy-7-methoxy-3,5-di-O-toluoyl-α,β-L-ribose as a result of interaction with acetic and hydrochloric acids (H. Urata et al., Nucleic Acids Res., 1992, 20(13):3325-3332; Zhang et al., Nucleosides and Nucleotides, 1999, 18 (11-12):2357).

Urata et al. also described obtaining 2'-deoxy-3',5'-di-O-para-toluoyl-L-thymidine of the 2-deoxy-3,5-di-O-para-toluoyl-α-L-iitrobotrepubblica and similarvideo thymine in the presence of chloroform with sleduyushim removing protection with the formation of 2'-L-deoxythymidine (H. Urata et al., Nucleic Acids Res., 1992, 20 (13):3325).

Intermediate 2,2'-anhydrous-1-paranoilise

It is shown that 2'-deoxy - and 2'-substituted nucleosides, and in particular, 2'-deoxy - or 2'-substituted nucleosides, which are pyrimidine bases, stabilize oligonucleotides from destruction by nucleases. The destruction of nucleases is a problem in the field of oligonucleotide therapeutics (Huryn et al., (1992), Chem. Rev. 92:1745-88; English et al., (1991), Angew. Chem. 30:613-722). To date, however, the modification of pyrimidine nucleosides in the 2'-position was carried out only in hard conditions and methods of synthesis that are inefficient, as a rule, with low yields of product (Verheyden et al., (1971), J. Org. Chem. 36:250-254).

Tronchet et al. described recovery Aksenovo derived from 2'-getorigin using BH3that mainly gives 2'-hydroxy - or 2'-aminonucleoside in arabino-configuration (Tronchet et al., (1990), Tetrahedron Lett. 31:351). This work Tronchet is one of the few attempts stereoselective synthesis of 2'-ribofuranosylpurine - or 2'-ribofuranosylpurine.

Early approaches to the synthesis of 2'-deoxy - or 2'-substituted pyrimidine nucleosides were focused on suitable protective groups for ribose, xylose and arabinose, which were the source reagents in synthesis. For example, there have been many p is dhody to the synthesis parallelomania ribofuranose as an intermediate product in ways to get nukes. Approaches include (i) 7-stage stereospecificity method, which started with D-ribose and gave β-D-2'-deoxyribofuranosyl with the release of the final product is about 40% (M. Jung, and Y. Xu, Tetrahedron Lett. (1997), 38:4199); ii) 3-stage method, starting with L-ribose and resulting yield of 56% (E. F. Recondo and H. Rinderknecht, Helv. Chim. Acta, (1959) 42:1171); (iii) 8-stage method using L-arabinose as a starting material and getting about 20% product yield (J. Du et al., Nucleosides and Nucleotides, (1999), 18:187); iv) 6-stage method, starting from L-xylose (% yield of the final product is not known) (Moyroud, E. and P. Strazewski, Tetrahedron (1999) 55:1277); and (v) multi-stage method, starting with D-ribose, which was first turned into three-O-acetylcytidine (U.S. patent No. 4914233).

In 1959 E. F. Recondo reported 5-stage method of obtaining toluoyl-, benzoyl and acetyl-protected ribofuranosyl with access about 70-80% of D-ribose (E. F. Recondo, Helv. Chim. Acta, (1959) 121:1171). Codington, Doerr and Fox described the synthesis of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine of β-D-thymidine by the interaction of β-D-thymidine with Fritillaria and pyridine for 24 hours at room temperature and then at about 70°C for 3 hours, in order to protect the 5'-OH β-D-thymidine; then through the interaction of the 5'-protected β-D-thymidine with mozillateam (TsCl and pyridine at 0°C, which gave tosyl-protected 2'-is the Rupp; and, finally, through the interaction of the 5'-trityl-O-protected 2'-tosyl-O-protected β-D-thymidine with sodium benzoate (NaOBz) and ndimethylacetamide at 100°C for 1 hour to obtain 2,2'-anhydrous-1-(5'-O-trityl-β-D-arabinofuranosyl)thymine with exit 61% (Codington et al., J. Org. Chem., (1963) 29:558-64).

It was reported enzymatic synthesis of β-D-thymidine usingE. coliand gipoksantina in the first stage and the interaction of the resulting 2-monophosphorylation connection ribofuranosyl with urediniospores and removing the desired product β-D-thymidine with the release of 45% using chromatography on a column (Zinchenko A.I., Chemistry of natural compounds, (1989), 4:587-88).

Another method of synthesis of nucleosides is the formation of intermediate 5-methyl-2,2'-angeborene of "open nuke". Open nucleoside formed by the reaction of an intramolecular nucleophilic substitution, which gives 2,2'-anhydrous-1-(β-D-arabinofuranosyl)nucleoside in the opening cycle 2,2'-anhydroglucose. In the application of Japan Kokai No. 8149398 (posted may 2, 1981) described the synthesis of anhydroglucitol, which requires as an intermediate product of an acid additive salt of the acylated aminoarabinose[1',2':4,5]oxazoline. On the application of available carbohydrate derived aminoquinoline as a predecessor of unhydrolysed with the Bali in 1971 (J. Mol. Biol., (1970) 47:537).

Rao et al. reported 6-stage synthesis, which used D-xylose as a starting reagent, forming a 1-β-D-xylopyranoside, which is then processed PhOCOOPh (diphenylcarbonate) and catalyst NaHCO3in the presence of DMF at 140-150°C for about 4 hours, getting 2,2'-anhydrous-1-(β-arabinofuranosyl)thymine with the release of 55% (A. V. Rama Rao et al., J. Chem. Soc. Comm., (1994), p.1255; EP 0683171 B1). And Schinazi et al. and Manfredi et al. described the synthesis, similar to the synthesis described by Rao et al., which used the same reagents, except for using 1-β-D-arabinofuranosyladenine instead of 1-β-D-xylopyranoside (Schinazi et al., J. Med. Chem., (1979) 22:1273; Manfredi et al., Bioorg. Med. Chem. Letters, (2001) 11:1329-32).

An early attempt formation of 3',5'-Dibenzoyl-protected 2,2'-anhydrous-1-(β-ribofuranosyl)thymine described Anton Holy et al. Holy et al. used β-D-ribofuranoside as the parent compound, he was subjected to interaction with 1.4 equivalent PhOCOOPh and catalyst NaHCO3in HMPA for about 20 minutes at a temperature of about 150°C to form 2,2'-anhydrous-1-(β-D-ribofuranosyl)thymine (5-methyluridine), which was subjected to interaction with PhCOCN in DMF to protect the 3'- and 5'-OH group through the formation of 2,2'-anhydrous-1-(β-3',5'-di-O-benzoyl)ribofuranoside with input about 87% (A. Holy et al., Collect. Czech. Commun., (1974), 39:3157-67). Holy et al. also reported an unsuccessful attempt p is vratiti 2-amino-β-D-arabinofuranose[1',2':4,5]-2-oxazoline in the O 2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine (ibid, 1377).

Fraser et al. improved the Holy way, using the same source reagent and exposing its interaction with 1.2 equivalent PhOCOOPh and catalyst NaHCO3in the presence of HMPA at about 150°C for about 2 hours, getting 2,2'-anhydrous-1-β-D-ribofuranosides. However, the method of Fraser et al. gave reduced the percentage yield of about 77%, compared with a yield of about 87%, obtained by synthesis according to the Holy et al. (Allister Fraser et al., J. Heterocycl. Chem., (1993) 30 (5):1277-88).

Yukio Aoyama et al. he described education silyl-protective cycle, which includes both 3'-and 5'-position of β-1-D-(2-Br-ribofuranosyl)thymine with the release of approximately 96% (Aoyama et al., Nucleosides and Nucleotides, (1996), 15 (1-3):733-8). 1-β-D-ribofuranoside used as the starting material and subjected to interaction with TPDSCl2and pyridine at room temperature, getting the 3', 5'-silyl-protected cyclic structure. Then silyl-protected structure was subjected to interaction with TfCl and DMAP in CH2Cl2at room temperature with the formation of intermediate 2,2'-amidopropyl and, finally, the intermediate 2,2'-amidopropyl were subjected to interaction with LiBr, BF3-OEt in 1,4-dioxane at about 60°C, receiving the final product, 1-β-D-2'-Br,3',5'-tri-O-di(dimethyl)silyl)ribofuranosides.

Mitsui Chemicals, Inc. reported : the Bach obtain 2,2'-anhydrous-1-(β-L-arabinofuranosyl)thymine and 2,2'-anhydrous-5,6-dihydrocyclopenta, which is applicable as intermediates in the synthesis of L-nucleic acids (PCT publication no WO 02/044194; EP 1348712 A1). 7-stage method according to Mitsui includes: (a) the interaction of L-arabinose with cyanamide to obtain L-arabinofuranosyladenine; (b) the interaction of L-arabinofuranosylcytosine derived from acrylic acid, with formation of a derivative of L-arabinofuranosylcytosine containing ester methylacrylate acid associated with the N-atom of the remainder of oxazoline; (c) the interaction of the product from step (b) with the same base, such as alkali metal, alkali metal alkoxide, carbonate, alkali metal bicarbonate, alkali metal, hydroxide of alkali metal, alkali metal hydride, an organic base, basic ion-exchange resin and the like, any of which forms when this tricyclic ring, which is a derivative of L-2,2'-anhydroglucitol acid; (d) isomerization derivative of L-2,2'-anhydroglucitol acid from step (c) to obtain 2,2'-anhydrous-1-(β-L-arabinofuranosyl)thymine; (e) 2,2'-anhydrous-1-(β-L-arabinofuranosyl)thymine from stage (d) expose any galogenirovannyie and subsequent protection, or the protection and subsequent galogenirovannyie or simultaneous galogenirovannyie and protection with the formation of halogenated 2'position derivative of L-thymidine; (f) dihalogenoalkane ha is generowania derivative of L-thymidine from step (e); and g) releasing the 3'- and 5'-positions of the product from step (f) to obtain L-thymidine. Although Mitsui reported good outputs the product as a result of this synthesis, it is desirable to have a method that requires fewer stages, to make it easier to adapt to large-scale industrial production.

The second closest method found in the prior art, is a method, Pfizer reported in EP 0351126 B1. How Pfizer is a new way of education O2,2'-anhydrous-1-(β-D-arabinofuranosyl)siminovich nucleosides (anhydroglucitol), which can be easily converted into a derivative of β-thymine. The method comprises the condensation reaction between 2-amino-β-D-arabinofuranose[1',2':4,5]-2-oxazoline or its 5'-trityl or silyl-protected form preferably methyl-2-formylpyridine in H2O and NaOH at pH 8.1 in for 48 hours at room temperature followed by treatment with an aqueous solution of acid to obtain O2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine with the release of approximately 42%. Alternatives to the use of methyl-2-formylpropyl include the use of methyl-3-Bromeliaceae in the presence of DMAP and Et3N approximately at 80°C for 4 days, which gave approximately 25% of the yield of the final product anhydration; the use of ethyl-2-formylpropyl in aqueous MeOH and Et3N at room temperature for PR is approximately 24 hours and then at about 60°C for 24 hours to exit anhydromannose product, approximately 8%; and the use of methyl-3-methoxynicotinate in DMSO at about 80°C for 4 days with getting anhydromannose product with a yield of approximately 32%.

The condensation reaction by Pfizer includes the use in the preferred embodiment, the main catalysts. Such catalysts are tertiary amines and inorganic salts, and preferred among them are dimethylaminopyridine, triethylamine, N-methylmorpholine and combinations thereof. Pfizer reported that the preferred method of transformation of O2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine in β-thymidine was the reaction of anhydration with HBr, followed by removal of Br as a result of interaction with BaSO4-poisoned Pd-catalyst. It is desirable to obtain industrially scalable synthesis, which would eliminate the necessity of using poisoned catalyst of this type.

Boehringer-Ingelheim Pharma GMBH reported a 4-stage method of obtaining β-L-2'-deoxythymidine to utilize L-arabinose as a starting material (PCT publication no WO 03/087118). The method included: (a) the interaction of L-arabinose with cyanamide in an aqueous or aqueous-alcohol solution or other polar solvent, such as DMF, pyridine or N-methylpyrrolidone, at a temperature of 80-100°C in the presence of a basic catalyst, such as NH3Et3N Il is triethylborane, the basic carbonate or carbonate dibasic, with the formation of a derivative of L-arabinofuranosylcytosine; (b) interaction of a derivative of L-arabinofuranosylcytosine from stage (a) 2-methyl-C-3-acid or its activated derivative in an inert solvent in the conditions of deposition by water, such as in the presence of DMF, DMSO, NMP, acetone, benzene, toluene or cyclohexane and catalyst in the form of a tertiary amine base or an inorganic salt, such DMAP, Et3N or N-methylmorpholine, at about 20-80°C; (c) the interaction of β-L-2,2'-angebotenen from stage (b) with a nucleophilic reagent, such as a halogen acid, like HCl, HI or HBr, toluensulfonate acid or teoksessa acid, the solvent is DMF or triperoxonane acid, to break the link C-O in the 2'-position; and d) the interaction of β-L-2'-gelegenheiten with a catalyst, preferably with either Pd or Raney Nickel to remove the group of halogen from 2'position and obtain β-L-thymidine in the quality of the final product.

Preferably before performing the stages of synthesis (a) or (b) any free hydroxyl group is protected, to prevent their interaction with derivative aminoquinoline or 2-methyl-C-3-acid.

In the specified method of synthesis by Boehringer preferred protective groups include benzyl, WPPT is ylmethyl, triphenylmethyl or silyl, where the three substituents on simile can be a C1-6-alkyl or phenyl, and phenyl groups optionally may be additionally substituted. Any protective groups can be removed at the final stage of the synthesis, and can also be added to the stage crystallization or purification.

Unfortunately, the first stage of the method disclosed Boehringer, required at least two stages of extraction, filtration and crystallization; the second stage of the method required the use of boiling cyclohexane and final purification by chromatography; and the fourth stage of the method required the use of a Pd catalyst or Raney Nickel. The reported output of the intermediate β-L-2,2'-anhydroerythromycin was approximately 49%. Thus, there is a need in the synthesis method, which avoids the use of a Pd catalyst or Raney Nickel, and which provides a higher percent yield intermediate 2,2'-angebotenen.

Holy and Pragnacharyulu et al. describe the use of L-arabinose as a starting substance, which is subjected to interaction with cyanamide, getting derived 1,2-oxazoline; derived oxazoline subjected to interaction with ethyl ether propionic acid, receiving the intermediate O2,2'-anhydrous-L-thymidine, which are benzoylation and recovery R is salenew or treated with hydrogen chloride, getting the required chlorine-sugar. (A. Holy, Coll. Czech. Chem. Commun. 1972, 37, 4072-4087).

Abushanab et al. reported the synthesis of chlorine-sugar, which lies in the interaction of ester methyloxycarbonyl acid oxazoline with intermediate O2,2'-anhydrous-L-thymidine (E. Abushanab and P. V. P Pragnacharyula, U.S. patent 5760208, June 2, 1998), while Asakura et al., Hirota et al. and A. Holy described the reaction ethylpropyl with oxazoline with getting O2,2'-anhydrous-L-uridine, which is then defended by its 3'- and 5'-positions and subjected to interaction with hydrogen chloride, obtaining 2'-deoxy-2'-chloro-sugar as an intermediate product (J.-I. Asakura and M. J. Robins, J. Org. Chem. 1990, 55, 4928-4933; J.-I. Asakura and M. J. Robins, Tetrahedron Lett. 1988, 29, 2855-2858; K. Hirota, Y. Kitade, Y. Kanbe, Y. Isobe and Y. Maki, Synthesis, 1993, 210, 213-215; and A. Holy, Coll. Czech. Chem. Commun. 1972, 37, 4072-4087).

In 2003 Abushanab and Pragnacharyulu reported a method of obtaining pyrimidine nucleosides, which involves the reaction of condensation type reactions Michael, between arabinofuranosylcytosine and substituted derivatives of apoximately; subsequent acylation of the condensed product processing revalorisation to place the group of chlorine in the 2'-position of thymidine; and, finally, dihalogenoalkane to remove chlorine Deputy, if the desired product is 2'-deoxythymidine (U.S. patent No. 6596859).

However, it is known that pualeilani syvaet disclosure of anhydrite and placement of the chlorine group in the 2'-position of thymidine, then require additional synthesis step to remove the chlorine group. It would also be useful to avoid the use of expensive reagent methyl-2-methylpyridine that Abushanab and Pragnacharyulu used in the condensation reaction in his way, as well as the use of acetonitrile used in the second stage of the method, and chromatographic separations that are required at each stage of the synthesis.

Pragnacharyulu et al. also reported the formation of 2,2'-anhydromannitol from L-arabinose by the interaction of L-arabinose with H2NCN, which provides the possibility of intramolecular elimination of one end OH and one H to obtain the intermediate 2,2'-anhydromannitol (Pragnacharyulu et al., (1995), J. Org. Chem. 60:3096-99).

Sawai et al. describe the stage of direct cyclization with the formation of 2,2'-anhydrous(arabinofuranosyl)thymine from D-arabinose. The synthesis of these authors included (1) obtaining D-arabinofuranosylcytosine from D-arabinose by methods known in the field; (2) interaction of D-arabinofuranosylcytosine with ethyl-α-(methyl bromide)acrylate in dimethylacetamide to obtain oxazoline-N-branched intermediate product with a yield of about 88%; and (3) the interaction of the intermediate product formed in stage (2), with KOtBu and tert-BuOH to obtain 2,2'-anhydrous(arabinofuranosyl)thymine with the release of about 30, or alternatively, the use of hydrogen iodide to disclose communications O2,2'-anhydrous-L-thymidine and then the interaction of the acyclic product with potassium iodide, to obtain di-O-benzoyl-2'-deoxythymidine (Sawai et al., (1994), Nucleosides and Nucleotides, 13 (6-7):1647-54; Sawai et al., Chem. Lett., 1994, 605-606). In this way advantageously avoid the use of catalysts, such poisoned Pd/BaSO4but the result is more low yields in %.

In U.S. patent No. 4914233 Freskos et al. described selective isolation of β-thymidine from a mixture of α - and β-anomers by the 5-stage method, which consists of formation of tri-O-acyl-β-ribothymidine and the transformation of 2,2'-anhydrous-β-thymidine 2'-halogen-2'-deoxy-5-methyluridine with subsequent transformation in the last β-thymidine.

In U.S. patent No. 5212293 Green et al. reported the synthesis of 2',3'-dideoxynucleosides by the interaction of protected anhydration with halogenerator agent, which contained alumoorganic connection for increased solubility of the reagent.

In U.S. patent No. 5596087 Alla et al. included the formation of 2,2'-angebotenen, which was subjected to bromirovanii and then recovered by methods known to experts in this field, to obtain β-thymidine.

In U.S. patent No. 6369040 Acevedo et al. described 3',5'-protected 2,2'-anydroreideon for the synthesis of the corresponding ar is Inositol.

