Method for synthesis of β-l-5-fluoro-2',3'-dideoxy-2',3'-didehydrocytidine (β-l-fd4c)

FIELD: organic chemistry, antiviral agents.

SUBSTANCE: invention relates to a method for synthesis of β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrocytidine (β-L-FD4C) that is used as an antiviral agent. This method can be used in the large-scale manufacture of β-L-FD4C and it displays the effectiveness, high economy and ecology acceptable.

EFFECT: improved method of synthesis.

23 cl, 1 tbl, 1 ex

 

The scope to which the invention relates

The present invention relates to the production of nucleoside analogues for their use as antiviral agents. In particular, the present invention relates to the synthesis of β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C).

Description of the prior

Acquired immunodeficiency syndrome (AIDS), called the human immunodeficiency virus (HIV)is a serious public health threat around the world. According to the world health organization worldwide by the end of 2001, there have been 40 million people are HIV-infected or AIDS patients. Approximately 5 million people became infected with HIV in 2001. HIV/AIDS ranked fourth on the causes of mortality worldwide, and only in 2001 from the disease died, 3 million people (Weekly Epidemiological Record 76:381-388 (2001)).

Another virus, posing a serious threat to human health, is the hepatitis b virus (HBV). In addition to acute hepatitis HBV can cause chronic infection, often leading to cirrhosis and cancer of the liver and death. According to the data available for the year 2000, the virus HBV infected 2 billion people (Fact Sheet WHO/204, World Health Organization (October 2000)).

Various synthetic nucleosides were ID neficiary as potential antiviral agents for the treatment of HIV and HBV. After the development of the 3'-azido-3'-deoxythymidine (AZT), used in anti-HIV therapy (Mitsuya et al., Proc. Natl. Acad. Sci. USA 82:7096-7100)(1985)) as a potential anti-HIV - and HBV infections were identified some 2',3'-dideoxy(dd)and 2',3'-didehydro-2',3'-dideoxy(D4)-nucleosides. So, for example, nucleoside analogs approved for clinical use as antiviral agents, are 2',3'-dideoxyinosine (ddI), 2',3'-dideoxycytidine (ddC) (Mitsuya et al., Proc. Natl. Acad. Sci. USA 83:1911-1915 (1986)) and 2',3'-didehydro-3'-deoxythymidine (D4T)(Mansuri et al., J. Med. Chem. 32:461-466 (1989)). Although these nucleoside analogues are used in the form of natural D-enantiomer, however, recent research conducted in this area, were also aimed at the assessment of some nucleoside analogues with unnatural L-configuration. So, for example, as a potential remedy for HIV and HBV therapy were identified, for example, β-L-5-fluoro-2',3'-dideoxy-3'-thiacytidine (FTC)(Jeong et al., J. Med. Chem. 36:181-195 (1993)), β-L-5-fluoro-2',3'-dideoxycytidine (β-L-FddC)(Lin et al., Biochem. Pharmacol. 47:171-174 (1994)) and β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C)(Lin et al., J. Med. Chem. 39:1757-1759 (1996)).

It was confirmed that β-L-FD4C is especially valuable antiviral treatment for HIV and HBV infections (Lin et al., J. Med. Chem. 39:1757-1759 (1996)). The currently used methods for the synthesis of β-L-F4C (Lin et al., J.Med. Chem. 39:1757-1759 (1996)) give a low yield of product, and therefore they are unsuitable for large-scale production. Thus, alternative methods have been proposed synthesis β-L-FD4C (U.S. patent No. 6005097). However, the need for the development of new synthesis methods, to provide an efficient, cost-effective and environmentally friendly industrial production connection β-L-FD4C, which can be used to prevent epidemics of HIV and HBV infections worldwide, is still relevant.

Brief description of the invention

The present invention aims at solving the above problems associated with the development of new synthetic methods suitable for large scale production. β-L-FD4C. Such methods give higher output and higher efficiency and are more cost-effective and does not adversely impact on the environment.

In one of its aspects the present invention relates to methods of synthesis of β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C). In accordance with these methods:

(a) L-xylose of formula I:

subjected to interaction with acetone in the presence of a first acid catalyst and dehydrating agent is obtaining diacetate formula II

(b) 2,3-acetal of diacetyl formula II hydrolyzing in the presence of a second acid catalyst to obtain the acetal of formula III

(C) an Alcohol group of the acetal of formula III acelerou in the presence of a basic catalyst to obtain a complex diapir formula IV

(d) Arealevel group complex diapir formula IV hydrolyzing in the presence of acid to obtain diol of formula V

(e) a Hydroxyl group of the diol of formula V are removed from the receiving glikas formula VI

(f) 5-fertilizin formula VII

put the protection in two positions with obtaining bis-protected 5-fertilizin formula VIII

where Z denotes a protective group.

(g) Glycol formula VI is subjected to reaction in combination with a bis-protected 5-fertilizing formula VIII in the presence of a halogenation agent to obtain halogenated casinomaha derivative of the formula IX

(h) Halogenated casinowe derivative of formula IX is treated with metallic zinc and acetic acid to obtain dideoxy,didehydrothymidine derivative of the formula X

(i) Ester group dideoxy,didehydrothymidine derivative of formula X hydrolyzing in the presence of a base to obtain β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C) formula XI

.

Brief description of drawings

Some embodiments of the invention described with reference to the drawing which is presented only for illustrative purposes and should not be construed as limiting the invention.

The drawing illustrates the method of synthesis β-L-FD4C in accordance with some of the options for implementing the present invention.

Detailed description of the invention

The present invention relates to new methods of synthesis β-L-FD4C and to intermediate compounds which can be used for the synthesis of β-L-FD4C. Such methods allow for efficient, cost-effective and environmentally friendly industrial production β-L-FD4C with high yield and purity. Issued patents, published patent applications, and literature cited here are entered into the present description by reference to the extent as if each was specifically and individually put into the description by reference. Any mismatch between these publications and the present invention should be R tresino in favor of the description of the present invention.

Used herein, the term "lower alkyl" means a straight or branched C1-C4alkyl group such as methyl, ethyl, isopropyl, tert-butyl, etc. Used herein, the term "acid catalyst" means any acid reagent that catalyzes the desired chemical reaction. Non-limiting examples of acid catalysts used in the here described methods of synthesis are inorganic acids such as sulfuric acid or hydrochloric acid, and cation exchange resin. Cation exchange resins are insoluble acid resin, including, but not limited to, sulfonated polystyrene resins, sulfonated poliforumleon resin and other cation-exchange resin based on polystyrene, dextran, agarose and the like, the Term "major catalyst" means any basic reagent, catalyzes the desired chemical reaction. Non-limiting examples of the basic catalysts used in the ways described here the synthesis, are pyridine, triethylamine and dimethylaminopyridine (DMAP). The term "halogenation agent" means any agent capable of halogenoalkane, i.e. the introduction of the halogen atom in the compound. The term "dehydrating agent" means any agent that promotes the removal of water. The term "protective group" means any group which the binding is to be in one or more reactive positions of the connection, thus preventing the passage of the reactions in these terms, and which can be removed from the specified provisions of the standard chemical methods. The term "derivative" or "analog" first connection means and a second compound having a chemical structure similar to the chemical structure of the first connection, but at the same time or not containing one or more functional groups or one or more of the substituents present in the first connection, or containing one or more additional functional groups or one or more additional substituents, missing in the first connection. Used herein, the term "dideoxy" means a nucleoside portion having a sugar group, in which each carbon atom in two positions instead of the hydroxyl group is hydrogen. Used herein, the term "didehydro" means a nucleoside portion having a sugar group, which contains a double bond. For example, β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C) 2'-and 3'-positions of the carbon atoms of the sugar part instead of hydroxyl groups contains hydrogen atoms and between carbon atoms is a double bond.

The drawing illustrates the synthesis of β-L-FD4C in accordance with some variations of the embodiment of the invention. β-L-FD4C 11 synthesize PU is eating venue 7-stage procedure using L-1 xylose as a starting material. In at least some embodiments of the invention β-L-FD4C synthesized from L-xylose 1 without the procedure of evaporation to dryness of any of the intermediate compounds 2-10.

