RU2039752C1 - Method of synthesis of substituted purine arabinosides or their pharmaceutically acceptable derivatives - Google Patents

Abstract

FIELD: organic chemistry. SUBSTANCE: product: substituted derivatives of purine arabinosides of the formula (I) where R₁ halogen, methoxy, ethoxy, propoxy, amino-group which is monosubstituted with methyl or C-C -cycloalkyl, or disubstituted with C-C-alkyl, or R₁ nitrogen-containing heterocycle; R₂ hydrogen, halogen or amino-group at condition that when R₂ hydrogen then R₁ does not methoxy, methylamino, dimethylamino, piperidino or pyrrolidino-group, and when R₂ amino-group then R₁ does not methylamino-group, and/or its pharmaceutically acceptable derivatives, except for 2,3,5-triacetate and 2,3,5-tribenzyl-derivatives of the formula (I) where R₁ chlorine or fluorine and R₂ - chlorine, fluorine, hydrogen or amino-group. Reagent 1: compound of the formula (II)

. Reagent 2: compound of the formula (III)

Description

 The invention relates to certain substituted purine arabinosides and their physiologically acceptable derivatives, in particular esters and their use for the treatment of certain DNA virus diseases.

 The Varicella herpes zoster virus (BBZ) virus, which causes syphilis in chicken and shingles, is a herpes DNA virus. Chicken pox (syphilis in chicken) is the main disease caused by VVZ in the host without an immune response; in the general case, it is a mild illness of young children, which manifests itself in the form of fever and frequency rash. Herpes zoster (shingles) is a recurrent form of the disease that occurs in adults who have previously been infected with the Varicella-zoster virus. The clinical symptoms of this infection are characterized by neuralgia and a vesicular skin rash, which has a unilateral and skin distribution. The spread of inflammation can lead to paralysis or convulsions, and in addition, if meningitis subsequently develops.

 In immunocompromised patients, this virus can spread, causing serious, often fatal diseases. The cause of immunodeficiency may be medications used in the treatment of patients with transplant or in the treatment of malignant neoplasms, or diseases such as AIDS, which destroy the immune system, as a result of which the patient becomes vulnerable to infections that otherwise would not be so dangerous.

 Cytomegalovirus (CMB) is another herpes virus. Infection can be acquired in childhood or in youth and in the fetus, and intrauterine infection is probably the most common form of infection, but up to 90% of congenital infections are asymptomatic during childbirth. Pre-infection of the mother during pregnancy is generally considered the greatest risk to an unborn baby, while re-activation of fetal infections is generally clinically rare. Clinical effects range from death and serious illness (microcephaly, enlarged liver or spleen, jaundice, mental retardation) through the cessation of the disease to its rapid development, susceptibility to pulmonary and ear infections, to the absence of obvious effects of the disease. In young people, an infection can go undetected or manifest as a fever caused by glandular inflammation, similar to a disease resulting from close physical contact.

 Serious infections can also occur as a result of reactivation of the dormant virus in patients with an ineffective immune system, as described for BBZ infections. Such infections lead to higher morbidity and mortality from inflammation of the lining of the eye, inflammation of the lungs and gastrointestinal disorders.

 It was found that some purine-arabino nucleosides, described in detail below, characterized by the presence of groups substituted at the 2- and 6-positions of the purine ring, are highly active against human viral infections, in particular those caused by Varicella zoster virus (BBZ) or cytomegalovirus (CMV).

 Some substituted purine-arabino nucleosides, in particular 9-β-D-arabinofuranosyl-6-methoxy-9H-puri, 9-β-D-arabinofuranosyl-6-pyrrolidino-9H-purine, 9-β-D-arabinofuranosyl-6 -methylamino-9H-purine and 9-β-D-arabinofuranosyl-6-dimethylamino-9H-purine, which are described below, when used to treat BBZ and CMV infections, are described previously in J. Org. Chem. Volume 27, 3274-9 (1962), Caner Jreatment Rep. 60 (10), 1567-84 (1976), Tetrahedron 40 (4), 709-13, (1984), Canada J. Biochem 43 (1), 1-15 (1965), J. Med. Chem. 12, 498-504, (1969), J. Biol. Chem. 251 (13), 4055-61 (1976), Ann. N.J. Acad. Sci, 284, 81-90, (1977), in European patent 002192, US patents 3 666 856, 4 371 613, 3758684.

(I) где R₁ представляет галоген, метокси, этокси, пропокси; амино-группу, монозамещенную метилом или С₃-С₆-циклоалкилом, или двузамещеную С_1-5-алкилом; или R₁ означает азотсодержащее гетероциклическое кольцо, содержащее 4-7 атомов углерода и связанное с пуриновым радикалом через атом азота; R₂ представляет водород, галоген или амино при условии, что когда R₂ означает водород, R₁ не означает метокси, метиламино, диметиламино, пиперидино или пирролидино; а когда R₂ означает аминогруппу, R₁ не означает метиламино и его приемлемые с физиологической точки зрения производные для лечения или профилактики вирусных инфекций у человека, вызванных ВВЗ или ЦМВ.Thus, in accordance with the invention, a method for producing a compound of formula I is provided:

(I) where R ₁ represents halogen, methoxy, ethoxy, propoxy; an amino group monosubstituted with methyl or C ₃ -C ₆ cycloalkyl, or disubstituted with C _1-5 alkyl; or R ₁ means a nitrogen-containing heterocyclic ring containing 4-7 carbon atoms and linked to a purine radical through a nitrogen atom; R ₂ represents hydrogen, halogen or amino, provided that when R ₂ means hydrogen, R ₁ does not mean methoxy, methylamino, dimethylamino, piperidino or pyrrolidino; and when R ₂ is an amino group, R _{1 is} not methylamino and its physiologically acceptable derivatives for the treatment or prophylaxis of human viral infections caused by BBH or CMV.

Preferred compounds of formula (I) are those in which:
(a) R ₂ is hydrogen, and
(b) R ₁ is C _1-5 alkoxy, especially methoxy for pharmaceutically acceptable derivatives or
(c) R ₁ is C _1-5 alkylamino, especially dimethylamino for pharmaceutically acceptable derivatives or
(d) R ₁ is halogen, especially iodine.

The following compounds are preferred compounds in accordance with the invention in view of their strong antiviral activity against BBZ or CMV:
1) 9-β-D-arabinofuranosyl-6-ethoxy-9H-purine,
2) 9-β-D-arabinofuranosyl-6-iodine-9H-purine,
3) 9-β-D-arabinofuranosyl-2-amino-6-iodopurine,
4) 9-β-D-arabinofuranosyl-2-chloro-6-methylamino-9H-purine,
5) 9-β-D-arabinofuranosyl-6-cyclopropylamino-9H-purine,
6) 9-β-D-arabinofuranosyl-6-ethylmethylamino-9H-purine,
7) 9-β-D-arabinofuranosyl-2-amino-6-methoxy-9H-purine,
8) 9-β-D-arabinofuranosyl-6-n-propoxy-9H-purine.

Physiologically acceptable derivatives of the compounds of general formula (I) other than 2 ′ 3 ′, 5 ′ triacetate and tribenzyl derivatives of the compounds of formula (I) in which R ₁ is chlorine or fluorine R ₂ is chlorine, fluorine, hydrogen or amino, are used in medical therapy, in particular for the treatment of human viral infections caused by VVZ or CMV.

 Pharmaceutically acceptable derivatives of the compounds of formula (I) are any pharmaceutically acceptable ethers, salts, esters or salts of such esters, or any other compound that, when applied to humans, is capable of producing (directly or indirectly) a compound of the formula (I) or an antiviral metabolite or residue thereof.

 Pharmaceutically acceptable esters of the aforementioned compounds of formula (I) are particularly preferred since they are able to provide high concentrations of the main compound in the patient’s plasma after dental use.

 Purine nucleosides of the formula (I) and their derivatives are hereinafter referred to as the compounds of the invention or active ingredients.

 In accordance with another preferred aspect of the invention, there is provided the use of a compound of the invention for the manufacture of a medicament for the treatment or prophylaxis of human viral infections caused by BBH or CMV.

 In addition, in accordance with the invention, there is provided a method for the treatment or prevention of VVZ and CMV infections in humans, which comprises applying to said person an effective amount of a compound of the invention.

 The method includes inhibiting the replication of BBZ or CMV viruses in a mammalian host cell, which comprises using an effective amount to inhibit virus replication of a compound of formula (I) or a pharmaceutically acceptable derivative thereof for infected cells.

 Examples of clinical diseases caused by BBZ and CMV infections that can be treated in accordance with the invention include the listed diseases.

 The compounds of formula (I) and their pharmaceutically acceptable derivatives (hereinafter collectively referred to as active ingredients) can be used by any method appropriate for the diseases, such methods include oral, rectal, nasal, topical (including buccal and under tongue), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal and epidural) methods. The preferred method may vary, for example, depending on the condition of the patient.

 For each of these uses and indications, the amount of active ingredient required (as defined above) will depend on various factors, including the severity of the disease to be treated and compatibility with the recipient, and will depend more on the decision of the attending physician. In general, for each of these uses and indications, the corresponding effective dose will vary from 0.1 to 250 mg per kg of recipient body weight per day, preferably in the range from 0.1 to 100 mg per kg of body weight per day, and in the most preferred embodiment, in the range from 1 to 20 mg per 1 kg of body weight per day; the optimal dose is about 15 mg per 1 kg of body weight per day (unless otherwise stated, all weights of the active ingredient are calculated for the main compound of formula (I): for its salts and esters, these numbers should be increased proportionally). The desired dose is preferably divided into two, three, four, or more sub-doses, which are applied at appropriate intervals throughout the day. Such sub-doses may be administered in unit dosage forms, for example, containing from 5 to 1000 mg, in a preferred embodiment from 20 to 500 mg, and in the most preferred embodiment, from 100 to 400 mg of active ingredient per unit dosage form.

 Although only active ingredients can be used, they are preferably used in pharmaceutical forms, the forms (compositions) according to the invention contain at least one active ingredient, as defined above, together with one or more acceptable carriers it, as well as possibly other therapeutic ingredients. The carrier (s) should be "acceptable" in the sense that it must be compatible with the other ingredients of the composition and should not adversely affect the recipient.

 These forms include forms suitable for oral, rectal, nasal, topical (including buccal and under the cheek) applications, vaginal or parenteral (including subcutaneous, intramuscular, intracutaneous and epidural) applications. Such forms may be prepared in unit dosage form and may be prepared by any of the well-known pharmaceutical techniques. Such techniques include the step of bringing the active ingredient into contact with a carrier, which includes one or more accessory ingredients. In general, such forms are obtained by uniformly and thoroughly mixing the active ingredient with liquid carriers or finely divided solid carriers, or both, and then giving the mixture the desired shape.

 Forms in accordance with the invention for dental use may take the form of discrete units, such as capsules, pills or tablets, each of which contains a predetermined amount of the active ingredient; like powder or granules; as a solution or suspension in an aqueous liquid or non-aqueous liquid; or as an oil in water liquid emulsion or a water in oil liquid emulsion. The active ingredient may also be in the form of a ball, medicinal porridge or paste.

 A tablet may be prepared by compression or molding, possibly using one or more accessory ingredients. Compressed tablets may be prepared by compression, using a suitable device, the active ingredient in a free-flowing form, such as powder or granules, may be mixed with a binding agent (e.g. povidone, gelatin, hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative an agent, a disintegrating agent (for example, sodium starch glycollate, crosslinked povidone, crosslinked sodium carboxymethyl cellulose), a surface active or dispersing agent. Molded tablets may be made by molding in a suitable device a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may be coated or provided with a groove and may be shaped to provide a slow or controlled release of the active ingredient, using, for example, hydroxypropyl methylcellulose in varying proportions so as to provide the desired release profile.

 When infecting the eye or other external tissues, such as the mouth and skin, the compositions are preferably used in the form of topical ointments or creams containing the active ingredient in an amount of, for example, from 0.075 to 20 w / v, in the preferred embodiment, from 0.2 to 15 / in a, in the most preferred embodiment, from 0.5 to 10 in / in. If the above form is an ointment, the active ingredients can be used together with either a paraffin or water-miscible ointment base. Alternatively, the active ingredients can be used to make an oil-in-water cream.

