Method of obtaining 2'-desoxy-2', 2'-difluorocytidine

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

 

The SCOPE of the INVENTION

The present invention relates to a method of stereoselective obtain 2'-deoxy-2',2'-diversityin.

DESCRIPTION PRECEDING the

2'-deoxy-2',2'-diversityin (gemcitabine) of the formula (I) has a nucleic acid base cytosine, stereochemical located in the β-orientation in the first position of the skeleton of the ribofuranose, and is effective for the treatment of various malignant tumors, such as non-small cell lung cancer (NSCLC), bladder cancer, breast cancer or ovarian cancer.

Gemcitabine may be obtained by conventional methods from lactolose connection, as shown in the reaction scheme 1, via the activated intermediate connection ribofuranose with a reactive leaving group.

The reaction scheme 1

where P1represents a protective group of hydroxyl and L represents a leaving group.

Examples of the activated intermediate ribofuranose for glycosylation are 1-sulfonate ribofuranose, such as α-methanesulfonate ribofuranose 1-halogenosilanes.

To conduct stereoselective glycosylation can be conducted by reaction of an α-methanesulfonate ribofuranose with nucleic base with obtaining demand is x β-nucleosides with a high yield (see U.S. patent No. 5371210, 5401838, 5426183, 5594124 and 5606048 and patent EP No. 577303). However, for α-methanesulfonate ribofuranose with a high ratio relative to β-methanesulfonate ribofuranose necessary cryogenic conditions below about -80°C, and thus, this method is not suitable for large-scale production.

Derivatives of 1-halogenosilanes can be easily obtained under mild conditions (for example, at room temperature), and for glycosylation can be conducted by reaction with anionic nucleic base (see U.S. patent No. 5744597 and patent EP No. 577304). However, glycosylation using the derived 1-halogenosilanes is not stereoselective (ie isomerizate 1 position), leading to a mixture of α - and β-nucleosides and ultimately to the low yield of the desired β-nucleoside.

In U.S. patent No. 5223608 describes the process for the selective extraction of β-anomer of citizenoriented from 1:1-mixture of α - and β-anomers of citizenoriented transformation mixture in the form of hydrochloride, by dissolving a mixture of hydrochloride in hot water, bringing the pH of the final solution to 8.2 and cooling and filtering the solution. However, this process also leads to a low yield of the β-anomer.

The authors of the present inventions have attempted to solve the problems of prior techniques and found the or, that isomerizate suppressed effectively removing halide compounds when his education in the process of glycosylation using the derived 1-halogenosilanes, and, thus, the stereoselectivity can be greatly increased.

The INVENTION

Thus, the main purpose of the present invention is the provision of an improved method of producing 2'-deoxy-2',2'-diversityin with a high degree of purity and with high yield by using a new stereoselective glycosylation reactions using 1-halogenosilanes.

In accordance with the present invention provides a method of obtaining 2'-deoxy-2',2'-diversityin formula (I), which includes stages:

(i) the interaction of compound 1-halogenosilanes formula (III) with nucleic base of the formula (IV) in a solvent to obtain a nucleoside of formula (II) with continuous removal of the formed during the reaction of sirgalahad formula (V) and

(ii) removing the protective group from a nucleoside of formula (II) to obtain 2'-deoxy-2',2'-diversityin formula (I):

where

R represents alkyl;

P1represents a protective group of hydroxy;

P2represents a protective group of amino and

X represents halogen.

TIMES THE first DESCRIPTION of the DRAWINGS

The above and other objects and features of the present invention will become clear from the following description of the invention in conjunction with the following accompanying drawings, on which:

Figure 1-3: chromatogram obtained using high-performance liquid chromatography (HPLC) compounds in accordance with example 4, comparative examples 1 and 2, respectively.

DETAILED description of the INVENTION

The method according to this invention is characterized by the fact that the compound of formula (I) can be efficiently obtained at a constant removal of sirgalahad formula (V), which is formed in the process of glycosylation.

Used herein, the term "enriched anomer" means that the content of a specific anomer mixture of anomers is more than 50%, including essentially pure anomer. Also, the term "isomerizate" means that the C1-position of the ribofuranose conducted epimerization ribofuranose essentially pure anomer or a mixture of α-anomer and β-anomer.

Used herein, the term "carrier" means a solvent used to remove sirgalahad generated in the process of glycosylation, the term "heating environment" means a solvent with a high boiling point, which can provide sufficient heating of the reaction system and support the controlling of the reaction mixture at a sufficiently high temperature to ensure the permanent removal of sirgalahad by distillation.

Used herein, the term "substituted" means substitution alone or in combination with at least one or more groups selected from hydrogen, cyano, halo, carbalkoxy, toluene, nitro, alkoxy and alkyl.

In accordance with the present invention stereoselective glycosylation is carried out, as shown in the reaction scheme 2.

The reaction scheme 2

Specifically, for glycosylation conduct the reaction enriched α-anomer 1-halogenosilanes formula (III) with nucleic base of the formula (IV) with a β-nucleoside of the formula (II) together with similaroriginal formula (V), which can act as a source of halide to conduct isomerization α-anomer. Thus, silicalite continuously removed as education simple distillation or by using an inert gas until the end of the glycosylation reaction. As a result, the duration of isomerization significantly reduced, and is vysokoselektivnye glycosylation in the direction β-anomer.

The distillation is carried out with simultaneous addition of drip in the reaction mixture for glycosylation carrier or mixture of carriers and the heating medium, which has a high boiling point.

Alternative inert gas is passed through a separate the felling of located in the reactor, to remove sirgalahad from the reaction mixture without affecting the glycosylation reaction. To remove sirgalahad inert gas is fed from the tube, which is set for (ozonation) in the reaction solution or (purging) of the reaction solution.

Enriched α-anomer 1-halogenosilanes formula (III)used as the starting material in the method according to this invention has a protective group of hydroxy and can be obtained by the method described in the patent application Korea No. 2004-59623. Illustrative protective hydroxy groups are formyl, acetyl, substituted acetyl, propionyl, butenyl, pialligo, benzoyl, biphenylene, substituted biphenylene, etoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl, phenoxycarbonyl, benzyl, diphenylmethyl, triphenylmethyl, tert-butyl, tetrahydropyranyl, allyl, N-phenylcarbamate, N-imidazolium, trialkylsilyl, isopropylalcohol, alkyldiethanolamine, triisopropylsilyl and tert-butyldimethylsilyl. Among them benzoyl, biphenylcarboxylic and substituted biphenylcarboxylic are more preferred.

