(-)-4-amino-5-fluoro-1-(2-hydroxymethyl-1,3-oxathiolan-5-yl)- (1h)-pyrimidine - 2-it, a mixture of its enantiomers, methods for their preparation and method of treatment


(57) Abstract:

The invention relates to new nucleoside analogues of 1,3-oxathiolane and their use for treating viral infections, HIV infection, hepatitis b, more specifically to the (-)-4-amino-5-fluoro-1-(2-oxymethyl-1,3-oxathiolan-5-yl)-(1H )-pyrimidine-2-ONU (I) and its pharmaceutically acceptable derivative containing pharmaceutical compositions. Describes a new mixture of (-)-4-amino-5-fluoro-1-(2-hydroxymethyl-1,3-oxathiolan-5-yl )-(1H) -pyrimidine-2-it (+)-4-amino-5-fluoro-1-(2-hydroxymethyl)-1,3-oxathiolan-5-yl)-(1H)-pyrimidine-2-it or their pharmaceutically acceptable derivatives, where (+)-enantiomer is present in an amount of not more than 5 wt.%. The method of obtaining the compound I, are that teach the selection (-)-enantiomer from a mixture of (-)- and (+)-enantiomers. The method of treatment is administered an effective amount of the compound (). 5 S. and 19 C.p. f-crystals, 3 tables.

The present invention relates to nucleoside analogues and their use in medicine. More specifically the invention relates to nucleoside analogues of 1,3-oxathiolane, including their pharmaceutical compositions and their use in the treatment of viral infections.

The only connection that midin (AZT, zidovudine, BW 509U). However, this compound has significant predisposition to side effects, and therefore may not be used or after the first attempts to use its further application is necessary to stop for a large number of patients. There remains an urgent need to develop compounds that were active against HIV, but would have a much better companion therapeutic index.

The compound according to formula (I)

< / BR>
is a racemic mixture of two enantiomers by formulas (I-1) and (I-2):

< / BR>
We unexpectedly found that (-)-enantiomer of compound (I) exhibits greater activity than (+)-enantiomer, although both enantiomers have unexpectedly low cytotoxicity. Thus, the first aim of the present invention is (-) (or levogyrate) enantiomer of compounds of formula (I) and its pharmaceutically acceptable derivatives.

(-)-enantiomer has the chemical name (-)-4-amino-5-fluoro-1-(2-oxymethyl-1,3-oxathiolan-5-yl)-(1H)-pyrimidine-2-he (hereinafter referred to as compound (A)). This enantiomer has an absolute stereochemical structure represented by formula (I-1).

The compound (A) primulina 5 wt.% (+)-enantiomer, more preferably no more than about 1 wt.%

By "pharmaceutically acceptable derivative" is meant a pharmaceutically acceptable salt, ester or salt of the ester compounds (A) or any other compound which upon appointment, the patient is able to identify (directly or indirectly) a compound (A), or a metabolite possessing antiviral activity, or radical.

For professionals it is obvious that the compound (A) may be modified, with the aim of obtaining its pharmaceutically acceptable derivatives, according to the functional groups as a fragment of the base, and oxymethylene group oxathiolane rings. Modifications on all of these functional groups are included in the scope of claims of the present invention. However, of particular interest are the pharmaceutically acceptable derivatives obtained by modification of the 2-oxymethylene group oxathiolane rings.

the preferred esters of the compounds (A) are derivatives in which the hydrogen atom in the 2-oxymethylene group substituted acyl group in which decarbonising fragment R of the ester is selected from a hydrogen atom, alkyl group with straight or branched chain (for example, methyl group, ethyl grueling group), aranceles group (e.g., benzyl group), aryloxyalkyl group (for example, phenoxymethyl group), aryl group (e.g. phenyl group optionally substituted by halogen atom (C1-C4) alkyl group or (C1-C4)alkoxygroup; a sulfonate such as alkyl - or Arakishvili group (for example, methanesulfonyl group); esters of amino acids (for example, L-valurile group or L-isoleucinol group) and mono-, di - or triphosphates.

With regard to the above esters, if not particularly specified, any alkyl fragment in them mainly contains from 1 to 16 carbon atoms, in particular from 1 to 4 carbon atoms. Any aryl fragment in these compounds is mainly a phenyl group.

In particular, the esters may be esters formed (C1-C16)alkyl groups, unsubstituted benzyl esters or benzyl esters, substituted by at least one halogen atom (bromine, chlorine, fluorine or iodine), (C1-C16)alkyl group, (C1-C6)alkoxygroup, or nitrogroup triptorelin group.

Pharmaceutically acceptable salts of the compounds (A) are salts derived from pharmaceutically acceptable inorganic and er is the one sulphuric, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, p-toluensulfonate, tartaric acid, acetic acid, citric acid, methanesulfonate acid, formic acid, benzoic acid, malonic acid, naphthene-2-sulfonic acid and benzosulfimide acid. Other acids, such as oxalic acid, which are not in themselves pharmaceutically acceptable, may be used to obtain the intermediate compounds in the synthesis of the compounds of the present invention and their pharmaceutically acceptable salts accession acids.

Salts formed from the corresponding bases are alkali metal salts (e.g. sodium), salts of alkaline earth metals (e.g. magnesium), ammonium salts and Quaternary ammonium salts NR4+ (where R is (C1-C4) alkyl group).

References in this description to the compound (A) according to the invention pertain to the compound (A) and its pharmaceutically acceptable derivatives.

Compounds of the present invention either possess antiviral activity, doubt the AI replication of retroviruses, including human retroviruses, such as human immunodeficiency virus (HIV) that causes AIDS.

Compounds of the present invention can also be used to treat animals, including humans, infected with hepatitis B.

The next target of the present invention is the use of compounds (A) or its pharmaceutically acceptable derivative as an active therapeutic agent, in particular antivertigo means, for example, in the treatment of retroviral infections or infections caused by hepatitis B.