 If necessary, the aqueous phase of the cream may include at least 30 iv of a polyhydric alcohol, namely, an alcohol containing two or more hydroxyl groups, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol, and mixtures thereof. Local forms may optionally contain a compound that increases the absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such skin penetration enhancing agents include dimethyl sulfoxide and related analogs.

 The oil phase of the emulsions in accordance with the invention can be formed from known ingredients using a known technique. Despite the fact that this phase may contain only an emulsifier (also known under another name emulsifier), in the preferred embodiment, it contains a mixture of at least one emulsifier with fat or oil, or with both. In a preferred embodiment, the hydrophilic emulsifier is included with the lynophilic emulsifier, which acts as a stabilizer. In a preferred embodiment, it also includes oil and fat together. Together emulsifier (s) with or without stabilizer (s) form the so-called emulsifying wax, and this wax together with oil and / or fat forms the so-called emulsifying base of the ointment, which forms the cream phase dispersed in the oil.

 Emulsifiers and emulsion stabilizers suitable for use in the form of the invention include Tween 60, Spen 80, ketostearyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodium lauryl sulfate.

 The choice of the appropriate oils or fats for the composition is based on the achievement of the necessary cosmetic properties, since the solubility of the active compound in most oils, such as those used in the form of pharmaceutical emulsions, is very low. Thus, the cream should preferably be a non-greasy, non-staining and rinse-off product with an appropriate consistency, which would avoid leakage from pipes or other containers. Mono- or dibasic straight chain or branched chain alkyl esters such as diisoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palminate, butyl stearate, palmethyl, 2-ethyl branched chain esters, known as Crodamol KAP, with the latter three being preferred esters. They can be used individually or in combination, depending on the required properties. Alternatively, high melting point lipids such as white soft paraffin and / or liquid paraffin or other mineral oils can be used.

Compositions suitable for topical application to the eyes also include eye drops in which the active ingredient is dissolved or suspended in an appropriate carrier, in particular an aqueous solvent for the active ingredient. The active ingredient is preferably contained in such forms in a concentration of from 0.5 to 20%, in a preferred embodiment, from 0.5 to 10%, for example, at a concentration of about 1.5 w / w
Compositions suitable for topical application in the oral cavity include lozenges containing the active ingredient on a palatable basis, generally in sugar and acacia or tragacanth; lozenges containing the active ingredient in an inert basis, such as gelatin and glycerin, or sugar and acacia; and oral texture containing the active ingredient in an appropriate liquid carrier.

 Compositions for rectal administration may be in the form of suppositories on a suitable basis containing, for example, coconut oil or salicylate.

 Compositions intended for use through the nose, in which the carrier is a solid substance, include coarse powder having a particle size, for example, in the region of 20 to 500 μm, which is applied using a method containing a container, namely by rapid inhalation through the nasal cavity from the powder container, which is held near the nose. Suitable compositions in which the carrier is a liquid for use, for example, in the form of a nasal spray or nasal drops, include aqueous or oily solutions of the active ingredient.

 Forms intended for vaginal use may take the form of pessaries, tampons, creams, gels, pastes, foams or aerosols containing carriers along with the active ingredient, the use of which is well known in the art.

 Formulations intended for parenteral use include aqueous and non-aqueous sterile injectable solutions, which may include antioxidants, buffering materials, bacteriostats, and dissolved substances that impart isotonic properties to the blood relative to the intended recipient; and aqueous and non-aqueous sterile suspensions, which may include suspending agents and thickening agents. These forms may take the form of a single dose or several doses, for example, sealed ampoules and vials, and may be stored under freeze-dried (lyophilized) conditions requiring the addition of only a sterile liquid carrier, for example, water for injection immediately before use. Untreated solutions and suspensions for injection can be obtained from sterile powders, granules and tablets of the specified type.

 Preferred unit dosage forms are those containing a daily dose or a single daily sub-dose, as defined above, or corresponding fractions of the active ingredient thereof.

 Along with the listed ingredients, the forms of the invention may include other agents, the use of which is well known depending on the type of form, for example, agents intended for dental use, may include flavoring agents.

(II) в которой R₁ и R₂ уже были определены, с соединением формулы (III):

X (III) в которой Х представляет пиримидиновое или пуриновое основание (отличное от соединения формулы II), и, возможно, последующее или одновременное с этим в том случае, когда полученным в результате соединением является соединение формулы (I), превращение его в его приемлемое с фармацевтической точки зрения производное, или в том случае, когда полученный в результате соединением является приемлемое с фармацевтической точки зрения производное, превращение его в другое, приемлемое с фармацевтической точки зрения, производное соединение формулы (I).The invention provides a process for preparing a compound of formula (I) or a pharmaceutically acceptable derivative thereof, in particular an ester thereof, comprising reacting a compound of formula (II)

(II) in which R ₁ and R ₂ have already been defined, with a compound of formula (III):

X (III) in which X represents a pyrimidine or purine base (other than a compound of formula II), and possibly subsequent or simultaneous in this case, when the resulting compound is a compound of formula (I), converting it to its acceptable from a pharmaceutical point of view, a derivative, or in the case where the resulting compound is a pharmaceutically acceptable derivative, converting it into another pharmaceutically acceptable derivative of a compound of the forms uly (I).

With regard to this method, X is preferably a uracil base. This reaction can be carried out, for example, by treating the compounds of formulas (II) and (III) with an enzyme, such as a phosphorylase enzyme, for example uridine phosphorylase or purine nucleoside phosphorylase, or a mixture thereof, in the preferred embodiment, in the presence of a phosphate salt at a pH in the region of 5 , 0-9,0 and a temperature of from 15 to 90 ^{° C,} preferably in the range from 40 to 60 ° ^C.

 Physiologically acceptable esters and salts of the compounds of formula (I) can be prepared using known techniques, for example, esters can be prepared by esterification of the basic compound with a suitable acyl halide or anhydride. Alternatively, esters can be prepared by substituting an appropriate displaceable group, for example a halide of a suitable carboxylic acid, or by breaking the corresponding anhydronucleoside of a basic compound with a suitable carboxylic acid or its salt.

 The following examples illustrate the invention, and therefore should not be construed as limiting the invention.

 PRI me R 1. 9-β-D-Arabinofuranosyl-6-methylamino-9H-purine.

6-Thiol-9-β-D-arabinofuranosyl-9H-purine (Reist E. J. et al. J. Org. Chem. 27, (1962), p. 3274-3279) (0.35 mmol, 100 mg) and 5ml of absolute methanol were combined and cooled to ^-10 ° C, while protecting from moisture. Chlorine gas was carefully bubbled through the suspension for 2 minutes. The resulting solution was stirred for 5 min at ¹⁰ ° C, then through the cold solution for 15 minutes, bubbled with dry nitrogen until until removed excess chlorine. The reaction mixture was added 2 ml of 40% aqueous methylamine solution, which was then heated in a stainless steel autoclave at ¹¹⁵ ° C, maintained for 4.5 hours. The reactor was cooled ^to 0 ° C and the contents were evaporated to dryness to provide 88% yield of compound from the title of an example. After recrystallization in water, the sample had a melting point of 201.5-202.5 ^about C.

Analysis results: calculated for C ₁₁ H ₁₃ N ₅ O ₄
Calculated, C 47.0; H 5.38; N, 24.9.

Found, C, 47.2; H 5.72; N 25.2
PRI me R 2. 9-β-D-Arabinofuranosyl-6-dimethylaminopurine
6-Dimethylaminopurin (Sigma Chemical Co., St. Louis, Mo.) (6.4 mmol, 1.04 g) and uracil arabinoside (Torrens, P.F. et al. J. Med. Chem. 22 (3), 1979 , pp. 316-319) (8 mmol, 1.96 g) were combined in 0.412 L of a 5 mM potassium phosphate solution, pH 8.0, with 0.02% potassium azide. Purified purine nucleoside phosphorylase (3260 units) and uridine phosphorylase (810 units), and the solution was stirred at 35 ° ^C. After 59 days the reaction was lyophilized. The residue was suspended in 250 ml of water and stirred at room temperature for 1 h. The solids were separated by filtration and the filtrate stored at 3 ° ^C.

 After 72 hours, the precipitate was collected and combined with the previous cake. All this was added in 100 ml of 95% ethanol in water, heated to boiling and filtered. The solvent was removed in vacuo and the residue was chromatographed on a BIORad P-2 type resin in 30% n-propanol (water o / o), once using a column with dimensions of 7.5 x 90 cm and two more times with a column of 5 x 90 cm As a result of this procedure, 0.12 g of 6-dimethylaminopurin-9-β-D-arabinofuranoside was obtained in the form of 0.5 hydrate.

Analysis results: calculated for C ₁₂ H ₁₇ N ₅ O ₄ ˙ 0.5 N ₂₀ .

Calculated, C 47.36; H 5.96; N, 23.01
Found, C, 47.23; H 5.59; N, 22.75
NMR and mass spectra are consistent with such a structure.

 PRI me R 3. 9-β-D-Arabinofuranosyl-6-methoxypurine.

6-Methoxypurin (Sigma Chemical Co., St. Louis, Mo.), (6.6 mmol, 1 g) and uracil arabinoside (Torrens, P.F. et al. J. Med. Chem. 22 (3), 1979 , (10.1 mmol, 2.45 g) were suspended in 575 ml of 10 mM potassium phosphate, 0.04% potassium azide solution, pH 7.8, containing 10% n-propanol (o / o). Purified uridine phosphorylase ( 560 m.u.) and purine nucleoside phosphorylase (10,000 m.u.) Krenitsky T.E. et al. Biochemistry, 20, p. 3615, 1981 and US patent N 4 381 444 and the solution was stirred at a temperature of 35 ^about C. Thirty days later, the reaction was filtered, and the filtrate was basified to pH 10.5 with ammonium hydroxide and and chromatographed on a column 2.5 x 7 cm containing resin Dowex-1-formate. The resin was eluted with 30% n-propanol / water (o / o).

 The fractions containing the product were combined and the solvent was removed in vacuo. The resin was dissolved in 30% n-propanol / water (o / o) and chromatographed on a Bio-Rad P-2 column (7.5 x 90 cm). Product fractions were combined and, after lyophilization, 0.922 g of 6-methoxypurin-9-β-D-arabinofuranoside was obtained as a dihydrate.

Analysis results: calculated for C ₁₁ H ₁₄ N ₄ O ₅ ˙ 2H ₂ O
Calculated, C 41.51; H 5.70; N, 17.70
Found, C 41.46; H 5.74; N, 18.13
NMR and mass spectra are consistent with such a structure.

 PRI me R 4. 9-β-D-Arabinofuranosyl-6-ethoxypurine.

6-Ethoxypurin (Sigma Chemical Co. St. Louis, Mo.) (3.05 mmol, 0.5 g) and uracil arabinoside (6.09 mmol, 1.48 g) were suspended in 100 ml of 10 mM potassium phosphate, 0 , 04% potassium azide solution at pH 7.4. Purified uridine phosphorylase (6000 moe) and purine nucleoside phosphorylase (8400 moe) were added by Krenitsky, T.E. et al. Biochemistry, 20, p. 3615, 1981 and US patent No. 4,381,444 and the suspension was mixed at a temperature of 35 ^about C.

 After 168 hours, another 18,000 units of uridine phosphorylase and 75,600 units of purine nucleoside phosphorylase were added. After seven days, the reaction was filtered, and the filtrate was chromatographed on a column containing Dowex-1-hydroxide resin (2.5 x 8 cm). The column was eluted with 90% methanol / water (o / o), the fractions containing the product were combined, and the solvent was removed in vacuo. The residue was dissolved in 30% n-propanol and water (o / o) and chromatographed on a column containing Bio Rad P-2 (5 x 90 cm). The product was eluted with 30% n-propanol / water (v / v). The product containing fractions were combined and the solvent was removed in vacuo to give 0.363 g of 6-ethoxypurin-9-β-D-arabinofuranoside, which was identified as 0.3 hydrate.