McGee and Murtiashaw, each reported receiving intermediate chloro-sugar L-arabinose as a starting material, which includes the formation of an intermediate O2,2'-anhydrous-L-thymidine was obtained from other compounds, reagents, other than compounds used or Holy Pragnacharyulu et al. (D. McGee, Boehringer Ingelheim Proposal to Novirio Pharmaceuticals, Inc., May 17, 2002; C. W. Murtiashaw, Eur. Patent, 0351126 B1, January 18, 1995).

McGee et al. described is a method of obtaining 2'-modified nucleosides in the reaction of intramolecular substitution (U.S. patent No. 6090932). McGee et al. announced the introduction of a substituent in the 2'-position 2,2'-angeborene with careful selection of the 3'-substituent, which can be activated, causing stereospetsifichno recovery in the 2'-position. The synthesis included the protection of the 5'-OH uridine via interaction with DMT with formation of 5'-O-(4,4'-dimethoxytrityl)uridine and gave the final product, 2'-deoxythymidin, with output of about 24%.

Although McGee et al. reported that their method can be scaled for industrial purposes, it is known that dioxane is highly flammable and prone to the formation of peroxide, and therefore is contraindicated for industrial purposes. In addition, McGee et al. silent about the fact whether their method D - or L-enantiomer of 2'-deoxythymidine, or require the separation of enantiomers.

Thus, there is a need for simple, cost effective the om and safe method of obtaining 2'-deoxynucleosides, their salts, analogs and prodrugs, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin, which makes it possible to avoid the use of dangerous, toxic, hazardous and/or trudnoponimaemyh processing reagents, which themselves are not suited for industrial production.

There is also a need in providing a synthesis for the preparation of 2'-deoxynucleosides, their salts, analogs and prodrugs, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin with using safe materials and reagents.

There is also a need in providing a synthesis for the preparation of 2'-deoxynucleosides, their salts, analogs and prodrugs, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin, under mild conditions the reactions.

There is also a need for efficient and cost-effective method for the synthesis of 2'-deoxynucleosides, their salts, analogs and prodrugs, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin, moderate reaction conditions.

There is also a need for synthesis, which is efficient, requiring a minimum number of stages.

There is also a need in the way that requires little or no stages requires separation of the products.

There is also a need to provide prom the industrial scalable method for the synthesis of 2'-deoxynucleosides, their salts, analogs and prodrugs, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin, which is cost effective and gives a final product with high yield.

There is also a need to provide industrial scale synthesis of β-2'-deoxynucleosides, their salts, analogs and prodrugs, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin, which gives the β-anomeric form the desired connection in excess compared to the α-anomeric form with good outputs.

There is also a need in providing a synthesis of amino acid prodrugs of 2'-deoxynucleosides, their salts and analogues, including β-L-2'-deoxynucleoside, such as β-L-2'-deoxythymidin.

The invention

The present invention discloses new and effective methods of synthesis for the preparation of 2'-, 3'- and/or 5'-substituted nucleosides and 2'-, 3'- and/or 5'-deoxynucleosides, such as 2'-substituted 2'-deoxynucleoside derived from natural and non-natural carbocyclic, heterocyclic and heteroaromatic nucleoside bases, and, in particular, β-L-2'-deoxythymidine (LdT) and its salts, prodrugs, stereoisomers and enantiomers. Also suggests ways of obtaining stereoisomeric, diastereoisomeric and enantiomeric forms of the compounds according to the present invention on the basis correspond to their original substances. The compounds obtained according to the present invention, can be used as intermediates in obtaining a wide range of other nucleoside analogues, or can be used directly as antiviral and/or antineoplastic funds.

In one embodiment, the 2'-deoxynucleosides and 2'-substituted nucleosides are naturally occurring pyrimidine nucleoside base. In a specific embodiment, the method is directed to the synthesis of β-L-2'-deoxythymidine (LdT). In another embodiment, 2'-deoxynucleosides and 2'-substituted nucleosides are not found in nature like a pyrimidine nucleoside base. In one particular variant not found in nature like a pyrimidine nucleoside base can be obtained by the synthesis method, disclosed in the present invention.

In one embodiment, the method according to the present invention does not require the separation of isomers and therefore is an improvement over the prior art.

In one embodiment, the introduction of functional groups at the 2'-position or the removal of such functional groups, to obtain 2'-deoxynucleoside, carried out through selective reactions using D-xylose, L-arabinose, L-ribose, D-galactose, D-gluconolactone, D-galactarate, D-glucose, D-hydroxyle the amino acid (for ribonolactone), alcohol or epoxied, isopropylbenzaldehyde or substituted dioxolane as a starting reagent.

In one specific embodiment of the invention the synthesis flow through the intermediate chloro-sugar. Thus, one specific intermediate product in the synthesis methods described in this description that does not contain intramolecular rearrangements, is the connection chloro-sugar.

In another specific embodiment of the invention the synthesis proceeds via an intramolecular nucleophilic substitution. Thus, one specific intermediate product of the synthesis methods described in this description is 2,2'-anhydrous-1-forensicmedicine ring.

In one embodiment of the invention one of the key intermediates get through recovery lactone such a reducing agent as Red-Al, as follows:

In one particular embodiment, the protective group of oxygen is toluol.

In another specific embodiment, the intermediate product is obtained in the following way:

Thus, in the embodiment of the present invention the method of synthesis involves the following stages:

Alternative synth is C according to the present invention for obtaining 2'-deoxythymidine includes the following stages of the method:

In yet another embodiment, the present invention proposes a method of obtaining 2'-deoxythymidine from D-xylose, which includes 2-deoxy-3,5-di-O-para-toluoyl-α-L-retroinformation as a key intermediate product.

In an alternative variant synthesis using mutilator intermediate product:

where P, P' and P” are independently mean H, alkyl or a suitable protective group oxygen. In one embodiment, P is methyl. In another embodiment, P' and P” combined with the formation of isopropylidene.

Thus, in one particular variant synthesis using mutilator intermediate product:

Alternatively, one of the key intermediates receive the following way:

Alternatively, one of the key intermediates receive CIS-alkene oxidation using a suitable oxidizing agent, capable of CIS-oxidation, such as OsO4, in the following way:

Thus, in one specific embodiment, the key intermediate product is obtained CIS-alkene oxidation using OsO4in the following way:

Alternatively, one of the key intermediates receive the following way:

Alternatively, one of the key intermediates receive the following way:

Alternatively, one of the key intermediates get through reaction with alcohol/acid one of the following ways:

where R is alkyl, preferably lower alkyl, such as methyl or ethyl and in particular methyl.

In one embodiment of the invention the alcohol is selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, Isobutanol, tert-butanol, s-butanol, pentanol, hexanol or mixtures thereof. In a particular embodiment, the alcohol is methanol or ethanol. In another specific embodiment, the alcohol is methanol.

Thus, in a particular embodiment of the invention, a key intermediate product is obtained by reaction with a solution of the alcohol/acid one of the following ways:

Another typical method according to the present invention is to use the reset is novices, such as Red-Al, in combination with a complexing agent such as 15-crown-5-ether, to cleave the cyclic intermediate 2,2'-anhydrous-1-paranoilise and get the desired nucleoside product.

Unexpectedly found that the use of complexing agent, such as 15-crown-5-ether, gives a higher yield in percent in the case when the selected protecting group is dimethoxytrityl, but a lower yield in percent in the case when the protective group used only trail. Therefore, in one embodiment, the invention features a method that includes a step of splitting the cyclic intermediate 2,2'-anhydrous-1-pyranoindoleacetic with the formation of the desired nucleoside product in the absence of a complexing agent. In a specific embodiment, the present invention features a method that includes a step of splitting the cyclic intermediate 2,2'-anhydrous-1-pyranoindoleacetic with the formation of the desired nucleoside product in the absence of complexing agent in the case, when the protective group is trityl.

Suggests ways of applying the appropriate nucleophilic agent, for example, ORGANOMETALLIC compounds (e.g., Grignard reagent or alkyllithium reagent), if you want Alkalyn is the first Deputy, to cleave the cyclic intermediate 2,2'-anhydrous-1-paranoilise with obtaining the desired 2'-substituted nucleoside product.

In one embodiment, the present invention relates to a method for producing 2'-deoxynucleoside or 2'-modified nucleoside, which includes: (a) do not necessarily protect one or more hydroxyl groups furanosyl rings, such as RIBO-, arabino or xylofuranosyl, through interaction with a protective group; (b) condensation furanosyl rings from step (a) optionally substituted natural or non-natural nucleoside base with the formation of the nucleoside; (c) interaction of nucleoside from step (b) with a condensing agent at an elevated temperature to obtain 2,2'-anhydrous-1-pyranoindoleacetic; (d) the interaction of 2,2'-anhydrous-1-pyranoindoleacetic from stage (c) with a reducing agent, such as Red-Al, and a complexing agent such as 15-crown-5-ether, preferably in a polar solvent at a low temperature, to obtain optionally protected 2'-deoxynucleoside or 2'-substituted nucleoside; and (e) removing the protecting optionally protected hydroxyl groups, if necessary or desirable, for example, by adding acid or an acidic resin at a temperature of about 50°C.

In another variant ways is obtaining 2'-deoxythymidine, which includes: (a) do not necessarily protect one or more hydroxyl groups furanosyl ring by interacting with a protective group; (b) the interaction is not necessarily protected furanosyl ring with cyanamide with education is not necessarily protected furanocoumarin; (c) the interaction is not necessarily protected furanocoumarin with collisuem or condensing agent to obtain optional protected 2,2'-anhydrous-1-pornositeleri; (d) the interaction is not necessarily protected 2,2'-anhydrous-1-forensicmedicine with a reducing agent, such as Red-Al, and a complexing agent such as 15-crown-5-ether, preferably in a polar solvent at a low temperature with getting optionally protected 2'-deoxythymidine; and (e) removing the protecting optionally protected 2'-deoxythymidine, if necessary or desirable, for example, by reaction with acids or acidic resin at about 50°C to obtain 2'-deoxythymidine.

In yet another embodiment, the present invention relates to a method for producing 2'-deoxythymidine, which includes stages (a)to(e)above, but does not include the use of complexing agent that is indicated for the stage (d).

In yet another embodiment, the present invention relates to a method of receiving the Oia 2'-deoxynucleoside or 2'-modified nucleoside, which includes: (a) do not necessarily protect one or more hydroxyl groups furanosyl rings, such as RIBO-, arabino or xylofuranosyl, through interaction with a protective group; (b) condensation furanosyl rings from step (a) optionally substituted natural or non-natural nucleoside base with the formation of the nucleoside; (c) interaction of nucleoside from step (b) with a condensing agent at an elevated temperature to obtain 2,2'-anhydrous-1-paranoilise; (d) the interaction of 2,2'-anhydrous-1-pyranoindoleacetic from stage (c) with a reducing agent, such as Red-A1, in the absence of a complexing agent, such as 15-crown-5-ether, preferably in a polar solvent at a low temperature with getting optionally protected 2'-deoxynucleoside or 2'-substituted nucleoside; and (e) removing the protecting optionally protected hydroxyl groups, if necessary or desirable, for example, by adding acid or an acidic resin at a temperature of about 50°C.

In another embodiment proposes a method of obtaining 2'-deoxythymidine, which includes: (a) do not necessarily protect one or more hydroxyl groups furanosyl ring by interacting with a protective group; (b) the interaction is not necessarily protected furanosyl ring with what yanagida education is not necessarily protected furanocoumarin; (c) the interaction is not necessarily protected furanocoumarin with collisuem or condensing agent to obtain optional protected 2,2'-anhydrous-1-pornositeleri; (d) the interaction is not necessarily protected 2,2'-anhydrous-1-forensicmedicine with a reducing agent, such as Red-Al, in the absence of a complexing agent, such as 15-crown-5-ether, preferably in a polar solvent at a low temperature, to obtain optionally protected 2'-deoxythymidine; and (e) removing the protecting optionally protected 2'-deoxythymidine, if necessary or desirable, for example, by reaction with acids or acidic resin at about 50°C to obtain 2'-deoxythymidine.

In the scope of the present invention are included the methods of obtaining 2'-modified nucleosides, phosphoramidite 2'-modified nucleosides, 3'- and 5'-mono-, di - and triphosphates of 2'-modified nucleosides and oligonucleotides that contain at least one nucleotide modified according to the method proposed in the present invention. Also included are methods of obtaining intramolecular functional groups, which include anhydroglucose in other positions other than 2'-position cycle furanose, for example, 3'- and/or 5'-position. The methods according to the present invention shall also include the modification of functional groups to obtain, for example, the corresponding 5'-diacylglyceride or 5'-dialkyldithiophosphate derivatives which can be used as prodrugs.

In the description and examples given in this description, additional variants of the present invention.

Brief description of schemes

Figure 1 is a diagram of a method according to the present invention for obtaining LdT from L-arabinose via intermediate mesilate.

Figure 2 is a diagram of a method according to the present invention for obtaining LdT from L-arabinose via an intermediate glycol.

Figure 3 is a diagram of a method according to the present invention obtain the LdT from L-arabinose via an intermediate glycol and recovery stage of elimination.

Figure 4 is a diagram of a method according to the present invention obtain the LdT from L-xylose via di-O-toluoyl-derived.

Figure 5 is a diagram of a method according to the present invention obtain the LdT from D-galactose.

6 is a diagram of a method according to the present invention obtain the LdT from D-gluconolactone.

Fig.7 is a diagram of a method according to the present invention obtain the LdT of D-galactorrhea.

Fig is a diagram of a method according to the present invention obtain the LdT from pornolation, euglenozoa achiral original substance.

Fig.9 is a diagram of a method according to the present invention obtain the LdT of ethyl-3,3-diethoxypropionate.

Figure 10 is a diagram of a method according to the present invention obtain the LdT from hydroxylamino acid.

11 is a diagram of a method according to the present invention obtain the LdT of the commercially available alcohol by epoxidation.

Fig is a diagram of a method according to the present invention obtain the LdT from apocopate.

Fig is a diagram of a method according to the present invention obtain the LdT of 1,2-O-isopropylidene-L-glyceraldehyde.

Fig is a diagram of a method according to the present invention obtain the LdT of 2-methyl bromide[1,3]dioxolane.

Fig is a diagram of a method according to the present invention obtain the LdT from glikas treated with acidic methanol.

Fig is a diagram of a method according to the present invention obtain the LdT from L-arabinose and cyanamide.

Fig is a diagram of a method according to the present invention obtain the LdT from L-arabinose by disclosing hydrogen chloride O2,2'communication connection.

Fig is a diagram of a method according to the present invention obtain the LdT from L-arabinose, as Fig, using alternative reagents for disclosure O2,2'communication connection.

Fig is a diagram of the of the procedure according to the present invention obtain the LdT from L-arabinose, as Fig, using hydrogen iodide for disclosure O2,2'communication connection.

Fig is a diagram of a method according to the present invention obtain the LdT from L-arabinose, which includes the interaction of ester 2-methyloxiran-2-carboxylic acid 1,2-oxazoline.

Fig is a diagram of a method according to the present invention obtain the LdT from L-arabinose via intermediate O2,2'-anhydrous-L-uridine.

Fig is a diagram of a method according to the present invention obtain the LdT from L-arabinose, as Fig passing through the intermediate 2'-deoxy-5-ethoxymethyl-L-uridine.

Fig is a diagram of a method according to the present invention obtain the LdT from D-xylose via the intermediate 2-deoxy-3,5-di-O-para-toluoyl-α-L-retroinformation.

Fig is a diagram of a method according to the present invention obtain β-L-deoxythymidine, in which the 5'-OH intermediate arabinofuranosylcytosine protect a group of trityl before formation of an intermediate 2,2'-anhydrous-1-(β-arabinofuranosyl)thymidine and its reductive cleavage using Red-Al and 15-crown-5-ether.

Fig is a diagram of a method according to the present invention obtain β-L-deoxythymidine, where protection of the 5'-OH residue of L-arabinofuranosyl occurs after the formation of the intermediate 2,2'-is gidro-1-(β-arabinofuranosyl)thymidine and its reductive cleavage of Red-Al and 15-crown-5-ether.

Fig is a diagram of a method according to the present invention obtain β-D-deoxythymidine of D-ribose, which includes the protection and destruction of the protection of the OH-protective group in the 2'-, 3'- and 5'-positions ribofuranosyl, and then use trityl as a protective group at the 5'-position separately before reductive splitting of Red-Al and 15-crown-5-ether.

Fig is a diagram of a method according to the present invention, in which the intermediate 2,2'-anhydrous-1-(β-ribofuranosyl)thymidine form directly from thymidine, then protect the 5'-OH trailvoy group and, finally, break down, using Red-Al and 15-crown-5-ether.

Fig is a diagram of a method according to the present invention, in which use L-ribose as the initial substance and which is realized by means of protection and removal to protect the hydroxyl groups of any suitable protecting group prior to the formation of an intermediate 2,2'-anhydrous-1-(β-ribofuranosyl)thymidine, which then protects the position of the 5'-OH before reductive splitting of Red-Al and 15-crown-5-ether.

Fig is a diagram of a method according to the present invention obtain β-D-deoxythymidine of 2,2'-anhydrous-1-β-D-arabinofuranosyladenine without the use of complexing agent during recovery.

Detailed description of the invention

Now the image is giving reveals new effective methods of synthesis for the preparation of 2'-, 3'- and/or 5'-substituted nucleosides and 2'-, 3'- and/or 5'-deoxynucleosides, such as 2'-substituted 2'-deoxynucleoside derived from natural and non-natural carbocyclic, heterocyclic and heteroaromatic nucleoside bases, and, in particular, β-L-2'-deoxythymidine (LdT), and their salts, prodrugs, stereoisomers and enantiomers. In the invention included methods of obtaining stereoisomeric, diastereoisomeric and enantiomeric forms of the compounds according to the present invention on the basis of appropriate starting materials. The compounds obtained according to the present invention, can be used as intermediates in obtaining a wide range of other nucleoside analogues, or can be used directly as antiviral and/or antineoplastic funds.

In one embodiment, the 2'-deoxynucleosides and 2'-substituted nucleosides are naturally occurring pyrimidine nucleoside base. In a specific embodiment, the method is directed to the synthesis of β-L-2'-deoxythymidine (LdT). In another embodiment, 2'-deoxynucleosides and 2'-substituted nucleosides are not found in nature like a pyrimidine nucleoside base. In one particular variant not found in nature like a pyrimidine nucleoside base can be obtained by the synthesis method, disclosed in the present the present invention.

In one embodiment, the method according to the present invention does not require the separation of isomers and therefore is an improvement over the prior art.

In one embodiment, the introduction of functional groups at the 2'-position or the removal of such functional groups, to obtain 2'-deoxynucleoside, carried out through selective reactions using D-xylose, L-arabinose, L-ribose, D-galactose, D-gluconolactone, D-galactarate, D-glucose, D-hydroxylamino acid (for ribonolactone), alcohol or epoxied, isopropylbenzaldehyde or substituted dioxolane as a starting reagent.

In one specific embodiment of the invention the synthesis flow through the intermediate chloro-sugar. Therefore, one specific intermediate product in the synthesis methods described in this description that does not contain intramolecular rearrangements, is the connection chloro-sugar.