As shown in figure 1, the procedure of synthesis begins with the conversion of L-xylose 1 1.2-protected acetal 3. This conversion is carried out by obtaining diacetate 2, followed by hydrolysis under mild conditions 3,4-acatalog part in connection 2 with the formation of acetal 3. To obtain diacetate 2, L-1 xylose unite with acetone, with a dehydrating agent and an acid catalyst. Dehydrating agent used in the synthesis procedure, illustrated in the drawing, is copper sulfate, which has been found to give excellent output. Non-limiting examples of alternative dehydrating agents are MgSO4and Na2SO4. The acid catalyst used in the procedure illustrated in the drawing, is a resin Amberlyst® 15 (macrostate cation exchange resin based on a strong acid with a functional group sulfonic acid, Rohm & Haas, Philadelphia, PA). Such a resin used as a catalyst, is the most appropriate because it provides an effective reaction product of sufficient purity, easy to handle, does not require significant economic the ski cost and can be easily removed by filtration. Non-limiting examples of alternative resins used as catalysts, are other cation-exchange resin such as sulfonated polystyrene resins, sulfonated poliforumleon resin and other cation-exchange resin based on polystyrene, dextran, agarose and other Alternative acidic catalysts, non-resins and used in this reaction include, but are not limited to sulfuric acid and hydrochloric acid.

Hydrolysis desasosiego connection 2 with the formation of acetal 3 is carried out by adding water in an organic solvent, such as, for example, lower alkilany alcohol, followed by the addition of an acid catalyst. In at least some embodiments of the invention, before carrying out this procedure, a solution of compound 2 obtained from L-xylose 1, neutralize to ensure that the subsequent replacement of the solvent (i.e. acetone on the lower alkilany alcohol or other organic solvent, will not lead to decomposition of the compound 2 due to the preliminary hydrolysis with the formation of compound 3 or even reverse transformation into xylose. The neutralization is carried out by replacing the solvent in the presence of a base, such as, for example, solid potassium carbonate. Replacement of solvent lets C who directly use diacetyl 2 in the subsequent hydrolysis with the formation of acetal 3, that leads to increased efficiency, as in this case there is no need for stage evaporation to dryness between the reaction stages. In some embodiments of the invention the solvent for the hydrolysis step is a lower alcohol, such as methanol, as shown in the drawing; the ethanol is less toxic; technical methylated spirit (IMS), which is a cost-effective alternative to absolute ethanol for industrial production; or a combination of both. A non-limiting example of an alternative solvent is toluene. The acid catalyst used at the stage of hydrolysis, as shown in the drawing, is a resin Amberlyst® 15. Alternative acid catalysts include, but are not limited to, other cation exchange resins, and inorganic acids such as sulfuric acid and hydrochloric acid. The use of other non-resin catalysts leads to decrease their impact on the environment due to the fact that this procedure allows to reduce the amount of solid waste. Unwanted xylose present in the resulting solution of the acetal 3, is removed by grinding. Non-limiting examples of solvents suitable for grinding, are tert-butyl methyl ether (T IS IU), toluene/TVME, toluene/ethyl acetate and dichloromethane (DHM)/ethyl acetate. Subsequent replacement of the solvent for grinding carried out without decomposition of the acetal 3 with the formation of xylose by replacing the solvent in the presence of a stabilizing base, such as sodium bicarbonate. The use of solvent for grinding, such as toluene/TME, which is also suitable for use in the next stage of the reaction, increases the efficiency of the reaction, since in this case the solution of the acetal 3 can be used directly in the next reaction stage after grinding.

The alcohol group in compound 3 acelerou with formation of the corresponding ester group in compound 4 by treatment with acid chloride, such as, for example, p-trouillard, and the main catalyst. In synthesis, illustrated in the drawing, as the basic catalyst used pyridine, and the solvent for the reaction using DHM. Non-limiting examples of suitable basic catalysts are pyridine, triethylamine, dimethylaminopyridine (DMAP), and combinations thereof. Solvents suitable for use in the acylation reaction are DHM, toluene, TWO and combinations thereof. The use of solvent increases the efficiency of the reaction; for example, the use of toluene, which is suitable for carrying out the subsequent stages of the reaction, avoids the need for stage evaporation to dryness between the reaction stages. In some embodiments of the invention, and in particular for large-scale synthesis, the use of more toxic compounds such as pyridine and dichloromethane, are limited due to their negative impact on the environment.

1,2-acetelene group of compounds 4 hydrolyzing the corresponding alcohol group in compound 5. The hydrolysis is carried out by introduction of acid, such as formic acid or triperoxonane acid in water. The drawing illustrates the hydrolysis using formic acid in water. Solvents suitable for carrying out hydrolysis step, include, but are not limited to, acetonitrile, toluene, and combinations thereof. In specific embodiments of the invention formic acid in water use, along with a mixture of toluene/acetonitrile to achieve adjustable homogeneous reactions and obtain delovogo 5 product of good purity. In some embodiments of the invention product 5 is cleaned by grinding, for example, in hexane/TME or in toluene/TBME/heptane. In some alternative embodiments of the invention replace rest is rites and precipitation from the solvent, such as, for example, isopropyl ether, is carried out to highlight delovogo product 5 having a higher degree of purity.

Diol 5 in turn halogenated 5-forcasino derivative 9 by carrying out two-stage reaction combinations. First diol 5 turn in glycol 6 by reaction of interaction with iodine, imidazole and triphenylphosphine, as shown in the drawing. A non-limiting example of a suitable solvent for this reaction is dichloromethane. The resulting glycol 6 stored in conditions that prevent decomposition. Non-limiting examples of such conditions are: store at a temperature below approximately 0°in the form of concentrated oil and store at a temperature of about 5 to 6°in DGM or TWO during the period of time up to about 3 days. In some embodiments of the invention the resulting solution of compound 6 are used directly at the stage of combination reaction, described below, which leads to increased efficiency of the process.

For reaction step combinations, where glycol 6 turn in halogenated 5-forcasino derivative 9, it is also necessary to use the bis-protected 5-fertilizin 8. The drawing illustrates defense reaction of 5-fertilizin 7 two trimethylsilyl (TMS) groups with obtaining bis-protected connection is in 8. Protection is carried out by contacting the compound 7 with 1,1,1,3,3,3-hexamethyldisilazane and catalyst. In some embodiments of the invention the specified catalyst is ammonium sulfate. Alternative protective groups are well known in the art, and such groups are, but are not limited to, dimethylhexylamine, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, triphenylmethyl. Non-limiting examples of solvents suitable for use in this reaction protection, are toluene, chlorobenzene, chloroform, dichloroethane, dichloromethane, isopropyl ether, and combinations thereof. In some embodiments of the invention use a solvent which is suitable for carrying out the subsequent reaction of the combinations described below, and a solution of bis-protected 5-fertilizin 8 injected directly into the reaction mix. The stage of protection in a solvent suitable for use in the subsequent stage combinations, for example in chlorobenzene, ethylene dichloride (EDC) or in dichloromethane (DHM), the efficiency of the process, and the yield and quality of the product, since this helps to avoid separation or evaporation to dryness unstable product 8. In some embodiments of the invention, and in particular when large scale production is e, the use of more toxic compounds, such as EDC and DHM, limited due to their negative impact on the environment.

Glycol 6 and bis-protected 5-fertilizin 8 is subjected to the reaction mix in the presence of a halogenation agent to obtain halogenated 5-fertilizing derivative 9. In some embodiments of the invention mentioned halogenation agent is N-iodosuccinimide (NIC), as illustrated in the drawing. In at least some embodiments, the reaction mix is carried out in a chlorinated solvent, such as, for example, DHM, EDC, chlorobenzene, and combinations thereof. In some embodiments, the use of EDC helps to reduce negative effects of this procedure on the environment. Product 9 separated from the chlorinated solvent by adding the lower Olkiluoto alcohol, such as ethanol, which leads to the deposition of the compound (9). Selection of 9 product without conducting stage evaporation to dryness helps reduce the time to receive and avoids lengthy stage of heating the compound (9), which leads to some degree of decomposition. Alternative product 9 allocate by trituration with ethanol. In some embodiments of the invention product 9 is dissolved, for example, a lower alkyl acetate such as, for example, noted the Etat or ethyl acetate, and used in the subsequent reaction stage. Add a solution of compound (9) to the remaining reagents used in the subsequent stage of synthesis, avoids the need to add solids in the reaction vessel.