Analysis results: calculated for C ₁₂ H ₁₆ N ₄ O ₅ ˙ 0.3 H ₂ O
Calculated, C 47.78; H 5.55; N, 18.57
Found, C, 47.99; H 5.54; N, 18.40.

 NMR and mass spectra are consistent with this structure.

 PRI me R 5. 9-β-D-Arabinofuranosyl-6-iodopurine.

6-Yodpurin (Sigma Chemical Co. St. Louis, Mo.) (4 mmol, 1 g) was dissolved in 15 ml of 1,2-dimethoxyethane with heating. 50 ml of a solution of uracil arabinoside (10.1 mmol) in 10 mM potassium phosphate, a 0.04% potassium azide solution, pH 7.4 were added. Purified uridine phosphorylase (6800 IU) and purine nucleoside phosphorylase (12,000 IU) and the reaction was stirred at 35 ° ^C. After 21 days, a further 4800 units of uridine phosphorylase and 20000 units of purine nucleoside phosphorylase. After ninety days, the reaction was filtered, and the solvent was removed in vacuo. The residue was dissolved in 100 ml of water, heated with steam, and then filtered. The filtrate was chromatographed on a column containing KSAD-2 resin (5 x 35 cm). This column was eluted with 2 L of water, and then 2 L of ethanol. The fractions containing the product were combined. and the solvent was removed in vacuo. The residue was dissolved in a mixture of 30% n-propanol / water (v / v) and subjected to chromatography on a column containing Bio-Rad P-2 (5 x 90 cm).

 The product was eluted with 30% n-propanol / water (v / v). The fractions containing the product were combined and the solvent was removed in vacuo. The residue was dissolved in 30% n-propanol / water (v / v) and chromatographed on a Sephadex G-10 column (5 x 90 cm). This column was eluted with 30% n-propanol / water (v / v). The product containing fractions were combined and, after removal of the solvent in vacuo, 0.253 g of 6-iodopurin-9-β-D-arabinofuranoside was obtained, which was identified as 1.5 hydrate.

Analysis results: calculated for C ₁₀ H ₁₁ JN ₄ O ₄ ˙1.5 H ₂ O
Calculated, C 29.65; H 3.48; N 13.83; J 31.32
Found, C, 29.43; H 3.53; N, 13.66; J 31,20
NMR and mass spectra are consistent with such a structure.

 PRI me R 6. 8-β-D-Arabinofuranosyl-2-amino-6-iodopurine.

2-amino-6-iodopurine (Sigma Chemical, St. Louis, Mo.) (25.5 mmol, 6.75 g) and uracil arabinoside (61.9 mmol, 15.1 g) were combined in 0.31 L of 10 mM potassium phosphate, pH 6.9, with 0.02% potassium azide. Purified purine nucleoside phosphorylase (17000 units) and uridine phosphorylase (2000 units), and the solution was stirred at ³⁷ ° C. After 18 days, a further 5700 units of uridine phosphorylase. After fifty-seven days, the reaction was filtered, and the filtrate was chromatographed on a column containing XAD-2 resin (8 x 11 cm). The product was eluted with a successive ethanol / water (v / v) gradient as follows: 0.35 L 10% 1 L 20% 1 L 50% 0.2 L 95% The fractions containing the product were combined and the ethanol removed in vacuo. The residue was dissolved in 30% n-propanol / water (v / v) and chromatographed on a column (7.5 x 90 cm) containing Bio-Rad P-2 resin. As a result, 1.1 g of 2-amino-6-iodopurin-9-β-D-arabinofuranoside was obtained as a 0.5 hydrate.

Analysis results: calculated for С ₁₀ Н ₁₂ N ₅ О ₄ ˙0.5 Н ₂ О
Calculated, C 29.87; H 3.26; N 17.41
Found, C, 29.86; H 3.29; N, 17.39.

 NMR and mass spectra confirmed this structure.

 PRI me R 7. 9-β-D-Arabinofuranosyl-6-pyrrolidinopurin.

6-Pyrrolidinopurin (Sigma Chemical Co. St. Louis, Mo.) (2.6 mmol), 0.5 g and uracil arabinoside (5.29 mmol, 1.29 g) were suspended in 100 ml of 10 mM potassium phosphate, 0 , 04% potassium azide solution at pH 7.4. Purified uridine phosphorylase (6000 IU) and purine nucleoside phosphorylase (8400 IU) Krenitsky, T.A. were added. et al. Biochemistry, 20, p. 3615, 1981 and US patent N 4,381,444) and the suspension was stirred at 35 ° ^C. Twenty days later the reaction was filtered and the filtrate was chromatographed on a column containing resin Dowex-1-hydroxide ( 2.5 x 8 cm). The product was eluted from the column using a 90% methanol / water (v / v) mixture. The fractions containing the product were combined and the solvent was removed in vacuo. The residue was dissolved in 50 ml of 30% n-propanol and water (v / v) and chromatographed on a Bio-Rad P-2 column (5 x 90 cm). The product was eluted using 30% n-propanol / water (v / v). The product containing fractions were combined and the solvent was removed in vacuo to give 0.573 g of 6-pyrrolidinopurin-9-β-D-arabinofuranoside.

Analysis results: calculated for C ₁₄ H ₁₉ N ₅ O ₄ .

 Calculated, C 52.33; H 5.96; N, 21.79.

Found, 52.60; H 6.09; N, 21.51
NMR and mass spectra confirm the structure.

 PRI me R 8. 9-β-D-Arabinofuranosyl-2-chloro-6-methylaminopurine.

A solution of 2,6-dichloro-2 ', 3', 5'-tri-0-benzyl-9- (β-D-arabinofuranosyl) purine (Keller F. et al. J. Org. Chem. 32, p. 1644 , 1967, Montgomery, J.E. and Hewson, C.J. J. Med. Chem. 12, p. 498, 1967 5.92 g, 10 mmol) in benzene (35 ml of a solution of 0.6 g of methylamine per 10 ml of benzene) was kept at room temperature in a sealed autoclave for 4 days. The reactor was ice-cooled, opened and the contents were filtered to remove methylamine hydrochloride. The solvent was removed in vacuo to give an oil which was combined with the oil reacting with the same amounts, but conducted at a temperature of 125 ^C. The total weight was 11.4 g As a result of thin layer chromatography (TLC) was adjusted that the material is mixture of starting material, mono- and dimethylamino compounds. The oil was chromatographed on 285 g of silica gel using 30% acetone and 70% cyclohexane (volumes). The components following the starting material (TLC, silica gel, 3 7, acetone cyclohexane) were collected and the solvent was removed in vacuo. Yield: 4.8 g of 2-chloro-6-methylamino-2 ′, 3 ′, 5′-tri-0-bezyl-9- (β-D-arabinofuranosyl) purine as an oil. A portion of 1.1 g of this material in 40 ml of 2-methoxyethanol was added to palladium chloride (0.87 g), which was previously reduced in a Parr apparatus. The mixture was hydrogenated at a pressure of 50 psi. inch (3,515 kg / cm ² ) for 30 minutes in a hydrogen atmosphere, which was replaced after the first 15 minutes. The catalyst was removed by filtration through a Celite pad and washed with methanol. The filtrate was neutrolized by the addition of Dowex-1 (HCO ₃ ). The resin was removed by filtration and washed with methanol. The filtrate was evaporated in vacuo and the residue was triturated with chloroform. The crude product was washed with hot water, dissolved in hot methanol, filtered, cooled and the solid was collected. Crystallization from boiling water gave the product as a hydrate.

Yield: 44.5 mg; melting point temperature: 224-225 ^о С.

Analysis results: calculated for С ₁₁ Н ₁₄ N ₅ О ₄ Сl˙Н ₂ О
Calculated, C 39.58; N, 20.98; H 4.84;
Found, C 39.27; N, 20.83; H, 5.16.

 PRI me R 9. 9-β-D-Arabinofuranosyl-6-cyclopropylaminopurine.

6-Cyclopropylaminopurine obtained by nucleophilic substitution of a chlorine group with 6-chloropurine (Sigma Chemical Co. St. Louis, Mo.) using cyclopropylamine in acetonitrile (2.85 mmol, 0.5 g) and uracil arabinoside Torrens, P. F. et al. J. Med, Chem. 22 (3), 1979, pp. 316-319 (5.71 mmol, 1.39 g suspended in 100 ml of 10 mM potassium phosphate, 0.04% potassium azide solution, pH 7.4. Purified uridine phosphorylase was added (6000 IU) and purine nucleoside phosphorylase (8 400 IU) Krenitsky et al. Biochemistry, 20, p. 3615, 1981 and US patent N 4 381 444 and the suspension was stirred at a temperature of 35 ^about C. After 120 hours the reaction filtered and the filtrate was chromatographed on a column containing Dowex-1-hydroxide resin (2.5 x 10 cm). The column was eluted with 90% methanol / water (v / v). The fractions containing the product were combined and the solvent was removed in vacuo. Osta the current was dissolved in 30% n-propanol and water (v / v) and chromatographed on Bio-Rad P-2 (5 x 90 cm). The column was eluted with 30% n-propanol / water (v / v). The reactions were recrystallized from hot methanol to give 0.0352 g of a compound that was identified as 6-cyclopropylaminopurin-9-β-D-arabinofuranoside monohydrate. After filtration, the filtrate was chromatographed on Bio-Rad P-2 resin (5 x 90 see) as described. The product containing fractions were combined from both columns and the solvent was removed in vacuo to give 0.342 g of 6-cyclopropylaminopurin-9-β-D-arabinofuranoside, which had the form of a 0.8 hydrate with 0.3 C ₃ H ₈ O .

Analysis results: calculated for С ₁₃ Н ₁₇ N ₅ О ₄ ˙1 Н ₂ О
Calculated, C 48.00; H 5.89; N, 21.53.

 Found, C, 48.05; H 5.89; N, 21.55.

 NMR and mass spectra confirm this structure.

 PRI me R 10. 9-β-D-Arabinofuranosyl-6-ethyl (methyl) aminopurine.

6-Ethyl (methyl) aminopurine was obtained by nucleophilic substitution of a chlorine group with 6-chloropurine (Sigma Chemical, St. Louis, Mo.) using 6-ethyl (methyl) amine in acetonitrile. 6-Ethyl (methyl) aminopurin (2.8 mmol, 0.5 g) and uracil arabinoside (5.6 mmol, 1.38 g) were suspended in 575 ml of 10 mM potassium phosphate, 0.04% potassium azide solution, pH 7 , 4 containing 10% n-propanol (o / o). Purified uridine phosphorylase (6,000 IU) and purine nucleoside phosphorylase (8,400 IU) were added (Krenitsky et al. Biochemistry, 20, p. 3615, 1981 and US Pat. No. 4,381,444), and the solution was mixed at ³⁷ ° C. After nineteen days the reaction was filtered and the filtrate chromatographed on a 2.5 x 13cm column containing resin Dowex-1-hydroxide. The resin was eluted with 90% methanol / water (v / v). The fractions containing the product were combined and the solvent was removed in vacuo. The residue was dissolved in 30% n-propanol / water (v / v) and chromatographed on a Bio-Rad P-2 column (7.5 x 90 cm). Product fractions were combined and, after lyophilization, 0.680 g of 6-ethyl (methyl) aminopurin-9-β-D-arabinofuranoside was obtained.

Analysis results: calculated for C ₁₃ H ₁₉ N ₅ O ₄
Calculated, C 50.48; H, 6.19; N, 22.64.

Found, C, 50.36; H, 6.25; N, 22.52
NMR and mass spectra are consistent with such a structure.

 PRI me R 11. 9-β-D-Arabinofuranosyl-2-amino-6-methoxypurine.