In another specific embodiment of the invention the synthesis proceeds via an intramolecular nucleophilic substitution. Therefore, one specific intermediate product of the synthesis methods described in this description is 2,2'-anhydrous-1-pyranoindoleacetic cycle.

In the first variant 2'-deoxythymidin derived from D-xylose as a starting material (figure 4). Specified Sint is C includes: (a) oxidation of D-xylose first aqueous solution of bromine, and then acetic acid and Hydrobromic acid with the formation of 2,5-dibromo-2,5-dideoxy-D-leksono-1,4-lactone (2); (b) interaction of the lactone ring of the product from step (a) with potassium iodide in triperoxonane acid (TN)to obtain the corresponding 5-iodine-compound with selective removal of the bromine atom at C-2 to obtain 5-iodine-2-deoxyactein (3); (c) effect on 5-iodine-2-detoxication aqueous solution of potassium hydroxide to obtain 4,5-epoxy derivative (4); (d) processing 4,5-epoxy derivative with an aqueous solution of acid to obtain the corresponding 2-deoxy-L-ribonolactone through stereospetsifichno inversion at C-4 (5); (e) protection of the C-3 and C-5 by interacting with any protective group, such as trouillard in TEA (6); (f) selective recovery of a protected 2-deoxy-L-ribonolactone reducing agent Red-Al to obtain the corresponding lactol (7); and (g) the transformation lactol from the stage (f) in the desired intermediate chloro-sugar (9).

In the second variant, an alternative synthesis for the preparation of 2'-deoxythymidine, which also use D-xylose as a starting substance, using alternative reagents and preferably eliminate three chromatographic purification using a highly polar water-soluble UV-inactive reagents (Fig). The method includes:(a) oxidation of D-xylose first bromine/water and potassium carbonate to obtain D-leksono-1,4-lactone (2); (b) interaction of the lactone from step (a) with acetic acid and Hydrobromic acid, for example, at 45°C for 1 hour and then at room temperature with stirring for approximately 1.5 hours, to obtain 2,5-dibromo-2,5-dideoxy-D-leksono-1,4-lactone (3); (c) the interaction of the lactone from step (b) with isopropylacetate and sodium iodide in TN, and, for example, heating the reaction mixture to about 85°C for about 1.5 hours with the formation of 5-bromo-2.5 dideoxy-D-thiopentone-1,4-lactone (4); (d) the interaction of the lactone from step (c) with potassium hydroxide and water and, for example, after 3 hours of heating the reaction mixture to about 80°C for 30 minutes, then cooling the mixture to room temperature with stirring overnight to obtain 2-deoxy-L-ribono-1,4-lactone (5); (e) adding toluylene protective groups to C-3 and C-5 through interaction of the lactone from step (d) with para-trouillard, for example with pyridine in DME (6); (f) the interaction of 2-deoxy-3,5-di-O-para-toluoyl-L-ribono-1,4-lactone with DIBAL and, for example, DME at about -60°C for approximately 1 hour to obtain 2-deoxy-3,5-di-O-para-toluoyl-L-ribose (7); (g) the interaction of the product from step (f) with dry HCl gas in acetic acid obtaining 2-deoxy-3,5-di-O-para-toluoyl-α-L-iitrobotrepubblica (8), which can then be subjected to interaction of ways, known and experts in this field, to obtain 2'-deoxythymidin as the final desired product.

In some embodiments, L-arabinose is used as the starting material to obtain 2'-deoxynucleosides, in particular 2'-deoxythymidine. These methods include the steps: (a) the conversion of L-arabinose in the appropriate methylglucoside, while protecting the hydroxyl group at C-3 and C-4 in the form of derivatives acetonide (2); (b) deoksigenirovanii hydroxyl group of C-2 by turning it in the appropriate mesilate group (3); and then (c) subjecting the intermediate mesilate reductive cleavage (5)using two additional stages of the way to get the key intermediate chloro-sugar (figure 1).

Alternative L-arabinose can be converted into the corresponding derived glikas through the recovery stage of elimination, see, for example, 2 and 3, stage (1) and (2) respectively, and the resulting intermediate glycol can then be converted into methyl-2-deoxyribofuranosyl, stage (4) and (5), respectively.

In other embodiments of the present invention as the initial substance use L-arabinose. Such methods include the steps: (a) the interaction of L-arabinose with cyanamide to obtain the intermediate 1,2-oxazoline (1), (b) interaction of the intermediate product from step (a) with lonefire derived 3-oxopropanoic acid or ethylpropylamine to obtain 2,2'-anhydrous-1-forensicmedicine cycle (2) and (c) the disclosure of the cycle from step (b) with the use of various reagents in different conditions the reaction to obtain LdT (Fig-22).

Alternative 2'-deoxynucleoside can also be formed from galactose as the starting material. In the case when the initial substance use D-galactose, it is subjected to oxidative cleavage and bromirovanii to obtain 2,5-dibromo-2,5-dideoxy-D-leksono-1,4-lactone, and the resulting lactone is subjected to selective hydrogenolysis, receiving 5-bromo-2-detoxication, which undergoes a series of transformations to obtain the key intermediate chloro-sugar (figure 5).

Similarly gluconolactone can serve as starting compounds for the synthesis of 2'-deoxynucleotides. Gluconolactone converted into 2,6-dibromo-2,6-dideoxy-D-mannono-1,4-lactone (1), successively treated with hydrazine and aqueous potassium hydroxide solution, acidified, causing inversion in position C-4 and C-5, which gives 2-deoxyactein (6), is subjected to rearrangement of epoxide Payne (5), is subjected to oxidative cleavage and recovery, getting lactone (7), which can be easily converted into the desired chlorine-sugar (11) (6).

Alternative 2'-deoxynucleoside can also be formed from galactarate as educt. In the case when the initial substance use Galaktionovich, it turned into acetylated Gibraltar (2), about abayat hydrazine and bromilow, getting 2-deoxyactein (3), which then deacetylase, is subjected to oxidative cleavage and restore using NaBH4getting 2-deoxy-L-ribono-1,4-lactone (5), and the obtained lactone protect via interaction with trouillard, is subjected to the recovery of Red-Al and chlorination, getting the final desired product chloro-sugar (9) (Fig.7).

The present invention also relates to additional methods of obtaining 2'-deoxynucleosides, and in particular, 2'-deoxythymidine from the original substances that are not carbohydrates (Fig)are DIOXOLANYL-derivative (Fig), acids, esters and aldehydes (figures 9, 10, 13), picalm (Fig) and alcohols (11 and 12). The details of these syntheses can be found in the examples in this manual are the preferred option (see figure 1-23).

Also suggests ways of applying a reducing agent, such as Red-Al, in combination with a complexing agent such as 15-crown-5-ether, in order to cleave the cyclic intermediate 2,2'-anhydrous-1-paranoilise and to obtain the desired 2'-deoxynucleosides product. Alternative ways of applying a reducing agent, such as Red-Al, in the absence of complexing agent to cleave the cyclic intermediate 2,2'-anhydrous-1-paranoilise in order to obtain the desired 2'-deoxynucleosides product. Alternative cyclic intermediate 2,3'-anhydrous-1-paranoilise can be used for the formation of the corresponding 3'-deoxynucleoside.

You can use any of the reducing agents known in the field, which provide the necessary chemoselective and regioselective recovery. Suitable reducing agents include Red-Al, Red-Al (bis[2-methoxyethoxy]alumoweld sodium), NaHTe, SmI2H2+Pd-phosphine catalyst and LiAl(OtBu)3H (tri-tert-butoxylated lithium).

The reaction disclosure cycle can be performed at any temperature at which achieved the desired results, for example, which is suitable for the reaction proceeded at a reasonable rate, not stimulating the collapse or excessive formation of by-products, preferably at low temperatures, such as about 0-5°C.

Can be selected any solvent in the reaction, which can reach the required temperature and which can dissolve the components of the reaction. Non-limiting examples, any polar aprotic solvent including, but not limited specified, dichloromethane (DHM) or dichloroethane, acetone, ethyl acetate, dithiane, THF, 1,2-dimethoxyethane (DME), dioxane, acetonitrile, diethyl ether, pyridine, dimethylformamide (DMF), dimethylsulfoxide (DMSO), dimethylacetamide or Liu is th combination of them although preferably THF and/or DME.

Alternative ways of applying a suitable nucleophilic agent, for example, ORGANOMETALLIC agent (e.g., Grignard reagent or alkyllithium reagent), if you want alkyl substituent to disclose cycle intermediate 2,2'-anhydrous-1-pyranoindoleacetic with obtaining the desired 2'-substituted nucleoside product. In another embodiment, the cyclic intermediate 2,3'-anhydrous-1-paranoilise or cyclic intermediate 2,5'-anhydrous-1-paranoilise can be used for formation of the desired 3'-substituted or 5'-substituted nucleoside product.

In particular, in one embodiment, the present invention relates to a method for producing 2'-deoxynucleoside or 2'-modified nucleoside, which includes: (a) do not necessarily protect one or more hydroxyl groups furanosyl rings, such as the cycle of RIBO-, arabino or xylofuranosyl, through interaction with a protective group (2); (b) condensation not necessarily protected furanosyl rings from step (a) optionally substituted natural or non-natural nucleoside base with the formation of nucleoside (3); (c) interaction of nucleoside from step (b) with a condensing agent at elevated temperatures to obtain 2,2'-anhydrous-1-furanosyl is leased (5); (d) interaction of 2,2'-anhydrous-1-pyranoindoleacetic from stage (c) with a reducing agent, such as Red-Al, and a complexing agent such as 15-crown-5-ether, preferably in a polar solvent at a low temperature, to obtain optionally protected 2'-deoxynucleoside or 2'-substituted nucleoside (8); and (e) removing the protecting optionally protected hydroxyl groups, if necessary or desirable, for example, by adding acid or an acidic resin at a temperature of about 50°C (9) (Fig).

In another embodiment proposes a method of obtaining 2'-deoxythymidine, which includes: (a) do not necessarily protect one or more hydroxyl groups furanosyl ring by interacting with a protective group (2); (b) the interaction is not necessarily protected furanosyl ring with cyanamide with education is not necessarily protected furanocoumarin (3); (c) the interaction is not necessarily protected furanocoumarin with collisuem or condensing agent to obtain optional protected 2,2'-anhydrous-1-furosimide (5); (d) the interaction is not necessarily protected 2,2'-anhydrous-1-forensicmedicine with a reducing agent, such as Red-Al, and a complexing agent such as 15-crown-5-ether, preferably in a polar solvent at low temperatures getting optionally protected 2'-deoxythymidine (8); and (e) removing the protecting optionally protected 2'-deoxythymidine, if necessary or desirable, for example, by reaction with acids or acidic resin at about 50°C to obtain 2'-deoxythymidine (9) (Fig).

In yet another embodiment, the present invention relates to a method for producing 2'-deoxynucleoside or 2'-modified nucleoside, which includes (a) condensation furanosyl ring optionally substituted natural or non-natural nucleoside base with the formation of the nucleoside; (b) interaction of nucleoside from step (a) with a condensing agent at an elevated temperature, receiving 2,2'-anhydrous-1-paranoilise (1); (c) the interaction of 2,2'-anhydrous-1-pyranoindoleacetic from stage (b) with a protecting agent such as triticina protective group, to protect 5'-position of the nucleoside (2); (d) adding a reducing agent, such as Red-Al, preferably in a polar solvent at a low temperature, to obtain optionally protected 2'-deoxynucleoside or 2'-substituted nucleoside (3); and (e) removing the protecting optionally protected hydroxyl groups, if necessary or desirable, for example, by adding acid or an acidic resin at a temperature of about 50°C (4) (Fig).

The preferred options are shown in figure 1-29.

Definitions

In the present invention, the term "isolated" refers to a nucleoside composition that includes at least 85 or 90 wt.%, preferably 95-98 wt.% and even more preferably 99-100 wt.% nucleoside, while the remaining part contains other chemical molecules or enantiomers.

The term "protected" as used in this description of the meaning, and unless otherwise stated, refers to a group, which is added to the oxygen atom, nitrogen or phosphorus, to prevent its further interaction or for other purposes. Specialists in the field of organic synthesis known to a wide variety of protective groups of oxygen, nitrogen and phosphorus.

Examples of suitable protective groups include, but are not limited to, benzoyl; substituted or unsubstituted alkyl groups, substituted or unsubstituted aryl group, a substituted or unsubstituted silyl group; a substituted or unsubstituted aromatic or aliphatic esters, such as aromatic groups such as benzoyl, toluoyl (for example, para-toluoyl), nitrobenzoyl, chlorbenzoyl; ether groups such as-C-O-aralkyl, -C-O-alkyl or-C-O-aryl; and aliphatic groups, such acyl or acetyl groups including any substituted or unsubstituted aromatic or aliphatic acyl, -(C=O)-aralkyl, -(C=O)-alkyl is whether -(C=O)-aryl; where the aromatic or aliphatic residue acyl group may be a straight or branched chain; all groups can also be optionally substituted by groups which do not affect the reaction, including improved synthesis (see Greene et al., Protective Groups in Organic Synthesis, John Wiley and Sons, 2ndEdition (1991)). For example, in one embodiment of the invention the protective group substituted by the groups are not affected by the selected reducing agent, such as Red-Al. In the case of use as a protective group of esters attention has been paid to the U.S. patent 6229008 Saischek et al., included in this description by reference, which says that the use of a simple ester as a protective group can provide significant benefits, especially in the 5'-position of pentofuranose, in relation to the stability of the reagents and process conditions. This gives the advantage for the separation, isolation and purification of the desired product and, therefore, in the interest of product yield.

Protective groups of hydroxyl sugar as non-limiting examples can be silyl, benzoyl, para-toluoyl, para-nitrobenzyl, para-chlorobenzoyl, acyl, acetyl, -(C=O)-alkyl, and -(C=O)-aryl, all of which can be unsubstituted or substituted by one or more groups are not affected by the selected reducing agent. In one embodiment, the protecting group hydro is the power of sugar is benzoyl. Protective groups of the amino acids are preferably BOC (butoxycarbonyl), -(C=O)-aralkyl, -(C=O)-alkyl or -(C=O)-aryl. In one embodiment of the invention a protective group of amino group is BOC (butoxycarbonyl).

The term "alkyl" used in this description of the meaning, and unless otherwise stated, includes saturated or unsaturated, straight chain, branched or cyclic, primary, secondary, or tertiary hydrocarbon, typically C1-C10and, in particular, includes methyl, trifluoromethyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, hexyl, isohexyl, cyclohexyl, cyclohexylmethyl, methylpentyl and dimethylbutyl. The term includes both substituted and unsubstituted alkyl group, alkylene, alkenyl, Alcanena, quinil and albinyana. The remains, which alkyl group may be substituted at one or more positions selected from the group consisting of halogen (including fluorine, chlorine, bromine or iodine), hydroxyl (for example, CH2OH), amino (e.g., CH2NH2CH2NHCH3or CH2N(CH3)2), alkylamino, arylamino, alkoxy, aryloxy, nitro, azido (for example, CH2N3), cyano (CH2CN), sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, either of which or all of which may be exposed and the and additionally protected if necessary, as is well known to specialists in this area and as indicated, for example, in Greene et al., Protective Groups in Organic Synthesis, John Wiley and Sons, 2ndEdition (1991).

The term "aryl" used in this description of the meaning, and unless otherwise stated, refers to phenyl, biphenyl or naphthyl. The term includes both substituted and unsubstituted residues. The aryl group may be substituted by one or more residues, including, without limitation, hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, either of which or all of which may be exposed or optionally protected as is well-known specialists in this field, as indicated, for example, in Greene et al., Protective Groups in Organic Synthesis, John Wiley and Sons, 2ndEdition (1991).

The term "acyl" includes a group-C(=O)-R, in which decarbonising the residue R represents, for example, having a straight chain, branched or cyclic alkyl or lower alkyl, alkoxyalkyl, including methoxymethyl, aralkyl, including benzyl, aryloxyalkyl, such as phenoxymethyl, aryl, including phenyl, optionally substituted with halogen, C1-C4-alkyl or C1-C4-alkoxy, sulphonate esters such as alkyl - or aralkylamines, including methanesulfonyl, mono-, di - or trifosfatnogo ester, trityl eliminasecurity, substituted benzyl, trialkylsilyl, such as, for example, dimethyl-tert-Boticelli or diphenylmethylsilane. Aryl groups in esters optimally contain a phenyl group. The term "lower acyl" refers to acyl group, in which decarbonising balance is lower alkyl.

The term pyrimidine nucleoside base includes a pyrimidine base or an analogue of pyrimidine bases. Examples of pyrimidine bases or analogues of pyrimidine bases include, but are not limited, thymine, cytosine, 5-fertilizin, 5-methylcytosine, 6-etherimide, including 6-azacytosine, 2 - and/or 4-mercaptopyrimidine, uracil, 5-halogenerator, including 5-fluorouracil, C5-alkylpyridine, C5-benzylpyrimidines, C5-halogenopyrimidines, C5-vinylpyridin, C5-acetylenic pyrimidine, C5-arylpyrimidine, C5-aminopyrimidine, C5-cyanopyrimidine, C5-nitropyrimidin, C5-aminopyrimidine, 5-azacitidine, 5-azauracil, triazolopyridines, imidazopyridines, pyrrolopyrimidine and pyrazolopyrimidines. Functional groups of oxygen and nitrogen bases can be protected, if necessary or desirable. Suitable protective groups are well known to specialists in this field and include trimethylsilyl, dimethylhexylamine, tert-butyldimethylsilyl and tert-BU is indefinitely, trityl, alkyl groups and acyl groups such as acetyl and propionyl, methanesulfonyl and para-toluensulfonyl. Alternative pyrimidine base or an analogue of pyrimidine bases optionally can be substituted so as to form a suitable prodrug, which can bein vivo. Examples of suitable substituents include acyl residue, amine or cyclopropyl (for example, 2-amino, 2,6-diamino - or cyclopropylamino).

Other reagents used in the method according to the present invention or in the prior art, is determined as follows: AIBN means azobis(isobutyronitrile); BSA (bis(trimethylsilyl)ndimethylacetamide); CAN mean nitrate ammonium-cerium; DIBAL means hydride diisobutylaluminum; TMSCl means chlorotrimethylsilane; TN means triperoxonane acid; TEA means triethylamine; TFAA means triperoxonane anhydride; TBDPSCl means tert-butyldiphenylsilyl; TBDMSCl means tert-butyldimethylsilyloxy; TBTN means hydride tri-n-butyanova; DET means diethyltartrate; TBS means tert-butyldimethylsilyl; DMTrCl means dimethoxytrityl; DME means 1,2-dimethoxyethane; "Pyr" is used as the reduction of pyridine; DMAP means 4-dimethylaminopyridine; DIBAL means hydride diisobutylaluminum; PhOCO2Ph means diphenylcarbonate; HMDS means hexamethylene Yazid; and DHM means dichloromethane.