As shown in the drawing, halogenated 5-forcasino derivative 9 is treated with metallic zinc and acetic acid to obtain dideoxy,didehydro-5-fortitudinous derived 10 through dehalogenase and removal of toluene acid. This reaction is carried out in the alcohol and alkyl acetate. So, for example, use a combination of lower Olkiluoto alcohol and a lower alkyl acetate such as methanol and ethyl acetate, as shown in the drawing. In some embodiments the invention, the complex of the interesterification reaction between the alcohol and the alkyl acetate can be avoided thanks to the use of alcohol and alkyl acetate having the same alkyl group, for example, methanol and methyl acetate, or ethanol and ethyl acetate. In some embodiments of the invention, the product 10 produce by rubbing with a solution of hexane/ethanol. An alternative to the solution add 10 acetone. The use of acetone allows you to remove trace amounts of the parent compound 9 and associated with tauola products and leads to USAID is the product 10, that enables the selection of the connection 10 without holding stage evaporation to dryness. In order to avoid losses of water-soluble product 10 after washing with water, which is carried out by reverse extraction, add acetone.

The ester group of compound (10 hydrolyzing and get the final product β-L-FD4C 11. Suitable solvents for the hydrolysis reaction include, but are not limited to, polar alcohols, such as methanol. In at least some embodiments of the invention, the hydrolysis is carried out using a base. This base is present in stoichiometric or catalytic amounts. Non-limiting examples of suitable bases are ammonia, sodium methoxide, potassium carbonate, 1,8-diazabicyclo[5.4.0]-7-undecene (DBU) and Isopropylamine. In synthesis, illustrated in the drawing, the hydrolysis is carried out using ammonia in methanol. In some embodiments of the invention the use of gaseous ammonia can reduce the risk that exists when working with toxic gases. Commercially available solutions of ammonia in methanol, in particular, are used in the subsequent selection of product β-L-FD4C 11 in the form of solids.

In at least some embodiments of the invention produce a product β-L-FD4C. So, e.g. the, the crude product 11 is cleaned by standard methods known from the prior art, such as grinding, crystallization and/or filtration through a layer of silicon dioxide. A non-limiting example of a suitable purification procedure is rubbing in ethyl acetate or in a mixture of ethyl acetate/methanol followed by column chromatography. Sometimes, namely when the purity of the crude β-L-FD4C 11 is below 95%, to obtain a product with the desired purity of conduct crystallization or filtration through silica gel despite the loss of a certain number of substances when carrying out such procedures. Using a cleaner source connection 10 also improves the quality of the product β-L-FD4C 11. The connection 10 is more volatile solubility than β-L-FD4C 11. So, for example, to improve the quality of the source material 10 can be made of various cleanup procedures over a wide range, including crystallization, grinding, and/or filtration through a layer of silicon dioxide, and thus may be indirectly improved the purity of the end connections β-L-FD4C 11.

In some alternative embodiments of the invention β-L-FD4C 11 highlight by adding a solvent, which causes its precipitation. Examples of solvents suitable for the initiation of deposition of the East β -L-FD4C 11 are ethyl acetate and isopropanol. The deposition of the final product β-L-FD4C 11 avoids the need for stage evaporation to dryness and subsequent cleaning procedures. No need for chromatography on silica gel is particularly attractive from the point of view of environmental protection, because it allows you to reduce the amount of used solvent and the amount of waste generated.

It should be noted that the methods of the present invention can be also applied to obtain the compounds, related β-L-FD4C. Such related compounds are nucleoside analogues, such as 2',3'-dideoxynucleoside or 2',3'-dideoxy-2',3'-didehydrothymidine having a purine or pyrimidine base, associated with ribose part. Pyrimidine is a heterocyclic compound of the General class containing compounds such as uracil, thymine, cytosine and related analogues. Purine is a heterocyclic compound of the General class containing such compounds, as gipoksantin, xanthine, adenine, guanine and their analogues. Non-limiting examples of analogs of purine or pyrimidine are the Foundation in which a CH group is replaced by a nitrogen atom, and the base having one or more substituents on the ring to which e can be introduced or removed, or they can be modified conventional substituents known to specialists, for example by halogen, hydroxyl, amino or1-C6the alkyl. The methods of the present invention can also be used for the synthesis of various synthetic intermediates described in this application, or their equivalents.

For detailed illustrations of some embodiments of the invention are the following non-limiting examples.

Example 1

β-L-FD4C received in accordance with the procedure illustrated in the drawing.

Getting acetal 3

L-1 xylose (1000 g, 6,66 mol, 1 EQ.), acetone (10 l), copper sulfate (1,33 kg, 8.3 mol, 1.25 EQ.) and the resin Amberlyst® 15 (1000 g) were combined in a 22-liter round-bottom flask, equipped with a mechanical stirrer, a temperature sensor and a device for intake/release nitrogen. The reaction mixture was stirred in nitrogen atmosphere at room temperature for 16 hours. TLC (100% ethyl acetate, visualization phosphomolybdenum acid (PMA)) indicated the absence of starting compound (Rf˜0,05). Then added 750 g of solid sodium carbonate and the mixture was stirred for 30 minutes. The solution was filtered through Celite® (diatomaceous earth, World Minerals Inc., Santa Barbara, CA) to remove solids. The filtrate was concentrated in vacuum and received 1.4 kg prozrachnoj the oil. 1H-NMR confirmed the absence of L-1 xylose. Then the oil was dissolved in 7 l of a solution of methanol/water 4:1. Then, at room temperature, stirring, was added 1.4 kg resin Amberlyst® 15. The solution was stirred at room temperature until until TLC (100% ethyl acetate, visualization PMA) showed the absence diamesinae xylose 2 (Rf˜0,75). The resulting solution was filtered and the filtrate is brought to pH 8 by adding solid sodium bicarbonate (approximately 20 g). The solvent was removed in vacuum at 50% and received 1160 g of a light oil. The resulting oil triturated with 10 l of a solution of dichloromethane/ethyl acetate, 3:2, and dried over sodium sulfate. Then the dried solution was filtered through Celite® and concentrated in vacuum to obtain 1055 g (83%) of compound 3.1H-NMR and TLC confirmed the structure.

Obtaining complex diapir 4

Acetal 3 (1050 g, 5,52 mol, 1 EQ.), pyridine (1800 ml, 23,18 mol, 4.2 EQ.) and dichloromethane (5,65 l) were combined in a 22-liter round-bottom flask equipped with addition funnel, a device for intake/release nitrogen, a temperature sensor, a mechanical stirrer and an ice bath. The solution was cooled to 5°C in an ice bath under nitrogen atmosphere. Then, through the addition funnel was added p-trouillard (1,76 kg, 11,48 mol, 2,08 equiv.) while maintaining the temperature below 25°C. the Solution was left for p is remesiana for 16 hours under nitrogen atmosphere. TLC (ethyl acetate/hexane, 1:1, PMA-visualization) indicated completion of the reaction. Then the solution was washed 1×5 l of water, 1×5 l 3h. HCl solution and 1×5 l of water and dried over magnesium sulfate for three hours. After filtering off the drying agent, the filtrate was concentrated in vacuum to obtain 2460 g (quantitative yield) of compound 4 in the form of a light oil. TLC and1H-NMR confirmed the structure of this product.

Getting diol 5

9,3 l of formic acid and 2 l of water were combined in a 22-liter round-bottom flask, equipped with a temperature sensor, a mechanical stirrer and a heating jacket. Complex fluids 4 (2320 g, 5,44 mol, 1 EQ.) was dissolved in 2.3 ml of acetonitrile and one portion was added to a solution of formic acid/water. The combined solution was brought to 50°and was stirred for 2.5 hours. TLC (ethyl acetate/hexane, 1:1, PMA-visualization) indicated absence of starting material. Then the solution was diluted with 6 l of saturated salt solution and was extracted with 2×8 l of dichloromethane (or 1,2-dichloroethane or chloroform). The combined dichloromethane layers were washed 2×6 l of water, 2×4 l of saturated sodium bicarbonate (to bring the pH to a value of 7-8, which was indicated pH paper to determine pH), 1×6 l of water and 1×10 l of saturated salt solution, and then dried over sodium sulfate. After deletion the of the desiccant, the solvent was removed in vacuum to obtain a 1.88 kg (89%) bright solids. Then the solid is triturated with a solution of hexane/tertiary of motivational ether, 4:0,5 (MTBE) for 16 hours. The obtained white solids were isolated by filtration, washed with 2 l of hexane and dried in a vacuum oven at 35°obtaining 1630 g (78%) of compound 5 as a pale brown solid. HPLC showed a purity of 85%and1H-NMR confirmed the structure.