Nucleic acid based formula (IV) has a protective group for amino groups, and it can be obtained using the methods described in U.S. patent No. 5371210, 5401838, 5426183, 5594124 and 5606048 and the patent EP No. 577303. The sludge is ultratune protective groups for the amino groups are silyl groups, such as trimethylsilyl, triisopropylsilyl, tributyrin, tert-butyldimethylsilyl and tert-butyldiphenylsilyl; carbamates, such as tert-butoxycarbonyl, benzyloxycarbonyl, 4-methoxybenzeneboronic and 4-nitrobenzenesulfonyl; formyl, acetyl, benzoyl and pivaloyl, methoxymethyl, tert-butyl, benzyl and tetrahydropyranyl. Among them (trimethylsilyl) is most preferred.

In the method according to this invention, nucleic acid based formula (IV) is used in amounts in the range from 5 to 50 molar equivalents, preferably from 10 to 30 molar equivalents, more preferably from 15 to 20 molar equivalents, based on 1-halogenosilanes formula (III).

Solvents suitable for use in the process of glycosylation, are benzene, substituted benzene, toluene, xylene, decalin, diglyme, 2-ethoxyethyl ether, diphenyl ether, substituted diphenyl ether, biphenyl, substituted biphenyl, C6-14alkane, substituted C6-14alkane and their mixture. Among them, preferred are toluene, C7-14alkane, diphenyl ether and a mixture thereof, and a mixture of diphenyl ether and heptane is preferred. The solvent is used in amounts in the range from 5 to 50 ml, preferably from 10 to 20 ml up to, based on 1 g of 1-halogenosilanes formula is (III).

The media used to facilitate the removal of sirgalahad formula (V) by distillation, must be inert under the reaction conditions glycosylation and preferably has a boiling point higher than the boiling point of sirgalahad. The media can be a benzene, substituted benzene, toluene, xylene, C6-14alkane, substituted C6-14alkane and their mixture. Among them, preferred are toluene, heptane, octane and Noonan, and heptane is preferred. The media used in amounts in the range from 50 to 1000 ml, preferably from 100 to 300 ml based on 1 g of 1-halogenosilanes formula (III).

In the method according to this invention, the heating medium with a high boiling point is 200°C or higher, in addition, can be used in the form of a mixture with a carrier so as to obtain a reaction system with a sufficient degree of heat to compensate for the loss of solvent in the distillation process. The heating medium should be inert under the reaction conditions glycosylation and preferably has a higher boiling point than the boiling point of the medium. The heating medium may be selected from the group consisting of decalin, diphenyl ether, substituted diphenyl ether, biphenyl, substituted biphenyl and mixtures thereof. Among them, diphenyl the new ether is most preferred. The heating medium is used in an amount in the range from 0.1 to 5% by vol., preferably from 0.5 to 3% vol., based on the number of media.

Preferably, the medium and the heating medium is continuously added to the reaction mixture at a constant speed until, until you have completed the glycosylation reaction, in order to ensure constant stereoselectivity.

In addition, to increase the removal of sirgalahad by distillation, the reaction mixture may optionally be added to the source silila, such as N,O-bis(trimethylsilyl)ndimethylacetamide (BSA) in the form of a mixture with a carrier. Source silila can be used in amounts in the range from 0.05 to 1.5% vol., preferably from 0.1 to 0.5% vol., based on the number of media.

Also to remove sirgalahad formula (V) may be used inert gas according to the present invention, such as nitrogen, helium, neon and argon, preferably nitrogen. The inert gas preferably is fed by a flow of 1 l/min or more, based on 100 g of compound 1-halogenosilanes formula (III). If inert gas is supplied to a flow rate of less than 1 l/min, the ratio of β-nucleosides and α-nucleosides is not more than 3.

Glycosylation in accordance with the present invention is carried out at a temperature in the range from 80 to 300°C, preferably from 100 to 200 is C, more preferably from 130 to 150°C for 4 to 24 hours.

The process of glycosylation can be checked by means of thin layer chromatography (TLC),1H nuclear magnetic resonance (1H-NMR) or high performance liquid chromatography (HPLC).

Removing the protective groups from enriched β-anomer nucleoside of formula (II) can be conducted in conventional manner. For example, most protective groups silila easy to split under the action of water or alcohol. The protective group of the aminoacyl, such as formyl, acetyl, pivaloyl and benzoyl are removed by hydrolysis with a strong base. Such bases include hydroxides of alkali metals such as sodium hydroxide or potassium hydroxide; alkoxides of alkali metals such as sodium methoxide or tert-piperonyl potassium; diethylamine, hydroxylamine, ammonia, hydrazine and the like, among them ammonia is preferred. Also protective acyl group may be removed using an acid catalyst, such as methanesulfonate acid, hydrochloric acid, Hydrobromic acid, sulfuric acid, or acidic ion exchange resins.

Enriched β-anomer nucleoside of formula (II) can be obtained in pure form by separation based on the differences in solubilities of mixtures enriched β-anomer nucleoside of formula (II) and not entered in the reactions is s cytosine, formed after removal of the protective group. The separation is preferably carried out using a solvent system consisting of methylene chloride and methanol, where enriched β-anomer nucleoside of formula (II) is vysokorazvitym, while unreacted cytosine is slightly soluble.

Thus, in accordance with stereoselective glycosylation of the present invention receive the product β-enriched nucleoside with a ratio of α:β equal to from 1:4 to 1:14.

β-nucleoside of the formula (I) can be isolated in the form of a hemihydrate or dihydrate with high purity 99.8% or more and a yield of 70% or more by a single recrystallization process, which involves the dissolution of a mixture of α/β anomers in water, heating the mixture to a temperature of from 40 to 60°C, cooling to a temperature of from 10 to 25°C and filtering solid matter deposited in the sediment during the cooling phase. This process can be carried out under stirring to obtain the form of the hemihydrate or without stirring to obtain the form of the dihydrate.

It was shown that the form of the hemihydrate or dihydrate β-nucleoside obtained by the present invention are resistant to change content of moisture in the conditions shown in table 1.

