DD255351A5 - PROCESS FOR THE PREPARATION OF CARBOCYCLIC PURE NUCLEOSIDES - Google Patents

Abstract

The present invention relates to a process for preparing a compound of the formula in which R is an optionally protected hydroxyl group and Y is an optionally protected purine base and salts thereof. The new compound can be used as an antiviral agent. formula

Description

Field of application of the invention

The present invention relates to nucleoside analogs having cyclopentane rings which can be used as substituents for purine nucleosides in the fields of biology, medicine or genetic manipulation and as antiviral agents.

Characteristic of the known state of the art

Derivatives of the compounds of the formula

wherein Y is guanin-9-yl or adenin-9-yl are examples of dideoxyan analogues to purine nucleosides used for the determination of base sequences in DNA [Proc. Nat. Acad. Be. USA, 74, 4563 (1977)]. However, 2 ', 3'-dideoxy analogues to purine nucleosides are so susceptible to acids that cleavage is very likely to occur in glycosyl formation, which is a great difficulty in their production.

Recently, it has been reported that 2 ', 3'-dideoxy analogs can act as purine nucleosides as inhibitors of virus origin reverse transcriptase, therefore these analogues have attracted attention as therapeutics in the treatment of diseases caused by the RNA virus [Chemical and Engineering News, January 27, No. 28 (1986)]. Although, as previously mentioned, dideoxynucleosides and their carbocyclic analogues have been studied to some extent, many issues remain to be elucidated. Therefore, it is important to prepare and evaluate various analogs.

Object of the invention

The aim of the invention is to provide new 2 ', 3'-dideoxycarbocyclic nucleosides which can be used as antiviral agents or for other purposes.

Explanation of the essence of the invention

The present invention is the result of the investigations made by the authors under the circumstances set forth above to obtain novel and useful purine nucleoside analogs. The invention therefore relates to a process for the preparation of the compounds of the formula I ₁

R 6 · 5'Y

• 2 · """ O '

wherein R represents an optionally protected hydroxyl group and Y represents an optionally protected purine base, and the salts thereof, which consists in that a compound of formula (II).

6'5 ¹ Y

1'

wherein R is an optionally protected hydroxyl group, either Ri and R _{2 is} a hydroxyl group, the other being hydrogen, and Y is an optionally protected purine base subjected to the reduction reaction of the 2'- or 3'-hydroxyl group.

The protective groups for the hydroxyl groups in the compounds of formula (I) or (II) are not particularly limited if they are those used as protective groups in nucleoside chemistry. According to the invention, protective groups which are relatively stable under alkaline conditions are preferably used, such as alkylsilyl groups having 3 to 10C atoms (eg tert-butyldimethylsilyl). Alkyl- or alkoxy-cyclic ethers having 4 to 10 C atoms (eg tetrahydrofuranyl and its derivatives having 4 to 7 C atoms, tetrahydropyranyl and its derivatives having 5 to 8 C atoms such as methoxytetrahydropyronyl), alkoxyalkyl groups having 3 to 10 C atoms (eg When the protecting group is an acyl group, the hydroxyl group may be in the form of an aliphatic acid ester (eg, a straight or branched one having 1 to 10 carbon atoms) or an arylcarboxylic acid ester (eg, methoxyethyl, methoxyethyl, and triethyl) and alkoxy-substituted trityl (eg, monomethoxytrityl, dimethoxytrityl) such with 5 to 30C atoms) are protected.

The purine bases, identified with the reference symbol Y, comprise various purine ring skeletal bases commonly used in the field of nucleic acid chemistry. Examples of such bases are adenine, hypoxanthine, guanine, isoguanine, xanthine, 3-deazaadenine, 7-deazaadenine, 8-azaadenine and 2,6-diaminopurine; they are linked via the nitrogen atom in the 9-position in the purine ring of the compound of the formula (I) or (II).

The protecting group of purine base is a compound of formula (I) or (II), d. That is, the amino-protecting group at the 2- or 6-position is any one common in the art of nucleoside chemistry. So z. For example, an arylcarboxylic acid residue (having 5 to 30C atomic acid) such as benzoyl as a protecting group for adenine and an aliphatic carboxylic acid residue (straight-chain or branched having 2 to TQC atoms) are preferably used as the guanine protecting group. ^

For the preparation of a compound of formula (I) from a compound of formula (II), the hydroxyl group in 2'- or 3'-position in a compound of formula (II) at 0 to 80 ⁰ C or preferably at room temperature thiocarbonyliert, followed by reduction mitTributylzinnchlorid in the presence of an equivalent amount or of an excess of a, a-azobisisobutyronitrile at 0 to 100 ⁰ C for 30 minutes to 2 hours, to obtain a 2 ', 3-dideoxy derivative of the formula (I). The thiocarbonylation can be conveniently carried out by using thiocarbonyldiimidazole for the thiocarbonylation and phenylchlorothiocarbonate for the phenoxythiocarbonylation. Further, S-methyldithiocarbonylation can also be carried out by means of a mixture of carbon disulfite and methyl iodide. After this reduction, the 6'-hydroxyl protecting group, e.g. For example, the 4,4'-dimethoxytrityl, in a simple manner under acidic conditions (eg., Treatment with acetic acid or 1 N hydrochloric acid at room temperature), the protective group of purine base can also under alkaline conditions (eg with concentrated ammonia water, 1 N Sodium hydroxide, 1N sodium ethylate).