The method according to the present invention is not limited to the use of the above as examples of nucleoside and reagents. Suitable alternative reagents can be used for the present invention instead of the reagents listed above. For example, DME (1,2-dimethoxyethane) can be replaced by any suitable polar aprotic solvent, such as THF (tetrahydrofuran) or any simple ether; and Red-Al (bis[2-methoxyethoxy]alumoweld sodium) in toluene can be replaced NaHTe, SmI2H2+Pd-phosphine catalyst or LiAl(OtBu)3H (tri-tert-butoxylated lithium), all of which provide chemoselective and regioselective recovery.

A detailed description of the stages of method

Cyclic intermediate anhydrous-1-paranoilise

One of the key compounds for the methods according to the present invention is 2,2'-anhydrous-1-paranoilise, for example, α - or β-, D - or L-, 2,2'-anhydrous-1-paranoilise General formula:

where:

each D is hydrogen or a suitable protective group of hydroxyl, such as substituted or unsubstituted alkyl, substituted or unsubstituted aryl, substituted or unsubstituted acyl, silyl or amino;

each R1and R1'regardless of oz is achet hydrogen, substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkenyl, substituted or unsubstituted lower quinil, substituted or unsubstituted aryl, alkylaryl, halogen (F, Cl, Br or I), NH2, Other5, NR5R5', NHOR5, NR5Other5', NR5NR5'R5”, OH, OR5, SH, SR5, NO2, NO, CH2OH, CH2OR5, CO2H, CO2R5, CONH2, CONHR5, CONR5R5'or CN;

each R3and R3'independently means a hydrogen or halogen (F, Cl, Br or I), OH, SH, OCH3, SCH3, NH2, NHCH3CH3C2H5CH=CH2, CN, CH2NH2CH2OH or CO2H;

each Y2means O, S, NH or NR6;

each Y3means O, S, NH or NR7; and

each R5, R5', R6and R7independently means hydrogen, substituted or unsubstituted lower alkyl, C1-C6arylalkyl or substituted or unsubstituted aryl.

2,2'-anhydrous-1-paranoilise can be purchased or synthesized by any method known in this field, including the synthesis of furnishers with 2'-hydroxyl using a standard way to associate sugar with subsequent condensation with the formation of 2,2'-anhydrous connection, or an alternative binding sugar with cyanamide to obtain split timing is th oxazoline, then you build the Foundation with the necessary ciclismo or condensing agent.

In specific embodiments of the present invention β - or α-, D - or L-, 2'-deoxy or 2'-substituted nucleoside get through 2,2'-anhydrous-1-paranoilise according to the following protocols.

From arabinofuranose

In one method according to the present invention can be used to furanose, such as L-furanose, in particular L-arabinose, as the starting material to obtain 2,2'-anhydrous-1-pyranoindoleacetic, which then restores according to the present invention, obtaining 2'-deoxynucleoside, such as β-L-2'-deoxynucleoside, in particular β-L-2'-deoxythymidin. Alternative 2,2'-anhydrous-1-paranoilise may be subjected to interaction with nucleophilic agent, such as ORGANOMETALLIC agent (for example, a Grignard reagent or alkyllithium reagent)to obtain the desired 2'-substituted nucleoside.

On Fig in the present invention as starting substances used L-arabinose (1to obtain β-L-2'-deoxythymidin during the 5-stage synthesis. L-arabinose (1initially subjected to interaction with cyanamide in the conditions described in the prior art, with the formation of intermediate L-arabinofuranosylcytosine (2) (see WO 02/44194).Then, 5'-OH arabinose remainder of the intermediate L-arabinofuranosylcytosine ( 2protect via interaction with Fritillaria (TrCl) and pyridine at a temperature of about 45°C (3). Adding protective group OH in the specified conditions well known to specialists in this area (Greene et al., Protective Groups in Organic Synthesis (1991), John Wiley and Sons, 2ndEdition).

Stage 3 of the method depicted in Fig, which shows the interaction in suitable conditions, which are indicated in the prior art, 5'-trityl-protected L-arabinofuranosylcytosine (3with collisuem or condensing agent, selected from the following groups:

For example, if the condensing or ciclismo agent use the structure (ii)shown above, the reaction is carried out in the presence of Na2CO3/H2O and subsequent isomerization is carried out with the addition of Pd/Al2O3/H2O (see WO 02/44194). However, when using condensing or cyclisme agent (i), the reaction is carried out in methyl-2-formylpyridine at a temperature of education phlegmy for 1 hour (see EP 0351126). Cyclization leads to the formation of 2,2'-anhydrous-1-(L-arabinofuranosyl)thymidine (4).

The next stage according to the present invention includes the recovery of 2,2'-anhydrous-1-(L-arabinofuranosyl)thymidine (4such a reducing agent, as Red-Al, and a complexing agent such as 15-crown-5-ether, in the presence of a polar solvent, such as THF and/or DME, preferably at low temperatures, such as about 0-5°C, to obtain the β-L-5'-trityl-2'-deoxythymidine (5).

The use of a complexing agent, such as 15-crown-5-ether, at this stage it is useful due to the increased solubility of 2,2'-anhydrous-1-(L-arabinofuranosyl)thymidine, which leads to a higher percentage yield of the product and avoids the use of such reagents as palladium catalysts, removal of which requires time-consuming effort. In addition, the use of 15-crown-5-ether allows you to dispense with the use of HBr to reveal agitations structure, requiring the use of H2with the poisoned catalyst Pd-BaSO4to remove the bromide (see EP 0351126), thus allowing to avoid the use of dangerous chemicals found in some methods of the prior art. Finally, the method according to the present invention avoids the use of dioxane as a reagent. This has the advantage, as dioxane flammable and is not suitable for synthesis on an industrial scale.

The final stage in the method shown in Fig is removing trailvoy protecting group from the 5'-put what I β-L-2'-deoxythymidine ( 5) processing 80% AcOH at a temperature of about 50°C to remove L-2'-deoxythymidine (6).

Alternative selective protection of L-arabinose (1in C-5-position using trityl possible by the interaction of L-arabinose (1with TrCl (Fritillaria) and pyridine at a temperature of about 45°C with the formation of 5-TrO-L-arabinose (structure not shown). Then 5-TrO-L-arabinose is subjected to interaction with cyanamide in the conditions described in the prior art, with the formation of intermediate 5-TrO-L-arabinofuranosylcytosine (3) (see WO 02/44194). The remaining stages of the method are the same as the stage shown in Fig for education structures (4), (5and (6).

On Fig shows the synthesis according to the present invention, which is similar to the synthesis depicted in Fig, but different stages in which protects OH intermediates. As Fig, L-arabinose (1) used as the starting material and is subjected to interaction with cyanamide, receiving L-arabinofuranosyluracil as an intermediate product (2). Then L-arabinofuranosyluracil (2) is subjected to interaction with any of cyclessa/condensing reagent, the above structures (i)to(ix), in suitable conditions, which are indicated in the prior art, produces the 2,2'-anhydrous-1-(L-arabinofuranosyl)thymidine ( 3). 2,2'-anhydrous-1-(L-arabinofuranosyl)thymidine (3) then subjected to interaction with TrCl and pyridine at a temperature of about 45°C, to protect the 5'-OH residue arabinose 2,2'-angelasweeney (4). The specified stage should be compared with the stage 2 on Fig, where a group of trityl added to the 5'-position arabinoside of arabinofuranosylcytosine to its interaction with collisuem or condensing reagent. The last 2-stage 5-stage method, shown in Fig, identical to the last 2 stages, shown in Fig, and give 5'-trityl-protected thymidine (5) and 2'-deoxythymidin with the remote protection (6).

The applicants concluded that the method described in Fig, is more effective than the method depicted in Fig, based on the fact that the accession trailvoy protective group at the 5'-position takes place at the stage, which is carried in the synthesis earlier than was done in the prior art.

From ribofuranose

In another method according to the present invention can be used to furanose, such as L-furanose, in particular L-ribose, as the starting material to obtain 2,2'-anhydrous-1-pyranoindoleacetic, which then restores according to the present invention, obtaining 2'-deoxynucleoside, such as β-L-2'-deoxynucleoside, in particular β-L-2'-de is oxicillin. Alternative 2,2'-anhydrous-1-paranoilise can be subjected to interaction with nucleophilic agent, such as ORGANOMETALLIC agent (for example, a Grignard reagent or alkyllithium reagent)to give the desired 2'-substituted nucleoside.

On Fig shows the 7-step synthesis to obtain 2'-deoxythymidine using D-ribose as the original substances. In the first stage of this method, all of the OH-group D-ribose protect such manner as, for example, acetyl group or benzoyl, as is well known to specialists in this field. Then secure D-ribose is subjected to interaction with thymine in the presence of, for example, SnCl4, HMDS and TMSCl, as is known in the prior art, receiving thymidine, which has a protective group at the 2'-, 3'- and 5'-positions of the nucleoside. The protective group is removed in stage 3 reagents and conditions suitable for removal of a particular related protective group. The intermediate product obtained in stage 3, is thymidine.

In stage 4, Fig, enter stage cyclization/condensation directly from thymidine, and not from pornographisation, as shown in Fig and 25. In this case thymidine subjected to interaction with PhOCOOPh and catalyst NaHCO3in the presence of DMF at a temperature of about 150°C, receiving 2,2'-anhydrous-1-(ribofuranosyl)thymidine.

Structure,2'-anhydrous-1-(ribofuranosyl)thymidine 5and6represent two separate variants according to the present invention, since the structure of5receive from the structure of thymidine4with protective groups in 3'- and 5'-positions of the RIBO-thymidine residue, and the structure of6receive from the structure of thymidine, in which the 3'- and 5'-position of the RIBO-thymidine residue is not protected. In any case, the 2'-OH thymidine should be free OH group, so he could take part in the interaction, giving the structure of 2,2'-anhydrous-1-(ribofuranosyl)thymidine. If the synthesis passes through the structure5in one embodiment, the 5'-protecting group is trityl; then can be carried out an additional step to remove the protective group from the 3'-position of the RIBO-residue before restoring such a reducing agent as Red-Al, and a complexing agent such as 15-crown-5-ether, with the formation of patterns (8).

In a variant according to the present invention, depicted in Fig, synthesis passes through the structure6where TrCl, pyridine subjected to interaction with 2,2'-anhydrous-1-(ribofuranosyl)thymidine at about 45°C, obtaining 5'-trailerbonnie 2,2'-anhydrous-1-(ribofuranosyl)thymidine, which is a structure (7). Then 5'-trailerbonnie 2,2'-anhydrous-1-(ribofuranosyl)thymidine restore through its interaction with such a reducing agent as Red-Al, and sets obrazuyuschim agent, such as 15-crown-5-ether, a polar solvent, such as THF and/or DME, preferably at a temperature of about 0-5°C. the Specified stage gives 5'-trailerbonnie 2'-deoxythymidine (8), which is then removed as a result of its interaction with 80% AcOH at about 50°C, receiving D-2'-deoxythymidine (9). The method shown in Fig, provides a way to obtain 2,2'-anhydroerythromycin directly from thymidine or protected thymidine without the participation of the intermediate pornographisation and accompanying stage condensation or cyclization.

On Fig shows the 5-stage method for L-2'-deoxythymidine, starting from L-ribose. In this synthesis of L-ribose is subjected to interaction with thymine and SnCl4in TMSCl and HMDS with the formation of thymidine (2). Then thymidine subjected to interaction with PhOCOOPh and catalyst NaHCO3in DMF at about 150°C, receiving L-2,2'-anhydrotetracycline (3). The obtained L-2,2'-anhydrotetracycline subjected to interaction with TrCl and pyridine at about 45°C, obtaining 5'-trityl-protected L-2,2'-anhydrotetracycline (4and it in turn restore through interaction with such a reducing agent as Red-Al, and a complexing agent such as 15-crown-5-ether, a polar solvent, such as THF and/or DME, preferably at t is mperature about 0-5°C, receiving 5'-trityl-L-2'-deoxythymidine (5). Finally, remove the secure connection (5) when interacting with 80% AcOH at about 50°C with the formation of L-2'-deoxythymidine (6). The specified synthesis is the effective number of required stages, and also allows you to avoid the formation of ribofuranosylthiazole.

On Fig depicts the 8-step method of obtaining 2'-deoxythymidine of L-ribose. First, L-ribose (1protect any protecting group under conditions suitable for the use of such protective group (2), which are known to the person skilled in the art. Protected L-ribose (2) is subjected to interaction with thymine and SnCl4in the presence of TMSCl and HMDS, on stage, known in the prior art to form thymidine, which has a protective group at the 2'-, 3'- and 5'-positions (3). Protected thymidine (3) is subjected to removal protection (4), using reagents and conditions suitable for removing the protective group, and unprotected thymidine (4) is subjected to interaction with PhOCOOPh and catalyst NaHCO3in the presence of DMF at about 140-150°C with the formation of 2,2'-anhydrous-1-ribofuranosylpurine (5or (6). It should be noted that if the resulting intermediate product is 2,2'-anhydrous-1-ribofuranosylpurine (5), additional with the adiya after the formation of the intermediate product ( 4), in which the 3'- and 5'-position of thymidine subjected to interaction, to put these provisions of the protective group. Triteleia group are preferred protective groups for this intermediate product. If you receive an intermediate product (6), it can be obtained directly from the structure of thymidine (4).

Then the intermediate product (5in the case of its use shall be subjected to the removal of protection in its 3'-position reagents and conditions suitable for the removal of the protective group from this position to obtain the 5'-trityl-protected 2,2'-anhydrous-1-ribofuranosylpurine (7). However, if use of the intermediate product (6), he may be subjected to interaction with TrCl and pyridine at about 45°C to obtain the 5'-trityl-protected 2,2'-anhydrous-1-ribofuranosylpurine (7).

Then 5'-trityl-protected 2,2'-anhydrous-1-ribofuranosylpurine (7) restore such a reducing agent as Red-Al, and a complexing agent such as 15-crown-5-ether, a polar solvent, such as THF and/or DME, preferably at a temperature of about 0-5°C, obtaining 5'-trityl-protected thymidine (8), which is then removed by reaction with 80% AcOH at a temperature of about 50°C, receiving L-RIBO-2'-deoxythymidine (9).

Synthesis, shows the output for Fig, avoids passing the reaction through the intermediate pornographisation, which requires an additional stage of condensation for the formation of the corresponding 2,2'-angelasweeney. It also provides a choice about what intermediate products must be protected at different stages of the method.

In another method according to the present invention the synthesis of the desired compounds can be carried out in the absence of complexing agent (see Fig). The use of complexing agent, such as 15-crown-5-ether, gives a higher percentage yield of the product in the case, when the protective group is dimethoxytrityl, however, in the case when the protective group used only trail, the use of complexing agent, such as, for example, 15-crown-5-ether, gives lower interest yield. Thus, in some embodiments of the invention the synthesis of the desired compounds can be carried out in the absence of a complexing agent, when the protective group used trail as Fig.

On Fig depicts the 3-stage method of obtaining 2'-deoxythymidine. The method includes:

(a) obtaining 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2) of 2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine (1) suspendirovanie,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine ( 1), for example, pyridine and DMAP, and adding portions of Fritillaria, for example, at room temperature. The reaction mixture can be maintained at room temperature or, if necessary, be heated, for example, the reaction mixture can be maintained at room temperature for about 1 hour and then heated to 45°C (internal temperature) for about 15 hours. The reaction can be controlled, for example, TLC (Rfthe original substance of 0.15; Rfproduct 0,43). Then, the reaction mixture can be quenched and purify the desired product, for example, cooled to about 0°C by slow addition of saturated aqueous NaHCO3within a 15-minute period of time without changes in the internal temperature. Immediately can be precipitated white solid from the solution, and the white slurry can be stirred for 30 minutes at room temperature. The solid can be distinguished by filtration through a Buchner funnel and then washed with water. The remaining solid can be placed in dichloromethane and stirred for about 30 minutes at room temperature. The remainder may be allocated by filtration through a Buchner funnel, washed with dichloromethane and dried in vacuum overnight to obtain 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2with approximately 73% in view of the white solids;

(b) obtaining 2'-deoxy-5'-O-trityl-β-D-thymidine (3) of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2) recovery of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2), for example, suspendirovanie (2) in anhydrous tetrahydrofuran and cooling the suspension to about 0-5°C in a bath with ice. In a separate flask, immersed in a bath of ice, 65 wt.% a solution of Red-Al in toluene may be diluted with a suitable solvent, for example, the addition of dry tetrahydrofuran. Then the obtained diluted solution of Red-Al can be cooled to about 0-5°C and added dropwise via syringe is added to a suspension of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2). The speed of adding drops of a solution of Red-Al is important for the reaction, and the addition can be completed in about 1 hour. The resulting clear solution was maintained at about 0-5°C for 1 hour, after this time TLC analysis showed the presence of educt (Rf0,34), the desired product (Rf0,47) and impurities (Rfof 0.42 and 0.26). The HPLC analysis showed the presence of the original substance (11,35 min, 36,5% AUC), product (12,60 min, 24%) and slightly basic impurities (11,7 min, 2.9 per cent). After about 2 hours in total at a temperature of about 0-5°C dropwise via a syringe was added an additional portion of "undiluted" 65 wt.% solution Re-Al in toluene over a period of time, approximately 20 minutes to the reaction mixture, which was maintained at about 0-5°C. after 1 hour the analyses TLC and HPLC showed the presence of the original substance (11,35 min, 3.2 per cent). Was added dropwise next portion 65 wt.% solution of Red-Al in toluene and the reaction mixture is maintained at about 0-5°C for 45 minutes. After a specified period of time TLC analysis showed only trace remaining of the original substance. The reaction was suppressed by addition of a saturated solution of NH4Cl and a layer of tetrahydrofuran decantation. The aqueous layer was extracted with isopropylacetate and the resulting emulsion was stratified by slow addition of 5 n HCl solution. The organic layer was separated, combined with a layer of tetrahydrofuran and washed with saturated solution of NH4Cl and then a saturated solution of salt. At this point the pH of the layer of saturated salt solution ranged from 6.5 to 7, and the organic layer was dried over Na2SO4, filtered and concentrated in vacuum, getting foamy solid. Coarse residue evaporated together with toluene, concentrated in vacuo and the resulting residue was collected in toluene by heating to about 45°C. the Mixture was cooled to room temperature and stirred at this temperature until a white solid began to fall in ocado is. Was added dropwise water and the resulting mixture was stirred at room temperature for about 3 hours. The solid was isolated by filtration and the filter cake was washed with water and toluene. The solid was dried at about 45°C in high vacuum for about 1 hour and then at room temperature under vacuum over night, getting a 2'-deoxy-5'-O-trityl-β-D-thymidine3with the release of approximately 41%;

(c) obtaining 2'-deoxy-D-thymidine (4from 2'-deoxy-5'-O-trityl-β-D-thymidine (3) (1,215 g, 2.5 mmol) suspendirovanie (3) in methanol and heating the reaction mixture to about 45°C in a water bath until dissolution (3). Then the flask was cooled to room temperature and concentrated. To the mixture was added HCl and stirred at room temperature. After about 25 minutes white solid began to precipitate from solution. After 1 hour, TLC analysis showed that the original substance remained (Rf0,53) and formed the main product (Rf0,21). To the reaction mixture was added portion of n-heptane and stirred at room temperature for about 15 minutes. White solid was isolated by filtration. The filtrate was divided into two layers and layer in methanol was extracted with n-heptane and then concentrated in vacuo to a volume of 2 ml of the remainder of the joint is issued with 405 mg of a white solid, suspended in TBME, and stirred at room temperature for 1 hour. White solid was isolated by filtration, washed with TBME and dried in a vacuum furnace, obtaining 2'-deoxy-D-thymidine (4with approximately 78%.