Table 1
Moisture content (%)
The hemihydrateThe dihydrate
The air1 day3,6the 11.6
7 days3,711,8
14 days3,411,7
40°C with relative humidity of 75%1 day3,712,1
7 daysthe 3.811,9
14 daysthe 3.811,7
The calculated moisture content in the gemcitabine: hemihydrate 3.3%, dihydrate 12,0%

Highly pure hemihydrate or dihydrate β-nucleosides can be used directly without further purification to obtain the pharmaceutically acceptable salt is hydrochloride with the degree of purification described on pp. 892-894 in the U.S. Pharmacopoeia (2004).

So, this is completed with the invention also relates to a method for obtaining hydrochloride 2'-deoxy-2',2'-diversityin, comprising the reaction of 2'-deoxy-2',2'-diversityin formula (I) or its hemihydrate or dihydrate with hydrochloric acid in an organic solvent.

The present invention is described in more detail based on examples. However, it should be understood that the present invention is not limited to the specific examples.

In the examples OCOBiPh or BiPhOCO - mean structure

.

Also each of the obtained product was analyzed by HPLC under two conditions: (1) column Bond RX-C8 (4,5x250 mm, 5 μm), NaH2PO4·H2O 13.8 g/H2PO4(pH 2,4-2,6) 2.5 ml, dissolved in 1 l of water, for the compounds of formula (I); and (2) column YMC hydrosphere Cl8 (4,6x150 mm, 5 μm), a mixture of 760 ml of methanol and 240 ml NaH2PO4·H2O 13.8 g/H2PO4(pH 2,4-2,6) 2.5 ml, dissolved in 1 l of water, for the compounds of formula (II).

Example

A method of obtaining a 1: Obtain 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate

Stage 1: Obtaining 2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate

13.5 g three-tert-butoxylated lithium was dissolved in 160 ml of tetrahydrofuran, was stirred at room temperature for 30 minutes and cooled to -40°C, and then added 20 g of D-Erythro-2-deoxy-2,2-differentforms-1-ulose-5-benzoyl-(4-phenyl), dissolved in 80 ml of tetrahydrofuran. The mixture was slowly heated to room temperature and allowed to react at this temperature for 2 hours. After completion of the reaction, to the reaction mixture was added 220 ml of 1 N. HCl and a layer of tetrahydrofuran was separated. The aqueous layer was extracted with 220 ml of a simple ester was combined with the previously separated by a layer of tetrahydrofuran, washed sequentially 220 ml of water, saturated sodium bicarbonate and brine, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure and the residue was purified column chromatography with silica gel to obtain 18.3 g specified in the title compound (yield: 91%) as a pale yellow syrup.

1H-NMR (300 MHz, CDCl3, δ); 3,89-3,91 (d, 1H), br4.61-to 4.81 (m, 2H), 5,31-of 5.92 (m, 2H), 7,26-of 7.70 (m, 10H), 8,05-8,16 (m, 4H).

Stage 2: Obtaining 2'-deoxy-2'2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate-1β-diphenylphosphate

of 18.3 g of 2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained in stage 1, was dissolved in 146 ml of toluene was added to 6.7 ml of triethylamine and drip was added to 12.4 ml diphenylchlorophosphine, diluted with 37 ml of toluene. After 4 hours to neutralize residual triethylamine to the reaction mixture was added 48 ml of 1 N. HCl, the toluene layer was separated and the aqueous layer was extrage is ovali 48 ml of ether. The ether extract was combined with the previously separated by a layer of toluene and then washed with water, saturated sodium bicarbonate and saline. The organic layer was separated, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure to obtain a mixture of α - and β-phosphate in the form of solids. The mixture was investigated by analysis1H-NMR and found that the ratio of the α-phosphate:β-phosphate was 1:10,6. β-phosphate selectively recrystallized from a mixture of isopropanol and water 3:1 (vol./about.) with the receipt of 26.5 g (yield: 87%) indicated in the title compound as a white solid.

1H-NMR (300 MHz, CDCl3, δ); 4,56-of 4.25 (m, 3H), 5,80 (m, 1H), 5,95 (t, 1H), 7,44-6,98 (m, 16H), 7,51 (d, 2H), EUR 7.57 (d, 2H), 7,89 (d, 2H), 8,01 (d, 2H).

Melting point: 101-103°C.

The HPLC purity (area %): anomer α-phosphate of 1.76%, anomer β-phosphate 98,24%.

Stage 3: Obtain 1-α-bromo-2'-deoxy-2'2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate

To 80,5% 30 ml of a mixture of HBr/acetic acid was added to 22.8 g of 2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate-lβ-diphenylphosphate obtained in stage 2, and the mixture was allowed to react at room temperature for 6 hours. The resulting solution was diluted with 400 ml of methylene chloride and slowly added to 500 ml of ice water. The water layer is dalali and the methylene chloride layer was washed successively with ice water, saturated sodium bicarbonate and saline. The methylene chloride layer was dried over magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain a mixture of α - and β-isomers in the form of solids. The mixture was investigated by analysis1H-NMR and found that the ratio of α-bromo:β-bromo amounted to 10.7:1. The compound β-bromo selectively recrystallized from isopropanol to obtain 17.0 g (yield: 82%) indicated in the title compound as a white solid.

1H-NMR (300 MHz, CDCl3, δ); 8,19 (d, 2H), of 8.06 (d, 2H), 7,73 (d, 2H), 7,63 (d, 2H), of 7.64-7,41 (m, 6H), 6,56 (d, 1H), ceiling of 5.60 (DD, 1H).

Melting point: 111-112°C.

The HPLC purity (area %): anomer α-bromo 99,74%, anomer β-bromo 0,26%.

A method of obtaining a 2: Obtain 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-3,5-di-(4-phenyl)benzoate

Stage 1: Obtaining 2'-deoxy-2'2'-debtor-D-ribofuranosyl-3,5-di-(4-phenyl)benzoate

8,66 g of tri-tert-butoxylated lithium was dissolved in 120 ml of tetrahydrofuran, stirred at room temperature for 30 minutes and cooled to -40°C, and then was slowly added to 15 g of D-Erythro-2-deoxy-2,2-debtor-pentofuranose-1-ulose-3,5-di-(4-phenyl), rastvorennogo in 100 ml of tetrahydrofuran. Then the mixture was heated to room temperature and allowed to react for 1 hour. To the reaction mixture to slow the drip was added 142 ml of 1 N. hydrochloric acid to break down the excess tri-tert-butoxylated lithium and the organic layer was separated. The aqueous layer was extracted with 150 ml of a simple ester was combined with the previously separated organic layer was sequentially washed with 220 ml of water, saturated sodium bicarbonate and brine, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure and the resulting solid was recrystallized from toluene to obtain a 13.4 g specified in the title compound (yield: 89%) as a white solid.