The compounds of formula (II) may, for. The compounds of formula (II) wherein Y is an optionally protected adenin-9-yl can also be prepared according to the Japanese Patent Application Laid-Open No. 62992/1975, Chemical & Parmaceutical Bulletin 24,2624 (1976) or Nucleic Acids Symposium Series IMr. 16, 141 (1985). So ζ. For example, according to the process described in Japanese Patent Application Laid-Open No. 62992/1975 or Chemical & Pharmaceutical Bulletin 24,2624 (1976), a compound wherein Y is adenin-9-yl in formula (II) is either R ₁ or R _{2 is} a hydroxyl group and the other group is hydrogen and R is a hydroxyl group, using aristeromycin as the starting compound; a compound in which Y in the formula (II) is N ⁶ -benzoyladenin-9-yl, R is a hydroxyl group protected with 4,4'-dimethoxytrityl, and Ri is hydrogen and R _{2 is} a hydroxyl group, may be prepared according to the method described in of the above-mentioned Nucleic Acids Symposium Series. A compound in which Y in formula (II) represents optionally protected guanin-9-yl or hypoxanthine-9-yl, R is an optionally protected hydroxyl group and the hydrogens are in the 2'-position and a hydroxyl group in the 3'-position may be prepared according to the method described in Japanese Patent Application No. 238,858 / 1985 and the corresponding US patent application Ser. No. 920,116 (see Reference Examples 1 to 8).

On the other hand, the compound of the formula (II) in which Y is 2,6-diaminopurine-9-yl, R _{1 is} hydrogen and R _{2 is} a hydroxyl group can be prepared as follows: The hydroxyl group of the compound wherein Y is adenine Protected by N'-oxidation with hydrogen peroxide or metachloroperbenzoic acid, then the amino group at 6-position is deaminated with nitrous acid and heated with phosphorus oxychloride (Japanese Patent Publication 4347/1967) to give the corresponding 2,6- To obtain dichloropurine-9-yl derivative. The 6-position chlorine is substituted by an amino group, and the deamination is carried out using sodium nitrite in aqueous acetic acid to obtain the 2-chloro-6-hydroxyl-9-yl product. The compound thus obtained is subjected to amidation in the 2-position, and after chlorination in the 6-position, the chlorine in this position is substituted by an amino group to obtain the desired compound.

The salts of the compounds of formula (I) according to the present invention include those having the amino group in the purine base and a mineral acid (eg hydrochloric acid, sulfuric acid, nitric acid), an organocarboxylic acid (eg acetic acid, lactic acid, tartaric acid, maleic acid, succinic acid ) or an organosulfonic acid (eg, methanesulfonic acid, etan-sulfonic acid, benzenesulfonic acid).

The compounds of formula (I) according to the invention are useful tools in cloning genes. That is, the analogs derived from the cyclopentane ring compounds of this invention are the carbocyclic analogues of Purrn 2 ', 3'-dideoxynucleotide and can be readily prepared because they have no glycosyl linkage and that the triphosphate derivatives thereof can be used as a means of disrupting DNA chain extension in the determination of DNA sequences.

On the other hand, the compounds of the formula (1) have antiviral activity against DNA viruses or RNA viruses. Examples of DNA viruses are the group of herpesviruses (eg herpes simplex type I or II, cytomegalovirus, Epstein-Barr virus), adenovirus (for example type III), hepatitis B virus or smallpox or Pox virus; as RNA virus as human immunodeficiency virus (HIV), which is a pathogen of acquired immunodeficiency syndrome (AIDS), vesicular stomatitis virus ,. Feline leukemia virus and infectious anemia in horses causing virus. The compounds of the invention have particularly potent antiviral activities against RNA viruses, in particular the HTLV-III (human T-cell lymphotropic virus, type III, HIV-HIV-III which is in one of HIV, as possible inhibitors of reverse transcriptase; ,

The compounds of the invention can therefore be used for the treatment of various viral infections. All such viral infections are acquired immunodeficiency syndrome, herpes simplex type I or II, varicella, zoster, keratitis, conjunctivitis or acute hepatitis; various opportunistic infections, malignant tumors or central nervous system symptoms caused by viral infections and immunodeficiency. The compounds of formula I and their pharmaceutically acceptable salts of the present invention may also be used as antiviral agents for the treatment of viral diseases in animals, particularly mammals (experimental animals such as rabbits, rats and mice, domestic animals such as dogs and cats, cattle such as cattle , Horses, sheep and pigs.).