2-amino-6-methoxypurine obtained by nucleophilic substitution of a chlorine group by 2-amino-6-chloropurine (Sigma Camicel, St. Louis, Mo.) using methanol with sodium hydride in tetrahydrofuran (6.4 mmol, 1, 05 g) was combined with 35 ml of a solution of uracil arabinoside (7.04 mmol 1.75 g) in 10 mM potassium phosphate and 7% n-propanol (o / o). pH was provided at a level of 6.75. Purified purine nucleoside phosphorylase (18000 units) and uridine phosphorylase (1020 units), and the solution was incubated at ³⁷ ° C. After 26 days the reaction was filtered and the filtrate chromatographed on a column of resin Dowex-1-formate (2 x 7cm ) after adjusting the pH to 10.5 using concentrated ammonium oxide hydrate. The column was eluted using a 7% n-propanol / water (v / v) mixture and the fractions containing the product were combined and the solvent was removed in vacuo. The residue was extracted with 25 ml of water and the filtrate was separated from the solid by centrifugation. The upper layer after keeping at ambient temperature gave crystals of 2-amino-6-methoxy-9-β-D-arabinofuranoside, which gave, after drying under vacuum, 0.327 g of the product.

Analysis results: calculated for C ₁₁ H ₁₅ N ₅ O ₅
Calculated, C 44.44; H 5.09; N, 23.56
Found, C, 44.49; H 5.13; N, 23.52.

 NMR and mass spectra confirmed this structure.

 PRI me R 12. 9-β-D-Arabinofuranosyl-6-n-propoxypurine.

6-n-Propoxypurin (5.6 mmol, 1 g), (Sigma Chemical, St. Louis, Mo.), was combined with 545 ml of a solution of uracil arabinoside (10.1 mmol) in 10 mM potassium phosphate and 7% n- propanol (o / o). Purified uridine phosphorylase (680 m. F.) And purine nucleoside phosphorylase (12,000 IU), and the reaction was stirred at 35 ° ^C. The reaction was filtered after 58 days and the filtrate stored at 3 ° ^C for 20 hours The resulting precipitate was collected by centrifugation, dissolved in 30% n-propanol / water (v / v) and chromatographed on a Dowex-1-formate column (2.5 x 5 cm) after the pH was adjusted to 10 , 5 using a concentrated solution of ammonium hydrate.

 The column was eluted with 30% n-propanol / water (v / v) and the fractions containing the product were combined and the solvent was removed in vacuo. The residue was dissolved in n-propanol and subjected to chromatography on a column containing Bio-Rad P-2 (5 x 90 cm). The column was eluted with 30% n-propanol / water (v / v). The content of the product fraction was combined and, after removal of the solvent under vacuum, the residue was dissolved in water and chromatographed on a column containing Bio-Rad P-2 (5 x 90 cm). The column was eluted with water. The product containing fractions were combined and, after lyophilization, 0.758 g of 6-n-propoxypurin-9-β-D-arabinofuranoside was obtained in the form of a monohydrate.

Analysis data: calculated for C ₁₃ H ₁₈ N ₄ O ₅ ˙ 1.0 N ₂ O
Calculated, C 47.56; H, 6.14; N 17.06
Found, C 47.63; H, 6.13; N, 17.11.

 NMR and mass spectra are consistent with this structure.

 PRI me R 13. 9- (5-0-Benzoyl-β-D-arabinofuranosyl) -6-methoxy-9H-purine.

9- (β-D-arabinofuranosyl) -6-methoxy-9H-purine of Example 2 (0.283 g, 1.0 mmol) was dissolved in anhydrous dimethylacetamide (5,0ml), was rapidly cooled ^to 4 ° C and benzoyl chloride (0.155 g, 1.1 mmol). The mixture was stirred for 24 hours under argon, then allowed to slowly warm to room temperature. Then, 1.01 equivalents of benzoyl chloride was added at room temperature and stirring was continued for another 24 hours. The reaction was quickly quenched by pouring on 50 ml of ice-water, extracted with CHCl ₃ (3 x 30 ml) and the organic extracts dried ( magnesium sulfate). The evaporation residue was purified by flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) with a successive gradient from CHCl ₃ to CHCl ₃ : acetone 1: 1. Fractions containing the product with Rf 0,21 (silica gel, CHCl _3: acetone 1: 1) were combined, to give 92 mg of the title compound: temperature of the melting point: 202-204 ° ^C.

Analysis results: calculated for C ₁₈ H ₁₈ N ₄ O ₆
Calculated, C 55.96; H 4.70; N 14.50
Found, C 56.04; H 4.74; N, 14.40.

 NMR and mass spectra confirm the above structure.

 PRI me R 14. 6-Methoxy-9- [5-0 - (- 4-methylphenylsulfonyl) -β-D-arabinofuranosyl] -9H-purine.

4-Toluenesulfonyl chloride (0.312 g, 1.63 mmol) immediately after recrystallization and 9- (β-D-arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.308 g, 1.09 mmol) were suspended in anhydrous acetonitrile (25 mL) cooled to 3 ° ^C in an ice bath, and pyridine (5.0 ml) to dissolve the nucleoside. The solution was stirred under argon at 3 ° ^C for 1 hour, then kept at ^-15 ° C for 42 hours. The reaction was quenched by adding 5% NaHCO ₃ (3 mL), evaporated to approximately 10 ml, then evaporated again with 95% ethanol. The residue was absorbed on a minimal amount of silica gel and added to the silica gel column for flash chromatography (25.0 g, 2.5 x 15 cm) in CH ₂ Cl ₂ : MeOH (15: 1). As a result of elution with such a solvent (450 ml), then CH ₂ Cl ₂ : MeOH (10: 1, 550 ml), three UV-absorbing materials were obtained. Fractions containing material with Rf 0.42 (silica gel, CH ₂ Cl ₂ : MeOH 10: 1) were collected and then purified on three consecutive silica gel plates for preparative chromatography, the first using CH ₂ Cl ₂ : MeOH (10 : 1) and acetone: CH ₂ Cl ₂ (1: 1) on the following two plates to obtain 88 mg of the target material in the form of a transparent glass: melting point: 177-181 ^about C.

Analysis results: calculated for С ₁₈ Н ₂₀ N ₄ O ₇ S˙ 0,15Н ₂ О
Calculated, C 49.23; H 4.66; N, 12.76.

 Found C, 49.28; H 4.71; N, 12.71.

 NMR and mass spectra are consistent with such a structure.

 PRI me R 15. 6-Methoxy-9- (5-0-methylsulfonyl-β-D-arabinofuranosyl) -9H-purine.

The 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.600 g, 2.13 mmol) was suspended in dry acetonitrile (40 ml) and anhydrous pyridine (8 ml) was added. The flask was cooled to -3 ^{° C} in an ice-salt bath and added methane sulfonyl chloride (0.16 mL, 2.13 mmol). After 25 min, the solution was quenched with water (3 mL) and concentrated to a volume of 10 ml, then repeatedly evaporated with several additions of ethanol, keeping the temperature in the region of below 38 ^o C during the entire time. Residual pyridine was removed by vacuum pump. The residue was applied to a flash chromatography silica gel column (25.0 g, 2.5 x 15 cm), equilibrated in CH ₂ Cl ₂ : MeOH (15: 1). This column was first eluted with 150 ml of this solvent, and then with CH ₂ Cl ₂ : MeOH (10: 1, 450 ml). Fractions containing material with Rf 0,34 (silica gel, CH ₂ Cl _2: MeOH 10: 1) to give 0.448 g of product (57%) as a white foam: temperature of the melting point: 177-181 ^C (dec.).

Analysis results: calculated for С ₁₂ Н ₁₆ N ₄ О ₇ Si˙0.5 Н ₂ О
Calculated, C 39.02; H 4.64; N 15.17
Found, C 39.02; H 4.66; N, 15.08.

 NMR and mass spectra confirm this structure.

 PRI me R 16. 9- [5-0- (4-Methylbenzoyl) - β-D-arabinofuranosyl] -6-methoxy-9H-purine.

 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3. (0.283 g, 1.0 mmol) was suspended in anhydrous acetonitrile (10.0 ml), pyridine (2.3 ml) was added to to obtain complete dissolution, then 4-methylbenzoyl chloride (0.170 g, 1.1 mmol) was added. The mixture was stirred at room temperature for 24 hours under argon, the reaction was quickly stopped by the addition of isopropanol (5 ml) and evaporated to dryness, and then evaporated again with ethanol (2 x 10 ml).

Next, the residue was purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) using a successive gradient from CHC ₃ to CHCl ₃ : acetone 1: 1. Fractions containing the product with Rf 0.41 (silica gel, CHCl ₃ : acetone 1: 1) were combined, whereby 132 mg of the target compound was obtained. Melting point: 127-128 ^о С.

Analysis results: calculated for С ₁₉ Н ₂₀ N ₄ O ₆ ˙0.5 Н ₂ О
Calculated, C 55.74; H 5.17; N 13.69
Found, C 55.48; H 5.36; N, 13.64.

 NMR and mass spectra confirmed the above structure.

 PRI me R 17. 9- [5-0- (4-Chlorobenzoyl) - β-D-arabinofuranosyl] -6-methoxy-9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.283 g, 1.0 mmol) was suspended in anhydrous acetonitrile (10.0 ml), then pyridine (2.3 ml) was added to to obtain complete dissolution, finally, 4-chlorobenzoyl chloride (0.193 g, 1.1 mol). The mixture was stirred at room temperature for 24 hours under argon, the reaction was quickly stopped by the addition of isopropanol (5 ml) and evaporated to dryness, then evaporated again with ethanol (2 x 10 ml). The residue was further purified by flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) using a consistent gradient from CHCl ₃ to CHCl ₃ : acetone (1: 1). Fractions containing the product with Rf 0,37 (silica gel, CHCl _3: acetone 1: 1) were combined to give 105 mg of the title compound: temperature of the melting point: 122-124 ° ^C.

Analysis results: calculated for C ₁₈ H ₁₇ CIN ₄ O ₆ ˙ 0.5 N ₂ O
Calculated, C 50.30; H, 4.22; N 13.04.

 Found, C, 50.20; H 4.28; N, 12.94.

 NMR and mass spectra confirmed the above structure.

 PRI me R 18. 9- [5-0- (4-Methoxybenzoyl) -β-D-arabinofuranosyl] -6-methoxy-9H-purine.

Example 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine (0.283 g, 1.0 mmol) was suspended in anhydrous acetonitrile (10.0 ml), pyridine (2.3 ml) was added to effect complete dissolution, then 4-methoxybenzoyl chloride (1.1 mmol) was added. The mixture was stirred at room temperature for 24 hours under argon and the reaction was quickly stopped by the addition of isopropanol (5 ml) and evaporated to dryness, then evaporated again with ethanol (2 x 10 ml). The residue was then purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) using a consistent gradient from CHCl ₃ to CHCl ₃ : acetone 1: 1. Fractions containing the product with Rf 0,30 silica gel, CHCl _3: acetone 1: 1 convenient connectivity to obtain 110 mg of the title compound: temperature of the melting point: 195-197 ° ^C.

Analysis results: calculated for С ₁₉ Н ₂₀ N ₄ O ₇ ˙ 0.25 Н ₂ О
Calculated, C 52.22; H 4.91; N, 13.31.

 Found, C 54.22; H 4.94; N, 13.30.

 NMR and mass spectra were consistent with such a structure.

 PRI me R 19. 5-Methoxy-9- (5-0-phenylacetal-β-D-arabinofuranosyl) -9H-purine.

The 9- (β-D-arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.300 g, 1.06 mmol) was suspended in acetonitrile (25 ml), then anhydrous pyridine (5 ml) was added. The solution was cooled to 3 ° ^C in an ice bath and added finilatsetil chloride (0.20 mL, 1.5 mmol). After stirring at 3 ° ^C for 1 h, the solution was cooled to ^-15 ° C and kept at this temperature for 40 hours. The reaction was quenched by adding 5% NaHCO ₃ (3 mL), concentrated to 10ml, and evaporated with the addition several times ethanol. The residue was taken up in a mixture of CH ₂ Cl ₂ : MeOH (15: 1) and applied to a flash chromatography silica gel column (25.0 g, 2.5x x 15 cm) located in the same solvent, 500 ml of the same solvent was eluted then with a mixture (15: 1) CH ₂ Cl _2: methanol (15: 1, 500 ml). Fractions with an Rf of 0.38 (silica, 10: 1, CH2Cl2: methanol) gave 0.128 g of crude product with impurities having higher Rf. As a result of subsequent purification by preparative thin layer chromatography using CH ₂ Cl ₂ : MeOH 10: 1, and then using a second plate using a mixture of acetone: CH ₂ Cl ₂ 1: 1, 0.102 g of the expected product is obtained in the form of a white foam: temperature melting points: 74-75 ^about C.