1H-NMR (300 MHz, CDCl3, δ); of 3.45 (s, 1H), 4,85-4,50 (m, 3H), 5,8-to 5.4 (m, 2H), 7,49-the 7.43 (m, 6H), 7,71-to 7.61 (m, 8H), 8,18-to 8.12 (m, 4H).

Melting point: 156-158°C.

Stage 2: Obtaining 2'-deoxy-2'2'-debtor-D-ribofuranosyl-3,5-di-(4-phenyl)benzoyl-1β-diphenylphosphate

13 g of 2'-deoxy-2'2'-debtor-D-ribofuranosyl-3,5-(4-phenyl)benzoate obtained in stage 1, was dissolved in a mixture of 130 ml of toluene and 100 ml of methylene chloride and added to 5.1 ml of triethylamine. To the mixture at room temperature drip was added to 7.6 ml diphenylchlorophosphine. After 5 hours the solvent was removed under reduced pressure, the obtained solid substance was dissolved in 130 ml of methylene chloride was added 65 ml of 1 N. HCl. The organic layer was separated, then washed with water, asystem sodium bicarbonate and brine, was dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure to obtain a mixture of α - and β-phosphate in the form of solids. The mixture was investigated by analysis1H-NMR, and found that the ratio of the α-phosphate:β-phosphate was 1:10,8. β-phosphate selectively recrystallized from isopropanol to obtain 15 g (yield: 83%) indicated in the title compound as a white solid.

1H-NMR (300 MHz, CDCl3, δ); 4,70-and 4.40 (m, 3H), 5,90 (m, 1H), between 6.08 (t, 1H), 7,70-was 7.08 (m, 24H), 8,15-of 8.04 (DD, 4H).

Melting point: 145-147°C.

The HPLC purity (area %): anomer α-phosphate of 1.29%, anomer β-phosphate 98,71%.

Stage 3: Obtain 1-α-bromo-2'-deoxy-2'2'-debtor-D-ribofuranosyl-3,5-di-(4-phenyl)benzoate

To 83,2 ml of a mixture of 30% HBr/acetic acid was added 13 g of 2'-deoxy-2',2'-debtor-D-ribofuranosyl-3,5-di-(4-phenyl)benzoyl-1β-diphenylphosphate obtained in stage 2, and the mixture was allowed to react at room temperature for 7 hours. In the reaction solution was slowly added 50 ml of ice water and the resulting solid was filtered. The filtered solid was a mixture of forms of α - and β-bromo, and analysis1H-NMR showed that the ratio of the forms α-bromo:β-bromo amounted to 10.9:1. The connection form α-bromo selectively recrystallized from ethanol to obtain 8,45 g (yield: 83%) specified the title compound as a white solid.

1H-NMR (300 MHz, CDCl3, δ); 4,89-4,22 (m, 3H), 5,62 (DD, 1H), 6,55 (d, 1H), 7,73-7,42 (m, 14H), 8,63-8,11 (DD, 4H).

Melting point: 151-153°C.

The HPLC purity (area %): anomer α-bromo 99,67%, anomer β-bromo 0,33%.

Example 1: 1-(2-Deoxy-2',2'-debtor-5-benzoyl-3-(4-phenyl)benzoyl-D-ribofuranosyl-4-aminopyrimidine-2-he

Example 1-1

of 44.5 g of cytosine, 252 ml hexamethyldisilazane and 252 mg of ammonium sulfate were mixed and boiled under reflux until a homogeneous solution, which was further heated under reflux for 1 hour. Was added 200 ml of ethyl acetate and heated to remove residual unreacted hexamethyldisilazane. To the resulting solution was added a mixture of 160 ml of heptane and 40 ml of diphenyl ether and 10.4 g of 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained by the method of obtaining 1. Spent the reaction mixture for 8 hours during dropwise addition of a mixture of diphenyl ether (40 ml)/heptane (4 l) and while holding distillation while maintaining the reaction temperature from 130 to 140°C. This process ensures the permanent deletion of trimethylsilylpropyne from the reaction mixture during the reaction. After completion of the reaction, to the reaction mixture were added 140 ml of heptane. The solution was cooled to 100°C, carefully extinguished 12 ml of water and the PE amasyali at room temperature. The obtained solid was filtered and washed with heptane to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:8,8. A solid substance containing a mixture of nucleosides and unreacted cytosine was added to a mixture of methylene chloride (200 ml) and methanol (40 ml)was boiled under reflux for 1 hour and filtered to remove cytosine. The person to distil the filtrate under reduced pressure, to the precipitate was added isopropyl simple ether, filtered and the filtrate was dried by a flow of warm air to obtain 10.8 g (yield: 98%) indicated in the title compound as a white solid.

1H-NMR (300 MHz, DMSO d-6, δ); and 8.1 (d, 2H), 7,9 (d, 2H), and 7.8 (d, 2H), 7.7 (d, 2H), and 7.6 (d, 2H), 7,5 to 7.4 (m, 7H), and 6.3 (t, 1H), and 5.8 (m, 1H), 5,7 (d, 1H), 4,7-4,6 (m, 3H).

The ratio of anomers (HPLC analysis): α-nucleoside/β-nucleoside = 1/8,8.