It should be understood that all methods of synthesis described in figure 1-29, and all examples are equally applicable to any stereochemical form, α - or β-, D - or L-, any of the original substance, and that the parent compound is not limited to ribose, xylose and arabinose specified herein, and also include 5 - and 6-membered cycles with S, N, or CH2instead O specified in non-limiting examples and figures 1-29.

The present invention is better described in the following non-limiting series of examples. Specific solvents, reagents and/or reaction conditions described in this description may be replaced by an equivalent, similar or suitable solvents, reagents and/or reaction conditions, without departing from the essence and not going beyond the scope of the invention.

EXAMPLES

Example 1

L-arabinose is transformed into the corresponding methylglucoside and 3 - and 4-hydroxyl group protected in the form of a derivative acetonide. The diagram below shows a simple way to desoxyribose 2-hydroxyl group of the compound2through its transformation into the corresponding mesilate group, and adversa specified intermediate mesilate to the conditions of reductive cleavage, to obtain 2-deoxy-intermediate product4. Cm. H. Urata, E. Ogura, K. Shinohara, Y. Ueda and M. Akagi, Nucleic Acids Res. 1992, 20, 3325-3332; and J. W. Pratt, N. K. Richtmyer and C. S. Hudson, J. Am. Chem. Soc. 1952, 74, 2200-2205.

Example 2

L-arabinose is transformed into the corresponding derived glikas through key stage reductive elimination and transformation of the resulting intermediate glikas in methyl-2-deoxyribofuranosyl. Cm. B. K. Shull, Z. Wu and M. Koreeda, J. Carbohydr. Chem. 1996, 15, 955-964; M. L. Sznaidman, M. R. Almond and A. Pesyan., Nucleosides, Nucleotides &Nucleic Acids 2002, 21, 155-163; and Z.-X. Wang, W. Duan, L. I. Wiebe, J. Balzarini, E. D. Clercq and E. E. Knaus, Nucleosides, Nucleotides &Nucleic Acids 2001, 20, 11-40.

Example 3

L-arabinose is transformed into the corresponding derived glikas through key stage reductive elimination and transformation of the resulting intermediate glikas in methyl-2-deoxyribofuranosyl. Cm. M. L. Sznaidman, M. R. Almond and A. Pesyan., Nucleosides, Nucleotides &Nucleic Acids 2002, 21, 155-163; and Z.-X. Wang, W. Duan, L. I. Wiebe, J. Balzarini, E. D. Clercq and E. E. Knaus, Nucleosides, Nucleotides &Nucleic Acids 2001, 20, 11-40; and R. V. Stick, K. A. Stubbs, D. M. G. Tilbrook and A. G. Watts, Aust. J. Chem. 2002, 55, 83-85.

PR is measures 4

D-xylose oxidized by bromine water and then the resulting 1,4-lactone is exposed to a mixture of HBr/acetic acid to obtain 2,5-dibromo-2,5-dideoxy-D-leksono-1,4-lactone2. Processing dibromoethane2potassium iodide in TN gives the corresponding 5-iodine-connection, and also leads to the selective removal of the bromine atom at C-2, giving 5-iodine-2-detoxication3. Exposing the specified 5-INTACTO3the impact of an aqueous solution of potassium hydroxide, get 4,5-epoxy derivative, which upon treatment with an aqueous solution of the acid gives the corresponding 2-deoxy-L-ribonolactone through stereospetsifichno inversion at C-4. Protected 2-deoxy-L-ribonolactone6selectively reduced to the corresponding lactol7using Red-Al. Then lactol7turn in the desired chlorine-sugar9. Cm. H.S. Isbell, Methods in Carbohydrate Research, 1963, 2, 13-14; K. Bock, I. Lundt, and C. Pedersen, Carbohydrate Research, 1981, 90, 17-26; K. Bock, I. Lundt, and C. Pedersen, Carbohydrate Research, 1982, 104, 79-85; and I. Lundt and R. Madsen, Topics in Current Chemistry 2001, 215, 177-191.

Example 5

D-galactose is subjected to oxidative cleavage, getting D-lexikalische, which is then subjected to bromirovanii to obtain 2,5-dibromo-2,5-dideoxy-D-xileno-1,4-lactone2. Selective hydrogenolysis2gives 5-bromo-2-detoxication3that then under the will eraut successive transformations, such transformations shown in example 7, receiving a key intermediate chloro-sugar8. Cm. K. Bock, I. Lundt, and C. Pedersen, Carbohydrate Research, 1981, 90, 17-26; K. Bock, I. Lundt, and C. Pedersen, Carbohydrate Research, 1979, 68, 313-319; K. Bock, I. Lundt, and C. Pedersen, Acta Chem. Scand. B 1984, 38, 555-561; and W. J. Humphlett, Carbohydrate Research, 1967, 4, 157-164.

Example 6

D-glucono-1,4-lactone converted into 2,6-dibromo-2,6-dideoxy-D-mannono-1,4-lactone1. Treatment of the lactone1with hydrazine and then with an aqueous solution of bromine gives 6-bromo-2,6-dideoxy-D-arabinogalactan-1,4-lactone3. Interaction3with an excess of an aqueous solution of potassium hydroxide, followed by acidification leads to inversion at C-4 and C-5, giving 2-deoxy-L-rebulican-1,4-lactone6. The specified transformation includes the opening cycle of the lactone by rearrangement Payne primary epoxide4in the secondary epoxide5. 2-deoxy-L-rebulican-1,4-lactone6is subjected to oxidative cleavage and subsequent reduction of the resulting aldehyde with getting lactone7that in turn required chlorine-sugar, using the sequence of reactions similar to the sequence of reactions shown in example 5. Cm. K. Bock, I. Lundt, and C. Pedersen, Carbohydrate Research, 1979, 68, 313-319; and K. Bock, I. Lundt, and C. Pedersen, Acta Chem. Scand. B 1984, 38, 555-561.

Example 7

D-galactono-1,4-lactone is transformed into acetylated Gibraltar2that when processed with hydrazine and subsequent bromirovanii gives 2-detoxication3. The lactone3will deacetylases and subjected to oxidative cleavage and subsequent reduction of the resulting aldehyde using NaBH4to obtain 2-deoxy-L-ribono-1,4-lactone5. Cm. K. Bock, I. Lundt, and C. Pedersen, Carbohydrate Research, 1979, 68, 313-319; and K. Bock, I. Lundt, and C. Pedersen, Acta Chem. Scand. B 1984, 38, 555-561.

Example 8

Pornolation, commercially available neugeborne and achiral compound, used as the starting material, to obtain chlorine-sugar. The key step in this method is asymmetric dihydroxypropane of ester 2Z-pentanoate2with the formation of a derivative of 2(R),3(R)-pentanoate3. The intermediate product3subjected to stereoselective cyclization, receiving 2-deoxy-L-sugar4. Connection4turn in the desired protected chloro-sugar through three transformations by direct synthesis. Cm. D. C. Liotta, and M. W. Hager, U.S. patent 5414078, may 9, 1995.

Example 9

Ethyl-3,3-diethoxypropane1is inexpensive neugeborne acyclic and achiral original substance. Connection1reduced to the corresponding aldehyde2using DIBAL. At the next stage, the aldehyde2converted into α,β-unsaturated ester, 5,5-diethoxy-2E-pentenoic using modified Horner-Emmons Wittig reaction, using aminobutiramida(etoxycarbonyl)methylphosphonate, and the resulting ester to restore derived 2E-penten-1-ol3. Received properly allyl alcohol3converted into the corresponding 2(S),3(S)-epoxied4using the conditions of asymmetric epoxidation on he sang sharpless. The resulting epoxied4protect and then subjected to acid hydrolysis, receiving a key intermediate product6. Connection6cyclist to the derived 2-deoxy-L-ribofuranose7, which is then converted into the desired L-chloro-sugar9. Cm. D. C. Liotta, and M. W. Hager, U.S. patent 5414078, may 9, 1995; and M. W. Hager and D. C. Liotta, Tetrahedron Lett. 1992, 33, 7083-7086.

Example 10

Hydroxylamino acid1subjected to cyclization, getting derived 2-deoxy-L-1,4-ribonolactone2; which then in turn require the th chloro-sugar 5in four stages. Cm. R. F. Schinazi, D. C. Liotta, C. K. Chu, J. J. McAtee, J. Shi, Y. Choi, K. Lee and J. H. Hong, U.S. patent 6348587B1, February 19, 2002; U. Ravid, R. M. Silverstein, and R. L. Smith, Tetrahedron 1978, 34, 1449-1452; and M. Taniguchi, K. Koga, and S. Yamada, Tetrahedron 1974, 30, 3547-3552.

Example 11

Commercially available alcohol1subject to the conditions of epoxidation on he sang sharpless, getting epoxide2. Epoxied2treated with benzyl alcohol in the presence of Ti(Oi-Pr)4getting diol3that turn in the corresponding acetonide derivative4. Connection4oxidized, using reaction conditions Walker to get aldehyde5which is treated with an aqueous solution of hydrochloric acid, receiving 5-O-benzyl-2-deoxy-L-ribofuranose6. Connection6turn in the desired chlorine-sugar9during the four simple stages. Cm. M. E. Jung and C. J. Nichols, Tetrahedron Lett. 1998, 39, 4615-4618.

Example 12

Epoxied1protect in the form of benzyl ester2and open the epoxide using benzilate sodium benzyl alcohol, then protect the resulting alcohol3receiving Tris-benzyl ether4. Transformation connection4in the aldehyde6(which is a protected 2-detox is-L-ribose) do using hydroporinae/oxidation of H2O2with subsequent oxidation by the Turn of the resulting alcohol5. Removal of the protecting benzyl ethers6using palladium hydroxide on coal gives a mixture of three ethyl-2-deoxy-L-riboside 7a, 7b and 7c in the ratio 2:2:1. Protection 7a and 7b using trouillard and processing the resulting getaway-derived hydrogen chloride gives the desired chlorine-sugar8. Cm. M. E. Jung and C. J. Nichols, Tetrahedron Lett. 1998, 39, 4615-4618.

Example 13

1,2-O-isopropylidene-L-glyceraldehyde1treated with allylbromide in the presence of zinc and an aqueous solution of ammonium chloride, receiving appropriate homoallylic alcohol2. A group of isopropylidene2removed using an aqueous solution of acetic acid, obtaining the intermediate product3. Subsequent ozonolysis and recovering the dimethyl sulfide compounds3gives 2-deoxy-L-ribofuranose4that in turn protected chloro-sugar7in three stages. Cm. J. S. Yadav and C. Srinivas, Tetrahedron Lett. 2002, 43, 3837-3839; and T. Harada and T. Mukaiyama, Chem. Lett. 1981, 1109-1110.

Example 14

In this prospect the advantage of 2-methyl bromide[1,3]dioxolane instead of allylbromide, which is used in example 13, which eliminates the need for ozonolysis and subsequent reduction of the intermediate product3. Cm. J. S. Yadav and C. Srinivas, Tetrahedron Lett. 2002, 43, 3837-3839; and T. Harada and T. Mukaiyama, Chem. Lett. 1981, 1109-1110.

Example 15

Glycol1(which can be obtained from L-ribose in four stages) treated with acidic methanol to obtain 2-deoxy-L-ribose3that in turn protected chloro-sugar5in two stages. Cm. H. Ohrui and J. J. Fox, Tetrahedron Lett. 1973, 1951-1954; W. Abramski and M. Chmielewski, J. Carbohydr. Chem. 1994, 13, 125-128; and J. C.-J. Cheng, U. Hacksell, and G. D. Daves, Jr., J. Org. Chem. 1985, 50, 2778-2780.

The original substance1can be obtained from L-ribose

Example 16

L-arabinose is subjected to interaction with cyanamide, getting derived 1,2-oxazoline1. If interoperability with ethyl ether of 2-methyl-3-oxopropanoic acid compound1gives O2,2'-anhydrous-L-thymidine2. Connection2are benzoylation and the resulting di-O-benzoyl-derived3subject to the conditions of reductive cleavage to obtain 3',5'-di-O-benzoyl-LdT4. Connection4amrabat who live methanol solution of sodium methoxide, getting LdT. Cm. A. Holy, Coll. Czech. Chem. Commun. 1972, 37, 4072-4087; and P. V. P. Pragnacharyulu, C. Vargeese, M. McGregor, and E. Abushanab, J. Org. Chem. 1995, 60, 3096-3099.

Example 17

In this example, use a different method of disclosure O2,2'connection connection3using hydrogen chloride. The resulting 2'-deoxy-2'-chloro-derived4process TBTH/AIBN, obtaining 2'-deoxy-protected nucleoside5that when desetilirovania gives LdT. Cm. P. V. P. Pragnacharyulu, C. Vargeese, M. McGregor, and E. Abushanab, J. Org. Chem. 1995, 60, 3096-3099.

Example 18

This example differs from example 17 that the derived 1,2-oxazoline1subjected to interaction with other compounds, to obtain the O2,2'-anhydrous-L-thymidine2. Cm. D. McGee, Boehringer Ingelheim Proposal to Novirio Pharmaceuticals, Inc. May 17, 2002; and C.W. Murtiashaw, European patent 0351126 B1, January 18, 1995.

Example 19

The hydrogen iodide is used for the disclosure of O2,2'connection connection3to obtain 2'-deoxy-2'-iodine-derived4. Connection4treated with potassium iodide, getting the 3',5'-di-O-benzoyl-2'-deoxy-L-thymidine5. Connection5exposed to a methanol solution IU is sodium oxide, getting LdT. Cm. H. Sawai, A. Nakamura, H. Hayashi and K. Shinozuka, Nucleosides &Nucleotides 1994, 13, 1647-1654; and H. Sawai, H. Hayashi and S. Sekiguchi, Chemistry Lett. 1994, 605-606.

Example 20

Ester 2-methyloxiran-2-carboxylic acid is subjected to the interaction of 1,2-oxazoline1receiving the derived O2,2'-anhydrous-L-thymidine2. Connection2treated with revalorisation to protect the 3'- and 5'-hydroxyl group, and split O2,2'connection and to obtain 2'-deoxy-2'-chlorocresol3. Connection3treated with thionyl chloride to eliminate hydroxyl group residue basis, and the resulting connection is restored using TBTH/AIBN, to remove 2'-chloro and get protected LdT5. Connection5treated with sodium methoxide in methanol, getting LdT. Cm. E. Abushanab and P. V. Pragnacharyula, U.S. patent 5760208, June 2, 1998.

Example 21

Ethylpropyl subjected to interaction with 1,2-oxazoline1getting O2,2'-anhydrous-L-uridine2. Connection2protect and the resulting 3',5'-di-O-benzoyl-derived3treated with hydrogen chloride, obtaining 2'-deoxy-2'-chlorocresol4. Connection4then treated TBTH/AIBN, to remove 2'-chloro, and then 2'-deoxy-derivative is subjected to the hcpa is setiu conditions iodization, to obtain 5-odnokletochnoe derived6. 5-iodine-group connections6replace the methyl group using AlMe3and (Ph3P)4Pd, receiving 3',5'-di-O-benzoyl-LdT7that when processed by sodium methoxide in methanol gives LdT. Cm. A. Holy, Coll. Czech. Chem. Commun. 1972, 37, 4072-4087; J.-I. Asakura and M. J. Robins, J. Org. Chem. 1990, 55, 4928-4933; J.-I. Asakura and M. J. Robens, Tetrahedron Lett. 1988, 29, 2855-2858; and K. Hirota, Y. Kitade, Y. Kanbe, Y. Isobe and Y. Maki, Synthesis, 1993, 210, 213-215.

Example 22

This example differs from example 21 only by way of introduction of a methyl group in position 5 of the derivative of 2'-deoxy-L-uridine5. Connection5treated with formaldehyde in an alkaline medium, receiving 5-hydroxymethyl-derived6which when exposed to acidic ethanol gives 2'-deoxy-5-ethoxymethyl-L-uridine7. Connection7gives LdT under conditions of catalytic hydrogenation. Cm. A. Holy, Coll. Czech. Chem. Commun. 1972, 37, 4072-4087.

Example 23

The synthesis of the key intermediate 2-deoxy-3,5-di-O-para-toluoyl-β-L-iitrobotrepubblica from D-xylose

Example 23(a)

D-xileno-1,4-lactone (2) from D-xylose (1) by oxidation with bromine

D-xylose1(100 is, 666,1 mmol) was dissolved in distilled water (270 ml) and cooled to 0°C under stirring on top in the three-neck round-bottom flask with a volume of 1 liter Portions was added potassium carbonate (of 113.2 g, 819,3 mmol), keeping the temperature below 20°C. Then was added dropwise bromine (39,4 ml, 766,0 mmol) at 0-5°C over a period of 2 hours while maintaining the temperature below 10°C. the Reaction mixture is maintained at a temperature of approximately 5-10°C for 30 minutes, then was heated to room temperature and stirred throughout the night. After about 8 hours TLC analysis (10% methanol in ethyl acetate, visualization using vanillin) showed the absence of the original substance (Rf0,0) and the presence of a new product (Rf0,3). The reaction mixture was mixed with formic acid (6.6 ml) for about 15 minutes and then concentrated in vacuum at 45°C to a volume of about 50 ml. Conducted joint evaporation of acetic acid (200 ml) and concentration under vacuum at about 45°C to a volume of 60 ml and the crude D-xileno-1,4-lactone2transferred for use in this form in the next stage.

The advantages of this synthesis are switching from BaCO3known in the prior art, for K2CO3that gave the best value when loading (50 g D-xylose1in 15 ml of water, compared with 400 ml in the case of BaCO 3); the lactone can be used without additional purification/removal of salt KBr at the next stage; residual KBr can be used for the next reaction; and joint evaporation with acetic anhydride to remove residual water, leads to the formation of less polar products when TLC analysis.