Another objective of the present invention is a method of treating viral infections, particularly infections caused by hepatitis B virus or retroviruses, such as HIV, in animals, including humans, which consists in the use of effective amounts of compounds (A) or its pharmaceutically acceptable derivative.

Another or alternative aim of the invention is the use of compounds (A) or its pharmaceutically acceptable derivative in the manufacture of medical preparations for the treatment of viral infections.

Compounds of the present invention the progressive total of lymph nodes, AIDS-related neurological conditions (such as dementia and trophic prepares), positive anti-HIV antibody and HIV-positive conditions, Kaposi Kaposi, thrombotic purpura and diseases caused by conditionally pathogenic microorganisms, for example, pneumocystis carinii.

Compounds of the present invention is also useful for preventing the development of disease in the clinical form in patients who show a positive response to anti-HIV antibody and HIV-antigen, and for prophylaxis after exposure to HIV.

The compound (A) or its pharmaceutically acceptable derivatives may also be used to prevent viral contamination of physiological fluids such as blood and semen in vitro.

Compounds of the present invention can also be used in the treatment of animals, including humans, infected with hepatitis B.

For professionals it is obvious that this method of treatment also extends to the prevention of disease and treatment of the identified infections or diseases with pronounced symptoms.

It should be emphasized that the number of connections Narodnogo connection but from the destination path, the nature of the disease, the age and General condition of the patient and is left to the discretion of the attending physician or veterinarian. However, in the General case, an acceptable dose is from about 0.1 to about 750 mg/kg body weight per day, preferably from 0.5 to 60 mg/kg / day, and most preferably from 1 to 20 mg/kg / day.

The desired dose may be a single dose or divided into several doses that are assigned after a certain period of time, such as two, three, four or more dosages per day.

The compound of the present invention is convenient to assign a standard dose containing, for example, from 10 to 1500 mg, preferably from 20 to 1000 mg, more preferably from 50 to 700 mg of active ingredient on the standard dose.

Ideally, the number assigned to the active ingredient must be such that its maximum concentration in plasma was approximately 1 to approximately 75 microns, preferably from about 2 to 50 μm, most preferably from 3 to about 30 microns. This is achieved, for example, intravenous injection, in particular, the salt solution with a concentration of from 0.1 to about 100 mg of the active ingredient. The desired concentration of drug in the blood can be maintained with a continuous infusion in the amount of from about 0.01 to about 5.0 mg/kg per hour or alternating infusions containing about 0.4 to about 15 mg/kg of the active ingredient.

Despite the fact that therapeutic purpose, the compound of the present invention may be administered in the form of a pure substance, it is preferable to introduce the active ingredients in the form of pharmaceutical compositions.

Thus, in the present invention hereinafter claimed pharmaceutical composition comprising the compound (A) or its pharmaceutically acceptable derivative with one or more pharmaceutically acceptable carriers and, optionally, other therapeutic and/or prophylactic methods. The carrier must be "acceptable" in the sense that they must be combined with each other in the composition and do not adversely affect patient who uses this structure.

Pharmaceutical compositions include pharmaceutical compositions suitable for oral, rectal, intranasal, local (including buccal or podlaczenia or in the form suitable for inhalation or insufflation of powdered drugs. The compositions, if possible, easier to prepare in the form of discrete doses that can be received by any of the means well known in pharmacy. All methods include mixing the active substance with a liquid carrier or thoroughly pulverized solid carrier or both of these media, and then giving the desired product form.

Pharmaceutical compositions that are convenient for oral purposes, can be in the form of standard doses, such as capsules, starch capsules or tablets, each of which contains a certain amount of the active ingredient; in the form of powders or granules; in the form of a solution, suspension or emulsion. The active ingredient may also be in the form of a ball, medicinal porridge or paste.

Tablets and capsules for oral use can contain conventional excipients such as binders, lubricants, disintegrating agents or wetting agents. Tablets can be coated by methods well known in the art. Liquid preparations for oral use can be in the form of, for example, suspensions in water or oil, solutions, emulsions, syrups or elixirs, PIM suitable liquid carrier. Such liquid preparations may include conventional additives, such as suspendresume agents, emulsifying agents, non-aqueous liquid media (which may include edible food oils or preservatives.

Compounds of the present invention can be prepared for parenteral use (for example, by injection or continuous infusion) and may be represented by a single dose in the form of ampoules filled syringes, containers for infusion of small amounts of drugs or containers that contain a large number of doses, which add preservatives. The compositions can be in the form of suspensions, solutions or emulsions in oil or aqueous media, and can contain auxiliary compounds such as suspendresume tools, stabilizers and/or dispersants. The active ingredient may be in powder form, obtained aseptic isolation, sterilization or by lyophilization from solution, and it is necessary before use to mix with a suitable carrier, in particular with sterile pyrogen-free water.

For local application on the layer of the epidermis of the compound of the present invention may be in the composition of ointments, creams or lotions, as well as in the group of suitable thickeners and/or gelling funds. Lotions can be prepared using water and oil, as well as to contain in the General case, one or more emulsifiers, stabilizers, dispersants, suspendida agents, thickeners or dyes.

Formulations suitable for oral assignments include tablets that contain the active ingredient in a flavored mass, usually sucrose, gum or resin tragakant; lozenges comprising the active ingredients in an inert carrier, such as gelatin and glycerol or sucrose and gum; liquid for rinsing the mouth, consisting of an active ingredient in a suitable liquid carrier.

Pharmaceutical compositions, convenient for rectal purpose in which the carrier is a solid compound, preferably prepared in the form of a suppository, containing the standard dose. Suitable carrier materials include peanut butter and other materials commonly used for these purposes, and they suppositories conveniently be prepared by mixing the active ingredients with the softened or melted carrier with subsequent cooling and casting in the form.

Formulations for vaginal destination can be presented with vaginal suppositories, tampons, for example, GE is their purpose well-known carriers.

Intranasal purpose of the connection of the present invention can be used in the form of liquid aerosol or sprayable powders in the form of drops.

Drops can be prepared using water and non-aqueous liquids together with one or more dispersing agents, soljubilizatorami and suspendresume agents. Liquid aerosols mainly served from cans under pressure.