Example 1-2

11.1 g of cytosine, 63 ml hexamethyldisilazane and 63 mg of ammonium sulfate were mixed and boiled under reflux for 2 hours. To the mixture was added 60 ml of toluene and heated to remove the remaining unreacted hexamethyldisilazane. To the resulting solution was added a mixture of 40 ml of octane and 20 ml of diphenyl EF the RA and 3.5 g 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate, obtained by the method of obtaining 1. Spent the reaction mixture for 10 hours at a dropwise addition of a mixture of diphenyl ether (10 ml)/heptane (1 l) and simultaneous carrying out distillation while maintaining a temperature of 140 to 150°C. This process ensures the permanent deletion of trimethylsilylpropyne from the reaction mixture during the reaction. After completion of the reaction, to the reaction mixture were added 50 ml of heptane. The solution was cooled to a temperature of from 80 to 100°C, gently drip was added 12 ml of water and the mixture was stirred at room temperature for 1 hour. The obtained solid was filtered and washed with heptane to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:5,6. A solid substance containing a mixture of nucleosides and unreacted cytosine was added to a mixture of methylene chloride (70 ml) and methanol (15 ml)was boiled under reflux for 1 hour and filtered to remove cytosine. The person to distil the filtrate under reduced pressure, to the precipitate was added isopropyl ether, filtered and the filtrate was dried by a flow of warm air from the receipt of 3.45 g (yield: 93%) indicated in the title compound as a white solid in the society.

Data H-NMR were the same as in example 1-1.

The ratio of anomers (HPLC analysis): α-nucleoside/β-nucleoside = 1/5,6.

Example 1-3

of 2.23 g of cytosine, and 12.6 ml hexamethyldisilazane and 12.6 mg of ammonium sulfate were mixed and boiled under reflux until a homogeneous solution, which is then further boiled under reflux for 1 hour. Was added 200 ml of ethyl acetate and heated to remove the remaining unreacted hexamethyldisilazane. To the resulting solution was added 0.26 g of 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained by the method of obtaining 1. Spent the reaction mixture for 6 hours during dropwise addition of a mixture of N,O-bis(trimethylsilyl)ndimethylacetamide (2 ml)/heptane (200 ml) while carrying out distillation while maintaining the reaction temperature from 125 to 140°C. This process ensures the permanent deletion of trimethylsilylpropyne from the reaction mixture during the reaction. After completion of the reaction the solution was cooled to 80°C, gently drip was added 1 ml of water and the mixture was stirred at room temperature for 1 hour. The obtained solid was filtered and washed with heptane to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture nuclease the s was investigated by HPLC analysis and found the ratio of α-nucleoside:β-nucleoside was 1:14.

Example 1-4

340 g of cytosine, 1.835 squa l hexamethyldisilazane and 1.84 g of ammonium sulfate were mixed and boiled under reflux until a homogeneous solution, which was further heated under reflux for 1 hour. To the resulting solution was sequentially added 1.2 l of heptane and 500 ml of diphenyl ether to reduce the temperature of the solution to 100°C. Then was added 100 g 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained by the method of obtaining 1. Spent the reaction mixture for 12 hours at room special reactor tube and a flow of nitrogen with a flow rate of 1.0 to 1.3 l/min, flow with maintaining the reaction temperature from 140 to 143°C. This process ensures the permanent deletion of trimethylsilylpropyne from the reaction mixture during the reaction. After completion of the reaction the solution was cooled to 80°C and carefully drip was added 100 ml of water. The mixture was stirred at room temperature for 1 hour. The obtained solid was filtered and washed with heptane to obtain a mixture of α - and β-isomers of nucleosides, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was :4,9.

Example 1-5

The method of example 1-4 was repeated except that nitrogen was applied to the pipe at a flow rate of from 3.0 to 3.5-6 l/min, to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:6,1.

Example 2: 1-(2'-deoxy-2',2'-debtor-3,5-di-(4-phenyl)benzoyl-D-ribofuranosyl-

4-aminopyrimidine-2-he

of 22.2 g of cytosine, 126 ml hexamethyldisilazane and 126 mg of ammonium sulfate were mixed and boiled under reflux for 2 hours and was added 100 ml of ethyl acetate to remove by distillation of unreacted hexamethyldisilazane. To the resulting solution was sequentially added 80 ml of heptane, to 5.93 g 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-3,5-di-(4-phenyl)benzoate obtained by the method of obtaining 2 and 20 M diphenyl ether. The resulting mixture was allowed to react for 9 hours during dropwise addition of 4 l of heptane and while conducting distillation while maintaining the reaction temperature from 130 to 140°C. This process ensures permanent removal from the reaction mixture of trimethylsilylpropyne during the reaction. After completion of the reaction, to the reaction mixture were added 160 ml of heptane. The solution of the OHL who waited up to 100°C and carefully drip was added 8 ml of water. The solution was stirred at room temperature and was filtered. The obtained solid was washed with heptane to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:5,4. A solid substance containing a mixture of nucleosides and unreacted cytosine was added to a mixture of methylene chloride (200 ml) and methanol (40 ml)was boiled under reflux for 1 hour and filtered to remove cytosine. The person to distil the filtrate under reduced pressure to obtain 4 g (yield: 64%) indicated in the title compound as a white solid.

1H-NMR (300 MHz, CDCl3, δ): a total of 8.74-7,27 (m, 19H), 6,38 (m, 1H), of 5.83 (m, 1H), 5,78 (d, 1H), 4,78 is 4.45 (m, 3H).

Melting point: 250-255°C.

The ratio of anomers (HPLC analysis): α-nucleoside/β-nucleoside = 1/5,4.

Example 3: 2'-Deoxy-2',2'-diversityin (Compound of formula (I)-1: gemcitabine)

Example 3-1: 2'-deoxy-2'2'-diversityin hemihydrate

To 86 ml of 7N mixture of ammonia in methanol was added 10.8 g of 1-(2'-deoxy-2',2'-debtor-5-benzoyl-3-(4-phenyl)benzoyl-D-ribofuranosyl-4-aminopyrimidine-2-she obtained in example 1-1, and was additionally added 216 ml of methanol. The mixture was stirred at room t is mperature within 12 hours and the solvent was removed under reduced pressure. To the mixture was added 120 ml of water and 80 ml of ethyl acetate under stirring. The aqueous layer was separated and the ethyl acetate layer was extracted with 40 ml of water. The aqueous layers were combined, washed with 40 ml of diethyl ether and person to distil under reduced pressure to remove water. To the obtained precipitate was added 25 ml of water, the mixture was heated to a temperature of from 45°to 50 ° C to dissolve the solids, cooled, and stirred at room temperature for 2 hours to ensure precipitation of solid substances. The solid was filtered, washed with water and acetone and dried by a flow of warm air throughout the night with the receipt of 3.99 g (yield: 76,9%) specified in the connection header in the form of pure white hemihydrate.