Example 23(b)

2,5-dibromo-2,5-dideoxy-D-lyxo-1,4-lactone (3) of the D-xileno-1,4-lactone (2)

D-xileno-1,4-lactone (2) (crude acetic acid, 666,08 mmol) was transferred to a flask with a volume of 3 l, using warm acetic acid (200 ml) and the stirred suspension was slowly added 30% HBr-AcOH (662 ml, 3330 mmol). The reaction mixture was heated to 45°C for 1 hour and then cooled and stirred for 1.5 hours at room temperature. After a specified period of time TLC analysis (1:1, ethyl acetate:hexane) showed the presence of two major products (Rf0,63 [3-O-acetyl-2,5-dibromo-2,5-dideoxy-D-lyxo-1,4-lactone3a] and Rf0,5 [2,5-dibromo - 2,5-dideoxy-D-lyxo-1,4-lactone3]), and some of the remaining source material (TLC analysis, 10% methanol in ethyl acetate, Rf0,0, rendering using vanillin). The reaction mixture was cooled at 0°C was added methanol (850 ml) for 1 hour, keeping the temperature below 20°C. Then the reaction mixture gave perhaps the th to warm to room temperature and was stirred overnight. After a specified period of time TLC analysis (1:1, ethyl acetate:hexane) showed the conversion of one product (Rf0,63) into another product (Rf0,44). The reaction mixture was filtered through a Buchner funnel to remove residual KBr (173,89 g), and then concentrated in vacuo and evaporated with water (2×250 ml). Added ethyl acetate (800 ml) and water (250 ml) and the layers were separated. Then the aqueous layer was extracted with ethyl acetate (2×300 ml) and the combined organic extracts were washed with saturated aqueous sodium hydrogen carbonate solution (400 ml) and water (100 ml). The layers were separated, the aqueous layer was extracted with ethyl acetate (2×300 ml) and the combined organic extracts were dried using sodium sulfate (125 g), filtered and concentrated in vacuum at 50°C. Before concentration to dryness of the organic extract was evaporated together with heptane (200 ml)to give a brown semi-solid substance. Rubbing the obtained solid substance was carried out using 20% heptane in isopropyl ether (100 ml heptane and 500 ml of isopropyl ether, and dried in vacuum at 30-35°C during the night, receiving 2,5-dibromo-2,5-dideoxy-D-lyxo-1,4-lactone3in the form of pure light brown solid (71,9 g, 40% over 2 stages).

TPL 92-94°C [Lit. 92-93°C]; δN(d6-DMSO, 400 MHz): 3,65 (1H, DD, J4,5'=8,1 Hz, J5,5'=to 10.7 Hz, H-5'), of 3.73 (1H, DD, J4,5=5,9 Hz, J 5,5'=to 10.7 Hz), 4,4 (1H, m, H-3 or H-4), TO 4.73 (1H, m, H-4 or H-3), 5,31 (1H, d, J2,3=4.4 Hz, H-2), 6,38 (1H, users, 3-OH); δN(CDCl3, 400 MHz): 3,65 (1H, DD, J5,5'=10.3 Hz, J4,5=5,9 Hz, H-5'), and 3.72 (1H, a-t, J5,5'=9,88 Hz, H-5), 4,63 (1H, m, H-3), 4,71 (1H, m, H-4), A 4.86 (1H, d, J2,3=4.4 Hz, H-2).

The advantages of this synthesis is the ability to carry out the reaction at 45°C, significantly reducing the reaction time in comparison with 24 hours, as it was known in the prior art, removal of KBr salt by filtration was possible after treatment with methanol, and this is important, as it provides a simple extraction of the product; and the precise control of reaction temperature damping is very important in order to prevent the formation of by-products.

Example 23(c)

5-bromo-2,5-dideoxy-D-thiopentone-1,4-lactone (4)

Method 1 - sodium iodide

2,5-dibromo-2,5-dideoxy-D-lyxo-1,4-lactone3(35 g, 127,8 mmol) was dissolved in izopropilazette (300 ml) was added sodium iodide (76,6 g, 511,2 mmol) and triperoxonane acid (14,8 ml) at room temperature. The reaction mixture was heated to approximately 85°C (internal temperature) for 1.5 hours. After a specified period of time TLC analysis (1:1, ethyl acetate:hexane) showed a small amount of the remaining original substance (Rf0,44) and the presence of a new product (Rf0,19). The reaction mixture ohla is given to room temperature and was stirred for 4 hours. The TLC analysis showed the absence of the original substance, therefore, the reaction mixture was concentrated in vacuo to 20 ml, to remove triperoxonane acid, and diluted with isopropylacetate (200 ml). The reaction mixture was washed with a saturated aqueous solution of sodium bicarbonate (200 ml) and the layers were separated. Then the aqueous layer was extracted with isopropylacetate (3×200 ml). The combined organic extracts were treated with an aqueous solution of sodium thiosulfate (48 g in 160 ml of water). The aqueous layer was extracted with isopropylacetate (2×200 ml) and the combined organic extracts were dried using sodium sulfate (20 g), filtered and concentrated in vacuum, obtaining 5-bromo-2,5-dideoxy-D-thiopentone-1,4-lactone4(16,38 g, crude yield 92%) as a brown oily residue. The obtained product was dissolved in water and used as such for further reaction. In other cases, isopropylacetate replaced with water and the aqueous solution was used as such for the reaction with KOH.

δH(D2O, 400 MHz): 2,64 (1H, d, J2,2'=18.3 Hz, H-2'), of 3.12 (1H, DD, J2,2'=18,0 Hz, J2,3=5,49 Hz, H-2), 3.45 points (0,125H, DD, H-5' and H-5 for iodide 4I); 3,70 (2H, a-d, J=of 6.71 Hz, H-5, H-5'), 4,74 (1H, a-t, H-3), to 4.87 (1H, m, H-4). δWith(D2O, 100 MHz): 27,1 (C-5), 39,0 (C-2), with 67.9 (C-3), and 84.6 (C-4), 178,8 (C-1)δN(d6-DMSO, 400 MHz): 2,34 (1H, a-d, J2,2'=and 17.1 Hz, J2',3=6.3 Hz, H-2'), 2,95 (1H, DD, J2,2'=and 17.1 Hz, J2,3=5.4 Hz, H-2) 3,39 (0,125H, DD, J=7,3 Hz, J=6,8 Hz, J=11.2 Hz, H-5' and H-5 for iodide 4I), of 3.60 (1H, DD, J5,5=to 10.7 Hz, J4,5'=8,3 Hz, H-5'), 3,70 (1H, DD, J5,5=to 10.7 Hz, J4,5=5.4 Hz, H-5), 4,39 (1H, m, H-3), 4,63 (1H, m, H-4), 5,61 (1H, d, J3=4.4 Hz, 3-OH); m/z (ES-ve): 253 (M+AcOH)-; Found: C, 30,69, N, 3,55, Br, 41,22%; With5H7Ged3requires, 30,80, N, 3,62, VG, 40,97%.

Method 2 - hydrogenation

2,5-dibromo-2,5-dideoxy-D-lyxo-1,4-lactone3(7.5 g, 27.6 mmol) was dissolved in ethyl acetate (120 ml) and stir the solution was added triethylamine (4 ml, 28.7 mmol). The reaction mixture was stirred at room temperature in hydrogen atmosphere (atmospheric pressure) in the presence of 5% anhydrous palladium on coal (1 g) for about 1.5 hours. After a specified period of time TLC analysis (1:1, ethyl acetate:hexane) showed the presence of a new product (Rf0,16), the residual of the original substance (Rf0,44). Therefore, the reaction mixture was purged with argon (three times) and then stirred in an atmosphere of hydrogen for 2 hours. After a specified period of time TLC analysis showed the presence of small amounts of the original substance, therefore, the reaction mixture was filtered through celite (ethyl acetate as eluent), washed with 4 M HCl (30 ml) and the aqueous layer was additionally extracted with ethyl acetate (2×20 ml). The combined organic extracts were dried using sulfate intothree is (10 g), was filtered and concentrated in vacuum, obtaining 5-bromo-2,5-dideoxy-D-thiopentone-1,4-lactone4(5,15 g, crude yield 96%) as a crude pale yellow oil.

δH(D2O, 400 MHz): 2,65 (1H, d, J2,2'=18.3 Hz, H-2'), to 3.09 (1H, DD, J2,2'=18.3 Hz, J2,3=5.8 Hz, H-2), 3,71 (2H, a-d, J=7.01 Hz, H-5, H-5'), 4,74 (1H, a-t, J=4.9 Hz, J=4.6 Hz, H-4), to 4.87 (1H, m, J=3,7 Hz, H-3); δWith(D2O, 100 MHz): 27,7 (C-5), 39,5 (C-2), and 68.5 (C-3), to 85.2 (C-4), 179,5 (C-1)δN(d6-DMSO, 400 MHz): 2,34 (1H, d, J2,2'=17,1 Hz, H-2'), with 2.93 (1H, DD, J2,2'=and 17.1 Hz, J2,3=5.4 Hz, H-2), of 3.60 (1H, DD, J5,5=to 10.7 Hz, J4,5'=8,3 Hz, H-5'), 3,70 (1H, DD, J5,5=to 10.7 Hz, J4,5=5.4 Hz, H-5), and 4.40 (1H, m, H-3)and 4.65 (1H, m, H-4), 5,58-5,64 (1H, users, 3-OH); δWith(d6-DMSO, 100 MHz): 29,6 (C-5), 39,5 (C-2), and 67.2 (C-3), or 83.2 (C-4), 175,4 (C-1).

These stages in the method provides the advantages associated with the reaction of hydrogenation, which required much less solvent for extraction, replacement of potassium iodide sodium iodide and heating of the reaction mixture was shortened reaction time from 6 hours to 2 hours, and the replacement of acetone on isopropylacetate improved extraction of the product from the aqueous layer.

Example 23(d)

2-Deoxy-L-ribono-1,4-lactone 5 out of 5-bromo-2,5-dideoxy-D-thiopentone-1,4-lactone 4

Potassium hydroxide (14.9 g, 230,7 mmol) was dissolved in water (124 ml) and cooled to 15°C. the resulting solution was added to the mix is the solution of 5-bromo-2,5-dideoxy-D-thiopentone-1,4-lactone 4(15 g, 76.9 mmol) in water (62 ml) at room temperature. After about 3 hours analysis of TLC (2% methanol in ethyl acetate) showed the absence of the original substance (Rf0,55) and the presence of a new product (Rf0,0). The reaction mixture was heated up to 80°C (internal temperature) for 30 minutes, cooled to room temperature and did not observe changes in the analysis of TLC. Added amberlite IR-120 plus an acidic resin (50 g) and the reaction mixture was stirred at room temperature for 30 minutes and measured at this point the pH was 3. Added additional amount of resin (40 g) and the reaction mixture was stirred at room temperature for 30 minutes and measured at this point the pH was 1. The reaction mixture was stirred at room temperature overnight, after a specified period of time TLC analysis showed the formation of a new product (Rf0,21). The resin was removed by filtration through a porous funnel (water as eluent, 200 ml) and concentrated in vacuum. Do not let dry, conducted joint evaporation of 1,2-dimethoxyethane (2×100 ml). The red residue was dissolved in 1,2-dimethoxyethane (200 ml) and stirred with MgSO4(10 g) for 40 minutes at room temperature. Filtration, washing with 1,2-dimethoxyethane (75 ml) and concentration in vacuum at 45°C gave 2-detox is-L-ribono-1,4-lactone 5(10,47 g, crude yield 90%) as a crude red solid. In other cases, the product was stored in DME and used as such for further reaction.

δN(d6-DMSO, 400 MHz): 2,22 (1H, DD, J2,2'=17.6 Hz, J2',3=2.0 Hz, H-2'), 2,80 (1H, DD, J2,2'=18,1 Hz, J2,3=6.3 Hz, H-2), a 3.50 (1H, DD, J5,5'=and 12.2 Hz, J4,5'=3,9 Hz, H-5'), of 3.54 (1H, DD, J5,5'=and 12.2 Hz, J4,5=4,2 Hz, H-5), 4.26 deaths (2H, m, H-3 and H-4), 4,7-5,0 (2H, users, HE).

Example 23(e)

2-deoxy-3,5-di-O-para-toluoyl-L-ribono-1,4-lactone 6 from 2-deoxy-L-ribono-1,4-lactone 5

A solution of 2-deoxy-L-ribono-1,4-lactone5(10,47 g, 76.9 mmol) and pyridine (31,1 ml, 384,4 mmol) in 1,2-dimethoxyethane (100 ml) was cooled to a temperature of from 0 to -5°C in argon atmosphere. From the dropping funnel was added para-trouillard (21,4 g, 138 mmol) over 20 minutes, maintaining the temperature between 0 and -5°C. After 3.5 hours, maintaining the temperature between 0 and -5°C TLC analysis (30% ethyl acetate in hexane) showed the presence of a new product (Rf0,76) and no remaining original substance (Rf0,36) and HPLC analysis showed that the reaction is complete. The reaction mixture was cooled (0°C) and extinguished solution of sodium bicarbonate (25 g in 300 ml). From the reaction mixture was separated by a brown oil, which gradually hardened under stirring at room temperature. After 1.5 hours of solid ve is estvo was collected by filtration, washed with water (150 ml) and the crude solid (25,84 g) was dried overnight. The crude lactone was dissolved in dichloromethane (150 ml) and stirred with MgSO4(10 g) for 1 hour. The solid is collected by filtration, washed with dichloromethane (50 ml) and the filtrate was concentrated at 40°C to about 50 ml TBME was Added (100 ml) and the mixture was concentrated at 40°C to about 50 ml. and Then the remaining concentrated solution was stirred at room temperature, and it formed a dense suspension. Added TBME (50 ml) and stirring continued at room temperature for 2 hours. The solid is collected by filtration, washed with TBME (50 ml) and dried in vacuum at 30-35°C during the night, getting 2-deoxy-3,5-di-O-para-toluoyl-L-ribono-1,4-lactone6(10,46 g, 37% for 3 stages) as a pale brown solid.

δN(CDCl3, 400 MHz): 2,42, 2,43 (2×s, 2×WithN3AG, 2×3H), 2,82 (1H, DD, J2,2'=18.7 Hz, J2',3=1.8 Hz, H-2'), and 3.16 (1H, DD, J2,2'=18.7 Hz, J2,3=7,3 Hz, H-2), br4.61 (1H, DD, J5,5'=and 12.4 Hz, J4,5'=3.3 Hz, H-5'), 4,71 (1H, DD, J5,5'=12.1 Hz, J4,5=3,7 Hz, H-5), of 4.95 (1H, m, H-4), 5,61 (1H, a-d, J=of 7.69 Hz, H-3), 7,25-7,28 (4H, m, 2×ArH), 7,86-7,93 (2×2H, 2×d, J=8,4 Hz, 2×ArH); δC(CDCl3, 100 MHz): 21,9, 35,3, 63,9, 71,8, 82,8, 125,1, 126,5, 129,4, 129,5, 129,6, 130,0, 130,4, 144,6, 145,0, 166,0, 166,1 (2×ArCO2), 174,2 (C-1).

The advantages of this stage of the common way was the fact, that toluoyltartaric removed from the process through processing TBME, and phase chromatography on a column was removed from the method.

Example 23(f)

2-deoxy-3,5-di-O-para-toluoyl-L-ribose 7 from 2-deoxy-3,5-di-O-para-toluoyl-L-ribono-1,4-lactone 6

A solution of 2-deoxy-3,5-di-O-para-toluoyl-L-ribono-1,4-lactone6(9.0 g, 24,42 mmol) in 1,2-dimethoxyethane (90 ml) was cooled to about -60°C in argon with the upper stirring. Dropwise through an addition funnel was added a 1 M solution of hydride diisobutylaluminum in toluene (32,4 ml, 32,4 mmol) for 15 minutes. The internal temperature was maintained at -60°C for 1 hour, and HPLC analysis showed complete reaction. The reaction mixture was suppressed by the addition of acetone (10 ml) for 2 minutes and then add 5 N. HCl (30 ml) for 5 minutes. The mixture was stirred at room temperature for 30 minutes and concentrated under vacuum at 35°C to about 30 ml. Remaining oil was combined with a saturated salt solution (24 g 60 ml) and was extracted with ethyl acetate (3×100 ml). The combined organic extracts were dried with sodium sulfate (10 g), concentrated to a volume of 50 ml and evaporated together with TBME, receiving 2-deoxy-3,5-di-O-para-toluoyl-α,β-L-ribose 7. Part concentrated in vacuum to dryness to1H-NMR analysis. The product received is not further characterized as it was used crude in the next stage.

δN(CDCl3400 MHz processor, the ratio of anomers is 0.75:1): 2,2-2,6 (m, 2×WithN3Ar and H-2/H-2' for α - and β-anomers), 4,4-of 4.75 (m, H-4 and H-5/H-5' for α - and β-anomers), 5,4-5,8 (4×m, H-1 and H-3 for a and7,18-8,02 (m, 7H, aromatic protons for a and b).

Example 23(g)

2-deoxy-3,5-di-O-para-toluoyl-β-L-retroinformation 8

Method 1: Directly from lactol 7

A solution of 2-deoxy-3,5-di-O-para-toluoyl-α,β-L-ribose7(about 7.0 g, 18,91 mmol) in TBME (30 ml) was diluted with TBME (15 ml) and was stirred for 20 minutes at room temperature. Was added acetic acid three portions of 1 ml with stirring until obtaining a clear brown solution. The solution was cooled to 0°C in argon and passed dry HCl gas constant stream for 25 minutes. After 10 minutes at 0°C aliquot extinguished anhydrous ethanol (1.2 ml) and left to stand at room temperature with periodic shaking for 10 minutes, thus obtaining a clear solution. The reaction mixture was stirred at 0-5°C and 65 minutes, the reaction mixture was filtered. The solid product washed with TBME (30 ml) and dried in vacuum for 5 hours, obtaining 2-deoxy-3,5-di-O-para-toluoyl-β-L-retroinformation8(4,79 g, 65%) as a white crystalline solid. TPL 118-121°C.

δN(CDCl3, 400 MHz): 2,41, 2,3 (2×s, 2×WithN3AG, 2×3H), 2,74 (1H, a-d, J2,2'=14.6 Hz, H-2'), 2,87 (1H, DDD, J2,2'=a 12.7 Hz, J2,3=7,3 Hz, J1,2=5.3 Hz, H-2), 4,59 (1H, DD, J5,5'=and 12.2 Hz, J4,5'=4.4 Hz, H-5'), and 4.68 (1H, DD, J5,5'=and 12.2 Hz, J4,5=3,4 Hz, H-5), a 4.86 (1H, m, J=3,4 Hz, H-3/H-4), to 5.56 (1H, a-DD, J=1,95 Hz, J=6.3 Hz, H-3/H-4), 6,47 (1H, d, J1,2=5.4 Hz, H-1), of 7.23-7,28 (4H, m, 2×AGN), 7,89, 7,99 (2×2H, 2×d, J=8,3 Hz, 2×AGN). [a]D25= -117 (C, 1.0 in l3) [CMS chemicals Ltd: [a]D20= -118,9 (C, 1 in DHM)].

Method 2: Through the methoxide 7-OMe

A solution of 1% HCl in methanol was obtained by adding acetylchloride (0.2 ml) to methanol (10 ml), previously cooled to 5°C. 2-deoxy-3,5-di-O-para-toluoyl-α,β-L-ribose7(470 mg, of 1.27 mmol) was dissolved in anhydrous methanol (9 ml) and cooled to approximately 10°C. was Added portion 1% HCl solution in methanol (1 ml) and the reaction mixture is kept at 10-15°C for 1.5 hours. After a specified period of time HPLC analysis showed the presence of unreacted educt, so I added an additional portion of 1% HCl in methanol (1 ml) and stirring continued at room temperature for 1.5 hours. The HPLC analysis showed that the reaction was near completion, and the reaction mixture was concentrated in vacuo at 30°C and evaporated together with TBME (10 ml). The residue was dissolved in TBME (10 ml) and observed suspended white solid. Added etilize is at (15 ml), to dissolve the suspension, and the solution was dried with sodium sulfate (2 g), filtered and concentrated in vacuum, obtaining methyl-2-deoxy-3,5-di-O-para-toluoyl-α,β-L-ribose 7-OMe (480 mg, crude yield 98%) as a brown oil.