In general, for the above-mentioned purposes, a suitable effective dose of the compounds of the present invention is within a range of 30 to 500 mg / kg of body weight per day, preferably within a range of TQO to 300 mg / kg of body weight per day. The desired dose is generally divided into two, three or four subsets

administered at appropriate intervals throughout the day. Administration may be by any suitable means including oral, rectal, nasal, topical (e.g., buccal, sublingual), vaginal and parenteral (e.g., subcutaneous, intramuscular, intravenous, intradermal). The preferred mode of administration may, for. B. vary with the condition and age of the recipient. Although the compounds of this invention may be administered generally, they are preferably administered as components of a pharmaceutical formulation. The formulations according to the invention comprise at least one of the compounds of the formula (I) together with one or more compatible carriers for the same and optionally further therapeutic ingredients.

The formulation may conveniently be administered in unit dosage form. Formulations containing the compound of the invention for peroral administration include single units such as capsules or tablets; Powder or granules; Solutions or suspensions; or a liquid oil-in-water emulsion, liquid water-in-oil emulsion or the like. on.

Tablets may be made by densification or molding, optionally with one or more additional ingredients (-zien). Compressed tablets may be prepared by compressing the compound of the invention in the form of a powder or granules in a suitable machine, optionally mixed with a binder (e.g.

Hydroxypropylcellulose), lubricants (e.g., magnesium stearate), inert diluents (e.g., starch), preservatives, surfactants, or dispersants.

Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers or bacteriostats; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents.

The formulations can be used in single-dose or multi-dose containers such. B. sealed ampoules and vials distributed and stored frozen; immediately prior to use, the sterile liquid carrier, e.g. As water for injections added. Improvised injection solutions and suspensions can be prepared from sterile powders, granules and tablets.

The formulations for topical administration include lozenges containing the compounds of the invention in a flavored base such as sucrose and gum arabic or tragacanth; Lozenges containing the compound of the invention in an inert base such as gelatin and glycerine or sucrose and gum arabic; and mouthwashes containing the ingredient to be administered in a suitable liquid vehicle.

Formulations for rectal administration may take the form of suppositories having a suitable base, such as cocoa butter.

Formulations for vaginal administration may be in the form of pessaries, tampons, creams, gels, pastes, foams or spray formulations containing, in addition to the compound of the invention, art-recognized conventional carriers.

Among the compounds of the formula (I), 2 ', 3'-dideoxyaristeromycin (the compound of Example 3) and 9 - [(1S, 4R) -4-hydroxymethylcyclopentan-1-yl] guanine (the compound of Example 4) especially the growth of the AIDS virus and are therefore particularly useful compounds.

The invention will be explained in more detail with reference to the following reference examples, examples and experimental examples.

Reference Example 1:

Preparation of 9 - [(1R, 5R, R) RM-Methyl-Hydroxyl-S] -tetraisopropyldisiloxanioxycyclopentan-i-yl-hypoxanthine The C-analog to inosine (10g, 37.5 mmol) was dissolved in 200 mL anhydrous DMF, then 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane (13 mL, 41 mmol) and imidazole (11.3 g, 165 mmol) were added and the mixture was stirred at room temperature for 2.5 hours Water was added dropwise, and the precipitate was filtered off, washed with water and then rapidly with diethyl ether and dried to give the title compound as a white powder (17.2 g) A portion of the product was recrystallized from dichloromethane to give crystals to obtain a mp of 135 to 138 ⁰ Czu

 Note: The C analog to inosine is known from Chemical & Pharmaceutical Bulletin 24,2624 (1976).

Reference Example 2:

Preparation of 9 - [(1R, 2S, 2R, 4R) -4-methyl-2-phenoxythiocarbonyloxy-3,6- (tetraisopropyldisiloxanyl) -dioxycyclopentan-1-yl] hypoxanthine

The compound (11.2 g, 22.3 mmol) obtained in Reference Example 1 was dissolved in 300 ml of anhydrous acetonitrile, then dimethylaminopyridine (15.8 g, 53.5 mmol) and phenoxythiocarbonyl chloride (5 g, 29 mmol) were added, and the mixture was added Room temperature for 7 hours. The solvent was removed under reduced pressure and the residue was dissolved in 250 ml of chloroform. The solution was washed with 0.5M potassium dihydrogen phosphate solution (250ml χ 2) and then with water (200ml), dried (anhydrous sodium sulfate) and concentrated at reduced pressure to give a yellow syrup. This was washed by silica gel chromatography (90 g, solvent: CHCl ₃ and CHCl ₃ / MeOH = 60/1) to obtain the title compound as a light yellow glassy substance (13.0 g).

NMR (60MHz, CDCl ₃ ) δ ppm: 1.0-1.23 (28H, m), 2.13-2.43 (3H, m, H4 ', H5'), 3.93-4.10 (2H, m, H ₆ '), 4, 80-5.20 (2H, m / H, ', H ₃ '), 6.00-6.20 (H, m, H ₂ '), 7.03-7.50 (5H, m), 7.87 (TH , s), 8.13 (1 H, s) s; Singlet, m; Multiple "

Reference Example 3:

Preparation of 9 - [(1R) -SRM-Methyl-Se-iteraisopropyldisiloxanyOdioxycyclopentan-i-yl-hypoxanthine Anhydrous toluene (30 ml) was added to the compound (13.0 g, 2OmMoI) obtained in Reference Example 2, followed by concentration under reduced pressure pressure. the residue was dissolved in 300 ml of anhydrous toluene, and the solution was for 20 minutes introduced nitrogen gas. After addition of tributyltin hydride (11 ml, 4OmMoI), the solution was heated to 80 ⁰ C, and crystalline α, α'-azobisisobutyronitrile (AIBN) (820 mg) was added in four aliquots at 15 minute intervals, and after stirring for 3 hours with stirring, the solvent was removed under reduced pressure, the resulting oil was purified by silica gel chromatography (80 g, solvent: CHCl ₃ and CHCl ₃ /). MeOH = 60/1 to 30/1) to obtain the title compound as a colorless glassy substance (10.4 g).