Moisture content: 3,4%.

1H-NMR (300 MHz, DMSO d-6, δ), and 7.7 (1H, d), 7,39 (1H, d)and 6.2 (1H, d)and 6.1 (1H, t), and 5.8 (1H, t), 4,2 (m, 1H), from 3.9 to 3.8 (m, 2H), and 3.7 (m, 1H).

Melting point = 198-202°C.

The HPLC purity (area %):

β-anomer - 99,97%

the α-anomer is less than 0.02%

cytosine - less than 0.01%.

Example 3-2: 2'-deoxy-2'2'-diversityin the dihydrate

The method of example 3-1 was repeated except that the solution was cooled without stirring during the precipitation of solid substances with getting 4,22 g (yield: 81,3%) specified in the connection header in the form of pure white dihydrate.

Moisture content: 11,5%.

Melting point = 220-224°C.

Data H-NMR were the same as in example 3-1.

The HPLC purity (area %):

β-anomer - of 99.98%.

the α-anomer is less than 0.01%

cytosine - less than 0.01%.

Example 4: 2'-deoxy-2',2'-diversityin (Compound of formula (I)-2: gemcitabine)

To 184 ml hexamethyldisilazane added to 32.2 g of cytosine and 184 mg of ammonium sulfate. The mixture was boiled under reflux for 1 hour and added 250 ml of heptane and heated to a temperature of from 135 to 140°C up until will be removed unreacted, hexamethyldisilazane. To the resulting solution was added 150 ml of heptane and 10.0 g of 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained by the method of obtaining 1, and then added of 38.7 ml of diphenyl ether. The resulting mixture was allowed to react for 10 hours during dropwise addition of 1.5 l of heptane and simultaneous carrying out distillation while maintaining the reaction temperature from 135 to 140°C. This process ensures permanent removal from the reaction mixture of trimethylsilylpropyne during the reaction. After completion of the reaction to the resulting solution was added 240 ml of heptane and was slowly added to 11.6 ml of water. The obtained solid was stirred, filtered, washed with heptane and dried at room temperature to obtain a mixture of α - and β-isomers of the nucleoside, including not entered in the district who share cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:6,1 (see Fig. 1). The solid is suspended in 300 ml of methylene chloride and 60 ml of methanol and boiled under reflux for 2 hours. The resulting mixture was filtered, the filtered solid was washed with a mixture of methylene chloride (150 ml) and methanol (30 ml) and person to distil under reduced pressure to obtain a mixture of α/β 1-(2'-deoxy-2',2'-debtor-5-benzoyl-3-(4-phenyl)benzoyl-D-ribofuranosyl-4-aminopyrimidine-2-it. Precipitated solids were added 200 ml of methanol and 83 ml of 7 n ammonia/methanol and stirred at room temperature overnight. After completion of the reaction the solvent was removed under reduced pressure and to the precipitate was added 80 ml of ethyl acetate and 90 ml of water. The aqueous layer was separated and the ethyl acetate layer was extracted with 40 ml of water. The aqueous layers were combined and washed with 40 ml of simple ether (x2). Water drove under reduced pressure up until the number of remaining water was not 5 times the calculated mass of the desired product, and the precipitate was heated to a temperature of from 50°to 55 ° C and cooled to room temperature under stirring for 2 hours to ensure precipitation of solid substances. Precipitated precipitated solid is shown is ovali, washed with water and acetone and dried by a flow of warm air throughout the night with the receipt of 3.69 g (yield: 72.6 per cent) specified in the connection header in the form of pure white crystal.

Moisture content: 3.5 per cent.

Data H-NMR and melting point were the same as in example 3-1.

The HPLC purity (area %):

β-anomer of 99.9%

the α-anomer is less than 0.01%

cytosine - less than 0.02%.

Example 5: 2'-deoxy-2',2'-diversityin (Compound of formula (I)-3: gemcitabine)

24 g of N-acetylcytosine and 126 ml hexamethyldisilazane and 126 mg of ammonium sulfate were mixed and boiled under reflux for 2 hours. To the mixture was added 100 ml of heptane and unreacted, hexamethyldisilazane was removed by distillation. To the resulting solution was added 50 ml of octane and 5 g of 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained by the method of obtaining 1. Spent the reaction mixture for 8 hours during dropwise addition of a solution of N,O-bis(trimethylsilyl)ndimethylacetamide (1.8 ml)/heptane (900 ml) and carrying out distillation while maintaining the reaction temperature from 135 to 140°C. This process ensures the permanent deletion of trimethylsilylpropyne from the reaction mixture during the reaction. After completion of the reaction to the resulting solution, which was cooled to 100°C, was added 60 ml of heptane and slowly added 12 m the water. The obtained solid was stirred at room temperature for 2 hours, filtered and washed with heptane to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:4,8. A mixture of nucleosides suspended in 108 ml of methanol and 45 ml of 7 n ammonia/methanol, the solvent was removed under reduced pressure and to the precipitate was added 50 ml of ethyl acetate and 60 water. The aqueous layer was separated and the ethyl acetate layer was extracted with 20 ml of water. The aqueous layers were combined and washed with 40 ml of simple ether (x2). Water drove under reduced pressure and to the precipitate was added 15 ml of water, was heated to a temperature of from 50°to 55 ° C and cooled to room temperature under stirring for 2 hours to ensure precipitation of solid substances. Precipitated precipitated solid was filtered, washed with water and acetone and dried by a flow of warm air throughout the night with getting to 32.2 g (yield: 63%) indicated in the title compound in the form of pure white crystals.

Data H-NMR and melting point were the same as in example 3-1.

The HPLC purity (area %):

β-anomer - 99,8%

the α-anomer is less than 0.02%

cytosine - less than 0.02%.

Example 6: hydrochloride 2'-detox the -2',2'-diversityin

Example 6-1

3.5 g of 2'-deoxy-2',2'-diversityin of hemihydrate (moisture content: 3.8 per cent)obtained in example 3-1 was dissolved in 35 ml of acetone and the drip was added 1.2 ml of concentrated hydrochloric acid. The resulting mixture was stirred at room temperature for 2 hours. The obtained solid was filtered, washed with acetone and dried by a flow of warm air from getting to 3.52 g (yield: 91,9%) specified in the connection header in the form of pure white crystals.