For inhalation the compounds according to the present invention can be obtained from the apparatus for injection, spray or an aerosol package or served with other appropriate means of delivery of aerosols. Aerosol can be used corresponding volatile compounds, such as DICHLORODIFLUOROMETHANE, Trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. In the case of aerosol packaging, standard dose set by valve, which measures a certain amount of drugs.

Upon appointment by inhalation or insufflation, the compounds of the present invention can be prepared in the form of a dry powder composition, for example, a mixture of powders of the connection and the necessary bases such as lactose or starch. See, for example, gelatin or plastic bags, of which the powder can be entered using the inhaler or apparatus for injection.

If necessary, the above-described compositions can be adapted for batch allocation of the active ingredient over an extended period of time.

The pharmaceutical compositions according to the present invention can also contain other active ingredients such as antibacterial agents and preservatives.

Compounds of the present invention can also be used with other therapies, for example, with other anti-infective agents. In particular, the compounds of the present invention can be used in conjunction with known anti-virus tools.

Thus, another objective of the present invention is a complex composition comprising the compound (A) or its physiologically acceptable derivative together with another therapeutic agent, in particular an anti-virus tool.

These complex structures can be prepared, for convenience in the form of pharmaceutical compositions, thus, another aim of the invention are pharmaceutical compositions, sostoyatelnimi means for use in such complex compounds are acyclic nucleosides, such as acyclovir or ganciclovir, interferons such as alpha-, beta - or gamma-interferon, inhibitors of the renal secretions, such as probenecid, an inhibitor of the transport of nucleosides, such as dipyridamole, 2',3'-dideoxynucleoside, such as AZT, 2', 3'-dideoxycytidine, 2',3'-dideoxyadenosine, 2',3'-dideoxyinosine, 2',3'-dideoxythymidine, 2',3'-dideoxy-2',3'-didehydrothymidine and 2',3'-dideoxy-2',3'-didehydrothymidine, immunomodulators such as interleukin-2 (IL-2) and granulocyte-macrophage stimulating factor (GM-CSF), erythropoietin, ampligen, thymomodulin, thymopentin, foscarnet, ribavirin and inhibitors of the binding of HIV to CD4 receptors, in particular soluble CD4, CD4 fragments, CD4 hybrid molecules, glycosylation inhibitors such as 2-deoxy-D-glucose, castanospermine and 1-deoxynojirimycin.

Individual components of such complex compounds may be assigned sequentially or simultaneously in separate or joint pharmaceutical compositions.

In the case where the compound (A) or its pharmaceutically acceptable derivative is used in conjunction with a second therapeutic agent that is active against the same virus, the dose of each compound may b the scarfing specialist.

The compound (A) and its pharmaceutically acceptable derivative can be obtained by any method that is known for the synthesis of compounds with similar structure, as, for example, described in European patent publication 0382526 A2.

For professionals it is obvious that in some of the following methods necessary Strahinjica structure of the compound (A) can be obtained either by using optically active starting compound, or by cleavage of the racemic mixture on one of the stages of the synthesis. In all cases, the optically pure target product can be obtained by splitting the final product of each reaction.

In one such method 1,3-oxathiolan by the formula (VIII)


where anomeric group L is a leaving group, communicates with the appropriate base.

Suitable groups L are the group OR in which R denotes alkyl group, in particular (C1-C6)alkyl group such as methyl group, or R represents (C1-C6)acyl group, in particular acetyl group, or a halogen atom such as iodine, bromine or chlorine.

The compound according to formula (VIII) is readily interacts with 5-fertilizing or relevant PR is as hexamethyldisilazane) in a suitable solvent, such as methylene chloride, in the presence of a Lewis acid such as titanium tetrachloride, trimethylsilyltriflate, attributively or a compound of tin (V), for example, SnCl4.

1,3-oxathiolane by the formula (VIII) can, for example, be synthesized by the reaction of the aldehyde of formula (VII) with mercaptoacetate by the formula (VI) in a suitable organic solvent, such as toluene, in the presence of an acid catalyst, for example, a Lewis acid such as zinc chloride.



Mercaptoacetate by the formula (VI) can be obtained by known methods, for example, G. Hesse and I. Jorder, Chem. Ber., Bd. 85, pp. 924-932 (1952).

Aldehydes by the formula (VII) can be obtained by known methods, for example, E. G. Halloquist and H. Hibbert, Can. J. Research, V. 8, pp. 129-136 (1933). The crude aldehyde (VII) it is convenient to clean, turning it into a crystalline adduct with bisulfite and subsequent release of free aldehyde.

In the second method, the compound (A) is obtained by inter-conversion in the presence of a base compound according to formula (IX)


where B denotes a group which is transformed under the action of bases in the 5-fertilizin.

This inter-conversion can be the tion in cytosine), or enzymatic transformations using deoxytransferase. These methods and conditions interconversion well known in the chemistry of nucleosides.

In the third method according to the formula (XI)


can be converted to compound (A) of the anomeric conversion of the amino group in the 5-forcasino base by methods well known in the chemistry of nucleosides.

Many of these reactions in terms of synthesis of nucleosides represented in surveys, for example, Nucleoside Analogs-Chemistry, Biology and Medical Applikations, R. T. Walker et al., Eds., Plenum Press, N. Y. (1979), pp. 165-192 and T. Ueda, Chemistry of Nucleosides and Nucleotides, Vol. I, L. B. Towmnsend Ed., Plenum Press, N. Y. (1988) on pages 165-192, which are given here for reference.

It is obvious that the implementation of the above reactions may require the parent substance having a protected functional group, so that the intermediate or final stages for the separation of target compounds may require removal of the specified protection. Protect and unprotect functional groups can be performed using conventional methods. So, for example, amino groups can be protected with a group selected from aranceles group (particularly benzyl group), acyl group, aryl group is W ill result in the desired moment by hydrolysis or hydrogenolysis under standard conditions. Actigraphy you can protect any groups that are used for this purpose, as described, for example, in Protective Groups in Organic Chemistry, J. F. W. McOmie, Ed., Plenum Press, N. Y. (1973) or T. W. Green, Protected Groups in Organic Synthesis, John Wiley and Sons, N. Y. (1989).