Recrystallization of a portion of the product from ethanol gave colorless needles with a mp. from 200 to 202 ° C.

NMR (60 MHz), CDCl ₃ ) δ ppm: 0.93-1.20 (28H, s), 1.97-2.53 (5H, m, H ₂ ', H ₄ ', H ₅ ' ), 3.80-4.07 (2H, m, H ₆ '), 4.43-5.27 (2H, m, H ₁ ', H ₃ '), 7.87 (1H, s ), 8.20 (1 H, s)

Reference Example 4:

Preparation of S-lilR.SS ^ Rl ^ -iMonomethoxytrityloxyl-methyl-S-hydroxylcyclopentan-1-yl-1-yl-methoxymethyl-hypoxanthine) The compound (9.8 g, 19.8 mmol) obtained in Reference Example 3 was dissolved in 240 ml of anhydrous dioxane, then was added rapidly to the solution of sodium hydride (880 g, 21.8 mmol) under ice-cooling and stirring, then the mixture was stirred at room temperature for 1.5 hours. Then, methoxymethyl chloride (2 ml, 21.8 mmol) was rapidly added to the mixture under ice-cooling. The whole mixture was stirred at room temperature for 3 hours.

The solvent was removed under reduced pressure, and the oily residue was dissolved in 200 ml of chloroform. The solution was washed with 0.1 M triethylammonium bicarbonate (TEAB) buffer (pH 7.5, 100 ml x 2) and further with water (200 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a syrup. Purification of the syrup by CiaSilica gel chromatography (0 5.3 x 7.0 cm, solvent: acetone-water, 55% to 80%) gave a colorless glassy compound (8.5 g).

This compound (8.0 g) was dissolved in 32 ml of tetrahydrofuran (THF), then tetrabutylammonium fluoride trihydrate (TBAF · 3H ₂ O) (10 g) was added, and the mixture was stirred at room temperature for 0.5 hour. The solvent was removed under reduced pressure, and the oily residue was dissolved in 100 ml of water. The solution was washed with diethyl ether (100χ2), the tetrabutylammonium salt was removed by treatment with Dowex-50 resin (pyridine form, 60 ml). The solution and wash (240 ml) were combined and concentrated, and the concentrate was azeotroped with three portions of pyridine. The residue was dissolved in 100 ml of pyridine and added with monomethoxytrityl chloride (MMTrCl) (5.4 g), and the mixture was stirred at 37 ° C for 4 hours. The solvent was removed under reduced pressure and the oily residue was partitioned between 0.1 M TEAB buffer (50 ml) and CHCl ₃ (100 ml), the organic layer was washed with water (100 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the concentrate was subjected to dehydration by azeotropic distillation with toluene to obtain a colorless syrup. The 0.1 M TEAB buffer fraction and the washing were combined and concentrated to give the non-monomethoxytritylated compound. The compound was purified on HP-20 resin (190 ml, solvent: water and 30% ethanol-water) and monomethoxytritylated after concentration and azeotropic distillation with pyridine as described above. Both yields of the title compound thus obtained were combined and purified by silica gel chromatography (80g solvent: CHCl ₃ / MeOH = 100 / 1.60 / 1.50 / 1) to give a colorless, glassy product (6.1). to obtain. A solution of a portion of this product in dichloromethane, when added dropwise to η-hexane, gave a white powder.

NMR (60MHz, CDCl ₃ ) δ ppm: 1.87-2.70 (5H, m, H ₂ ', H ₄ ', H ₆ '), 3.20-3.40 (2H, m, H ₆ ' ), 3.43 (3H, s, CH ₃ OCH _2), 3.80 (3H, s), 4.30 to 4.57 (1 H, m, H ₃ '), 4.87 to 5.10 (1H, m, H ₁ '), 5.47 (2H, s, CH ₃ OCH ₂ -N), 6.73-6.97 (2H, m), 7.17-7.53 (12H, m), 7.73 (1H, s), 7.98 (1H, s)

Reference Example 5:

Preparation of 1 - [(1R, 3S, 4R) -4- (monomethoxytrityloxy) methyl-3-hydroxycyclopentan-1-yl] - (4-carbamoyl-5-aminoimidazole)

The compound obtained in Reference Example 4 (6.1 g, 10.7 mmol) was dissolved in 490 ml of ethanol, then a heated 5M sodium hydroxide solution (130 ml) was added rapidly under reflux. Reflux was continued for a further 40 minutes. Then the solvent was removed under reduced pressure. The oily residue was dissolved in 200 mL chloroform and washed with water (100 mL χ 2), then 0.1 M TEAB buffer (100 mL χ 2) and further with a saturated aqueous sodium chloride solution (100 mL), dried over anhydrous sodium sulfate and concentrated Concentrated to obtain a syrup. Purification of this syrup by silica gel chromatography (90g, solvent: CHCl ₃ / MeOH = 100/1 to 20/1) gave a colorless glassy product (3.2g). A solution of a portion of this product in chloroform gave dropwise to n-pentane with stirring a white powder.