Analysis results: calculated for С ₁₉ Н ₂₀ N ₄ О ₆ ˙0.1 Н ₂ О
Calculated C 56.74; H 5.06; N, 13.93.

 Found, 56.74; H, 5.11; N, 13.90.

 NMR and mass spectra are consistent with the above structure.

 PRI me R 20. 6-Methoxy-9- (5-0-phenyloxyacetyl-β-D-arabinofuranosyl) -9H-purine.

The 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.322 g, 1.14 mmol) was suspended in dry acetonitrile (25 ml), then anhydrous pyridine (5 ml) was added. The solution was cooled to 3 ° ^C in an ice bath and added phenoxyacetyl chloride (0.24 ml, 1.7 mmol). After stirring for 2 hours at 3 ° C, ^the reaction was quenched with 5% aqueous NaHCO ₃ (2 mL) and concentrated to a volume of 10 ml and evaporated after several additions of ethanol. The residue was subjected to flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) in a mixture of CH ₂ Cl ₂ : methanol 15: 1). Elution with 400 ml of this solvent followed by 10: 1 CH ₂ Cl ₂ : methanol (500 ml) and 9: 1 CH ₂ Cl ₂ : methanol (200 ml) gave 0.213 g of crude product. This material was recrystallized from methanol and gave 0.101 g (20%) of a white crystalline material: thermal melting point 193-195 ° ^C.

Analysis results: calculated for С ₁₉ Н ₂₀ N ₄ O ₇ ˙ 0.05 Н ₂ О
Calculated, C 54.69; H 4.85; N, 13.43.

 Found, C 54.69; H 4.89; N, 13.40.

 NMR and mass spectra confirm this structure.

 PRI me R 21. 6-Methoxy-9- (5-0-methoxyacetyl-β-D-arabinofuranosyl) -9H-pyrine.

The 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.300 g, 1.06 mmol) was suspended in acetonitrile (25 ml), and then anhydrous pyridine (5 ml) was added. The solution was cooled to -3 ^{° C} in an ice-salt bath and methoxyacetyl chloride was added (0,10ml, 1.1 mmol). After stirring in an ice-salt bath for 2 hours, the reaction was stopped with 5% NaHCO ₃ (2 ml), the volume was concentrated to 10 ml, and then evaporated after several additions of ethanol. The residue was applied to silica gel column for operational chromatography (25.0 g, 2.5 x 15 cm) contained in a 10: 1 mixture of CH ₂ Cl ₂ : MeOH. As a result of elution with 500 ml of this solvent, then with a 9: 1 mixture of CH ₂ Cl ₂ : MeOH (300 ml), 0.086 g of crude product was obtained. This material was recrystallized from MeOH and water, after which the obtained 0.035 g (9.3%) of white crystals: the temperature of the melting point: 137-139 ° ^C.

Analysis results: calculated for C ₁₄ H ₁₈ N ₄ O ₇ ˙ 0.5 N ₂ O
Calculated, C 46.28; H 5.27; N, 15.42.

Found, C 46.33; H 5.27; N 15.37
NMR and mass spectra confirm this structure.

 PRI me R 22. 9- [5-0- (4-Nitrobenzoyl) - β-D-arabinofuranosyl] -6-methoxy-9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.283 g, 1.0 mmol) was suspended in anhydrous acetonitrile (10.0 ml), pyridine (2.3 ml) was added to effect complete dissolution, then 4-nitrobenzoyl chloride (0.205 g, 1.1 mmol). The mixture was stirred at room temperature for 24 hours under argon, the reaction was quickly quenched by the addition of isopropanol (5 ml) and evaporated to dryness, then evaporated two more times with ethanol (2 x 10 ml). The residue was then purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) and a sequential gradient of CHCl ₃ to CHCl ₃ : acetone 1: 1). Fractions containing the product with Rf 0,37 (silica gel, CHCl _3: acetone 1: 1) were combined, yielding 105 mg of the title compound: melting point temperature of 202-203 ° ^C.

Analysis results: calculated for C ₁₈ H ₁₇ N ₄ O ₈ .

 Calculated, C 50.12; H 3.97; N, 16.24.

Found, C, 50.21; H 4.02; N, 16.16
NMR and mass spectra confirm this structure.

 PRI me R 23. 6-Methoxy-9- (5-0-pentanoyl-β-D-arabinofuranosyl) -9H-purine.

The 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.852 g, 3.01 mmol) was suspended in dry acetonitrile (75 ml) and dry pyridine (15 ml) was added. The solution was cooled in an ice-water bath and pentanoyl chloride (0.4 ml, 3.31 mmol) was added. The reaction was quickly stopped after 2 hours by the addition of 3 ml of methanol and evaporation to a clear, viscous oil. The residue was purified using flash chromatography on silica gel and a sequential gradient of CHCl ₃ to CHCl ₃ : MeOH 9: 1. The 330 mg product contained impurities of the corresponding 2'-ester (see Example 32) and unknown material with lower Rf. Then the residue was purified using reverse phase preparative liquid chromatography (high performance) (HPLC) (Alltech C ₁₈ , 10 x 25 mm, particle size 10 μm 70% H ₂ O: 30% CH ₃ CN, 4.0 ml / min), preparing a solution of 10 mg / ml in the same solvent with an injection volume of 1.0 ml. The residue was again evaporated with acetone to yield 260 mg of a white brittle foam (24%): melting point temperature: 85-95 ^° C

Analysis results for C ₁₆ H ₂₂ N ₄ O ₆ x x 0.05 (CH ₃ ) ₂ CO˙0.55 H ₂ O.

 Calculated, C 51.16; H, 6.22; N, 14.78.

 Found, C, 50.98; H 6.03; N, 14.63.

 NMR and mass spectrum confirm the indicated structure.

 PRI me R 24. 9- [5-0- (4-Aminobenzoyl-β-D-arabinofuranosyl)] - 6-methoxy-9H-purine.

9- [5-0- (4-Nitrobenzoyl) - β-D-arabinofuranosyl] -6-methoxy-9H-purine of Example 22 (0.350 g, 0.81 mmol) was suspended in ethanol (100.0 ml) and 10 was added % palladium on carbon (0.100 g). After alternately pumping and filling the system with hydrogen, the reaction was shaken at a Parr installation under a pressure of 50 psi (3.515 kg / cm ² ) for 3 hours. Then the mixture was filtered through Celite and the filtrate was evaporated to dryness. The residue after evaporation was suspended in methanol and filtered to give the title compound as a white solid (0.285 g, 88%): melting point temperature: 198% -200 ^o C.

Analysis results for C ₁₈ H ₁₉ N ₅ O ₆ x x 0.3 H ₂ O
Calculated, C 53.15; H 4.86; N, 17.22.

Found, C, 52.96; H 4.64; N 17.07
Mass and NMR spectra confirm this structure.

 PRI me R 25. 6-Methoxy-9- (5-0-propionyl-β-D-arabinofuranosyl) -9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.847 g, 3.0 mmol) was suspended in anhydrous acetonitrile (30.0 ml) and pyridine (9 ml) was added to allow complete dissolution . After cooling ^to 5 ° C was added propionyl chloride (0.305 g, 3.3 mmol) and the mixture was stirred overnight under argon while warming to 13 ^C. After the reaction terminated with isopropanol (5 ml) and evaporation to dryness, additional evaporation with ethanol (2 x 10 ml), the residue was purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) and a successive gradient from CHCl ₃ to CHCl ₃ : acetone 1: 1. Fractions containing a product with Rf 0.38 (silica gel, CHCl ₃ : acetone 1: 1 were combined and this material was again purified using silica gel chromatography under medium pressure (tandem columns, 1.5 x 25.0 cm and 1.5 x100,0 x cm; CHCl _3: acetone 3: 1) to give 0.114 g of the title compound as a white solid: melting point temperature: 62-64 ^° C

Analysis results: calculated for C ₁₄ H ₁₈ N ₄ O ₆ ˙ 0.5 CH ₃ OH
Calculated, C 50.68; H 5.76; N, 15.25.

 Found, C, 50.72; H 5.80; N, 15.28.

 NMR and mass spectra confirm this structure.

 PRI me R 26. 9- (5-0-Butanoyl-β-D-arabinofuranosyl) -6-methoxy-9H-purine.

9- (β-D-arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.847 g, 3.0 mmol) was suspended in anhydrous acetonitrile (30.0 ml) and pyridine (9.0 ml) was added to effect complete dissolution. After cooling ^to 5 ° C was added butyryl chloride (0.352 g, 3.3 mmol) and the mixture was stirred overnight under argon while warming to 13 ^C. After the reaction terminated with isopropanol (5 ml) and evaporation to dryness, the subsequent evaporation with ethanol (2 x 10 ml), the residue was purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) and a sequential gradient of CHCl ₃ to CHCl ₃ : acetone 1: 1. Fractions containing the product with Rf 0.38 (silica gel, CHCl ₃ : acetone 1: 1) were combined. This material was purified again using chromatography under a pressure of medium on silica gel (tandem columns, 1.5 x 25.0 cm and 1.5 x 100.0 cm; CHCl ₃ : acetone 2: 1), whereby 267 mg of the expected compounds in the form of a white solid: melting point 108-110 ^about C.

The results of the analysis for C ₁₅ N ₂₀ N ₄ O ₆
Calculated C 51.13; H 5.72; N 15.90
Found, C 51.21; H 5.73; N, 15.81.

 NMR and mass spectrum confirmed the indicated structure.

 PRI me R 27. 9- [5-0- (2,2-Dimethylpropionyl) -β-D-arabinofuranosyl] -6-methoxy-9H-purine.

The 9- [β-D-Arabinofuranosyl] -6-methoxy-9H-purine of Example 3 (0.850 g, 3.01 mmol) was suspended in dry acetonitrile (75 ml) and pyridine (15 ml) was added. The solution was cooled to 3 ^° C in an ice bath and 2,2-dimethylpropionyl chloride (0.41 ml, 3.3 mmol) was added. After stirring at 3 ° ^C for 6 hours, the reaction was stopped quickly using MeOH (2 mL) and concentrated to a volume of 10 ml, and then repeatedly evaporated after several additions of ethanol. The residue was applied to a flash chromatography column on silica gel (20.0 g, 2.5 x 12.0 cm) placed in CH ₂ Cl ₂ : MeOH 10: 1. Elution with 300 ml of this solvent gave 0.464 g of the starting material. material and 0.553 g of crude product.

As a result of another purification by flash chromatography (silica, 2.5 x 10.0 cm), eluting with a gradient from CH ₂ Cl ₂ to MeOH in CH ₂ Cl ₂ 1: 9, 0.419 g (38%) of clear glass was obtained : melting point temperature: 73-76 ^about C.

Analysis results for C ₁₆ H ₂₂ N ₄ O ₆ x x 0.5 N ₂ O
Calculated C 51.19; H, 6.18; N, 14.93.

Found, 51.43; H, 6.06; N 14.91
NMR and mass spectrum are consistent with this structure.

 PRI me R 28. 9- (5-0-Acetyl-β-D-arabinofuranosyl) -6-methoxy-9H-purine.