1H-NMR (300 MHz, DMSO d-6): for 9.95 (s, 1H), 8,81 (s, 1H), 8,05 Od, 1H), x 6.15 (d, 1H), 5,96 (m, 1H), 4,14-a 4.03 (m, 1H), 3,79 (d, 1H), 3,70-3,51 (m, 2H).

Melting point: 287-292°C.

Example 6-2

3.5 g of 2'-deoxy-2',2'-diversityin dihydrate (moisture content: 11,5%)obtained in example 3-2 was dissolved in 35 ml of acetone and the drip was added 1.2 ml of concentrated hydrochloric acid. The resulting mixture was stirred at room temperature for 2 hours. The obtained solid was filtered, washed with acetone and dried by a flow of warm air from the receipt of 3.23 g (yield: 91.5 per cent) specified in the connection header in the form of pure white crystals.

Data H-NMR and melting point were the same as in example 6-1.

Comparative example: Obtaining 2'-Deoxy-2',2'-diversityin without distillation to remove Seeley is the halide

Comparative example 1

To 184 ml hexamethyldisilazane added to 32.2 g of cytosine and 184 mg of ammonium sulfate. The mixture was boiled under reflux for 1 hour and added 250 ml of heptane and heated to a temperature of from 135 to 140°C for distillation of unreacted hexamethyldisilazane. To the resulting solution was added 150 ml of heptane and 10.0 g of 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained in accordance with the method of obtaining 1, and then added of 38.7 ml of diphenyl ether. The resulting mixture was allowed to react for 10 hours while boiling under reflux and maintaining the reaction temperature from 135 to 140°C. After completion of the reaction to the resulting solution was added 240 ml of heptane and was slowly added to 11.6 ml of water. The obtained solid was stirred, filtered, washed with heptane and dried at room temperature to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:1,4 (see Fig. 2). The solid is suspended in 300 ml of methylene chloride and 60 ml of methanol and boiled under reflux for 2 hours. The resulting mixture was filtered, the filtered solid substances which has been washed with a mixture of methylene chloride (150 ml) and methanol (30 ml) and person to distil under reduced pressure to obtain a mixture of α/β 1-(2'-deoxy-2',2'-debtor-5-benzoyl-3-(4-phenyl)benzoyl-D-ribofuranosyl-4-aminopyrimidine-2-it. To the mixture was added 200 ml of methanol and 83 ml of 7 n ammonia/methanol and stirred at room temperature overnight. After completion of the reaction the solvent was removed under reduced pressure and to the precipitate was added 80 ml of ethyl acetate and 90 ml of water. The aqueous layer was separated and the ethyl acetate layer was extracted with 40 ml of water. The aqueous layers were combined and washed with 40 ml of simple ether (x2). Water drove under reduced pressure as long as there was a quantity of water, 5 times more calculated mass of the desired product, and the precipitate was heated to a temperature of from 50°to 55 ° C and cooled to room temperature under stirring for 2 hours to ensure precipitation of solid substances. Precipitated precipitated solid was filtered, washed with water and acetone and dried by a flow of warm air throughout the night with the receipt of 1.80 g (yield: 35.5 per cent) specified in the connection header in the form of pure white crystals.

Moisture content: 3.7 per cent.

Data H-NMR and melting point were the same as in example 3-1. The ratio of anomers (HPLC analysis): α-nucleoside/β-nucleoside = 1/1,4.

Comparative example 2

To 184 ml hexamethyldisilazane added to 32.2 g of cytosine and 184 mg of ammonium sulfate. The mixture was boiled under reflux for 1 hour and added 250 ml of heptane and heated to a temperature of from 135 to 140°C for the distillation of the unreacted hexamethyldisilazane. To the resulting solution were added 10.0 g of 1-α-bromo-2'-deoxy-2',2'-debtor-D-ribofuranosyl-5-benzoyl-3-(4-phenyl)benzoate obtained by the method of obtaining 1,, 36.3 ml of anisole. The resulting mixture was allowed to react for 10 hours while boiling under reflux and maintaining the reaction temperature from 135 to 140°C. After completion of the reaction to the resulting solution was added 240 ml of heptane and was slowly added to 11.6 ml of water. The obtained solid was stirred, filtered, washed with heptane and dried at room temperature to obtain a mixture of α - and β-isomers of the nucleoside, including unreacted cytosine, in the form of a white solid. The mixture of oligonucleotides was investigated by HPLC analysis and found that the ratio of α-nucleoside:β-nucleoside was 1:1,3 (see Fig. 3). The solid is suspended in 300 ml of methylene chloride and 60 ml of methanol and boiled under reflux for 2 hours. The resulting mixture was filtered, the filtered solid was washed with a mixture of methylene chloride (150 ml) and methanol (30 ml) and person to distil under reduced pressure to obtain a mixture of α/β 1-(2'-deoxy-2',2'-debtor-5-benzoyl-3-(4-phenyl)benzoyl-D-ribofuranosyl-4-aminopyrimidine-2-it. To the mixture was added 200 ml of methanol and 83 ml of 7 n ammonia/methanol and stirred at room temperature overnight. After completion of the reaction RA is the solvent was removed under reduced pressure and to the precipitate was added 80 ml of ethyl acetate and 90 ml of water. The aqueous layer was separated and the ethyl acetate layer was extracted with 40 ml of water. The aqueous layers were combined and washed with 40 ml of simple ether (x2). Water drove under reduced pressure until such time as the amount of water got in 5 more calculated mass of the desired product, and the precipitate was heated to a temperature of from 50°to 55 ° C and cooled to room temperature under stirring for 2 hours to ensure precipitation of solid substances. Precipitated precipitated solid was filtered, washed with water and acetone and dried by a stream of warm air during the night to obtain 1.64 g (yield: 32.3 per cent) specified in the connection header in the form of pure white crystals.

Moisture content: 3.5 per cent.

Data H-NMR and melting point were the same as in example 3-1.

The ratio of anomers (HPLC analysis): α-nucleoside/β-nucleoside = 1/1,3.

The results of glycosylation and removal of the protective group in accordance with example 4 and comparative examples 1 and 2 are summarized in table 2.