Obtaining halogenated 5-fertilizing derivative 9

Bis-protected 5-fertilizin 8:Compound 8 was prepared as follows. 5-Fertilizin 7 (605 g, 4,69 mol, 1.0 EQ.), 1,1,1,3,3,3-hexamethyldisilazane (5 l, with 23.7 mol, 5 EQ.) and ammonium sulfate (24 g, catalyst) were combined in a nitrogen atmosphere in a clean, dry, 12-liter round bottom flask fitted with mechanical stirrer, heating jacket, temperature sensor, a refrigerator and a device for intake/release nitrogen. After boiling under reflux for 30 minutes, the solids were dissolved, and the solution boiled under reflux for a further 2 hours. The resulting solution was left to cool to about 70°and was transferred to a nitrogen atmosphere at a rotary evaporator. The solvent was removed in vacuum at 85°and subjected to azeotropic distillation with 2×2 l of anhydrous xylene to obtain compound 8 as a white crystalline solid fuel is Dogo substances. This crystalline solid was dissolved in 6 l of dichloromethane and got a solution, which was stored prior to the reaction step combinations.

Glycol 6:Compound 6 was prepared as follows. Dichloromethane (25 l), iodine (1985, 7.82 mol, 2 equiv.) triphenylphosphine (2051 g, 7.82 mol, 2 EQ.) and imidazole (1170, 17,18 mol, 1.4 EQ.) were combined in a 100-liter reactor equipped with a cooling coil, a device for intake/release nitrogen, a temperature sensor, a mechanical stirrer and addition funnel. As you add imidazole color of the solution changed from purple to yellow and the temperature was increased to about 30°C. the Solution was cooled to 15°C in nitrogen atmosphere. Compound 5 (1510, 3,91 mol, 1 EQ.) was dissolved in 10 l of dichloromethane and portions were added to a 100 l reactor while maintaining the temperature below 20°C. After adding the total number of connections 5 the solution was stirred at room temperature for at least 2 hours. After 2.5 hours, TLC (ethyl acetate/hexane, 1:1, PMA-visualization) indicated the absence of starting compound (Rf˜0,5), weak spot at Rf˜0,8 (intermediate compound) and product glycol 6 (Rf˜to 0.9). The solution was suppressed 20 l of 20%aqueous sodium thiosulfate solution and stirred for about 20 minutes. The layers were separated and the organic layer was washed 1×20 l of water, 1×20 l is Assenova of the salt solution and dried over magnesium sulfate, at least for 1.5 hours. After removal of the drying agent, the solution was concentrated in vacuum to obtain oil and this oil is triturated with 4 liters of tertiary motivational ether (MTBE). Solid (triphenylphosphine oxide) was removed by filtration and washed with 3 l of MTBE. The filtrate was concentrated in vacuum to obtain oil and kept in an argon atmosphere at -10°prior to the reaction step combinations.

Combination:Stage combination was carried out as follows. The solution resulting from the reaction of protection 5-fertilizin, was added in nitrogen atmosphere in a clean, dry 22-liter round-bottom flask equipped with a mechanical stirrer, temperature sensor, an ice bath, and a device for intake/release nitrogen. Glycol 6 was dissolved in 7 l of dichloromethane and added to a 22-liter round bottom flask. The combined solution was stirred in nitrogen atmosphere while adding portions of N-iodosuccinimide (NIS, 1100 g, 4,88 mol, 1.25 EQ.). The temperature was maintained below 15°using an ice bath. After adding NIS solution was stirred at room temperature for at least 2 hours. After 4 hours TLC (ethyl acetate/hexane, 1:1, PMA-visualization) pointed to the lack of glikas 6 (Rf˜0,9), after which the reaction was suppressed h l of 20%aqueous sodium thiosulfate. The solution was left to stir at which Erno for 20 minutes. As the formation of significant amounts of solid substances, they were left to settle for 24 hours to facilitate filtering. Solids were removed by filtration through the filter bag 1 micron on the centrifuge. The filtrate was again placed in a 100 l reactor and the layers were separated. The organic layer was washed 1×20 l of water and 1×20 l of saturated salt solution, and then dried over sodium sulfate. After removing the desiccant, the dichloromethane was removed in vacuum to obtain a dark oil. The resulting oil was washed with 4 l of ethanol at 20°C and stirred 16 hours. The solids were isolated by filtration, washed with 4 l of ethanol and dried in a vacuum oven for 16 hours at 35°obtaining 1259 g (53%) of compound (9) in the form of not-quite-white solid. The purity of the product was 98%, as it was shown by HPLC and its structure was confirmed1H-NMR.

Getting dideoxy,didehydrothymidine derived 10

Two separate series of reactions was carried out as follows.

Series 1:Compound 9 (935 g 1,544 mol, 1 EQ.), ethyl acetate (8,4 l), methanol (1 l) and acetic acid (93 ml, 1,544 mol, 1 EQ.) was added in a clean, dry 22-liter round bottom flask. The solution was stirred for 10 minutes and one portion was added zinc (200 g, is 3.08 mol, 2.0 EQ.). The temperature was increased from 15 to 23°C for 15 mine is. TLC (ethyl acetate/methanol, 9:1, PMA - and UV visualization) did not show any reaction. Then one portion was added one equivalent of zinc (100 g), after which the temperature was increased to 41°C for 15 minutes. TLC indicated completion of the reaction after 30 minutes. The reaction mixture was left for 16 hours (overnight) at room temperature for mixing. The zinc was removed by filtration and the filtrate was washed 1×10 l of water and 1×10 l of 10% solution of ammonium chloride. Then the ethyl acetate was concentrated under vacuum to 1.5 l and the resulting suspension was left overnight at room temperature for mixing. The solids were isolated by filtration, washed with 1 l of ethyl acetate and dried in a vacuum oven for 16 hours at 35°obtaining 275 g (52%) of compound 10 as a white solid. The purity was 98%, as it was shown by HPLC and the structure of the obtained product was confirmed1H-NMR. Attempts to obtain additional number of connections from the mother liquor did not succeed.

Series 2:The original connection 9 for this series was kept at -10°C for approximately 3 months. TLC did not point to any decomposition. In a clean, dry 22-liter round bottom flask was added ethyl acetate (5.5 l), methanol (600 ml), acetic acid (61 ml, 1.01 mol, 1 EQ.) and zinc (195 g, 3,01 mol, 3 EQ.). The solution premesis is whether within 20 minutes. Then one portion was added compound 9 (615 g, 101 mol, 1 EQ.) and the temperature maintained below 30°using an ice bath. TLC (ethyl acetate/methanol, 9:1, UV and PMA-visualization) indicated completion of the reaction after 3 hours. The solution was left for 16 hours (overnight) at room temperature for mixing. The solids were filtered off and the filtrate was washed 1×4 l of water, 1×6 l of 10% ammonium chloride and 1×6 l of 10% potassium carbonate solution, saturated sodium chloride and then dried over sodium sulfate. After removing the desiccant, the solvent was removed in vacuum to obtain 382 g (quantitative yield) of a brownish solid. This solid is triturated with a solution of hexane/ethanol, 9:1 for 16 hours. The solids were isolated by filtration, washed with 500 ml of the above solution and dried in a vacuum oven for 16 hours at 35°obtaining 226 g (65%) of compound 10 in the form of a brownish powder. The purity was 95%, as it was shown by HPLC, and1H-NMR indicated the presence in this structure of small amounts of impurities. Elemental analysis indicated the content of 8%ash content, which was removed by filtration through Celite® in the next stage.

Getting β-L-FD4C 11

Two separate series of reactions was carried out as follows.

Series 1:Compound 10, obtained as described above in series 1 (273 g of 0.79 mol, 1 EQ.), and anhydrous methanol (3 l) were combined in a clean, dry 22-liter round-bottom flask, equipped with a mechanical stirrer, ice bath, the temperature sensor and the tube with a gas dispersion medium. Under stirring and maintaining the temperature at 25°in the flask for 1 hour was barbotirovany anhydrous gaseous ammonia. Then the flask was tightly closed and left for 24 hours (overnight) at room temperature for mixing. TLC (ethyl acetate/methanol, 9:1, PMA-visualization) indicated completion of the reaction. The solution was filtered through Celite® and the filtrate was concentrated in vacuum to obtain 180 g of bright solids. The solid is triturated with 2 l of ethyl acetate for 16 hours, were isolated by filtration and dried in a vacuum oven for 16 hours at 35°to obtain 151 g (84%) β-L-FD4C 11 in the form of a white solid. Purity was 99.7 per cent, as it was shown by HPLC.1H-NMR,13C-NMR, MS, elemental analysis and optical rotation confirmed the structure and purity of the compounds obtained.