δN(CDCl3400 MHz processor, the ratio of anomers is 1:1): 2,2-2,6 (m, 2×WithN3AG and H-2/H-2' for α - and β-anomers), to 3.36 (s, 3H, och3of 3.42 (s, 3H, och34,4-4,6 (m, H-4 and H-5/H-5' for α - and β-anomers), 5,19 (d, 1H, H-1 forto 5.21 (DD, 1H, H-1 for5,41 (m, 1H, H-3 for5,59 (m, 1H, H-3 for7,18-8,02 (m, 8H, aromatic). δWith(CDCl3, 100 MHz): 21,9, 39,5, 55,3, 55,4, 64,5, 65,3, 74,8, 75,6, 81,2, 82,1, 105,3, 105,8, 127,1, 127,2, 127,3, 127,4, 129,3, 129,3, 129,9, 130,0, 130,0, 143,9, 144,0, 144,1, 144,2, 166,3, 166,5, 166,6, 166,7.

Methyl-2-deoxy-3,5-di-O-para-toluoyl-α,β-L-ribose 7-OMe (480 mg, 1.25 mmol) was dissolved in TBME (3 ml) and acetic acid (1 ml) and the solution was cooled to 0°C in argon. The resulting solution was barbotirovany dry HCl gas for 15 minutes and the reaction mixture was allowed to mix at 0-5°C for 10 minutes. The HPLC analysis showed the presence of the remaining original substance even after the reaction continued for 1 hour and 10 minutes. White solid, which crystallized from actionnow mixture, collected by filtration, washed (TBME) and dried in vacuum, obtaining 2-deoxy-3,5-di-O-para-toluoyl-β-L-retroinformation8(228 mg, 47% for stage 3) in the form of a white crystalline solid. A dedicated connection in all respects was identical to the connection shown above in the case of method 1.

Example 23(h)

2'-deoxy-3',5'-di-O-para-toluoyl-L-thymidine 9

A mixture of thymine (1.0 g, a 7.92 mmol), HMDS (1.66 g, 10,28 mmol) and ammonium sulfate (100 mg, from 0.76 mmol) was heated at about 145°C for 2 hours, and at this time thymine was dissolved. After another 4 hours at 145°C, the reaction mixture was concentrated in vacuum at 60°C. the thus Obtained sililirovany thymine (7,92 mol) is suspended in anhydrous chloroform (15 ml) and cooled to 15°C. Portions were added 2-deoxy-3,5-di-O-para-toluoyl-β-L-retroinformation8(1.48 g, 3.8 mmol) in 2-3 minutes and the reaction mixture (yellow solution) was stirred at room temperature in argon atmosphere. After 2 hours, HPLC analysis showed that starting material was absent. The reaction mixture was cooled to approximately 5°C and extinguished 90% ethanol (0.25 ml). Was the loss of the white solid precipitate, and the reaction mixture was stirred at room temperature for 20 minutes. The reaction mixture was filtered through celite (10 g) and about ivali dichloromethane (60 ml). The filtrate was washed with water (2×25 ml) and left to stand to the emulsion stratified. Then the organic layer was washed with an aqueous solution of sodium bicarbonate (2 g in 25 ml water) and a saturated solution of salt (10 g NaCl in 30 ml of water). The organic layer was dried with sodium sulfate (5 g) and filtered through celite (8 g). The filtrate was concentrated in vacuum at 40°C and the residue is suspended in hexane (20 ml)which was stirred at room temperature for 1.5 hours to obtain a homogeneous dispersion. The mixture was filtered, the filter cake was washed with hexane (10 ml) and was briefly dried under vacuum, obtaining white solid. The obtained solid was stirred in ethanol (30 ml) at 60°C for 40 minutes, concentrated in vacuo (removed 15 ml) and the remaining suspension was cooled to room temperature. The suspension was filtered, washed with cold ethanol (10 ml) and TBME (5 ml) and dried in vacuum at 55°C during the night, getting a 2'-deoxy-3',5'-di-O-para-toluoyl-L-thymidine9(1,38 g, 76%) as a white solid. HPLC: the ratio of α:β=268:1.

δN(CDCl3, 400 MHz): to 1.61 (3H, s, 5-Me), 2,31 (1H, m, H-2”), 2,42, 2,43 (2×3H, 2×s, WithN3AG), a 2.71 (1H, DD, J=4,8 Hz, J2',2"=14,3 Hz, H-2'), to 4.52 (1H, m, H-4'), with 4.64 (1H, DD, J4',5"=3.3 Hz, J5',5"=12,5 Hz, H-5”), of 4.77 (1H, DD, J4',5'=2,6 Hz, J5',5”=12,5 Hz, H-5'), 5,64 (1H, a-d, J=6,6 Hz, H-3'), 6,47 (1H, DD, J1,2=5.5 Hz, J1,2=8,8 Hz, H-1'), 7,2-7,3 (5H, m, H-6 and ArH), to $ 7.91-of 7.96 (4H, m, Ar-H), and 8.6 (1H, users, NH); δC(CDCl3, 100 MHz): 12,3, 21,5, 21,9, 38,2, 64,4, 75,1, 83,0, 85,0, 111,9, 126,3, 126,7, 129,5, 129,7, 129,7, 130,0, 134,6, 144,8, 150,6, 163,8, 166,2, 166,3.

Example 23(i)

2'-deoxy-L-thymidine 10

Stir a suspension of 2'-deoxy-3',5'-di-O-para-toluoyl-L-thymidine9(500 mg, 1.05 mmol) in anhydrous methanol (6 ml) was cooled to 5°C in argon. One portion was added sodium methoxide (64 mg, 1,19 mmol). After 5 minutes the cooling bath was removed and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was heated to 45-50°C for 1 hour, and it remained in the form nearly insoluble suspension. Added anhydrous tetrahydrofuran (4 ml) and received a clear solution. The temperature was maintained at 45-50°C for 30 minutes, at which point HPLC analysis showed the presence of the remaining source material. So after another 30 minutes (only 2.5 hours) added an additional portion of sodium methoxide (31 mg, or 0.57 mmol) and the reaction mixture was stirred at 45-50°C. After 2.5 hours (5 hours) before HPLC analysis showed that the reaction was not completed, so even after 1 hour (6 hours) added an additional portion of sodium methoxide (25 mg, 0.46 mmol) and the reaction mixture was stirred overnight at 40-45°C. After a specified period of time HPLC and TLC analysis (10% methanol in ethyl acetate) pok who had complete metamorphosis educt (R f0,73) in the product (Rf0,15) [sample prepared for HPLC and TLC analysis = an aliquot of resin Dowex-H+the resin was diluted with methanol, filtered and analyzed]. The reaction mixture was cooled to room temperature and was added ion-exchange resin DOWEX 50W × 2-200(H) (pre-washed with methanol (3×10 ml). After stirring for 30 minutes at room temperature the pH was equal to 3, and the reaction mixture was filtered using a porous glass funnel, washed with methanol (5 ml) and the filtrate was concentrated in vacuum at 45°C. the Remaining methanol is evaporated together with a mixture of TBME and dichloromethane (1:1, 10 ml) and the residue was dissolved in TBME (10 ml). The solids were dispersively at 40-45°C for 1 hour, cooled to room temperature and collected by filtration. The resulting solid is washed with TBME (5 ml) and dried in vacuum, obtaining 2-deoxy-L-thymidine10(225 mg, 88%) as a white solid.

Found that the compound obtained was identical in all respects to an authentic sample of 2-deoxy-L-thymidine10.

Example 24

2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine (2) of 2,2'-anhydrous-1-β-D-arabinofuranosyl)thymine (1)

To pre-cooled (0-5°C) a mixture of 2,2'-anhydrous-1-β-D-arabinofuranosyl)thymine1(2,40 is, 10.0 mmol) and DMAP (111 mg, 0.9 mmol) in anhydrous pyridine (15 ml) portions was added 4,4'-dimethoxytrityl (of 3.56 g, 10.5 mmol) over a period of time equal to 3 minutes. The resulting mixture was stirred at 0-5°C in an argon atmosphere, and after 1.5 hours, TLC analysis (plates silica, methanol:dichloromethane 1:9) showed no remaining source materials. The reaction mixture was concentrated in vacuum at 45°C. the Residue was collected in dichloromethane (50 ml) and saturated sodium bicarbonate solution (20 ml). After stirring at room temperature for 10 minutes, the layers were separated, the organic layer was washed with distilled water (2×20 ml) and dried with anhydrous sodium sulfate. The reaction mixture was concentrated in vacuum at 50°C and the residue was evaporated together with toluene (2×10 ml). The resulting crude residue is triturated in dichloromethane (5 ml) and TBME (25 ml). After stirring at room temperature for 1 hour, the solid was collected by filtration under reduced pressure and washed with TBME (15 ml). The yellow solid was dried in vacuum, obtaining 2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine2(5.3g, the yield of 97.8%, 91% AUC (area under the curve) by HPLC analysis).

δN(d6-DMSO, 400 MHz): 1,78 (3H, s, Me), was 2.76 and 2,90 (2H, ABX, H-5' and H-5”), and 3.72 (6N, s, 2×OMe), 4,22-to 4.28 (2H, 2×m, H-3' and H-4'), to 5.17 (1H, d, J1',2'=5,9 Hz, H-2') 5,93, (1H, d, J3',OH=4.4 Hz, 3'-OH), 6,30 (1H, d, J1',2'=5,9 Hz, H-1'), 6,79-7,28 (13H, m, Ar-H), to 7.84 (1H, s, H-6).

Example 25

2'-deoxy-5'-O-dimethoxytrityl-β-D-thymidine (3) of 2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine (2)

Example 25 illustrates the comparison between the percentage yield 5'-DMTrO-protected 2'-deoxythymidine (3)obtained by method 1, which uses Red-Al in toluene, with output in % of product (3)obtained by method 2, which uses Red-Al in combination with 15-crown-5-ether in DME. Method 1 was carried out using known methods (for example, U.S. patent No. 6090932), and received the product (3with 21%. Method 2 was carried out according to the method proposed in the present invention, in which use Red-A1 in combination with 15-crown-5-ether in DME, which gave the product (3with 35%.

Method 1: Interaction of Red-Al with 2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine 2 in toluene

To pre-cooled (0-5°C) solution of 2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine2(1.08 g, 2.0 mmol) in anhydrous toluene (50 ml) was added dropwise Red-Al (65 wt.% in toluene, of 0.90 ml, 3.0 mmol) within a time period of 10 minutes. The mixture was kept under stirring at 0-5°C in an argon atmosphere. The reaction was controlled by TLC (diox the d silicon, 5:95 methanol in dichloromethane) and HPLC analysis. After stirring for 2 hours at 0-5°C to the reaction mixture was added an additional amount of Red-Al (0.5 EQ., 65 wt.% in toluene, of 0.30 ml, 1.0 mmol). After stirring for 45 minutes took an aliquot from the reaction mixture in THF, net for HPLC (approximately 1 ml), was suppressed by adding dropwise distilled water, and were injected with the device for HPLC. The result showed a ratio of 1:1 product (37,4% AUC) to the original substance (36% AUC). The reaction was suppressed by the addition of saturated salt solution (30 ml) at 5°C. After stirring for 30 minutes the mixture was filtered through a pad celite and washed with ethyl acetate (60 ml). The filtrate was distributed in a separating funnel. The organic layer was washed with a saturated aqueous solution of NH4Cl (30 ml) and saturated salt solution (2×25 ml) and dried using anhydrous sodium sulfate. The reaction mixture was concentrated in vacuum at 40°C. the crude residue (1.01 g, yellow foamy solid) was purified by chromatography on a column (silica gel, 5% methanol in dichloromethane)to give 2'-deoxy-5'-O-dimethoxytrityl-β-D-thymidine3(0,23 g, yield 21%) as a pale yellow solid.

δH(d6-DMSO, 400 MHz): USD 1.43 (3H, s, Me), 2.14 and 2,22 (2×1H, 2×m, H-2' and H-2”), 3,18 (2H, m, H-5 and H-5'), and 3.72 (s, 6H, 2×OMe), a 3.87 (1H, m, H-4'), 4,30 (1H, m, H-3'), 5,32 (1H, d, J3',OH=4.4 Hz, 3'-OH), to 6.19 (1H, m, H-1'), 6,85-7,39 (13H, the, DTr), to 7.50 (1H, s, H-6), 11,38 (1H, s, NH); MS (ESI+, M+1=545,3, M+Na+=567,3).

Method 2: Interaction of Red-Al with 2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine 2 in DME in the presence of 15-crown-5

To pre-cooled (0-5°C) solution of 2,2'-anhydrous-1-(5-O-dimethoxytrityl-β-D-arabinofuranosyl)thymine2(120 mg, 0.22 mmol) and 15-crown-5 (65 μl, 0.33 mmol) in anhydrous DME (5 ml) was added dropwise Red-Al (65 wt.% in toluene, 0.10 ml, 0.33 mmol) over a period of time of 4 minutes. The reaction mixture was stirred at 0-5°C in an argon atmosphere. The reaction was monitored by HPLC analysis. After stirring for 3.5 hours the reaction mixture was added an additional amount of Red-Al (0.5 EQ., 65 wt.% in toluene, 0,030 ml, 0.10 mmol). After stirring for 30 minutes the results of HPLC analysis showed that the ratio of product:starting material was increased from 1.7:1 to 2.4:1. The reaction mixture was allowed to warm to room temperature and kept at room temperature for 16 hours. The HPLC analysis showed that the ratio of the product to the original substance increased slightly to 2.8:1. Then the reaction mixture was cooled to 0-5°C and the reaction mixture was added 0.5 equivalent of Red-Al (65 wt.% in toluene, 0,030 ml, 0.10 mmol). After stirring at 0-5°C for 1 hour the HPLC results showed that the ratio of the product to the original substance which was srastello to 4.0:1 (62,7% AUC of product compared with 15.8% of the AUC of the original substance). Further addition of 15-crown-5 and Red-A1 did not lead to the improvement of the education product. To the reaction mixture was added a small amount of acetone (about 0.1 ml). After stirring for 10 minutes the reaction mixture was concentrated in vacuum at 40°C and the residue was evaporated together with isopropylacetate (10 ml). The residue was distributed between isopropylacetate (20 ml) and distilled water (5 ml). The organic layer was washed with a saturated aqueous solution of NH4Cl (5 ml) and saturated salt solution (5 ml) and dried with anhydrous sodium sulfate. After concentration in vacuo at 40°C crude residue (120 mg, yellow foamy solid) was purified by chromatography on a column (silica gel, 5% methanol in dichloromethane)to give 2'-deoxy-5'-O-dimethoxytrityl-β-D-thymidine3in the form of a light yellow solid (45 mg, yield 35%). Range1H-NMR corresponds to the structure obtained according to method 1.

Example 26

2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (4) (2,2'-anhydrous-1-β-D-arabinofuranosyl)thymine (1)

2,2'-anhydrous-1-β-D-arabinofuranosyl)thymine1(500 mg, of 2.08 mmol) was dissolved in anhydrous pyridine (5 ml) and stir the solution was added DMAP (12.5 mg, 0.1 mmol). Portions were added trailhead (1.28 g, to 2.29 mmol) for 3 minutes at room temperature. Receiving the reduction in the reaction mixture was stirred at room temperature for 2 hours and then at 40°C overnight in an argon atmosphere. After a specified period of time TLC analysis (plate of silica, methanol:dichloromethane 2:8) showed that the original substance remained (Rf0.3) and produced a new product (plate of silica, methanol:dichloromethane 1:9, Rf0,17). The reaction mixture was cooled to 0°C, using a bath of ice, portions was slowly added to a saturated solution of NaHCO3(15 ml), and white solid precipitated from solution in the sediment. The resulting suspension was stirred at room temperature for 30 minutes, the white solid was collected by filtration and then washed with distilled water (25 ml). The crude solid (3 g) was collected in TBME (18 ml) and stirred at room temperature for 30 minutes. White solid was collected by filtration, washed with TBME (8 ml) and then dried in vacuum, obtaining 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine4(844 mg, 84%) as a white solid.

δH(d6-DMSO, 400 MHz): or 1.77 (3H, s, Me), 2,73 (1H, DD, J4',5”=7,4 Hz, J5',5”=10,2 Hz, H-5”), of 2.92 (1H, DD, J4',5'=4,3 Hz, J5',5”=10,2 Hz, H-5'), 4,24-4,28 (2×1H, 2×m, H-3' and H-4'), 5,16 (1H, d, J1',2'=5,86 Hz, H-2'), 5,94 (1H, d, J3'HE=4,23 Hz, 3'-OH), of 6.29 (1H, d, J1',2'=the 5.45 Hz, H-1'), 7,2-7,27 (15H, m, Tr), 7,83 (1H, users, H-6).

Example 27

2'-deoxy-5'-O-trityl-β-D-thymidine (5) of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)Tim is on (4)

To pre-cooled (0-5°C) a mixture of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine4(241 mg, 0.5 mmol) and 15-crown-5 (0.15 ml, 0.75 mmol) in anhydrous THF (10 ml) was added dropwise Red-Al (65 wt.% in toluene, to 0.23 ml, 0.75 mmol) over a period of time of 5 minutes. The mixture was maintained at 0-5°C in an argon atmosphere. The reaction was monitored by TLC analysis (silica, 5:95 methanol in dichloromethane) and HPLC. After stirring at 0-5°C for 1 hour from the reaction mixture were taken of the sample in THF, net for HPLC (approximately 1 ml), was suppressed by the addition dropwise of distilled water and were injected with the device for HPLC. The results showed that only 8% of the AUC (area under the curve) of the original substance and was attended by 70.8% of the product. The reaction was suppressed by the addition of saturated aqueous solution of NH4Cl (5 ml) at 5°C and was stirred for 15 minutes. After a specified period of time, the layers were separated and the aqueous layer was then extracted with isopropylacetate (10 ml). The organic layers were combined, washed with saturated salt solution (5 ml) and dried using anhydrous sodium sulfate. After concentration in vacuo at 40°C crude residue (287 mg, white foamy solid) was purified by chromatography on a column (silica gel, 5% methanol in dichloromethane)to give 2'-deoxy-5'-O-trityl-β-D-timide the 5(106 mg, yield 44%) as a white solid.

δN(d6-DMSO, 400 MHz): 1,45 (3H, s, Me), to 2.15 (1H, m, H-2”), 2,22 (1H, m, H-2')and 3.15 (1H, DD, J4',5”=2,6 Hz, J5',5”=10.5 Hz, H-5”), up 3.22 (1H, DD, J4',5'=4,8 Hz, J5',5”=10.5 Hz, H-5'), a 3.87 (1H, m, H-4'), or 4.31 (1H, m, H-3'), 5,33 (1H, d, J3=a 4.83 Hz, 3'-OH), to 6.19 (1H, a-t, J=6,6 Hz, J=7,0 Hz, H-1'), 7,25-7,39 (15H, m, Tr), 7,49 (1H, users, H-6), 11,35 (1H, s, NH).