Table 2
The HPLC analysis (area is the area, %)Ratio
β/α
General
output
β-anomerα-anomer
Example 484,9113,946,0to 72.6%
(3,69 g)
Comparative
tion example 1
57,6040,901,435,3%
(1,80 g)
Comparative
hydrated example 2
56,1742,551,332,3%
(1,64 g)
The retention time for the peak of the β-anomer: 10,08~of 10.09
The retention time for the peak of the α-anomer: 8,23

As can be seen from table 2, in accordance with the present invention the β-anomer obtained with significantly higher yield as compared with comparative examples 1 and 2.

Although this invention is described in relation to specific embodiments, it should be considered that the experts in this field can be made of various modifications and changes of this invention, which will also be ahadiths in the scope of this invention, as defined by the attached claims.

1. Method for producing enriched β-anomer of 2'-deoxy-2',2'-diversityin formula (I), which involves the following stages:
(i) the interaction enriched α-anomer of the compound 1-halogenosilanes formula (III) with nucleic base of the formula (IV) in a solvent to obtain enriched β-anomer nucleoside of formula (II) with continuous removal of the formed during the reaction of sirgalahad formula (V) by distillation using media or by passing an inert gas through the reaction mixture; and
(ii) removing the protective group from enriched β-anomer nucleoside of formula (II):





where R represents a C1-C3alkyl;
R1represents a protective group of hydroxy;
R2represents a protective group of amino;
X represents halogen.

2. The method according to claim 1, where the nucleic acid based formula (IV)used in stage (i), present in an amount in the range from 5 to 50 molar equivalents, based on 1-halogenosilanes formula (III).

3. The method according to claim 1, where the solvent used in stage (i)selected from the group consisting of benzene, substituted be the ash, toluene, xylene, decalin, diglyme, 2-ethoxyethyl ether, diphenyl ether, substituted diphenyl ether, biphenyl, substituted biphenyl,6-14alkane, substituted C6-14alkane, and mixtures thereof.

4. The method according to claim 1, where silicalite formula (V) represents trimethylborane.

5. The method according to claim 1, where the carrier is selected from the group consisting of benzene, substituted benzene, toluene, xylene,6-14alkane, substituted C6-14alkane, and mixtures thereof.

6. The method according to claim 5, where the carrier is a heptane.

7. The method according to claim 1, where the media is used in amounts in the range from 50 to 1000 ml based on 1 g of 1-halogenosilanes.

8. The method according to claim 1, where the media is used together with the heating medium or N,O-bis(trimethylsilyl)ndimethylacetamide (BSA).

9. The method of claim 8, where the heating medium is selected from the group consisting of decalin, diphenyl ether, substituted diphenyl ether, biphenyl, substituted biphenyl and mixtures thereof.

10. The method according to claim 9, where the heating medium is a diphenyl ether.

11. The method of claim 8, where the heating medium is used in an amount in the range from 0.1 to 5 vol.%, based on the number of media.

12. The method of claim 8, where N,O-bis(trimethylsilyl)ndimethylacetamide (BSA) is used in amounts in the range from 0.05 to 1.5 vol.%, based on the number of media.

13. The method according to claim 1 where the inert gas is selected from the group consisting of nitrogen, helium, neon and argon.

14. The method according to claim 1 where the inert gas is fed in the form of ozonation or blowing.

15. The method according to claim 1 where the inert gas is fed with a flow rate of 1 l/min or more, based on 100 g of 1-halogenosilanes formula (III).

16. The method according to claim 1, where the stage (i) is conducted at a temperature in the range from 80 to 300°C.

17. A method of obtaining a hydrate enriched β-anomer of 2'-deoxy-2',2'-diversityin formula (I), which involves the following stages:
(i) the interaction enriched α-anomer of the compound 1-halogenosilanes formula (III) with nucleic base of the formula (IV) in a solvent to obtain enriched β-anomer nucleoside of formula (II) with continuous removal of the formed during the reaction of sirgalahad formula (V) by distillation using media or by passing an inert gas through the reaction mixture; and
(ii) removing the protective group from enriched β-anomer nucleoside of formula (II) obtaining enriched β-anomer nucleoside of the formula (I);
(iii) the dissolution of the nucleoside of the formula (I) in water;
(iv) heating the resulting solution to a temperature of from 40 to 60°C;
(v) cooling the solution to a temperature in the range from 10 to 25°C with stirring or without him, and without change of pH; and
(vi) filtrowanie is precipitated in the sludge solids:





where R represents a C1-C3alkyl;
R1represents a protective group of hydroxy;
R2represents a protective group of amino;
X represents halogen.



 

Same patents:

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.

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EFFECT: enhanced and valuable medicinal properties of agent.

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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.

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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)

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

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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: chemistry; pharmacology.

SUBSTANCE: invention refers to method of 6-O-[β-D-(2,3,4,6-tetra-O-acetyl)glucopyranosyl]-d,1-α-tocopherol (1) or 6-O-[β-D-(2,3,4,6-tetra-O- acetyl)galactopyranosyl]-d,1-α-tocopherol (2), consisting in interaction of α-tocopherol and α- or β-anomer of related D-gluco- or D-galactopyranose pentaacetates with added catalyst systems: BF3·OEt2 ionic liquid [bmim]PF6 at mole ratio α-tocopherol:sugar pentaacetate: BF3·OEt2:[bmim]PF6 = 1:1:2.5:0.3-5, in methylene chloride within 3 h at room temperature.

EFFECT: given compounds are precursors of related deacetylated glycosides with observed antiallergic and anti-inflammatory activity.

1 cl, 8 ex

FIELD: chemistry; pharmacology.

SUBSTANCE: invention refers to method of 6-O-[β-D-(2,3,4,6-tetra-O-acetyl)glucopyranosyl]-d,1-α-tocopherol (1) or 6-O-[β-D-(2,3,4,6-tetra-O- acetyl)galactopyranosyl]-d,1-α-tocopherol (2), consisting in interaction of α-tocopherol and α- or β-anomer of related D-gluco- or D-galactopyranose pentaacetates with added catalyst systems: BF3·OEt2 ionic liquid [bmim]PF6 at mole ratio α-tocopherol:sugar pentaacetate: BF3·OEt2:[bmim]PF6 = 1:1:2.5:0.3-5, in methylene chloride within 3 h at room temperature.

EFFECT: given compounds are precursors of related deacetylated glycosides with observed antiallergic and anti-inflammatory activity.

1 cl, 8 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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