Series 2:Compound 10 (226 g, 0,665 mol, 1 EQ.) and anhydrous methanol (3 l) were combined in a clean, dry 22-liter round-bottom flask. Under stirring and maintaining the temperature below 25°in the flask for 1 hour was barbotirovany of betwedn the th gaseous ammonia. Then the flask was tightly closed and left for 24 hours (at night) for mixing. TLC indicated completion of the reaction, and the solution was filtered through Celite® removal of suspended nerastvorimogo substances. The filtrate was concentrated in vacuum to obtain a brown-yellow solid. The solids were crushed with mortar and pestle, and mixed with 3 l of ethyl acetate for 3 days (weekend days). The solids were isolated by filtration, washed with 1 l of ethyl acetate and dried in a vacuum oven for 16 hours at 35°obtaining 147 g (98%) β-L-FD4C 11 in the form of a brown solid. Purity was 96,5%, as it was shown by HPLC. After conducting some research, 147 g of untreated β-L-FD4C 11 triturated with 10 ml/g of the solution of ethyl acetate/ethanol, 1:1 for 16 hours. The solids were isolated and dried to obtain 105 g (70,6%) β-L-FD4C 11 in the form of a brown solid. HPLC indicated at 98%purity. However,1H-NMR showed a certain amount of impurities, and elemental analysis found 8%ash content.

Example 2

β-L-FD4C received on the experimental equipment in accordance with the procedure illustrated in the drawing, but using several alternative solvents and reagents described below. All mass and volume of what we are rated (conditional) if it is not specifically mentioned. The number of source and final product purity and outputs are presented below in Table 1.

Getting acetal 3

Into a reaction vessel were loaded L-xylose (1.0 wt., 1,0 mol. EQ.) and acetone (7.9 wt., 10,0.). The obtained suspension was intensively mixed and added to the anhydrous copper sulfate (to 1.33 wt.), maintaining the reaction temperature below 25°C. were Then added to the resin Amberlyst® 15 (1.00 wt.), maintaining the reaction temperature below 25°C. the resulting mixture was intensively stirred at 20-25°until then, while the reaction with the formation of compound 2 was not supposedly completed, as was recognized by1H-NMR (<1 mol.% the residual L-1 xylose, typically about 16 hours).

The resulting mixture was filtered through Celite® and the filtrate is transferred through a 1 μm filter to the second vessel. The filter cake was washed with acetone (2×1,58 wt., 2×2,0 about.) and the combined filtrate was transferred into a second vessel containing sodium carbonate (0.5). The resulting mixture was intensively stirred for 30-40 minutes, after which it was confirmed that the supernatant in water had a pH >7. The mixture is then concentrated to about 5 vol. by vacuum distillation at temperatures of up to 30°C. Then, the vessel was loaded technical methylated spirit (IMS 2,02 wt., 2,5 about.) and the resulting mixture was concentrated in vacuum at 25-3° From about to about 5. Then the vessel was loaded another portion of IMS (2,02 wt., 2,5 about.) and the resulting mixture was subjected to vacuum distillation at 25-35°From about to about 5. The boot routine IMS and the distillation was repeated one more time.1H-NMR was used to confirm that the sample contained less than 1 mol.% acetone relative to ethanol. Then the mixture was filtered and the filter cake was washed with IMS (1,62 wt., 2,0 about.). The combined filtrate and washing was heated to 25-30°and added 1,2M HCl solution (0.34 in.), maintaining the reaction temperature at 25-30°C. At this stage the pH was ≤ 1. The reaction mixture was stirred until then, while the reaction to unprotect the formation of compound 3 was not supposedly completed, as was recognized by1H-NMR (>94% product 3, usually about 4 hours).

The resulting mixture was transferred to a separate vessel containing a suspension of sodium bicarbonate (1.0 wt.) in IMS (1,61 wt., 2,0 about.), while maintaining the temperature of 20-23°C. the resulting mixture was stirred for 30-40 minutes, after which it was confirmed that the supernatant in water had a pH of ≥7. The mixture was concentrated to about 4 vol. by distillation under vacuum at temperatures up to 35°C. Then, the vessel was loaded toluene (or 4.31 wt., 5,0 about.) and the resulting mixture was concentrated in vacuum at 25-35°From about to about 4. Then the vessel was loaded complement is inuu portion of toluene (or 4.31 wt., 5,0 about.) and the contents of the vessel was concentrated in vacuum at 25-35°With obtaining just about 4 about.1H-NMR was used to confirm that this sample contained <1 mol.% ethanol (IMS) with respect to toluene. After that the vessel was loaded tertiary butyl methyl ether (TVMA)(2,96 wt., 4,0 about.) and have1H-NMR analysis which confirmed that the molar ratio of toluene/TME was 1:1. The resulting mixture was stirred for 30-40 minutes, filtered and the filter cake washed TWO (1,48 wt., 2,0.).

Obtaining complex diapir 4

In the reaction vessel was loaded dimethylaminopyridine (DMAP)(to 0.025 wt.). Then the vessel was added a solution of acetal 3 (1.0 wt., 1,0 molecu.) in toluene/TME (total volume of about 10), obtained as described above, and the content was stirred, after which was added triethylamine (2,11 wt., 2,9 about., 4,0 molecu.). Then the vessel was loaded, TWO (0,74 wt., 1,0 about.) for washing. The mixture was cooled to 0-5°and the vessel was loaded p-trouillard (1,79 wt., 1,53 about., 2,2 molecu.), while maintaining the temperature at 0-10°With at least within 30 minutes. The container was rinsed, TWO (0,74 wt., 1,0 about.) in the vessel. Then the mixture was heated to 20-25°C for 30-40 minutes and was stirred until then, while the reaction with the formation of compound 4 presumably was not completed, as it was shown by HPLC (<05% square monoarylamino intermediate compounds, approximately 4 hours). The vessel was loaded 3M HCl solution (4,0 about.), while maintaining the reactor at a temperature below 25°C, after which it was confirmed that the aqueous phase had a pH of < 1. The reaction mixture is left to separate. The organic layer was washed with water (2×2,0 about.), and then with a solution of sodium bicarbonate (1.0 in.). It was confirmed that the aqueous phase had a pH >7. The organic phase was washed with purified water (2×2,0 about.). After washing the mixture was concentrated to about 4.5 to about. by vacuum distillation at 30-35°C.

Getting diol 5

To a solution of complex diapir 4 (1.0 wt., 1,0 molecu.) in toluene, obtained as described in the previous stage (approximately 2 vol.), added acetonitrile (1,57 wt., 2,0 about.), and then the purified water (1.0 in.) and formic acid (4,88 wt., 4,0 about.). The obtained two-phase mixture was heated to 40-45°and mixed up until reaction with the formation of diol 5 was not completed, as it was shown by HPLC analysis (<7% of starting compound 4; usually 12-16 hours).

The reaction mixture was cooled to 20-25°and to this mixture was added 30 wt./wt.% saturated salt solution (3.0 R.), and then TWO (0,74 wt., 1,0.). The layers were separated and the aqueous layer washed TWO (of 1.85 wt., 2,5 about. and at 2.59 wt., 3,5 about.). The combined organic layers were washed with water (2×30 vol.), after this mixture, 1:1, n is sydeny salt solution:5% (wt./about.) the sodium bicarbonate solution (4,0 about.), and then 5% (wt./about.) a sodium bicarbonate solution (2×about 30.) and finally, purified water (3,0 about.). The resulting organic solution was concentrated to approximately 4 rpm. by vacuum distillation at temperatures of up to 35°and determined the water content. If the water content was >3 wt.%, it was added toluene (of 3.46 wt., 4,0 about.) and water was removed by vacuum distillation at temperatures of up to 35°C. Then determined the content of the water, and if necessary, azeotropic distillation with toluene was repeated. After that, the solution was osvetleni, the filter was washed with toluene (1,73 wt., 2,0 about.) and the solution was concentrated to about 2 vol. by vacuum distillation at temperatures of up to 35°C. If necessary, the solution was brought up to 30-35°and slowly added isopropyl ether (IPE, 4,35 wt., 6,0 vol.), maintaining the temperature of 30-35°C. the resulting solution was cooled to 0-5°and kept for 3-4 hours after which the solid was isolated by centrifugation. The solid residue washed with IPE (2×1.45 wt., 2×2,0 about.) and the obtained solid was dried in vacuum at temperatures up to 35°C.