Elerrientaranalyse (%) for C ₃₀ H ₃₂ N ₄ O ₄ · 0.5H ₂ O, molecular weight 521,616

Calculated: C, 69.08; H, 6.38; N, 10,74

found: C, 69.14; H, 6.09; N, 10.54

NMR (100 MHz, CDCl ₃ ) δ ppm: 1.36-2.52 (5H, m), 3.00-3.40 (3H, m, H ₆ ', OH), 3.77 (3 H, s), 4,12-4,60 (2H, m, H-, ', H ₃ '), 4,80-5,28 (2H, bs, NH ₂ ), 5,64-6, 44 (2H, bs, NH ₂ ), 5.64-6.44 (2H, bs, NH ₂ ), 6.76-6.94 (3H, m), 7.14-7.48 (12H, m ) bs; wide singlet

Reference Example 6:

Preparation of 1 - [(1R 1 -S) RM-iMonomethoxytrityloxy-methyl-S-hydroxycyclopentanilo-K-carbamoyl-6-iN-benzoyl-S-methylisothiocarbamoyoaminoimidazole]

The compound (0.88 g, 1.7 mmol) obtained in Reference Example 5 was dissolved in 25 ml of anhydrous acetone, then a solution of benzoylisothiocyanate (260 μL, 1.9 mmol) in acetone (8 ml) was added dropwise in 10 minutes while refluxing. followed by 50 minutes of refluxing. The solvent was distilled off under reduced pressure, and the resulting light yellow glassy substance was purified by silica gel chromatography (15 g, solvent: CHCl ₃ / MeOH = 50/1 to 30/1) to obtain a light yellow glassy compound (0.87 g) receive. A small amount of acetone was added to this compound (0.84 g, 1.2 mmol) and the resulting syrup was converted to a homogeneous solution by adding 12.5 ml of 0.2N NaOH and sonication. Then dimethylsulfate (130μΙ, 1.4 mmol) was added with stirring and then vigorously stirred at room temperature for 1 hour. The mixture was mixed for distribution with CHCl ₃ (15 mL χ 2), the organic layer was washed with 0.1 M TEAB buffer (15 mL χ3) and then with saturated aqueous sodium chloride solution (20 mL) over anhydrous sodium sulfate dried and concentrated at reduced pressure; the residue was purified by silica gel chromatography (15 g, solvent: CHCl ₃ / MeOH = 100/1 to 60/1). To the obtained glassy substance was added a small amount of dichloromethane, the mixture was added dropwise to hexane, and the resulting precipitate was collected by centrifugation and dried to obtain the title compound as a powder (400 mg).

Elemental analysis (%) for C ₃₉ H ₃₉ N ₅ O ₅ S, molecular weight 689,835

calculated: C, 67.90; H, 5.70; N, 10.15;

Found: C, 67.45; H, 5.45; N, 9.89;

NMR (10OMHz, CDCl ₃ ) δ ppm: 1.34-2.60 (5H, m), 2.52 (3H, s, SCH ₃ ), 3.04-3.44 (2H, m, H ₆ ' ), 3.79 (3H, s, OCH _3), 4.08 to 4.44 (1 H, m, H ₃ '), 4.60-5.00 (1H, m, H _1'), 5.64 (1H, bs, NH ₂ ), 6.72-6.94 (3H, m), 7.12-7.52 (15H, m), 7.80-7.96 (2H, m ), 11.35 (1H, bs, NH)

Reference Example 7:

Preparation of 9 - [(1R) -SRJ 1 -Monomethoxytrityloxymethyl-S-hydroxycyclopentane-i-ynguanine The compound (360 mg, 0.53 mmol) obtained in Reference Example 6 was added to heated 6N sodium hydroxide solution (18 ml), and Mixture was refluxed for 1 hour The product was extracted from the reaction mixture by CHCl ₃ , and the extract was washed with 0.1 M TEAB buffer (30 ml) and then with saturated aqueous sodium chloride solution (30 ml) over anhydrous Dried over sodium sulfate and subjected to silica gel chromatography (8 g;

CHCl ₃ / MeOH = 40/1 to 6/1). The obtained glassy substance was added with a small amount of acetone, the mixture was dropwise added to benzene, and the resulting precipitate was collected by centrifugation and dried to obtain the title compound as a powder (210 mg).