Examples of suitable groups for protection actigraphy are groups selected from alkyl groups (for example, methyl group, tert-butilkoi group or methoxymethyl group), aranceles group (e.g. benzyl group, diphenylmethylene group or triphenylmethyl group), heterocyclic groups such as tetrahydropyranyl group, acyl groups (for example, acetyl group or benzoline group) and a silyl group such as trialkylsilyl group (e.g. tert-butyldimethylsilyl group). Protection actigraphy can be removed by conventional means. So, for example, alkyl group, acyl group and heterocyclic group can be removed by solvolysis, for example by hydrolysis in an acidic or alkaline environment. Kalkilya groups, such as triphenylmethyl group can also be removed by solvolysis, for example by hydrolysis in an acidic environment. Kalkilya groups such as benzyl group can be removed at a desired stage of the synthesis, for example, by treatment with efratom traktorist Tetra-p-butylammonium.

In the above methods, the compound (A) is usually obtained as a mixture of CIS - and TRANS-isomers, of which the compound of interest is a CIS-isomer.

These isomers can be divided by physical methods, e.g. by chromatography on silica gel or by fractional recrystallization directly or in the form of a suitable derivative, in particular, in the form of acetates (obtained, for example, with acetic anhydride), followed (after separation of isomers) reverse conversion to the desired product (in particular, by deacetylation of methanolic solution of ammonia).

Pharmaceutically acceptable salts of the compounds of the present invention can be obtained by the method described in U.S. Patent 4383114, which is reproduced here for reference. For example, when you want to get from the compound (A) salt accession acid, the product through one of the above methods is possible in the normal way to turn into a salt by processing the selected free base corresponding acid in the presence of, if necessary, a suitable solvent, such as an ester (e.g. ethyl acetate) or an alcohol (e.g. methanol, ethanol or ISO-propanol). Salts of inorganic onalmost, methanol). Pharmaceutically acceptable salts can also be obtained from other salts, including other pharmaceutically acceptable salts of the compounds (A), using conventional methods.

The compound (A) can be converted into pharmaceutically acceptable phosphates or other esters interaction with fosforiliruyusciye agents such as phosphorus oxychloride, or suitable tarifitsiruemye agents such as allalone or anhydride. Ester or salt of the compound (A) can, for example, to translate in the original compound by hydrolysis.

Cleavage of the final product, intermediate product or the ingredients, you can spend any known way: see, for example, E. L. Elies, Stereochemistry of Carbon Compounds, McGraw Hill (1962) and S. H. Wilen, Tables of Resolving Agents.

For example, the compound (A) can be distinguished using chiral liquid chromatography high resolution, using a suitable stationary phase, for example acetylated beta-cyclodextrin or cellulose triacetate and an appropriate solvent, e.g. an alcohol, such as ethanol, or an aqueous solution, for example, acetate of triethylamine. Connections can be split also using stimulated by enantioselective enzyme inhibition appropriate farmcreated. In that case, if the splitting spend enzyme, the enzyme may be contained in the solution or, more conveniently, be present with immobilized form. Immobilization of enzymes can be carried out by known methods, for example by applying a resin such as Eupergit S..

The invention is further illustrated by the following examples, which in no case do not limit the present invention. All temperatures are given in degrees Celsius.

The intermediate connection 1.


(i) 2-Benzoyloxymethyl-5-acetoxy-1,3-ostiole.

Benzyloxyacetaldehyde (216,33 g, 1,32 mol) dissolved in pyridine (373 ml, br4.61 mol) and the resulting solution was added 1,4-dition-2,5-diol (100,31 g, 9,66 mol). The heterogeneous mixture was stirred in nitrogen atmosphere at a temperature of 65oC for 1 hour. By this time, the dissolution is completed. To the reaction mixture was added dichloromethane (650 ml) and the mixture cooled to 0oC in the bath with ice. Dropwise at 0-5oC for 1.5-2 hours is added acetyl chloride (281 ml, of 3.95 mol). Stirred at 0-5oC for 30 minutes and gently pour the mixture into a cold (0oC) a saturated solution bitici unite, wash with saturated sodium bicarbonate solution (3 200 ml) and salt (200 ml), dried over sodium sulfate and evaporated in vacuum. The remains of the pyridine is removed by azeotropic distillation with benzene. Get 320,79 g crude product which is purified by distillation or filtration through a short column with silica gel (eluent - hexane/ethyl acetate, 3/1).

(ii) CIS - and TRANS-2-benzoyloxymethyl-5-(N'-acetyl-5'- fertilizin-1'-yl)-1,3-oxathiolan

5-ferritin (4,30 g, 33.3 mmol), hexamethyldisilazane (25 ml) and ammonium sulfate (120 mg) is refluxed until complete dissolution of cytosine (3 hours), and then boiled for 2 hours. Hexamethyldisilazane distilled off in vacuo and to the residue was added toluene (100 ml) was evaporated to solvents. The resulting solution of bis(trimethylsilyl)fortitudine in dichloromethane (40 ml) was added in an argon atmosphere to a solution of 2-benzoyloxymethyl-5-acetoxy-1,3-oxathiolane (8,537 g, 30.3 mmol) in dry dichloromethane (100 ml) containing molecular sieves (4A, 2 g), which are pre-incubated in an atmosphere of argon and cooled to 0oC for 20 minutes.

To the mixture was added at 0oC [(trifloromethyl) oxy]trimethylsilane (6 ml, 31 mmol) and the resulting solution was stirred PR is the thief of salt and once with distilled water. The organic layer is dried over magnesium sulfate, filtered and evaporated to dryness. Get a crude derivative of 5-fortitudine (10.1 g). Rf= 0,57 (EtOAc:MeOH, 9:1).