The 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (0.850 g, 3.01 mmol) was suspended in dry acetonitrile (75 ml), then anhydrous pyridine (15 ml) was added. The solution was cooled to 3 ^° C in an ice bath and acetyl chloride (0.24 ml, 3.4 mmol) was added. After stirring in an ice-bath, the reaction was interrupted for half an hour with methanol (2 ml), concentrated to a volume of 10 ml, then evaporated several times after several additions of ethanol. The residue was purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm), eluting with a mixture of CH ₂ Cl ₂ : MeOH (10: 1, 300 ml) to give 0.710 g of crude product. The second column (25.0 g, 2.5 x 15 cm) after elution with a mixture of CH ₂ Cl ₂ : MeOH (15: 1) gave 0.610 g of clear glass, which nevertheless contained a small amount of impurities with higher Rf. Subsequent purification of the product using preparative thin layer chromatography using CH ₂ Cl ₂ : MeOH 9: 1, then by loading flat layers onto an on-line chromatography column (20.0 g 2.5 x 12 cm) in CH ₂ Cl ₂ and eluting the gradient of MeOH in CH ₂ Cl ₂ received the target product in the form of a white foam (0.308 g, 31%): melting point: 64-67 ^about C.

Analysis results for C ₁₃ H ₁₆ N ₄ O ₆ x x 0.5 N ₂ O
Calculated, C 46.85; H 5.14; N 16.81
Found, C, 47.04; H, 5.12; N, 16.72.

 NMR and mass spectra are consistent with the structure.

 PRI me R 29. 6-Methoxy-9- [5-0- (2-methylpropionyl) -β-D-arabinofuranosyl] 9H-purine.

Example 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine (0.500 g, 1.77 mmol) was suspended in dry acetonitrile (25 ml), anhydrous pyridine (5 ml) was added. The solution was cooled to 3 ^° C in an ice bath and isobutyryl chloride (0.21 ml, 2.0 mmol) was added. After stirring at 3 ° ^C for 3 hours, the reaction was stopped quickly using MeOH (2 mL), concentrated to 10ml, and then evaporated several times after several additions of ethanol. The residue was applied to a flash chromatography column on silica gel (25.0 g, 2.5 x 15 cm) placed in CH ₂ Cl ₂ : MeOH 10: 1. After elution with 300 ml of this solvent, 0.167 g of starting material and 0.221 g of crude product were obtained. As a result of subsequent purification of the product using preparative thin layer chromatography with CH ₂ Cl ₂ : MeOH 9: 1, subsequent loading of flat layers onto a column for flash chromatography on silica gel (16.0 g, 2.5 x 10 cm), placed in CH ₂ Cl ₂ . The column was eluted with a gradient of MeOH in CH ₂ Cl ₂ to give 0.127 g of a clear glass (20%): melting point temperature of ^about 68-71 C.

Analysis results: calculated for C ₁₅ H ₂₀ N ₄ O ₆ ˙ 0.25 H ₂ O 0.05 MeOH
Calculated, C 50.43; H 5.82; N, 15.63.

 Found, C, 50.49; H 5.83; N, 15.62.

 NMR and mass spectra confirmed the indicated structure.

PRI me R 30. 6-Methoxy-9- [2-0- (2,2-dimethylpropionyl) -β-D-arabinofuranosyl] 9H-purine
9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (298.5 mg, 1.06 mmol), 4-dimethylaminopyridine (1 mg, 10 mmol), acetonitrile (20 ml) and pyridine ( 5 ml) was added to a three-necked round-bottom flask equipped with a thermometer, an argon supply valve, a magnetic stirrer, a refrigerator and a jacket. Was added trimethylacetic anhydride (0.22 mL, 1.06 mmol) and the solution was heated to a temperature of 40 ^{° C} to 26 hours. The reaction was quenched at this point by adding 2ml of water and evaporating to a clear oil. The residue was purified by flash chromatography on silica gel using a gradient sequence of CHCl ₃ to CHCl ₃ MeOH 9: 1. This material was further purified using preparative thin layer chromatography and CHCl ₃ MeOH 9: 1, and then evaporated with carbon trichloride and acetone, resulting in 90 mg of a white brittle foam.

Analysis results: calculated for C ₁₆ H ₂₂ N ₄ O ₆ ˙ 0.05 (CH ₃ ) ₂ CO ˙ 0.45 CCl ₄ . Calculated, C 45.47; H 5.13; N, 12.78.

Found, C 45.67; H 5.27; N, 12.67
NMR and mass spectra confirm the above structure.

 PRI me R 31. 6-Methoxy-9 - [(2,3,5-tri-0-acetyl) -β-D-arabinofuranosyl] -9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (1.009 g, 3.57 mmol), 4-dimethylaminopyridine (0.0098 g, 80 mol), triethylamine (0.59 ml, 4 , 2 mmol) and anhydrous acetonitrile (52 ml) were added to a three-necked round-bottomed flask equipped with a thermometer, argon inlet, magnetic stirrer and dry ice-acetone bath. After white solution was cooled to ^-20 ° C, was added acetic anhydride (2.4 mL, 25.4 mmol). The solution instantly became transparent. After 5 minutes, the reaction was stopped by the addition of MeOH (5 ml) and evaporated to a clear viscous oil. As a result of processing using flash chromatography on silica gel with a gradient of CHCl ₃ and MeOH, the product was obtained as a clear, viscous oil. By dissolving this material in a minimum amount of acetone, diluting with water and lyophilization, 1.03 g (71%) of the product was isolated in the form of a white powder.

Analysis results for C ₁₇ H ₂₀ N ₄ O ₈
Calculated C 50.00; H 4.94; N 13.72
Found, C, 49.80; H 5.09; N 13.50
NMR and mass spectra confirmed the above structure.

 PRI me R 32. 6-Methoxy-9- (2-0-pentanoyl-β-D-arabinofuranosyl) -9H-purine.

Method 1. 9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (283 mg, 1.0 mmol), acetonitrile (20 ml) and pyridine (5 ml) were added to a round neck three-necked flask. equipped with a thermometer, a valve for supplying argon and a magnetic stirrer. Valerian anhydride was added (0.2 mL, 1.0 mmol) and the solution was stirred ^at 20 ° C for 3 hours. The reaction was quenched with water (2 ml) and evaporated to a clear oil, then purified by flash chromatography on silica and gradient sequences from CHCl ₃ to CHCl ₃ : MeOH 9: 1. As a result, 110 mg of a 2'-ester with impurities of a 5'-ester was obtained (see Example 23), a material with a lower Rf. This material was further purified by preparative thin-layer chromatography on silica with CHCl ₃ : MeOH 9: 1 and coevaporation with acetone, whereby 70 mg of 2'-ester was obtained in the form of a clear glass.

Analysis results for C ₁₆ H ₂₂ N ₄ O ₆ x x 0.5 (CH ₃ ) ₂ CO.

 Calculated, C 53.16; H 6.37; N, 14.17.

 Found, C, 52.89; H 6.37; N, 13.96.

 NMR and mass spectra are consistent with this structure.

Method 2.5. 5-Methoxy-9- [3,5-0- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D-arabinofuranosyl] -9H-purine of Example 40 (2 5 g, 4.8 mmol) was added to a round-bottomed flask with a capacity of 250 ml together with 4-dimethylaminopyridine (0.06 g, 0.47 mmol). Dry acetonitrile (30 ml) and triethylamine (2.65 ml) were added, and acetonitrile (20 ml) and pentanoic anhydride (1.13 ml, 5.7 mmol) were charged through a funnel for continuous supply. The reaction mixture was cooled to 3 ° ^C in an ice bath under argon flow. The pentanoic anhydride solution was slowly added over 2 hours. The reaction mixture was then treated with methanol (10 ml) and concentrated under reduced pressure. The residue was taken up in CHCl ₃ (200 ml) and extracted with water (2 x x50 ml). The combined aqueous layers were again extracted with CHCl ₃ (25 ml) and the combined organic layers were dried (magnesium sulfate), filtered and concentrated to give 3.44 g of crude product.

The crude 6-methoxy-9- [2-0-pentanoyl-3,5-0- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D-arabinofuranosyl] -9H-purine was dissolved in tetrahydrofuran (120 ml) and water (3 ml). The solution was cooled to 3 ^° C in an ice bath and a 1.0 M solution of tetrabutylammonium fluoride in tetrahydrofuran (6.0 ml) was added. After 1.5 hours, a saturated solution of ammonium chloride (2 ml) was added and the reaction mixture was directly passed to a silica gel purification column (5.0 x 8.0 cm) in chloroform. The column was eluted with chloroform (200 ml), then with a 1: 1 mixture of acetone: chloroform (400 ml) and all fractions containing the product were combined and concentrated. Final purification was carried out on a flash chromatography column on silica gel (5.0 x 15 cm), eluting with a successive gradient of acetone in CHCl ₃ , whereby 1.17 g (67%) of a clear, sticky glass with pentanoic acid impurities was obtained.

Analysis results for C ₁₆ H ₂₂ N ₄ O ₆ x x 0.2 C ₅ H ₁₀ O ₂ .

Calculated, C 52.79; H, 6.25; N, 14.48
Found, C, 52.97; H 6.38; N, 14.60.

 PRI me R 33. 9- (2-0-Butanoyl-β-D-arabinofuranosyl) -6-methoxy-9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (283 mg, 1.0 mmol), acetonitrile (20 ml) and dry pyridine (5 ml) were added to a three-necked round-bottomed flask equipped with thermometer, argon inlet, magnetic stirrer, and cooled to 4 ° ^C was added butyric anhydride (170 l, 1.0 mmol) and the solution was stirred at ⁵ ° C for 6 hours under argon. At this point, the reaction was stopped by adding 2 ml of water and evaporated to a clear oil. The reaction mixture was purified by flash chromatography using a sequential gradient of CHCl ₃ to 1: 1 CHCl ₃ acetone. This material was further purified by preparative thin-layer chromatography on silica gel using a 9: 1 CHCl ₃ : MeOH mixture to give 90 mg of the 2'-ester as a clear oil.

Analysis results for C ₁₅ H ₂₀ N ₄ O ₆ x x 0.3 H ₂ O.

 Calculated, C 50.36; H 5.80; N, 15.66.

 Found, 50.36; H 5.81; N, 15.66.

 NMR and mass spectra corresponded to the indicated structure.

 PRI me R 34. 6-Methoxy-9- [2-0- (2-methylpropionyl) -β-D-arabinofuranosyl] -9H-purine.

9- (β-D-arabinofuranosyl) -6-methoxy-9H-purine of Example 3 (290.0 mg, 1.03 mmol), acetonitrile (20 ml) and pyridine (5 ml) were added to a three-necked round-bottom flask, equipped with a thermometer, argon input, a magnetic stirrer, a refrigerator and a shirt. Was added isobutyric anhydride (170 l, 1.0 mmol) and the solution was heated to a jacket temperature of 30 ^{° C} for 4 hours. The reaction was quenched at this point by adding 2ml of water and evaporating to a clear oil. The residue was purified by flash chromatography on silica gel using a gradient sequence from CHCl ₃ to CHCl ₃ MeOH 9 1. Fractions containing the product with Rf 0.54 (silica gel, CHCl ₃ MeOH 20: 1), then purified by preparative thin layer silica gel chromatography with CHCl ₃ MeOH 9: 1. As a result, 90.0 mg of 2'-ester was obtained in the form of a clear glass.

Analysis results for C ₁₅ H ₂₀ N ₄ O ₆ x x 0.1 C ₃ H ₆ O
Calculated, C 51.31; H 5.80; N, 15.64.

 Found, C, 51.41; H 5.81; N, 15.72.

 NMR and mass spectra confirmed the indicated structure.

PRI me R 35. 9- [3-0-Benzoyl-β-D-arabinofuranosyl (6-methoxy-9H-purin-9) 2,3-anhydro-β-D-face sofuranosyl] -6-methoxy - 9H-purine (prepared according to Example 36) (0.264 g, 1.0 mmol) was dissolved in anhydrous ethanol (20.0 ml), heated to reflux and ammonium benzoate (0.209 g, 1.5 mmol) was added under argon. An additional 1.5 mmol of ammonium benzoate was added after 24 hours and 35 hours. The reaction was evaporated to dryness after 48 hours under reflux. The evaporation residue was purified using flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) and a mixture of CHCl ₃ acetone 3: 1. Fractions containing the product with Rf 0.52 (silica gel, CHCl ₃ : acetone 1: 1) were combined, resulting in 138 mg of the target compound. The temperature of the melting point is 180-182 ^about C.