Elemental analysis (%) for C ₃ iH ₃₁ N ₅ O ₄ 1 H ₂ O, molecular weight 555,633

Calculated: C, 67.01; H, 5.99; N, 12,60

found: C, 67.01; H, 5.69; N, 12,42

NMR (100MHz, DMSO-d ₆ ) δ ppm: 1.50-2.60 (5H, m).

3.01 (2H, bs), 3.98-4.20 (1H, m), 4.70-4.96 (2H, m), 6.37 (2H, bs, NH ₂ ), 6, 82-7.46 (14H, m), 7.68 (1H, s, H ₈ ), 10.60 (1H, bs, NH)

Reference Example 8:

Preparation of 9 - [(1R, 3S, 4R) -4-hydroxymethyl-3-hydroxycyclopentan-1-yl] guanine The compound (180 mg, 0.33 mmol) obtained in Reference Example 7 was dissolved in 10 ml of 80% acetic acid, and the solution was stirred at 40 ° C for 4.5 hours. The solvent was removed under reduced pressure, then the azeotropic distillation with water was carried out twice. Then 10 ml of water were added, the mixture was washed with ether (10 ml χ 2) and the water was removed under reduced pressure. In this way the named compound was obtained as colorless crystals (41 mg), mp. 246 to 248 ° C. [a] g ⁵ = + 7.7 ° (C = 0.5, DMF), ^ (nm): (H ₂ O); 255,278 (sh)

(H "); 257,282

(OH "); 256 (sh), 273

Elemental analysis (%) for CnH ₁₆ N ₅ O ₃ .0.5 H ₂ O. 0.1 C ₂ H ₆ OH, molecular weight 278.886 calculated: C, 48.24; H, 6.00; N, 25.11 found: C, 48.61; H, 6.41; N, 25.40

Example 1:

N ⁶ -benzene-6'-O- (4,4'-dimethoxytrityl) -3'-0 - [(imidazol-1-yl) thiocarbonyl] -2'-deoxyaristeromycin *

N ⁶ -benzoyl-6'-O- (4,4'-dirnethoxytrityl) -2'-deoxyaristeromycin (2.5 g) was dissolved in 10 mL of dry dichloromethane, then 8.0 g of thiocarbonyldiimidazole was added and allowed to stand for 20 hours stirred at room temperature. The reaction mixture was concentrated to dryness and subjected to purification by silica gel chromatography (Kieselgel 60, Merck Co., 50 g, solvent, ethyl acetate) to obtain a pale yellow glassy substance (yield 2.2 g). NMR (90MHz, CDCl ₃ ) 6ppm: 3.80 (6H, S, 2CH ₃ O-), 8.35 (1H, S, H ₈ ), 8.76 (1H, S, H ₂ ).

Example Z:

N ⁶ -Benzoyl-6'-O- (4,4'-dimethoxytrityl) -2 ', 3'-dideoxyaristeromycin

The 3'-thiocarbonyl derivative (2.0 g) obtained in Example 1 was dissolved in 20 ml of dry dioxane, then a solution of tributyltin hydride (4.5 g) in 10 ml of dry dioxane was added dropwise while refluxing. In the meantime, 500 mg of crystals of α, α'-azobisisobutyronitrile were added in small portions. The dropwise addition was complete in 20 minutes and refluxing was continued for a further 2 hours. The solvent was evaporated under reduced pressure, and the oily residue was purified by silica gel chromatography (40 g) (solvent: CHCl ₃ ) to obtain a colorless powder (1.1 g).

NMR (90MHz, CDCl ₃ ) ppm: 3.80 (6H, S, 2CH ₃ O-), 4.80-5.20 (1H, m, H ₁ ), 3.15 (2H, d, 2H ₆ ), 8.76 (1H, S, H ₂ ), 9.10 (1H, S, -NhJ ₁ -C-).

= 0

Example 3:

2 ', 3'-Didesoxyaristeromycin

The compound (1.0 g) obtained in Example 2 was dissolved in a small amount of pyridine and then added with 50 ml of concentrated ammonia water and heated in a pressure-resistant flask at 60 ^° C. for 5 hours. The reaction mixture was concentrated wurdezur dryness and treated with 100 ml of 80% acetic acid and heated to 60 ⁰ C for 2 hours, followed by concentration to dryness under reduced pressure. The residue was dissolved in 100 ml of water and washed twice with water. The aqueous layer was concentrated to dryness, and the residue was made into a powder in ether to obtain 0.23 g of 2 ', 3'-dideoxyaristeromycin.

Elemental analysis (%) for Ci ₁ H ₁₅ N ₅ O.H ₂ O molecular weight 251.29

Calculated: C, 52.57; H, 6.82; N, 27.87

found: C, 52.83; H, 6.95; N, 27.54

The thus-obtained 2 ', 3'-dideoxyaristeromycin was dissolved in the equivalent amount by weight of 1N HCl. After concentration

the solution was treated with ethanol and concentrated to dryness. This process was repeated several times. This

The process was repeated several times. The residue was recrystallized from hot ethanol to obtain the crystals of the hydrochloric acid salt. Mp 173 to 175 ° C.