At the next stage, the product received without further purification will acetimidoyl. The crude product is dissolved in an atmosphere of argon in dichloromethane (120 ml) in a round bottom flask with a capacity of 500 ml. To the resulting solution was added triethylamine (12,7 ml, to 91.1 mol) and dimethylaminopyridine (111 mg, 0.9 mmol). The flask under argon was placed for 1 hour in a bath with ice. Into a cooled flask inject acetic anhydride (4.5 ml, 45 mmol), distilled over sodium acetate. Leave the mixture under stirring overnight, then carefully decanted into a flask of Erlenmeyer containing a saturated solution of sodium bicarbonate. The product is washed with distilled water and then a saturated solution of salt. Portions of a solution in methylene chloride, dried and evaporated to dryness in high vacuum, obtaining the acetylated mixture of alpha/beta isomers as a colourless foam, which weight after drying is 9.6, Thin-film evaporation chromatography of this substance, using as eluent ethyl acetate:methanol (9:1) obtain 3.1 g (7.8 mmol, 46%) of pure TRANS - and 3.5 g (8.9 mmol, 30%) of pure CIS-and the>ax: 309 nm

An NMR spectrum ( M. D. in CDCl3): 8,77 (1H, Shir., C'4)-N-Ac), of 8.06 (2H, mult. , arene. ), of 7.70 (1H, duplicates., C'6- JCF= 6.3 Hz), a 7.62 (1H, mult., arene.), 7,49 (2H, mult. arene.), 6,51 (1H, duplicates. doublets, C5- , 5,91 (1H, duplicates. doublets, C2- ), 4,48 (2H, duplicates. doublets, C2-OOCOC6H5), 3,66 (1H, duplicates. doublets, C4- , 3,34 (duplicates. doublets, C4- , of 2.56 (3H, sing., NH-COC ).

CIS-isomer: Rf= 0,58 (ethyl acetate:methanol, 9:1)

UV spectrum (MeOH) max: 309 nm

An NMR spectrum ( M. D. in CDCl3): 8,72 (1H, Shir., C'4N - Ac), of 8.06 (2H, mult. , arene. ), 7,87 (1H, duplicates. C'6-H, JCF= 6.3 Hz), 7,60 (1H, mult., arene.), 7,49 (2H, mult., arene.), 6,32 (1H, duplicates. doublets, C5- vs. 5.47 (1H, duplicates. doublets, C2- ), to 4.73 (2H, duplicates. doublets, C2-C OCOC6H5), 3,62 (1H, duplicates. doublets, C4- 3,19 (duplicates. doublets, C4is 2.55 (3H, single., NH-COCH3).

(iii) (+)-CIS-oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolan

1.2 g (3,05 mmol) of CIS-2-benzoyloxymethyl-5-(N'-acetyl-5'-fertilizin-1'-yl)- 1,3-oxathiolane stirred for 2 hours at 0oC in 30 ml of methanolic ammonia solution and allowed to mix overnight at room temperature. The mixture is evaporated under reduced pressure and the residue triturated twice (2 x 30 ml) with absolute ether. Hard mod is inane:

so PL 204 - 206oC; Rf= 0,21 (ethyl acetate:methanol, 9:1). The target compound was characterized using H and C-NMR spectroscopy and UV spectroscopy

UV-spectrum (in water):max= 280,9 nm

Range1H-NMR ( M. D. in DMSO-d6: by 8.22 (1H, duplicates., C'6-H, JCF= 7,2 Hz), to 7.84 (2H, duplicates. C'4)-N , 6,16 (1H, triple., C5- 5,43 (1H, triple., C2-CH2-O 5,19 (1H, triple., C2- of 3.77 (2H, mult., C2-C-OH), to 3.35 (1H, duplicates. doublets C4-

Range13C-NMR (DMSO-d6): 153,46 (C'6), 158,14 (C'2,2JCF= 14,0 Hz), 134,63 (C'4I , JCF= 24,1 Hz), 126,32 (C'5I , JCF= 32,5 Hz), 86,82 (C5), 86,80 (C4), 86,77 (C2), 62,32 (CH2OH).

Example 1. (-)-4-Amino-5-fluoro-1-(2-oxymethyl-1,3-oxathiolan-5-yl)-(1H)-pyrimidine-2-it.

(i) Monophosphate (+)-CIS-2-oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolane.

To a stirred mixture of Intermediate 1 (500 mg, 2,024 mmol) in dry trimethylphosphate (10 ml), cooled to 0oC, are added dropwise phosphorus oxychloride (1,22 ml of 13.1 mmol). The reaction mixture was stirred at the same temperature for one hour and the reaction is interrupted, adding ice to the ox. Bring the pH of the cooled mixture to 3 by addition of 1N aqueous sodium hydroxide solution, carry on to ammonia ratio (10:10:1). Fractions that contain monophosphate, is separated and evaporated, and then transferred to a column filled with 15 g of DEAE Sephadex A25 (form HCO3). Spend gradient elution with water (300 ml), 0.1 M solution of ammonium bicarbonate (300 ml) and 0.2 M solution of ammonium bicarbonate (100 ml). Evaporation of the appropriate fractions after dilution with water (30 ml) allows you to select monophosphate (+)-CIS-2-oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolane in the form of a colorless substance, Rf= 0,5 (n-D: NHOH, 6: 4), the output 612 g (1.77 mmol, 87.9 per cent).

Range1H-NMR ( M. D. in D2O): of 8.27 (1H, duplicates., C'6-H, JCf= 6,47 Hz), 7,33 (1H, duplicates. doublets, C5(H) vs. 5.47 (1H, triple., C2(H) 4,84 (2H, mult. C2- CH2-OH), 3,63 (1H, duplicates. doublets, C4(H) 3,30 (1H, duplicates. doublets, C4-H).

Purity according to liquid chromatography high resolution (IHVR) is not less than 99%.

(ii) (+)-CIS-2-Oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolan.