Analysis results calculated for C ₁₈ H ₁₈ N ₄ O ₆ .

Calculated, C 55.96; H 4.70; N 14.50
Found, C, 55.90; H 4.71; N, 14.44.

 NMR and mass spectra confirmed the above structure.

 PRI me R 36. 9- (2,3-Anhydro-β-D-lixofuranosyl) -6-methoxypurine.

Triphenylphosphine (5.902 g, 22.5 mmol) was dissolved in 1,4-dioxane (138 ml) and heated to 70 ^C. To the solution was added 9- (β-D-arabinofuranosyl) - 6-methoxy-9H-purin Example 3 (4.234 g, 15.0 mmol), the mixture was stirred for 10 minutes and diethyl azodicarboxylate (3.919 g, 22.5 mmol) in 1,4-dioxane (50 ml) was added dropwise over 10 minutes. The reaction was stirred at ⁷⁰ ° C for 1 h, cooled to room temperature and evaporated to a brown high viscosity oil. This material was purified by flash chromatography on silica gel (188.0 g, 5.0 x x 21.0 cm), eluting with a mixture of CHCl ₃ acetone (5 1, 3.6 L) and CHCl ₃ acetone (3: 1, 2, 0 l). Fractions containing materials with Rf 0.24 (silica, CHCl ₃ acetone, 1: 1) were combined and evaporated to dryness. The residue was purified using flash chromatography on silica (250.0 g, 5.0 x 28.0 cm), eluting with E OAc to give 2.452 g (62%) of the target compound as a white foam: point temperature melting point 144-145.5 ^about C.

Analysis results calculated for C ₁₁ H ₁₂ N ₄ O ₄ ˙ 0.25 C ₃ H ₆ O ˙ 0.05 CHCl ₃ . Calculated, C 49.78; H 4.80; N, 19.68.

 Found, C, 49.80; H 4.95; N, 19.87.

 NMR and mass spectra confirmed the indicated structure.

PRI me R 37. 6-Methoxy-9- [2-0- (4-methoxybenzoyl) -β-D-arabinofuranosyl] 9H-purine. 6-Methoxy-9- [2- (4-methoxybenzoyl) -3.5-0-] 1,1,3,3-tetraisopropyldisiloxane-1, 3-diyl] -β-D-arabinofuranosyl] -9H-purine ( obtained according to example 43) (0.86 g, 1.3 mmol) was dissolved in THF (40 ml) and water (1 ml) was added. The solution was cooled to 3 ^C and added 1.0 M tetrabutylammonium fluoride solution in THF (6.3 mL). After 30 min, an additional 5 ml of water was added, the reaction was reduced by half, and passed directly through a flash chromatography silica gel column (25.0 g, 2.5 x 15 cm) located in CH ₂ Cl ₂ . The column was eluted with CH ₂ Cl ₂ (200 ml), then with a mixture of CH ₂ Cl ₂ : MeOH (20: 1, 400 ml) and all fractions containing material with Rf 0.20 (silica, CH ₂ Cl ₂ MeOH 20: 1) were combined, whereby 0.570 g of a white foam was obtained. Final purification was carried out on a column for operational chromatography on silica gel (5.0 x 15 cm) located in CHCl ₃ .

After sequential elution gradient of acetone in chloroform to give 0.325 g (60%) of product as a white foam: thermal melting point 71-75 ° ^C.

Analysis results calculated for C ₁₉ H ₂₀ N ₄ O ₇ ˙ 0.3 CHCl ₃ ˙ 0.25 C ₃ H ₆ O Calculated C 51.60; H 4.71; N, 12.00.

Found, C, 51.56; H 4.70; N 11.95
NMR and mass spectra are consistent with the indicated structure.

 PRI me R 38. 6-Methoxy-9 - [(2-0- (4-methylbenzoyl)] - β-D-arabinofuranosyl] -9H-purine.

6-Methoxy-9- [2- (4-methylbenzoyl) -3.5-0- [1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) -β-D-arabinofuranosyl] -9H-purine (prepared according to example 42) (0.85 g, 1.3 mmol) was dissolved in THF (40 ml) and water (1 ml). The solution was cooled to 3 ^C and added 1.0 M tetrabutylammonium fluoride solution in THF (5.0 mL). After 30 min, an additional 5 ml of water was added, the reaction volume was reduced by half, and passed directly through a flash chromatography silica gel column (20.0 g, 2.5 x 12 cm) located in CH ₂ Cl ₂ . The column was eluted using CH ₂ Cl ₂ (200 ml), then with a mixture of CH ₂ Cl ₂ MeOH (15: 1, 400 ml) and all fractions containing material with Rf 0.20 (silica, CH ₂ Cl ₂ : MeOH 20 : 1) connected.

The crude product was applied onto a second flash chromatography silica gel column (25.0 g, 2.5 x 18 cm) located in acetone CH ₂ Cl ₂ (1: 1). Elution with the same solvent gave 0.633 g of the product, which was then purified using preparative thin-layer chromatography (silica, acetone CH ₂ Cl ₂ 1: 1) and flash chromatography (25.0 g, 2.5 x 15 cm). The column was eluted with acetone gradient sequence in CHCl ₃ to give 0.243 g (47%) of a clear stekla: thermal melting point 69-73 ° ^C.

Analysis results calculated for C ₁₉ H ₂₀ N ₄ O ₆ ˙ 0.4 C ₃ H ₆ O ˙ 0.25 CHCl ₃ Calculated C 54.17; H 5.03; N, 12.36
Found, C 54.00; H 5.08; N, 12.30
NMR and mass spectrum confirmed the indicated structure.

 PRI me R 39. 9- [2-0- (4-Chlorobenzoyl) - β-D-arabinofuranosyl] -6-methoxy-9H-purine.

9- [2-4-Chlorobenzoyl- [3,5-0- (1,1,3,3-tetra-isopropyl disiloxane-1,3-diyl) -β-D-arabinofuranosyl] -6-methoxy-9H β-purine (prepared according to example 44) (1.07 g, 1.7 mmol) was dissolved in THF (40 ml) and water (1 ml). The solution was cooled to 3 ° ^C and 1.0 M solution of tetrabutylammonium fluoride in THF (4.5 mL). After 30 min, another 5 ml of water was added and the reaction mixture was passed directly through a flash chromatography silica gel column (20.0 g, 2.5 x 12 cm) located in CH ₂ Cl ₂ . This column was eluted with CH ₂ Cl ₂ (200 ml), then with a mixture of CH ₂ Cl ₂ : MeOH (20: 1, 400 ml) and all fractions containing material with Rf 0.20) silica, CH ₂ Cl ₂ : MeOH 20 : 1) connected. The crude product was applied onto a flash chromatography silica gel column (30.0 g, 2.5 x 18 cm) located in a mixture of acetone: CHCl ₃ (1: 1). After elution with the same solvent, 0.269 g of product was obtained, which was further purified using preparative thin-layer chromatography (silica, acetone: CHCl ₃ (2: 3) and flash chromatography (25.0 g, 2.5 x 15 cm). eluted sequence acetone in CHCl ₃ gradient analytical sample was prepared using the product transfer in anhydrous ether and precipitating with petroleum ether (40-60 ° ^C) and lyophilizing the precipitate to give 0.082 g (12%) of a white powder:. point temperature melting point 76-81 ^about C.

The results of the analysis calculated for C ₁₈ H ₁₇ N ₄ O ₆ Cl ˙1.20 H ₂ O ˙ 0.35 C ₃ H ₆ O Calculated, C 49.45; H 4.68; N, 12.11
Found, C 49.78; H 4.38; N, 11.88,
NMR and mass spectra confirm this structure.

 PRI me R 40. 6-Methoxy-9- [3,5-0- (1,2,3,3-tetraisopropyl-1,3-disiloxane-1,3-diyl) - β-D-arabinofuranosyl ] -9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine according to Example 3 (1.0 g, 3.54 mmol) and imidazole (0.965 g, 14.2 mmol) were dissolved in dry dimethylformamide (10 ml) and the solution cooled to 3 ^° C was added 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane (1.35 mL, 3.90 mmol) and the mixture was stirred under argon for 3 hours. Then the reaction was stopped by adding water (1 ml) and concentrated under reduced pressure. The residue was partitioned between ethyl acetate (150 ml) and water (50 ml), and the ethyl acetate layer was dried (magnesium sulfate), filtered and concentrated. The crude product was transferred into ethyl acetate and purified by flash chromatography on silica gel (25.0 g, 2.5 x 15 cm) in ethyl acetate. From the fraction with Rf 0.70 (silica, ethyl acetate), 1.48 g (80%) of the expected material are obtained in the form of a white solid.

UV _max. EtOH 248.4 nm.

 NMR and mass spectra confirmed the above structure.

 PRI me R 41. 6-Methoxy-9- [2-0- (2-aminobenzoyl) -β-D-arabinofuranosyl] -9H-purine.

9- (β-D-Arabinofuranosyl) -6-methoxy-9H-purine according to Example 3 (0.283 g, 1.0 mmol), acetonitrile (15 ml), p-carboxyanthranyl anhydride (0.189 g, 1.1 mmol) and sodium bicarbonate (84 mg, 1.0 mmol) was added to a three-necked round-bottomed flask equipped with a thermometer, an argon inlet, a magnetic stirrer, a refrigerator and a heating jacket. The suspension was refluxed for 3.5 hours, and then a second equivalent of p-carboxyanthranilic anhydride was added. After refluxing for another 1 h, the reaction was cooled to room temperature, filtered and the filtrate was evaporated to a foamy glass. The residue was purified using flash chromatography on silica gel and a successive gradient of CHCl ₃ to 9: 1 CHCl ₃ : MeOH. The result was 53.1 mg of 2'-ester in the form of brittle glass: melting point 91-93 ^about C.

Analysis results calculated for C ₁₈ H ₁₉ N ₅ O ₅ ˙ 0.3 H ₂ O ˙ 0.2 CH ₄ O
Calculated, C 52.90; H 4.98; N, 16.95.

 Found, C, 53.23; H 4.98; N, 17.21.

 NMR and mass spectra are consistent with this structure.

 PRI me R 42. 6-Methoxy-9- [2-0- [4-methylbenzoyl (-3.5-0-) 1,1,3,3-tetraisopropyl-disiloxane-1,3-diyl] β-D-arabinofuranosyl] -9H-purine.

6-Methoxy-9- [3,5-0- (1,1,3,3-tetraisopropyl-disiloxane-1,3-diyl) -β-D-arabino furanosyl] -9H-purine according to Example 40 (1, 0 g, 1.9 mmol) was added to a 100 ml three-neck round-bottom flask with 4-dimethylaminopyridine (0.02 g, 0.16 mmol), acetonitrile (25 ml) and triethylamine (0.4 ml) and the solution was cooled in ice bath to a temperature of 3 ° ^C for 5 min. Toluolhlorid added (0.30 mL, 2.3 mmol) and the mixture was stirred for 3 hours at a temperature of 3 ^° C was added an additional amount of triethylamine (0.5 ml) and toluene chloride (0.5 ml) and the mixture was heated to room temperature. After 7 hours, the mixture was treated with methanol (2 ml), concentrated under reduced pressure, and the residue was applied to a flash chromatography silica gel column (2.5 x 15 cm) in CH ₂ Cl ₂ . The column was eluted with a mixture of CH ₂ Cl ₂ MeOH (10: 1, 400 ml), whereby 0.90 g (74%) of the expected product was obtained.

Analysis calculated for C ₃₁ H ₄₆ N ₄ O ₇ Si ₂
Calculated, C 57.92; H, 7.21; N, 8.71.

Found, C 57.69; N, 7.43; N, 8.40
NMR and mass spectrum confirm the indicated structure.

 PRI me R 43. 6-Methoxy-9- [2-0- [4-methylbenzoyl (-3.5-0-) 1,1,3,3-tetraisopropyl-disiloxane-1,3-diyl] β-D-arabinofuranosyl] -9H-purine.