Obtaining halogenated 5-fertilizing derivative 9

Glycol 6:Compound 6 was prepared as follows. Into a reaction vessel were loaded iodine (1,447 wt.) and dichloromethane (DHM, 7,30 wt., 5,5 about.), and then added trifinio the fin (1.50 wt.) in DHM (of 5.84 wt., 4,4 about.), while maintaining a temperature of 20-30°C. Then were rinsed with dichloromethane (1.46 wt., 1,1.). In the suspension was loaded imidazole (0.85 wt.) in DHM (of 5.84 wt., 4,4 about.), maintaining the temperature 20-30°C. Then spent washing DHM (1.46 wt., 1,1 about.) and the suspension was cooled to 0-10°C. then slowly added compound 5 (1.00 wt., 1,0 molecu.) in DHM (of 5.84 wt., 4,4 about.), maintaining the temperature <10°C, and then washed DHM (1.46 wt., 1,1.). The resulting mixture was brought up to 5-10°and mixed up until reaction with the formation of compound 6 was not completed, as it was shown1H-NMR (disappearance of compound 5, usually 30 minutes). The reaction mixture was unstable and began to decompose after 3 hours.

To the reaction mixture, keeping the temperature <10°C, was added a 20% solution of sodium thiosulfate (11,0 about.) and a two-phase mixture was intensively stirred at 5-10°C for 15 minutes. The organic layer was analyzed for iodine content, then the layers were separated and the organic layer was washed with purified water (11,0 about.) at a temperature of 5-10°C. the organic phase was added magnesium sulfate (to 0.55 wt.) and the mixture was stirred for 2 hours at 5-10°C. the Dried solution was filtered and the solid is washed DHM (2,92 wt., 2,2 about.). The resulting organic solution was concentrated to about 5 vol. in vacuum in temp is the temperature up to 20° And added TWO (8,14 wt., 11,0.). After this, the solution was again concentrated to about 5 vol. and added TWO (8,14 wt., 11,0.). The solution was again concentrated to about 5 vol. and have1H-NMR analysis which confirmed TWO:DHM ≥ 10:1. The solid was filtered (precipitate on the filter with about 3 vol.) and washed TWO (2×1.63 wt., 2×2,2 about.). Then estimated the water content in the combined filtrate and the filtrate was concentrated to about 2 vol. at temperatures up to 20°C. To the concentrate was added DHM (7,30 wt., 5,5 about.) and the solution of compound 6 was kept at <5°until its use in the subsequent reaction stage combination. The product 6 was unstable and began to decompose after 48 hours at >5°C.

Bis-protected 5-fertilizin 8:Compound 8 was prepared as follows. The ratio of wt./about. means the mass of the parent compound 5 when the connection 6, as described above; mol eq (molecu.) refers to 5-forcasino 7. 5-Fertilizin 7 (0.4 wt.) and ammonium sulfate (0,016 wt., 0,04 molecu.) was loaded into a clean dry vessel, purged with nitrogen. Then was added chlorobenzene (2.2 wt., 2,0 about.) and the resulting suspension was subjected KF-analysis (usually <0.01 wt.%). After this was added, hexamethyldisilazane (1.08 wt., 1,42 about., 2,17 molecu.) and the resulting white suspension was heated to 110-115°and was stirred at e the th temperature for 16 hours. Obtained a clear, colorless solution of bis-protected 5-fertilizin 8 was cooled to 25-30°C, were analyzed using1H-NMR (usually 90-100 mol.% dicillo, 0-10 mol.% monosilane) and kept at room temperature before use in the subsequent stage combination.

The reaction mix:Stage of combination reaction was carried out as follows. The ratio of wt./about. means the mass of the parent compound 5 when the connection 6, as described above. In this stage used a clean dry jar. TVMA/DHM solution of compound 6 obtained as described above (volume Priam,7)were loaded into the reaction vessel containing chlorbenzoyl solution of compound 8 obtained as described above (about 4.4.), while maintaining the temperature <20°C. the Solution was cooled to 0-10°and for 50 minutes at a temperature <10°added 5 equal portions NIS (to 0.80 wt.). The reaction mixture was stirred at 5-10°until then, while the reaction with the formation of compound 9 was not supposedly completed, as was recognized by1H-NMR (usually 1 hour).

To the reaction mixture, keeping the temperature <20°C, was added 10% sodium thiosulfate solution (11,0 about.) and the resulting biphasic mixture was osvetleni to remove suspended solids. Then the layers were separated and the organic layer washed with purified water is th (7,7 about.). The organic layer was evaluated on the content of iodine and stirred with magnesium sulfate (1.1 wt.) for 2 hours at 20-25°C. the Dried organic layer was filtered and the solid is washed DHM (2,92 wt., 2,2 about.). To the combined organic extracts were added ethanol (8,64 wt., 11,0 about.) and the solution was cooled to 0-5°C for 2 hours. The obtained solid was filtered and washed with ethanol (2×1,73 wt. 2×2,2 about.) and the isolated product 9 was dried in vacuum at <20°C.

Getting dideoxy,didehydrothymidine derived 10

The ratio of wt./about. means the mass of the original compound (9), adjusted for the ethanol from the previous stage. The vessel was loaded with zinc powder (to 0.33 wt.), and then the acetate (3.5 in.), methanol (0,97 about.) and acetic acid (0,09 about.). Received the grey suspension was stirred for 30 minutes and then was heated up to 25-28°C. Compound 9 (1.0 wt.) was dissolved in ethyl acetate (about 4,5.) at 20-25°With in a separate vessel and slowly added to the zinc suspension, keeping the temperature <30°With (approximately 30-60 minutes). Washing with acetate (1 vol.) was carried out at 25°after that carried out the reaction with the formation of compound 10 and this reaction mixture was stirred at 25-30°C for 1 hour, and then took a sample for HPLC analysis (this analysis confirmed the presence of compound (9), <0.5% of p is osadi). The reaction mixture was filtered through the filter layer, the filter cake = 0,08 about.) and subjected prior to its clarification. The filter cake was washed with a solution of the acetate:methanol, 9:1, (2×1,0 about.) at 25-30°C. the combined filtrates were washed with 25 wt./wt.% ammonium chloride (5 vol.) and the layers were separated. The aqueous layer was washed with a mixture of methyl acetate:methanol, 9:1, (3,0 about.) at 25°and the layers were separated. The combined organic layers were washed with a mixture of saturated salt solution (30 wt./wt.%):sodium carbonate (20 wt./wt.%), 1:1 (5 vol., pre-mixed) and the layers were separated. The aqueous layer was washed with a mixture of methyl acetate:methanol, 9:1, (3,0 about.) at 25°and the layers were separated. The combined organic layers were dried over magnesium sulfate (1.0 wt.%). The dried solution was filtered and the filter cake was washed (with replacement) a mixture of methyl acetate:methanol, 9:1, (2×2,0 about.) at 25-30°and the combined filtrate was subjected to distillation at a temperature up to 30°to obtain about 3. Then acetone was added (4 vol.), the resulting solution was subjected to distillation at a temperature up to 30°with 3 about. This cycle is repeated up until the methanol content was <4 mol.% relative to the acetone. The obtained white suspension was cooled to 0-5°and kept for 1-2 hours, and then filtered. The filter cake was washed with acetone at 0-5° (2×Ob.) and dried in vacuum at temperatures up to 35° C.

Getting β-L-FD4C 11

The ratio of wt./about. means the mass of the parent compound 10. The vessel was loaded compound 10 (1.0 wt.), and then methanol (10.0 g.). The obtained suspension was stirred and added ammonia (6M in methanol, 5 vol.). Then the mixture was heated to 30-35°and intensively stirred until then, while the reaction with the formation of β-L-FD4C 11 has not been fully completed, as it was shown by HPLC (typically 24-32 hours). The temperature is brought up to 45-50°and the solution was osvetleni and washed with 1 volume of ethanol at 45-50°C. To the mixture was added isopropyl alcohol (IPA, 5 vol.). The resulting mixture was subjected to distillation under reduced pressure at temperatures up to 35°up until the molar ratio of IPA/methanol was not in the range from 1.5:1 to 2:1 (about 15-20. in the reaction vessel). The obtained white suspension was cooled to 0-5°C, filtered and the filter cake washed with IPA (2×2 vol.), then the product β-L-FD4C 11 was dried in vacuum at temperatures up to 35°C.