Elemental analysis (%) for C n H ₁₅ N ₅ O · 1 / 2H ₂ O molecular weight 278.73 Calc: C, 47.40; H, 6,15; N, 25.12; Cl, 12.72 found: C, 47.98; H, 6.06; N, 24.87; Cl, 12.71 [a] g ⁶ = -6.79 (C = 0.61, H ₂ O)

Example 4:

The compound (2.5g) obtained in Reference Example 8 was treated in a similar manner as described in Examples 1, 2 and 3 to obtain a crystalline powder of 9 - [(1S, 4R) -4-hydroxymethylcyclopentan-1-yl]. guanine (0.3g), mp. 269 ° C, to obtain.

UVX P ^ (nm): 255,280 (shoulder); UVA ^ 1 ° (nm): 253,270 (shoulder); UV ^ i ⁰ (nm): 258 (shoulder), elemental analysis (%) for CnHi ₆ O ₂ N ₅ molecular weight 249.27 calcd. C, 53.00; H, 6.07; N, 28.10 found: C, 52.81; H, 5.86; N, 27.83 [a] g ⁶ = -4.74 (C = 0.57, DMF) g-MIS ^ RM-hydroxymethylcyclopentan-i-yl hypoxanthine is obtained when a hypoxanthine derivative is substituted for N ⁶ -benzoyl-6 ' -0- (4,4'-dimethoxytrityl) -3'-0 - [(imidazol-1-yl) thiocarbonyl] -2'-deoxyaristeromycin in Example 1 is treated in a similar manner as described in Examples 1 to 3.

Elemental analysis (%) for C _n H ₁₄ N ₄ O ₂ Molecular weight 234.25 calculated: C, 56.40; H, 6.02; N, 23.92 found: C, 56.81; H, 6,33; N, 24.25

Example 5:

9 - [(1S, 4R) -4-hydroxymethyl-cyclopentan-1-yl] guanine

(1) Preparation of 9 - [(1R, 5S) RM-hydroxymethyl-S-hydroxylcyclopentan-1-yl hypoxanthine

The compound (12.4 g, 20 mmol) obtained in Reference Example 3 was dissolved in 200 ml of toluene, to which was added tetrabutylammonium fluoride (10.46 g, 40 mmol), followed by heating at 75 ° C for 2 hours. The reaction solution was concentrated to dryness and dissolved in water. The solution of the desalination with 30 g of activated carbon was subjected to the crude product was recrystallized from a mixture of methanol and ethyl ether to give colorless crystals (4.6 g), mp 170 ⁰ C to.

Elemental analysis (%) for CnHi ₄ N ₄ O ₃ .H ₂ O molecular weight 268.27 ^:

calculated: C, 49.25; H, 6.01; N, 20.88 found: C, 49.08; H, 5.86; N, 20.81

(2) Preparation of 9 - [(1R, 5S) RM-monomethoxytrityloxymethyl-S-hydroxylcyclopentan-1-yl hypoxanthine

The crystalline compound (2.3 g, 9.2 mmol) obtained in (1) was dissolved in 100 ml of pyridine, followed by addition of monomethoxytrityl chloride (3.1 g, 10 μmol), followed by stirring at room temperature for 5 hours. The reaction solution was purified by silica gel chromatography (80 g, solvent: CHCl ₃ / MeOH = 40/1 to 6/1) to obtain the desired product in powder form (4.3 g). A portion of the product was recrystallized from a mixture of chloroform and ether (mp, 244-246 ° C).

Elemental analysis (%) calculated for C ₃₀ N ₃ IH ₄ O ₄ · H ₂ O molecular weight 531.60: C, 70.04; H, 5.88; N, 10.54 found: C, 70.39; H, 5.77; N, 10.38

(3) Preparation of 9 - [(1S) RM-Monomethoxytrityloxymethyl-cyclopentan-1-yl-hypoxanthine

The compound obtained in (2) (4.32 g, 8.27 mmol) was dissolved in 70 ml of toluene, followed by the addition of thiocarbonyldiimidazole (2.2 g, 12.4 mmol), and stirred at room temperature for 5 hours. The reaction solution was concentrated to dryness and the residue was purified by silica gel chromatography (80 g, solvent: CHCl ₃ / MeOH = 100/1 to 60/1) to obtain a pale yellow powder (5.2 g). The product thus obtained was dissolved in 90 ml of toluene and reacted with tributyltin hydride (3.4 ml, 12.4 mmol) and α, α'-azobisisobutyronitrile (270 mg, 1.6 mmol) in a manner similar to that described in Reference Example 3, followed by purification by means of Silica gel chromatography (100 g, solvent: ethyl acetate / MeOH = 9/1) to give the desired product (1.63 g). A portion of the product was recrystallized from a mixed solution of methanol and ethyl ether. Mp 175-177 ° C. Elemental analysis (%) for C ₃ ! H ₃₀ N ₄ O ₃ 1 / 2H ₂ O molecular weight 515.60 calculated: C, 72.21; H, 6.06; N, 10.87 found: C, 72.69; H, 5.88; N, 10.92

(4) Preparation of 9 - [(1S, 4R) -4-hydroxymethylcyclopentan-1-yl] guanine

The desired compound can be prepared in a similar manner as in Reference Examples 4 to 8 using the compound obtained in the above step (3).