To a solution of monophosphate (+)-CIS-2-oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolane (100 mg, 0.29 mmol) in 3 ml of buffer solution on the basis of glycine [glycine (52,6 mg) and magnesium chloride (19 mg) in water (10 ml)] rapidly added 5'-nucleotidase (Sigma, 3.5 mg activity 29 u/mg). The mixture was incubated term the glycoprotein alpha-acids, eluent to 0.2 M solution of sodium phosphate with a pH of 7, the feed rate of 0.15 ml/min) through different periods of time. After 2.5 hours, the observed presence of only the (+)-enantiomer. Add more a certain amount (2 mg) enzyme and the mixture incubated for another 3 hours. Analysis according to GHUR clearly indicates selective and complete hydrolysis of (+)-enantiomer. The resulting mixture was transferred to a column filled with DEAE Sephadex A25 (form HCO). Spend gradient elution with water (155 ml), and then 0.1 M and 0.2 M solution of ammonium bicarbonate (100 ml each). The appropriate fractions containing the first suirvey nuke, combined and evaporated. The remaining solid is purified on a short column of silica gel, using as eluent a mixture of ethyl acetate and methanol (4,5:0,5), and then share with GHUR (in the same conditions as described earlier). Get pure (+)-CIS-2-oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolane (23 mg, 0,093 mmol, 32%) as a white solid.

[]2D1+123o(With, of 1.00, MeOH); so pl. 185oC.

Range1N-YN ( M. D. in DMSO): compared to 8.26 (1H, duplicates., C'6-H, JHFin 5,22 Hz), 7,87 (1H, sing., NH2D2O, currency), 7,63 (1H, single., NH2D2O, currency), of 6.20 (1H, DM is), 3,50 (1H, duplicates. doublets, C4(H) 3,37 (1H, duplicates. doublets, C4-H).

(iii) (-)-CIS-2-Oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolan. The appropriate fractions after column with Sephadex containing the second suirvey nuke, the receipt of which is described in stage (ii), are combined and evaporated under reduced pressure. The residue is dissolved in 2 ml of water and treated with alkalinebattery (Sigma, 1 ml of activity of 60 u/mg) followed by a temperature control for 1.5 hours at a temperature of 37oC. the Solvent is evaporated and the residue purified column chromatography on silica gel, elwira a mixture ethylacetate (4:1) followed GHUR (in the same conditions as mentioned previously). So pure (-)-CIS-2-oxymethyl-5-(5'-fertilizin-1'-yl)-1,3-oxathiolane (20 mg, of 0.081 mmol, 28%) with T. pl. 190oC (with decomp.): Rf= 0,21 (EtAc:MeOH, 4:1).

UV-spectrum (in water):max= 279,1 mm

Range1H-AMN ( M. D. in DMSO-d6): of 8.25 (1H, duplicates., C'6-H, JHF= 7,26 Hz), 7,88 (1H, Shir. WITH4'-NH2D2O, currency) a 7.85 (1H, Shir., C4'-NH2; D2O, currency), of 5.24 (1H, triple. C2(H) a 3.83 (2H, mult., C2C-OH), 3,19 (1H, duplicates. doublets, C4-H) and 3.15 (1H, duplicates. doublets, C4-H).

Intermediate compound 2 and Pimentel-5R-(5'-fertilizin-1"-yl)-1,3-oxathiolan-2S-carboxylate.

To a suspension of 5-fortitudine (155 mg, 1.2 mmol) in methylene chloride (1 ml) at room temperature in an argon atmosphere was added sequentially 2,4,6-kallidin (MX 0.317 ml, 2.4 mmol) and tert-butylmethylether-triftorbyenzola (0,551 ml, 2.4 mmol). The resulting mixture is stirred for 15 minutes to obtain a transparent solution. Add a solution of (1'R, 2'S, 5'R)-Menthyl-5R-acetoxy-1,3-oxathiolan-2S-carboxylate (330 mg, 1 mmol) in methylene chloride (0.5 ml), and then attributively (0,156 ml, 1.1 mmol). Stirring is continued for 3 hours. Diluted with a mixture of methylene chloride (20 ml) and washed sequentially with an aqueous solution of sodium bisulfite, water, saturated salt solution and evaporated. The residue is transferred into a mixture of ether-hexane (1: 1, 10 ml) and saturated sodium bicarbonate solution (2 ml) and stirred at room temperature for 15 minutes. The aqueous layer was separated and the organic layer is subjected to centrifugation and get a solid white color, which was washed with hexane (3x5 ml) and dried in vacuum. Thus obtained (+)-(1'R, 2'S, 5'R)-Menthyl-5R-(5"-fertilizin-1"-yl)-1,3-oxathiolan-29-carboxylate (350 mg, 88%) contains about 6% (1'R, 2'S, 5'R)-Menthyl-5S-(5"-fertilizin-1"-yl)-1,3-oxathiolan-2S-carboxylate (NMR). Received prophetic>/P>[]2D0+22o(With, at 0.19, MeOH); so pl. 216-218oC

Range1H-AMN ( M. D. in CDCl3): 0,78 (3H, duplicates., J = 7 Hz), of 0.91 (6H, triple. , J = 7,3 Hz), and 1.00 (2H, mult.), 1,39 - 2,04 (7H, mult.), of 3.12 (1H, duplicates. of doublets, J = 6,6 Hz, 6,1 Hz), 3,52 (1H, duplicates. of doublets, J = 4,7 Hz and 6.1 Hz), 4,79 (1H, duplicates. of doublets, J = 4,6 Hz, 4.3 Hz), 5,46 (1H, single.), of 5.75 (1H, Shir. single., currency), 6.42 per (1H, triple., J = 5.0 Hz), 8,10 (1H, Shir. single., currency), 8,48 (1H, duplicates., J = 6,6 Hz).

Example 2. 2S-oximeter-5R-(5'-fertilizin-1'-yl)-1,3-oxathiolan.