δWith(d6-DMSO, 100 MHz): 11,7, 54,9, 70,4, 83,7, 85,4, 86,4, 109,6, 127,2, 128,0, 128,3, 135,7, 143,5, 150,4, 163,7.

Removed the protection of the obtained compound (5using acetic acid at about 50°C, obtaining 2'-deoxy-D-thymidine in the quality of the final product, which was identical in all respects to the authentic sample of the known 2'-deoxy-D-thymidine.

Example 28

The formation of 2'-deoxy-D-thymidine (4) of 2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine (1)

Example 28(a)

2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2) of 2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine (1)

2,2'-anhydrous-1-(β-D-arabinofuranosyl)thymine (1) (10.0 g, 41,62 mmol) suspended in pyridine (100 ml) and DMAP (254 mg, of 2.08 mmol) and portions were added trailhead (25,48 g, 91,56 mmol) at room temperature. The reaction mixture was stirred at room temperature for about 1 hour and then heated to about 45°C (the internal temperature). After about 5 hours, TLC analysis (10% methanol in dichloromethane, rendering using 1% KMnO4and UV) showed the presence of significant amounts of the original substance (Rf0,15) and formation of product (Rf0,43). Therefore, the reaction mixture was stirred at about 45°C for about 15 hours (during the night). After a specified period of time TLC analysis showed that the original substance remained (Rf0,15). The reaction mixture was cooled to approximately 0°C and slowly for 15 minutes was added a saturated aqueous solution of NaHCO3(320 ml) (changes in the internal temperature did not occur). From the solution immediately precipitated white solid, and the white suspension was stirred for about 30 minutes at room temperature. The solid was isolated by filtration through a Buchner funnel and washed with water (3×100 ml). The remaining solid substance was collected in dichloromethane (150 ml) and was stirred for about 30 minutes at room temperature. The residue was isolated by filtration through a Buchner funnel, washed with dichloromethane (20 ml) and dried in vacuum over night, getting 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2) (14,66 g, 73%) as a white solid.

δN(d6-DMSO, 400 MHz): or 1.77 (3H, s, 5-Me), was 2.76 (1H, DD, J5',5”=10.3 Hz, J4',5”=7.8 Hz, H-5”), to 2.94 (1H, DD, J5',5”/sub> =10.3 Hz, J4',5'=3,9 Hz, H-5'), 4.26 deaths (1H, m, H-4'), the 4.29 (1H, m, H-3'), to 5.17 (1H, a-d, J=5,9 Hz, H-2'), 5,98 (1H, users, 3-OH), 6,30 (1H, d, J1',2'=lower than the 5.37 Hz, H-1'), 7,2-7,27 (15H, m, Tr), 7,83 (1H, s, H-6); δC(d6-DMSO, 125 MHz): 13.5cm (5-Me), 63,2 (C-5'), 74,8 (C-3'), 85,9 (TrC), To 86.7 (C-4'), At 88.1 (C-2'), 89,9 (C-1'), 116,9 (-6), 127,0, 127,7, 127,9, 128,0 (Tr), 132,1 (C-5), Br143.3 (Tr), 158,8 (C-2), Is 171.3 (C-4).

Example 28(b)

2'-deoxy-5'-O-trityl-β-D-thymidine 3 of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine 2

2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2) (4,30 g, 8,91 mmol) suspended in anhydrous tetrahydrofuran (43 ml) and cooled to about 0-5°C, using a bath of ice. In a separate flask, immersed in a bath of ice, at about 0-5°C, 65 wt.% diluted solution of Red-Al in toluene (3,26 ml, 10,69 mmol) is added to dry tetrahydrofuran (21,5 ml). The obtained diluted solution of Red-Al was cooled to about 0-5°C was added dropwise via syringe to a suspension of 2,2'-anhydrous-1-(5-O-trityl-β-D-arabinofuranosyl)thymine (2) in tetrahydrofuran. The speed of adding dropwise the solution of Red-Al is important for the reaction, and the addition was completed in about 1 hour. The resulting clear solution was kept at about 0-5°C for 1 hour, after a specified period of time TLC analysis (10% methanol in dichloromethane) showed the presence of educt (Rf0,34), the desired product R f0,47) and impurities (Rfof 0.42 and 0.26). The HPLC analysis showed the presence of the original substance (11,35 min, 36,5% AUC), product (12,60 min, 24%) and a small amount of basic impurities (11,7 min, 2.9 per cent). After only about 2 hours at a temperature of about 0-5°C, dropwise via a syringe over a period of time of about 20 minutes, adding an additional portion of "undiluted" 65 wt.% solution of Red-Al in toluene (1,63 ml, to 5.35 mmol) to the reaction mixture, which was kept at about 0-5°C. After about 1 hour analysis of TLC and HPLC showed the presence of the original substance (11,35 min, 3.2 per cent). Was added dropwise next portion 65 wt.% solution of Red-Al in toluene (of 0.26 ml, 0.85 mmol) and the reaction mixture was stirred at about 0-5°C during 45-minute period. After a specified period of time TLC analysis showed the presence of only trace amounts of the remaining source material. The reaction was suppressed by addition of a saturated solution of NH4Cl (40 ml) and a layer of tetrahydrofuran decantation. The aqueous layer was extracted with isopropylacetate (50 ml) and the resulting emulsion was subjected to separation by slow addition of 5 n HCl solution (15 ml). The organic layer was separated, combined with a layer of tetrahydrofuran and washed with saturated solution of NH4Cl (30 ml) and then with saturated salt solution (30 ml). At this point the pH of the saturated layer Rast is ora salt ranged from 6.5 to 7, and the organic layer was dried using Na2SO4, filtered and concentrated in vacuum, getting foamy solid (4.4 g). The crude residue was evaporated together with toluene (30 ml), concentrated in vacuo and the resulting residue was collected in toluene (25 ml), heating to about 45°C. the Mixture was cooled to room temperature and stirred at the same temperature until they began to precipitate white solid. Was added dropwise water (8.5 ml) and the resulting mixture was stirred at room temperature for about 3 hours. The solid was isolated by filtration and the filter cake washed with water (5 ml) and toluene (3 ml). The solid was dried at about 45°C in high vacuum for about 1 hour and then at room temperature under vacuum over night, getting a 2'-deoxy-5'-O-trityl-β-D-thymidine (3) (1.77 g, 41%) as a white solid.

δN(d6-DMSO, 400 MHz): 1,45 (3H, s, Me), to 2.15 (1H, m, H-2”), 2,22 (1H, m, H-2')and 3.15 (1H, DD, J4',5”=2,6 Hz, J5',5”=10.5 Hz, H-5”), up 3.22 (1H, DD, J4',5'=4,8 Hz, J5',5”=10.5 Hz, H-5'), a 3.87 (1H, m, H-4'), or 4.31 (1H, m, H-3'), 5,33 (1H, d, J3'HE=a 4.83 Hz, 3'-OH), to 6.19 (1H, a-t, J=6,6 Hz, J=7,0 Hz, H-1'), 7,25-7,39 (15H, m, Tr), 7,49 (1H, users, H-6), 11,35 (1H, s, NH). δWith(d6-DMSO, 100 MHz): 11,7, 54,9, 70,4, 83,7, 85,4, 86,4, 109,6, 127,2, 128,0, 128,3, 135,7, 143,5, 150,4, 163,7.

2'-deoxy-D-thymidine (4) from 2'-deoxy-5'-O-trityl-β-D-thymidine (3)

2'-deoxy-5'-O-trityl-β-D-thymidine (3) (1,215 g, 2.5 mmol) suspended in methanol (9.6 ml) and the reaction mixture was heated to about 45°C in a water bath until dissolution. Then the flask was cooled to room temperature, to the mixture was added concentrated HCl (200 μl, 2.5 mmol) and stirred at room temperature. After about 25 minutes white solid began to precipitate from solution. After about 1 hour, TLC analysis (10% methanol in dichloromethane, visualization vanilla and UV) showed that starting material remained (Rf0,53) and formed the main product (Rf0,21). To the reaction mixture was added portion of n-heptane (10 ml) and stirred at room temperature for about 15 minutes. White solid was isolated by filtration (405 mg solids). The filtrate was divided into two layers, the methanol layer was extracted with n-heptane (10 ml) and then concentrated in vacuo to a volume of 2 ml, the Residue was combined with 405 mg of a white solid, suspended in TBME (10 ml) and stirred at room temperature for about 1 hour. White solid was isolated by filtration, washed with TBME (3 ml) and dried in vacuum furnace, obtaining 2'-deoxy-D-thymidine (4) (471 mg, 78%). Received the initial product was identical in the analysis of 1H-NMR and HPLC authentic sample.

The invention is described with reference to various specific and preferred options and techniques. However, it should be understood that many changes and modifications will be obvious to experts in this field on the basis of the above detailed description of the invention and can be implemented without departing from the essence and not going beyond the scope of the invention.

1. The method of obtaining 2'-deoxy-(β-L-thymidine, including
a) obtaining 5'-O-trityl - or 5'-O-dimethoxytrityl - protected 2,2'-anhydrous-1-β-L-arabinofuranosyladenine;
b) interaction of the 5'-O-trityl - or 5'-O-dimethoxytrityl - protected 2,2'-anhydrous-1-β-L-arabinofuranosyladenine from stage (a) with a reducing agent RedAl and complexing agent 15-crown-5-ether in a polar solvent 1,2-dimethoxyethane (DME) or tetrahydrofuran (THF) to give 5'-O-trityl - or 5'-O-dimethoxytrityl - protected 2'-deoxy-β-L-thymidine; and
c) removing the protection of the product from step (b), if necessary or desirable.

2. The method according to claim 1, wherein in stage (C) removing the protection is implemented by adding an acid or an acidic resin at a temperature of about 50°C.

3. The method according to claim 1, wherein in stage (b) the reaction temperature is about 0-5°C.

4. The method according to claim 1, wherein said 5'-O-trityl - or 5'-O-dimethoxytrityl - protected 2,2'-is gidro-1-β-L-arabinofuranoside get through the following stages:
a) protection of L-ribofuranose by interacting with dimethoxytrityl or trailvoy protecting group;
b) condensation of the product from step (a) with thymine with the formation of nucleoside and
(C) interaction of nucleoside from step (b) with a condensing agent with formation of 5'-O-trityl - or 5'-O-dimethoxytrityl - protected 2,2'-anhydrous-1-β-L-arabinofuranosyladenine.

5. The method according to claim 4, in which in stage (b) the condensation is carried out in the presence of a solvent and optionally a catalyst.

6. The method according to claim 4, in which stage (s) of the condensing agent is a dialkyl - or varikont in the presence of a base and an organic solvent.

7. The method according to claim 6, in which the condensing agent is PhOCOOPh/NaHCO3and an organic solvent is DMF.

8. The method according to claim 4, in which in stage (C) the interaction occurs at elevated temperatures.

9. The method according to claim 8, in which the temperature is about 140-150°C.

10. The method of obtaining 2'-deoxy-β-L-thymidine, including
a) the interaction of L-arabinose with cyanamide with the formation of L-arabinofuranosylcytosine;
b) interaction of the product from step (a) with collisuem or condensing agent with the formation of 2,2'-anhydrous-1-β-L-arabinofuranosyladenine;
c) interaction of 2,2'-anhydrous-1-β-L-arabinofuranosyladenine with a reducing agent RedAl and to mlekoobraznymi agent 15-crown-5-ether in a polar solvent 1,2-dimethoxyethane (DME) or tetrahydrofuran (THF) to obtain 2'-deoxy-β-L-thymidine, moreover, L-arabinofuranosyluracil can be protected by trition or dimethoxytrityl in position 5' before or after interaction with collisuem or condensing agent; and
d) removing the protecting optionally protected 2'-deoxy-β-L-thymidine, if necessary or desirable.

11. The method according to claim 10, in which in stage (d) removing the protection is implemented by adding an acid or an acidic resin at a temperature of about 50°C.

12. The method according to claim 10, in which in stage (b) cyclisme or condensing agent selected from the group consisting of:
,,
,,,
,and

13. The method according to claim 10, in which at the stage (C) the reaction temperature is about 0-5°C.



 

Same patents:

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
The invention relates to the synthesis of nucleosides and relates to an improved method for the preparation of 3'-azido-2',3'-dideoxythymidine with the ability to suppress the reproduction of human immunodeficiency virus and finds application in medical practice for the treatment of AIDS

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

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: 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: medicine.

SUBSTANCE: method of obtaining of the agent possessing anti ulcerous activity is carried out by an extraction with the water cleared from waste of essential-oil manufacture of a medicinal chamomile in the ratio 1:20 at 80°C within 1.5 hours on a water bath, twice, filtration of extracts, processing by 90% ethanol, separation of a deposit, washing with ethanol and drying in vacuum. A waste of essential-oil manufacture of a medicinal chamomile is used for reception of biologically active agent stimulating reparative processes at a peptic ulcer of stomach and duodenum.

EFFECT: use of the received agent dilates indications to prescription at stomach and duodenum peptic ulcer.

FIELD: biotechnologies.

SUBSTANCE: invention refers to biotechnology, and namely to analysis method of heparins or low-molecular heparins, and can be used when controlling samples, and for standardising Lovenox method and obtaining homogeneous products. Method of quantitative determination of heparins or low-molecular heparins is implemented by means of the following stages: sample is depolymerised with heparinases, then the obtained depolymerisate is reduced, and after that the analysis is performed by means of high-efficiency liquid chromatography.

EFFECT: this invention allows to clearly differentiate Lovenox from other low-molecular heparins not containing "1,6-anhydro" derivatives.

7 cl, 2 dwg, 1 tbl, 6 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: 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: pharmacology.

SUBSTANCE: claimed invention relates to pyrazole derivatives, which are represented by general formula (I), as well as theirpharmacologically acceptable salts, which have inhibiting activity against human SGLT1, to pharmacological composition, inhibitor of human SGLT1 and based on them medications, to their application for producing pharmacologic composition and to intermediate compounds for their obtaining. where R1 represents H, hydroxy(C2-6)alkyl group, one of Q and T represents group, which is presented by general formula: or group, which is presented by general formula while another presents C1-6alkyl group; R2 represents hydrogen atom, C1-6alkyl group or group of formula: -A-R8, where A represents oxygen atom, and R8 represents C6hetherocycloalkyl group, containing oxygen atom as heteroatom; X represents simple bond or oxygen atom, Y represents C1-6alkylene group or C2-6alkylene group; Z represents carbonyl group or sulphonyl group; R4 and R5 are similar or different, and each represents hydrogen atom or C1-6alkyl group, which can have similar or different 1-3 substituents, selected from substituents (i) Values of sunstituents (i) are iven in invention formula.

EFFECT: obtaining of pyrazole derivatives and based on them medications.

28 cl, 3 tbl, 197 ex

FIELD: biology.

SUBSTANCE: present invention relates to biotechnology, more specifically to obtaining nucleoside-5'-triphosphates, labelled with phosphorous-32 (phosphorous-33) in the alpha-position, and can be used for analysis in molecular biology, genetics and medical biochemistry. The method is realised through treatment of labelled nucleosidephosphate in a buffer solution with a mixture of deoxyribonucleoside monophosphate kinase of bacteriophage T5 and pyruvate kinase with subsequent chromatographic purification of the target product.

EFFECT: simple method of obtaining nucleoside-5'-triphosphates and stable output of the target product.

4 ex

FIELD: medicine.

SUBSTANCE: invention relates to method of obtaining gemcitabine hydrochloride, characterised by the following: 2,2-dimethyl-[1,3]-dioxolane-4-carbaldehyde is subjected to interaction with ethyl bromodifluoracetate in presence of zinc in organic solvent medium processing reaction mixture with ultrasound for 5-60 minutes, obtained ethyl 3-hydroxy-2,2-difluoro-3-[2,2-dimethyl-[1,3]dioxolane-4-yl]propionate is subjected to hydrolysis and cyclisation by means of ion-exchange resin in water-alcohol medium obtaining (4R,5R)-4-hydroxy-5-hydroxymethyl-3,3-difluorodihydrofuran-2(3H)-on, which is processed with solution of trimethylchlorosilane in dichloromethane obtaining (4R,5R)-4-trimethylsilyloxy-5-((trimethylsilyloxy)methyl)-3,3-difluorodihydrofuran-2(3H)-on, which is subjected to reduction by means of lithium diisopropylalumohydride in organic solvent medium at cooling to -70°C obtaining (4R,5R)-2-hydroxy-4-(trimethylsilyloxy)-5-((thrimethylsilyloxy)methyl)-3,3-difluorotetrahydrofurane, which is converted into (4R,5R)-2-methylsulphonyloxy-4-(trimethylsilyloxy)-5-((trimethylsilyloxy)methyl)-3,3-difluorotetrahydrofurane by processing with methane sulphonylchloride in solvent medium at cold, obtained (4R,5R)-2-methylsulphonyloxy-4-(trimethylsilyloxy)-5-((trimethylsilyloxy)methyl)-3,3- difluorotetrahydrofurane after optic isomer separation is processed with bis-trimethylsilylacetylcytozine in water-free dichlorethane and boil with trifluoromethane sulphonyloxymethylsilane with further cooling and separation of obtained gemcitabine in form of base or hydrochloride, as well as method of gemcitabine hydrochloride purification by its re-crystallisation from water solution with processing with ultrasound.

EFFECT: invention results in increase of ratio 3-(R)-hydroxy-isomer to 3(S)-hydroxy-isomer.

6 cl, 2 dwg, 4 ex

FIELD: chemistry; pharmacology.

SUBSTANCE: invention refers to derivatives of olivomycin I antibiotic of aureolic acid group with anticancer activity by structural formula as follows, where R5 represents hydrogen, C3-C10-cycloalkyl or C1-C4-alkyl with straight or branched hydrocarbon chain, optionally substituted with one or more hydroxyls. Additionally, invention concerns method of production of the specified derivatives, consisting in selective modification of 2'-carbonyl group of olivomycin 1 by reaction with aminooxyacetic acid, followed by amidation reaction of produced intermediate 2'-(carboxymethoxime)olivomycin 1 and related amines condensing agent added.

EFFECT: method of production of antibiotic derivatives with anticancer activity.

2 cl, 5 tbl, 6 ex

FIELD: organic chemistry, biochemistry, medicine.

SUBSTANCE: invention relates to phosphoramidates of nucleoside analogs comprising 2',3'-dideoxy-2',3'-didehydrothymidine 5'-phosphodimorpholidate of the formula (I) and phosphoramidates of 3'-azido-3'-deoxythymidine of the formula (II) and the formula (III) that inhibit activity in reproduction of human immunodeficiency virus (HIV). Compounds are resistant to effect of dephosphorylating enzymes and able to penetrate into cells and elicit the selective activity in inhibition of DNA biosynthesis catalyzed by HIV-reverse transcriptase.

EFFECT: valuable medicinal and biochemical properties of nucleoside analogs.

4 dwg, 1 tbl, 5 ex

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