Example 6:

Tablets for peroral administration of the antiviral composition of the invention are prepared as described below:

200 mg of 2 ', 3'-dideoxyaristeromycin, 300 mg of lactose, 50 mg of starch and 2 mg of magnesium stearate are mixed in methanol, and after removal of the methanol by heating, the mixture is formed into tablets.

Example 7:

An injection of the antiviral composition is prepared as described below:

500 mg of 2 ', 3'-dideoxyaristeromycin are dissolved in 10 ml of sterilized water, the pH is adjusted to 6.0 with sodium hydroxide solution. The solution is filtered with sterilized filter and sealed in a vial.

Experimental Example 1: -

Materials and Methods *

* Antimicrob. Agents Chemother. 30, No. 6, Dec. 1986, 933-937

Cells: In this study, one cell line MT-4, HTLV type I carrier, and one HIV _H tlv-iii producing cell line, Molt-4 / HIVHTLv-iii- were used. The cells were maintained in RPM11640 medium were added to the 10% fetal calf serum, 100 IU penicillin per ml and 100μρ streptomycin per ml at 37 ⁰ C in a C0 ₂ incubator. Virus and viral infection: mV _HTLV . | Ii was obtained from culture supernatants of Molt-4 / HIVHTLv-iii ^w ' ^e described above [Virology 146,272 (1985)]. The titer of this virus preparation was 6 × 10 ⁴ PFE / ml. Infection of the ΜΤ-4 cells with HIVhtlv-iii was done with a challenge factor of 0.002. The cells were mixed with virus solution and cultured at 37 ⁰ C for one hour. After adsorption, the infected cells were washed and resuspended in fresh medium to a concentration of 3 x 10 ⁵ cells per ml. The concentration was cultured with and without the presence of various concentrations of carbocyclic 2 ', 3'-dideoxynucleosides in a C0 ₂ incubator at 37 ⁰ C for 6 days.

Analysis of HIV _H TLv-Induced Cytopathic Effect: The HIVm-w-IH-induced cytopathic effect was analyzed by measuring the decrease in viable cells. The viable cells were counted by the trypan blue exclusion staining procedure.

Analysis of HIVHTLv-non-antigen expression: HTLV-III-infected ΜΤ-4 cells with virus-specific antigens were counted by an indirect [immunofluorescence (IF)] method. Methanol-fixed cells were cultured with diluted anti-HTLV-III-positive human serum for 30 minutes at 37 ° C. The preparations were then washed for 15 minutes with phosphate buffered saline. The cells were then cultured with fluorescein isothiocyanate-conjugated rabbit anti-human immunoglobulin G (Dakoppatts A / S, Copenhagen, Denmark) for 30 minutes and washed again with phosphate buffered saline. More than 500 cells were counted under a fluorescence microscope and the percentage of IF-positive cells was determined. By the above-described analysis, it was confirmed that the compounds of the present invention apparently have anti-HIV _H TLv-nr activity. In the case of 2 ', 3'-dideoxyaristeromycin, the effective concentration was 50 to 100μΜ, and the cytotoxicity was observed at 500 to 1000μΜ.

Claims (8)

1. A process for the preparation of a compound of the formula I. R ⁶ " ^{5 Y} wherein R represents an optionally protected hydroxyl group and Y represents an optionally protected purine base, and salts thereof, characterized in that a compound of formula II. wherein R and Y are as defined above and either R ₁ or R _{2 is} a hydroxyl group and the other is hydrogen, which subjects the reduction reaction to 2 'or 3' deoxidation. 2. The method according to claim 1, characterized in that the hydroxyl group denoted by R ₁ or R ₂ in a compound of formula IL 5 '
R "I ' (II) wherein R, Y, R ₁ and R ₂ are as defined above, thiocarbonylated, followed by reduction with trityl tin hydride in the presence of α, α'-azobisisobutyronitrile and, if necessary, removal of all protecting groups of R and Y. 3. The method according to claim 1 and 2, characterized in that in the formula IR is a hydroxyl group and Y adenine-9-yl, hypoxanthine-9-yl, guanine-9-yl, isoguanine-9-yl, xanthine-9 -yl, 3-deazaadenin-9-yl, 7-deazaadenin-9-yl, 8-azaadenin-9-yl or 2,6-diaminopurine-9-yl. 4. The method according to claim 1 and 2, characterized in that 2 ', 3'-Didesoxyaristeromycin is produced. 5. The method according to claim ^ and 2, characterized in that 9 - [(1 S, 4R) -4-hydroxymethylcyclopentan-1-yl] -guanine is produced. 6. The method according to claim 1 and 2, characterized in that 9 - [(1 S, 4R) -4-hydroxymethylcyclopentan-1-yl] hypoxanthine is produced. 7. The method according to claim 1 and 2, characterized in that in the formula I R is protected with a monomethoxytrityl or dimethoxytrityl hydroxy group. 8. The method according to claim 1 and 2, characterized in that in the formula I Y is a protected with benzoyl, isobutyryl or acetyl purine base.