Table 1
The original connectionProduct
Stage synthesisPartyWeight (kg)Purity (LC-square%) PartyWeight (kg)Purity (LC-square%)Outlet
% no claim to any particular theorywt./wt.%
1→3And50efficiency of 99.7853,4-84,3107
In50D45,76-73,391,5
B199,94
B2at 99.95
B3efficiency of 99.78
B499,68
3→5**53,4-E50,5896,246,694,7
Dof 45.7 -F60,798,1765,3to 132.6
5→9E50,5896,2G57,799,5872,3114,1
F58,698,17H48,4299,2152,882,6
9→10G+H80I20,9498,934626,2
Gof 36.7699,58
H43,2499,21
10→11I20,9498,93J11,499,7682,754,4
* Means the conversion of compounds using known compounds, are indicated by numbers in figure 1

** Output connections 4 was theoretically accepted as 100%, so specify the total yield of compound 5 from compound 3

For a better understanding of the present invented what I presented above detailed description, however, for every person it is obvious that it can be made various changes without going beyond the scope defined in the following claims.

1. The method of synthesis β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C), including

(a) reaction of interaction of L-xylose of formula I

with acetone in the presence of a first acid catalyst and a dehydrating agent to obtain diacetate formula II

(b) hydrolysis of 2,3-acetal of diacetyl formula II in the presence of a second acid catalyst to obtain the acetal of formula III

(C) acylation of an alcohol groups of the acetal of formula III in the presence of a basic catalyst to obtain a complex diapir formula IV

(d) hydrolysis acatalog group complex diapir formula IV in the presence of acid to obtain diol of formula V

(e) removing the hydroxyl groups of the diol of formula V with getting glikas formula VI

(f) protection in two positions 5-fertilizin formula VII

obtaining bis-protection of the military 5-fertilizin formula VIII

where Z denotes a protective group,

(g) the reaction mix of glikas formula VI with bis-protected 5-fertilizing formula VIII in the presence of a halogenation agent to obtain halogenated casinomaha derivative of the formula IX

(h) processing of halogenated casinomaha derivative of the formula IX metallic zinc and acetic acid to obtain dideoxy, didehydrothymidine derivative of the formula X

(i) hydrolysis of the ester group of dideoxy, didehydrothymidine derivative of formula X in the presence of a base to obtain β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C) formula XI

2. The method according to claim 1, further comprising selecting a product β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C).

3. The method according to claim 2, where the specified allocation provides processing solvent, which causes deposition β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C).

4. The method according to claim 3, where the specified solvent is ethyl acetate or isopropanol.

5. The method according to claim 1, where the removal of hydroxyl groups on the stage (f) involves treatment with iodine, triphenylphosphine and imide what zolom.

6. The method according to claim 1, where the reaction of the protection specified connection in two positions on the stage (f) provides for the processing of 1,1,1,3,3,3-hexamethyldisilazane and a catalyst.

7. The method according to claim 6, where the specified catalyst is ammonium sulfate.

8. The method according to claim 1, where the specified protective group Z in each position of the compounds of formula VIII is trimethylsilyl.

9. The method according to claim 1, where these first and second acid catalysts in the stages (a) and (b) independently selected from the group consisting of sulfuric acid, hydrochloric acid and cation exchange resins.

10. The method according to claim 1, where the specified dehydrating agent at the stage of (a) selected from the group consisting of copper sulfate, magnesium sulfate and sodium sulfate.

11. The method according to claim 1, where the main catalyst in stage (C) is selected from the group consisting of pyridine, triethylamine, dimethylaminopyridine and their combinations.

12. The method according to claim 1, where the specified acid in stage (d) is formic acid or triperoxonane acid.

13. The method according to claim 1, where the specified base on the stage (i) is selected from the group consisting of ammonia, sodium methoxide, potassium carbonate, 1,8-diazabicyclo[5.4.0]-7-undecene (DBU) and Isopropylamine.

14. The method according to claim 1, where the specified halogenation agent in stage (g) is N-iodosuccinimide (NIS).

15. The method according to claim 1, where the stage (b) is carried out in a solvent such as a lower alkilany alcohol.

16. The method according to claim 1, where the stage (C) is carried out in a solvent selected from the group consisting of dichloromethane, toluene, tert-butyl methyl ether and combinations thereof.

17. The method according to claim 1, where the stage (d) is carried out in a solvent selected from the group consisting of acetonitrile, toluene, and combinations thereof.

18. The method according to claim 1, where the stage (e) is carried out in dichloromethane.

19. The method according to claim 1, where the stage (f) is carried out in a solvent selected from the group consisting of toluene, chlorobenzene, chloroform, dichloroethane, dichloromethane, isopropyl ether, and combinations thereof.

20. The method according to claim 1, where the stage (g) is carried out in a solvent selected from the group consisting of dichloroethane, dichloromethane, chlorobenzene and their combinations.

21. The method according to claim 1, where the stage (h) is carried out in the presence of lower Olkiluoto alcohol and a lower alkyl acetate.

22. The method according to claim 1, where the stage (i) is carried out in methanol.

23. The method according to claim 1, where β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrothymidine (β-L-FD4C) synthesized from L-xylose without holding stage evaporation to dryness of any of the intermediate compounds of formulas II-X.



 

Same patents:

FIELD: organic chemistry, biochemistry.

SUBSTANCE: invention relates to oligomer comprising at least one nucleoside analogue of L-ribo-CNA of the general formula (Ia) wherein X represents -O-; B represents nitrogen base; P means radical position in an internucleoside linkage followed by monomer or 5'-terminal hydroxy-group; P* means an internucleoside linkage with precede monomer or 3'-terminal hydroxy-group; R2* and R4* mean in common biradical -(CH2)0-1-O-(CH2)1-3-(CH2)0-1-S-(CH2)1-3- or -(CH2)0-1-NR-(CH2)1-3- wherein R means hydrogen atom, alkyl or acyl; R1*, R2, R3*, R5 and R5* mean hydrogen atom. Also, invention proposes nucleoside analogues used in preparing oligomers. Proposed oligomers elicit the enhanced affinity to complementary nucleic acids and can be used as a tool in molecular-biological investigations and as antisense, antigen agents of agents activating genes.

EFFECT: valuable properties of analogues.

15 cl, 3 tbl, 4 dwg, 17 ex

FIELD: organic chemistry, medicine, oncology, pharmacy.

SUBSTANCE: invention relates to new stable crystalline forms of derivative of pyrimidine nucleoside of the formula (I) eliciting the excellent anti-tumor activity. Also, invention relates to pharmaceutical composition eliciting an anti-tumor effect, applying crystalline form for preparing medicinal agent and to a method for prophylaxis or treatment of tumor diseases.

EFFECT: improved method for prophylaxis and treatment, valuable medicinal properties of derivative.

10 cl, 2 tbl, 4 dwg, 9 ex

The invention relates to derivatives of 5’-deoxycitidine General formula

where R1denotes a hydrogen atom or a group which is easily hydrolyzed under physiological environments; R2denotes a hydrogen atom or a group-CO-OR4where R4denotes a saturated hydrocarbon group with a straight or branched chain, containing from one to fifteen carbon atoms, or a group of the formula -(CH2)n-Y, where Y denotes a cyclohexyl or phenyl, and n is an integer from 0 to 4; R3denotes a hydrogen atom, bromine, iodine, cyano, C1-C4alkyl group which may be substituted atom (s) halogen, vinyl or etinilnoy group which may be substituted atom (s) halogen, C1-C4by alkyl; provided that R2and R3may not simultaneously denote a hydrogen atom

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

The invention relates to bicyclerelated analogues of General formula (1) and the oligonucleotides on the basis thereof, containing one or more structural units of the General formula (1A), where R1represents a hydrogen atom, a protective group, the group of phosphoric acid, etc., R2is sidegroup or amino group which may be substituted, represents a purine-9-ilen, or 2-oxo-1,2-dihydropyrimidin-1-ilen group which may be substituted by one or more substituents

The invention relates to nucleoside of the formula (I) having bicyclic sugar group containing 2',4'-bridge connection, and the oligonucleotides on the basis of their

The invention relates to a derivative copernicia and their pharmaceutically acceptable salts of General formula I

< / BR>
where R1represents a methyl group, R2represents a methyl group, R4represents a hydroxy-group and X represents a methylene group; R1represents a methyl group, R2represents a hydrogen atom, R4represents a hydroxy-group and X represents a methylene group; R1represents a methyl group, R2represents a methyl group, R4represents a hydrogen atom and X represents a methylene group; R1represents a hydrogen atom, R2represents a hydrogen atom, R4represents a hydroxy-group and X represents a methylene group; or R1represents a methyl group, R2represents a methyl group, R4represents a hydroxy-group and X represents a sulfur atom

The invention relates to substituted ammonium salts of 5'-H-phosphonate 3'-azido-3'-deoxythymidine General formula (1), where RR'R N are L-alanine, ethanolamine, triethanolamine, 6-aminocaproic acid, pyridoxine or dimethylaminoethanol, which are selective inhibitors of the production of human immunodeficiency virus HIV-1 and HIV-2

FIELD: organic chemistry, medicine, oncology, pharmacy.