To a suspension of socialogical (10 mg, 0.54 mmol) in THF (1 ml) is slowly added in an argon atmosphere at room temperature a solution of (+) - (1'R, 2'S, 5'R)-Menthyl-5R-(5"-foreclosing-1"-yl)-1,3-oxathiolan-2S-carboxylate (54 mg, is 0.135 mmol) in THF (2 ml). Stirred the reaction mixture for 30 minutes and interrupt the reaction, adding an excess of methanol (2 ml), and then add silica gel (3 g). The resulting mass of purified column chromatography on silica gel (ethyl acetate-hexane-methanol, 1:1:1) and get retinoid solid, which was dried by azeotropic distillation with toluene and release of 20.7 mg (63%) of white solid.

[]2D0+114o(With, of 0.12, MeOH)

Range1H-NMR ( M. D. in DMSO-d6): 3,14 (1H, duplicates. of doublets, J = 4,3 , the Il. mult., currency), 7,83 (1H, Shir. mult., currency), 8,29 (1H, duplicates., J = 7,66 Hz).

Example 3. Biological activity.

(i) Antiviral activity.

Antiviral activity of the compounds of example 1 defined relative to HIV-1 in the following cell lines.

Cells S, T-lymphoblastoid cell line human infected with HIV-1 strain RF.

Cells MT-4 cell line T-leukemia cells infected with HIV-1 strain RF.

Antiviral activity in cells Is determined by the inhibition of the formation of society (Tochikura et al. , Virology, Vol. 164, 542-546), and in cells MT-4 inhibition of transformation of formazan [Baba et al., Biochem. Biophys. Res. Commun., Vol 65, pp. 128-134 (1987); Mossman, J. Immun. Meth. Vol. 65, pp. 55-57 (1983)]. Antiviral activity determined by analyzing the number of HIV antigens p24, synthesized in the presence and in the absence of enantiomers.

The results are presented in tables 1 and 2.

(ii) Cytoxicity

Cytoxicity compounds of example 1 and the racemic mixture (Intermediate compound 1) define two cell lines CD4: H9 and CEM.

The compounds serially diluted from the original concentration of 100 μg/ml to 0.3 μg/ml (article in the control cell, which do not contain medication. Thermostatic for 5 days at a temperature of 37oC and count the number of viable cells by separation of the cell suspension and count the number of cells, with the exception of trypanosome, hemocytometer.

The results are presented in table 3.

1. (-)-4-Amino-5-fluoro-1-(2-hydroxymethyl-1,3-oxathiolan-5-yl)-(1H)-pyrimidine-2-one or its pharmaceutically acceptable derivative.

2. A mixture of (-)-4-amino-5-fluoro-1-(2-hydroxymethyl-1,3-oxathiolan-5-yl)-(1H)-pyrimidine-2-it (+)-4-amino-5-fluoro-1-(2-hydroxymethyl-1,3-oxathiolan-5-yl)-(1H)-pyrimidine-2-it or their pharmaceutically acceptable derivatives, where (+)-enantiomer is present in an amount of not more than 5 wt.%.

3. The mixture according to p. 2, where (+)-enantiomer is present in an amount of not more than 2 wt.%.

4. The mixture according to p. 2, where (+)-enantiomer is present in an amount of not more than 1 wt.%.

5. A method of obtaining a connection on p. 1, characterized in that the spending allocation (-)-enantiomer from a mixture of (-)- and (+)-enantiomers.

6. The method of obtaining a mixture of enantiomers on PP.2 to 4, characterized in that the spending allocation (-)-enantiomer of the first mixture of (+)- and (-)-enantiomers, (+)- enantiomer is present if the is a racemic mixture.

8. The method according to PP.5 to 7, characterized in that the separation is carried out using chiral high-performance liquid chromatography (HPLC).

9. The method according to p. 8, characterized in that for HPLC as stationary phase using acetylated beta-dextrin or cellulose triacetate.

10. The method according to PP.5 to 7, characterized in that the separation is carried out using stimulated by enantioselective enzyme inhibition.

11. The method according to p. 10, characterized in that the enzyme is used in an immobilized form.

12. The method according to p. 10 or 11, characterized in that the enzyme is tsitidindeaminazy.

13. The method according to p. 10 or 11, characterized in that the enzyme is 5'-nucleotidase.

16. Connection on p. 1 having therapeutic activity.

17. The mixture according to any one of paragraphs.2 to 4, has a therapeutic activity.

18. Connection on p. 1 having antiviral activity.

19. The mixture according to any one of paragraphs.2 to 4, having antiviral activity.

20. Connection on p. 1 having activity against HIV.

21. The mixture according to any one of paragraphs.2 to 4, with activity against HIV.

22. A connection on p is Rotel infection of hepatitis C.

24. A method of treating a mammal, including man, suffering from or susceptible to a viral infection, wherein the mammal is administered an effective amount of the compounds according to paragraphs.1 to 4.


Same patents:

The invention relates to new substituted cyclic derivatives of 1,3-oxathiolane having pharmacological activity, to processes for their preparation and intermediate compounds used for their production, to containing these derivatives, pharmaceutical compositions and to the use of these derivatives for the treatment of viral diseases in mammals

The invention relates to medicine

The invention relates to a new method of treatment of patients, such as people with benign prostatic hyperplasia (BPH), which includes treatment by assigning a therapeutically effective amount of an inhibitor 5- reductase in combination with blocker1- adrenergic receptor
The invention relates to medicine, Nephrology

The invention relates to the class of pyrimidine compounds, which are suitable for the treatment of diseases and disorders of the Central nervous system (CNS), for example to prevent cerebral ischemic lesions, containing their pharmaceutical compositions and to methods for their preparation

The invention relates to 1,4-disubstituted the piperazines of General formula (I), which means the group-CO - or-CH2-OCO; D - heteroaryl selected from a range including 1, 3, 5-triazinyl, pyrimidinyl and pyridinyl, possibly substituted by one or two substituents selected from a range, including mono-(C1-C6)-alkylamino, mono-(C3-C7)- alkynylamino-, di-(C1-C6)-alkylamino-,

(C1-C6)-alkyl-(C3-C7)-alkylamino and pyrrolidin-I-yl group; Raand Rbis a hydrogen atom or (C1-C3)-alkyl; n is an integer from 1 to 4; their enantiomers, racemic mixtures and their salts with pharmaceutically acceptable acids and bases