SUBSTANCE: invention relates to new stable crystalline forms of derivative of pyrimidine nucleoside of the formula (I) eliciting the excellent anti-tumor activity. Also, invention relates to pharmaceutical composition eliciting an anti-tumor effect, applying crystalline form for preparing medicinal agent and to a method for prophylaxis or treatment of tumor diseases.

EFFECT: improved method for prophylaxis and treatment, valuable medicinal properties of derivative.

10 cl, 2 tbl, 4 dwg, 9 ex

FIELD: organic chemistry, biochemistry.

SUBSTANCE: invention relates to oligomer comprising at least one nucleoside analogue of L-ribo-CNA of the general formula (Ia) wherein X represents -O-; B represents nitrogen base; P means radical position in an internucleoside linkage followed by monomer or 5'-terminal hydroxy-group; P* means an internucleoside linkage with precede monomer or 3'-terminal hydroxy-group; R2* and R4* mean in common biradical -(CH2)0-1-O-(CH2)1-3-(CH2)0-1-S-(CH2)1-3- or -(CH2)0-1-NR-(CH2)1-3- wherein R means hydrogen atom, alkyl or acyl; R1*, R2, R3*, R5 and R5* mean hydrogen atom. Also, invention proposes nucleoside analogues used in preparing oligomers. Proposed oligomers elicit the enhanced affinity to complementary nucleic acids and can be used as a tool in molecular-biological investigations and as antisense, antigen agents of agents activating genes.

EFFECT: valuable properties of analogues.

15 cl, 3 tbl, 4 dwg, 17 ex

FIELD: organic chemistry, antiviral agents.

SUBSTANCE: invention relates to a method for synthesis of β-L-5-fluoro-2',3'-dideoxy-2',3'-didehydrocytidine (β-L-FD4C) that is used as an antiviral agent. This method can be used in the large-scale manufacture of β-L-FD4C and it displays the effectiveness, high economy and ecology acceptable.

EFFECT: improved method of synthesis.

23 cl, 1 tbl, 1 ex

FIELD: chemistry; medicine.

SUBSTANCE: invention pertains to nucleoside derivatives with general formula (I) , where R1 represents hydrogen; COR5, where R5 is chosen from C1-18 alkyl, phenyl, CH2OPh and CH2Ph; C(=O)OR5, where R5 represents C1-18 alkyl; or COCH(R6)NHR7, where R6 represents C1-5 alkyl, and R7 represents R5OCO, where R5 is C1-18 alkyl, R2 represents hydrogen; COR5 , where R5 is chosen from C1-18 alkyl, C1-18 alkenyl, phenyl or CH2OPh; C(=O)OR5, where R5 is chosen from C1-18 alkyl, C1-18 alkenyl, substituted with low alkyl; C(=O)NHR5, where R5 represents C1-18 alkenyl; or COCH(R6)NHR7, where R6 is chosen from side-chains of natural amino acid and C1-5alkyl, and R7 is chosen from hydrogen and R5OCO, where R5 is C1-18 alkyl; R3 and R4 are the same and are chosen from hydrogen; COR5, where R5 is chosen from C1-18 alkyl, or phenyl; C(=O)OR5, where R5 is C1-18 alkyl, or R3 and R4 together represent C(CH3)2; or their pharmaceutical acidic additive salts; under the condition that, at least one of R1, R2, R3 or R4 is not hydrogen. The invention also relates to pharmaceutical compositions, which have antiviral activity to HCV.

EFFECT: obtaining of a range of new biologically active substances.

15 cl, 2 tbl, 9 ex

FIELD: chemistry.

SUBSTANCE: invention refers to anhydrous polymorphic salt representing hemisulphate 1-[4(S)-azido-2(S),3(R)-dihydroxy-4-(hydroxymethyl)-1-(R)-cycloamyl]- cytosine of formula (Ia) possessing extra stability and improved physical properties that facilitates production, transportation of compound and preparation of compositions on the basis of compounds of formula I . Invention also refers to various crystal forms of compounds of formula (Ia), characterised by certain values of lattice constant D (interplanar spacing), to methods of their production from compounds of formula (I), to pharmaceutical compositions on the basis of compounds of formula (Ia) and to method of disease treatment caused by C hepatitis virus, including injection if such treatment is required, of therapeutically effective amount of compound of formula (Ia).

EFFECT: production of compound possessing extra stability and improved physical properties that facilitates production, transportation of compound and preparation of compositions on its basis.

14 cl, 5 dwg, 1 tbl, 9 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: medicine.

SUBSTANCE: invention refers to new compounds of formula (IX) or to their pharmaceutically acceptable salts having inhibitory activity to hepatitis C, to the related pharmaceutical composition and to their application for making a medical product. In compound of formula (IX) , R1 and R2 independently represent H, phosphate or acyl; X represents O; base* represents pyrimidine base; R12 represents C(Y3)3; Y3 represents H and R13 represents fluorine.

EFFECT: higher efficiency of the composition and treatment method.

31 cl, 14 dwg, 26 ex

FIELD: chemistry.

SUBSTANCE: disclosed are α- and β-crystalline forms of 5'-desoxy-N4-carbopentyloxy-5-fluorocytidine of formula (III) , their preparation method through crystallisation of the raw product from a suitable solvent and pharmaceutical compositions based on the said compounds, having anti-cancer activity. The solvent used when preparing the α-modification is an ester or a mixture of ester-containing solvents. The solvent used when preparing the β-modification is a mixture of water and alkanol or a mixture of tetrahydrofuran and diethyl ether or carbon tetrachloride.

EFFECT: obtaining compounds and pharmaceutical compositions based on the said compounds, having anticancer activity.

10 cl, 2 dwg, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to phosphoramidite derivatives of general formula where Bx denotes adenine, guanine, cytosine, thymine or uracil, where the amine group of adenine, guanine and cytosine can be optionally protected by a protective group selected from acetyl and phenoxyacetyl; R1 is a substitute of general formula in which R11, R12 and R13 are identical or different, and each denotes hydrogen or alkoxy; R2a and R2b are identical or different, and each denotes alkyl; and WG1, WG2 denote a cyano group. The invention also pertains to a multistep method of producing the said compounds. The invention also relates to intermediate compounds of the said method, namely: an intermediate ether compound of general formula where L is a halogen or a C1-C5alkylthio group; WG1 is a cyano group; an intermediate compound of general formula where Bx denotes adenine, guanine, cytosine, thymine or uracil, where the amine group of adesine, guanine and cytosine can be optionally protected by a protective group selected from an acetyl group and a phenoxyacetyl group; and WG1 denotes a cyano group; an intermediate compound of general formula where Bx is as described above; R1 is a substitute of general formula (2); an intermediate compound of general formula where Bx is as described above; A is a silicon-containing substitute of general formula or where R6 denotes alkyl and WG1 denotes a cyano group. The invention also relates to a method of producing an oligonucleotide of general formula where each B independently denotes adenine, guanine, cytosine, uracil or thymine; each R independently denotes H or hydroxyl and at least one of R denotes hydroxyl; Z denotes H or a phosphate group; and n is an integer between 1 and 100, involving steps A-G, characterised by use of said phosphoramidite derivatives as a monomer compound of nucleic acid at step B.

EFFECT: high yield.

7 cl, 1 dwg, 21 ex

FIELD: chemistry.

SUBSTANCE: method enables to obtain 4-amino-1-((2R,3R,4R,5R)-3-fluoro-4-hydroxy-5-hydroxymethyl-3-methyl-tetrahydro-furan-2-yl)-1H-pyrimidin-2-one of formula (IV), which is a strong inhibitor of NS5B polymerase of hepatitis C virus (HCV).

EFFECT: high yield.

2 cl, 4 ex

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