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to an intermediate compound, i. e. tert.-butyl-(E)-(6-{2-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]-pyrimidine-5-yl}-(4R,6S)-2,2-dimethyl[1,3]dioxane-4-yl]acetate that can be used in synthesis of compound of the formula (IV)

eliciting inhibitory effect on activity of HMG-CoA-reductase and, therefore, can be used for preparing pharmaceutical agents for treatment, for example, hypercholesterolemia, hyperproteinemia and atherosclerosis. Also, invention relates to a method for preparing indicated intermediate compound by reaction of the new parent compound - diphenyl-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)amino]pyrimidine-5-ylmethyl]phosphine oxide with tert.-butyl-2-[(4R,6S)-6-formyl-2,2-dimethyl-1,3-dioxane-4-yl]acetate in the presence of a strong base in simple ether or aromatic solvents or their mixtures at temperature in the range from -200C to -900C. Also, invention relates to a method for preparing of compound of the formula (IV) wherein R1 means hydrogen atom or pharmaceutically acceptable cation and to a method for preparing intermediate compounds of the formula (VI):

wherein each P1 and P2 represents independently (C1-C4)-alkyl or group:

and wherein P3 represents (C1-C8)-alkyl. Applying new intermediate compounds and proposed methods provide enhancing quality and yield of compounds.

EFFECT: improved preparing methods.

9 cl, 1 tbl, 8 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of 5-phenylpyrimidine or their pharmaceutically acceptable acid-additive salts that elicit properties of antagonists of neuropeptide receptor neurokinin-1 (NK-1). This allows their applying for treatment of such diseases as Alzheimer's disease, cerebrospinal sclerosis, attenuating syndrome in morphine withdrawal, cardiovascular alterations and so on. Compounds of invention correspond to the general formula (I):

wherein R1 means hydrogen or halogen; R2 means hydrogen, halogen atom, (lower)-alkyl or (lower)-alkoxy-group; R3 means halogen atom, trifluoromethyl group, (lower)-alkoxy-group or (lower)-alkyl; R4/R4' mean independently hydrogen atom or (lower)-alkyl; R5 means (lower)-alkyl, (lower)-alkoxy-group, amino-group, hydroxyl group, hydroxy-(lower)-alkyl, -(CH2)n-piperazinyl substituted optionally with lower alkyl, -(CH)n-morpholinyl, -(CH2)n+1-imidazolyl, -O-(CH2)n+1-morpholinyl, -O-(CH2)n+1-piperidinyl, (lower)-alkylsulfanyl, (lower)-alkylsulfonyl, benzylamino-group, -NH-(CH2)n+1N(R4'')2, -(CH2)n-NH-(CH2)n+1N(R4'')2, -(CH2)n+1N(R4'')2 or -O-(CH2)n+1N(R4'')2 wherein R4'' means hydrogen atom or (lower)-alkyl; R6 means hydrogen atom; R2 and R6 or R1 and R6 in common with two ring carbon atoms can represent -CH=CH-CH=CH- under condition that n for R1 is 1; n means independently 0-2; X means -C(O)N(R4'')- or -N(R4'')C(O)-. Also, invention relates to a pharmaceutical composition.

EFFECT: valuable medicinal properties of compounds.

15 cl, 4 sch, 86 ex

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention describes a method for preparing 1,3-oxathiolan nucleosides or a method for preparing derivatives of 1,3-oxathiolanyl-5-one that involve effective methods for formation of 1,3-oxathiolan ring followed by condensation of 1,3-oxathiolan with pyrimidine or purine base. Using indicated methods these compounds can be synthesized as separate enantiomers with high selectivity.

EFFECT: improved preparing methods.

27 cl, 3 dwg, 16 ex

FIELD: medicine.

SUBSTANCE: method involves administering typical neuroleptics according to titration scheme and tricyclic antidepressants. Neuroleptics are applied according to titration scheme in the morning and tricyclic antidepressants are introduced as intravenous drop-by-drop infusion in the evening in combination with per os application of atypic neuroleptic risperidon. After having given 12-14 intravenous infusions, strategic supporting risperidon psychopharmacotherapy in combination with tricyclic antidepressants during 4-6 months.

EFFECT: enhanced effectiveness in overcoming pharmacological resistance; accelerated schizo-affective syndrome relief.

FIELD: medicine.

SUBSTANCE: method involves applying eradicative anti-helicobacterial therapy comprising Omeprazol administration at a dose of 20 mg twice a day and Ximedone at a dose of 500 mg twice a day in 12 days long course.

EFFECT: enhanced effectiveness of eradication; reduced adverse side effects risk.

FIELD: organic chemistry, medicine, pharmacy, pharmacotherapy.

SUBSTANCE: method involves administration in mammal the effective dose of 6-hydroxy-8-[4[4-(2-pyrimidinyl)piperazinyl]butyl]-8-azaspiro[4,5]-7,9-dione or its pharmaceutically acceptable salt of acid addition or its hydrate. Method expands arsenal of medicinal agents used for suppression of fear sensation.

EFFECT: valuable properties of agent.

3 tbl, 6 dwg, 4 ex

FIELD: medicine, narcology.

SUBSTANCE: invention relates to hepatoprotective and anti-encephalopathic agent used for reducing alcoholic intoxication. Invention comprises components based on succinic, fumaric, glutamic acids and, additionally, at least one vitamin of B group. Agent can comprise additionally vegetable extracts or their mixture, L-carnitine, glycine, L-arginine, taurine and/or their mixture, methylsulfonylmethane, dihydroquercitin, dimethylsulfoxide or their mixture, nicotinamide or nicotinic acid or their mixture, energy source and sweetening agent. Also, invention proposes a method for reducing alcoholic intoxication, prophylaxis and removing withdrawal syndrome, liver protection, among them, in non-alcoholic intoxication and protection against encephalopathy. Invention provides described effects without qualifying medical control.

EFFECT: valuable medicinal properties of agent.

16 cl, 3 tbl