6-Methoxy-9- [3,5-0- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D-arabinofuranosyl] -9H-purine according to Example 40, ( 1.0 g, 1.9 mmol) was added to a 100 ml round bottom flask with 4-dimethylaminopyridine (0.02 g, 0.16 mmol), acetonitrile (25 ml) and triethylamine (0.4 ml), then added anosoyl chloride (0.35 ml, 2.5 mmol). After 4 hours at room temperature, the reaction mixture was treated with methanol (2 ml) and concentrated under reduced pressure. The residue was applied to a flash chromatography silica gel column (25.0 g, 2.5 x x15 cm) in CH ₂ Cl ₂ : MeOH (20: 1). Elution with the same solvent gave 0.930 g (74%) of the expected product.

Analysis calculated for C ₃₁ H ₄₆ N ₄ O ₈ Si ₂ ˙ 0.35 N ₂ O
Calculated, C 55.97; H 7.08; N, 8.42.

 Found, C 56.02; H 7.01; N, 8.32.

 PRI me R 44. 9- [2-0- (4-Chlorobenzoyl) -3.5-0- (1,1,3,3-tetraisopropyl disiloxane-1,3-diyl) - β-D-arabinofuranosyl ] -6-methoxy-9H-purine.

6-Methoxy-9- [3,5-0- (1,1,3,3-tetraisopropyldisiloxane-1,3-diyl) -β-D-arabinofuranosyl] -9H-purine according to Example 40 (1 0 g, 1.9 mmol) was added to a 250 ml round bottom flask with 4-dimethylaminopyridine (0.02 g, 0.16 mmol), acetonitrile (25 ml) and triethylamine (0.4 ml) and the solution was cooled in ice bath at 3 ° ^C for 5 min. 4-Chlorobenzoyl chloride (0.31 ml, 2.5 mmol) was added and the reaction mixture was removed from the ice bath. After 8 hours at room temperature, the solution was treated with MeOH (3 ml) and concentrated under reduced pressure. The residue was taken up in CH ₂ Cl ₂ and loaded onto a flash chromatography silica gel column (25.0 g, 2.5 x 15 cm) in the same solvent. The column was eluted with CH ₂ Cl ₂ (200 ml), then with a mixture of CH ₂ Cl ₂ : MeOH (20: 1, 400 ml), whereby 1.05 g (82%) of product was obtained, the uniformity of which was confirmed using a thin layer chromatography (silica, CH ₂ Cl ₂ : MeOH 20: 1, Rf 0.44).

Analysis calculated for C ₃₀ H ₄₃ N ₄ O ₇ ClSi ₂
Calculated, C 54.32; H 6.53; N, 8.45
Found, C 54.48; H 6.72; N, 8.28.

 PRI me R 45. 5'-Monophosphate ester of 9-β-D-arabinofuranosyl-6-dimethylamine-9H-purine.

 The 9'-β-D-arabinofuranosyl-6-dimethylamine-9H-purine 5'-monophosphate ester was synthesized by transferring the terminal phosphate group of 5'-adenosine triphosphate (ATP) to the 5'-OH-group of 9-β D-arabinofuranosyl- 6-dimethylamine-9H-purine catalyzed by thymidine kinase (TC) isolated from Varicella zoster virus (BBZ) (Fife, Molecular Pharmac. 21, p. 432, 1982).

In 0.5 ml of 0.02 M Tris-hydroxymethyl / aminomethane-HCl buffer, pH 7.5, 50 nmol of 9-β-D-arabinofuranosyl-6-dimethylamine-9H-purine, 55 nmol of ATP (Sigma Chemicals, St. Louis, MO), 55 nmol MgCl ₂ and 0.005 m.u. TK VVZ. The solution was incubated at 21 ^{° C} for 2.5 hours and then filtered through a "Centricon 10" (YM membrane with a 10K molecular weight cutoff obtained from Amicon Inc. Fira. Danvers, MA) to remove protein. The product was analyzed for the absorption range in ultraviolet light using anion exchange thin layer chromatography, which varied with Rf common for monophosphate.

 PRI me R 46. Arabinoside 5'-monophosphate-6-methoxypurine.

A nucleoside, 6-methoxypurin arabinoside, 1.5 g (4.9 mmol) was dissolved in 15 ml of triethyl phosphate and 4.8 ml of triethylamine. This solution was cooled to -10 ° ^C. With stirring, 0.91 ml of phosphorus oxychloride. After 10 min, an additional amount (0.3 ml) of phosphorus oxychloride was added. After 10 minutes, the reaction was interrupted by adding 100 ml of ice-cold 0.5 M solution of triethylamine in water. All of these reagents were obtained from Aldrich.

 The components of the final reaction mixture were separated by ion exchange chromatography on QAE-Sephadex (Pharmacy). The components were eluted with a gradient of 50 mM-1M ammonium bicarbonate, 5'-monophosphate-6-methoxy-purinarabinoside accounted for only about 10% of the material eluted from the column. To remove inorganic orthophosphates from the compound, ion-exchange chromatography on BioRAD-AG-1X8 resin with a gradual gradient of 100 mM-750 mM ammonium bicarbonate was used. The fractions containing this compound were evaporated once in vacuo to remove excess ammonium bicarbonate, then lyophilized.

 6-Methoxy arabinoside-5'-monophosphate was obtained in 3% yield (0.14 mmol, 50 mg). The UV spectrum (max. 250 nm and min. 221 nm at pH 7) and the base / phosphate ratio (1.00 / 1.08) were in agreement with the structure of the compound. It can be cleaved into a nucleoside using alkaline phosphatase and 5'-nucleosidase.

 PRI me R 47. 6-Methoxypurin arabinoside 5'-triphosphate.

The 5'-monophosphate-6-methoxypurin arabinoside of Example 46 (36 mg, 93 mmol) was dissolved in 2 ml of hexamethylphosphoramide. Carbonyldiimidazole (82 mg, 506 mmol) was added and the mixture was stirred for 2 hours at room temperature. Methanol (0.032 ml, 800 mmol) was added and the mixture was stirred for 20 minutes. Tributylammonium pyrophosphate (220 mg, 470 mmol from Sigma) was added, which was previously dissolved with 1.5 ml of hexamethylphosphoramide, and the mixture was stirred for 18 hours at room temperature. The reaction was stopped by adding 50 ml of water. All reagents mentioned were obtained from Aldrich, unless otherwise indicated. 5'-triphosphate was purified using ion exchange chromatography on QAE Sephadex (Pharmacy), eluting with a successive gradient of 50-800 mM ammonium bicarbonate. Fractions containing the compound were evaporated once in vacuo to remove excess ammonium bicarbonate then re-dissolved in water and kept frozen at -20 ° ^C.

 5'-triphosphate-6-methoxypurinarabinoside was obtained in 60% yield (57 mol, 30 mg), UV spectrum (max. 250 nm and min 223 nm at pH 7) and base / phosphate ratio (1.00 / 3 , 10) were in agreement with the structure of the compound. It can be cleaved into nucleoside using alkaline phosphatase and monophosphate using phosphodiesterase 1.

 PRI me R 48. 6-Dimethylamino-9 [(2-0-valeryl) -β-D-arabinosyl] -9H-purine.

6-Dimethylaminopurine arabinoside (0.501 g, 1.67 mmol) was dissolved in 20 ml of pyridine and 20 ml of DMF. The solution was brought ^to 4 ° C and added slowly valerian anhydride (0.43 mL, 2.17 mmol, Aldrich Kemikel Co. Milwaukee, WI) dissolved in 5 ml of pyridine and 5 ml of DMF. The temperature of the solution was maintained at 40 ^° C and after 2 hours the solvent was removed in vacuo. The residue was treated on a flash chromatography silica gel column, 4.8 x 20 cm with 150 ml of each of the following dichloromethane mixtures: methanol (v / v): 100: 0, 98: 2, 96: 4, 94: 6, 92 : 8, 90:10. Fractions containing the product were collected and, after lyophilization, 0.195 g of 6-dimethylamino-9- (2-0-valeryl-β-D-arabinosyl) -9H-purine was obtained: TCX Rf 0.55 / silica gel, 9: 1 (dichloromethane: methanol).

Analysis for C ₁₇ H ₂₅ N ₅ O ₅ ˙ 0.5 H ₂ O
Calculated, C 52.57; H 6.75; N 18.03
Found, C, 52.61; H 6.77; N, 17.99.

 NMR and mass spectra are consistent with this structure.

 PRI me R 49. 6-Dimethylamino-9- (2,3,5-triacetyl-β-D-arabinosyl) -9H-purine.

Dimetilaminopurinarabinozid-6 (3.00 g, 10.0 mmol) was dissolved in 20 ml acetonitrile and equilibrated at 4 ^C. 45 ml of pyridine. Acetyl chloride (2.85 ml, 40.08 mmol, Aldrich Chemical Co. Milwaukee, VI) in 7 ml of acetonitrile was added dropwise to the reaction mixture. After 5 hours, the solvent was removed under vacuum at a temperature below ⁴⁵ ° C, and the residual material is treated with a high vacuum pump for 24 hours The dried residue was further subjected to flash chromatography on a column of silica gel, 4.8 x 25 cm using a mixture of 9.: 1 (dichloromethane methanol). The product containing fractions were collected and, after lyophilization, 0.445 g of 6-dimethylamino-9- (2,3,5-triacetyl-β-D-arabinosyl) -9-purine was obtained.

 TLC Rf 0.61 (silica gel, 9: 1 dichloromethane methanol).

Analysis calculated for C ₁₈ H ₂₃ N ₅ O ₇
Calculated, C 51.30; H 5.50; N, 16.62.

 Found, C, 51.40; H 5.55; N, 16.54.

 NMR and mass spectra confirm this structure.

 PRI me R 50. Compositions in the form of tablets.

 The following compositions A and B were obtained by wet granulating the ingredients with a solution of posidone, then magnesium stearate was added and the mixture was pressed (see table 1).

 Tablets were prepared from the aforementioned ingredients (C) by wet granulation and subsequent compression. In the alternative preparation povidone B.F. can be replaced with polyvylpyrrolidone.

 The resulting compositions D and E were obtained by direct compression of the mixed ingredients. Lactose in composition E is of direct compression type (Dairy Crest "Zeparox" company).

Composition D mg / capsule: Active ingredient 250 Pre-gelatinized starch NF15 150
____
400
Composition E, mg / capsule: Active ingredient 250 Lactose 150 Avicel 100
____
500
Composition F (for controlled release of the composition).

The composition was obtained by wet granulation of the ingredients with a solution of povidone, and then magnesium stearate was added and pressed, mg / tablet: a) Active ingredient 500 b) Oxypropylmethyl cellulose 112 (Metocel K4M Premium) c) Lactose B.F. 53 d) Povidone B.F. 28 e) Magnesium stearate 7
____
700
The release of the drug takes place within about 6-8 hours and ends after 12 hours.

PRI me R 51. The composition is in the form of capsules
Composition A.

 A capsule composition was prepared by mixing the ingredients of composition 1 in Example 50 and filling in a two-part hard gelatin capsule mixture. The composition was obtained in a similar manner.

Composition, B, mg / capsule: a) Active ingredient 250 b) Lactose B.F. 143 c) Sodium glycolate starch 25 d) Magnesium stearate 2
_____
420
Composition C, mg / capsule: a) Active ingredient 250 b) Macrogol 4000 B. F. 350
___
600
Capsules were prepared by melting a macrogol 4000 B.F. dispersing the active ingredient in the melt and filling the melt in hard gelatin capsules, consisting of two parts.

Composition J, mg / capsule: Active ingredient 250 Lecithin 100 Peanut butter 100
___
450
Capsules were prepared by dispersing the active ingredient in lecithin and peanut oil and filling the dispersion with soft, elastic gelatin capsules.

 Composition E (controlled release).

 The following controlled release capsule composition was prepared by extruding ingredients a, b and using an extruder, then pellets were prepared from the resulting material and dried. The dried beads were then coated with a release control shell (d), which was placed in a two-piece hard gelatin capsule.

Composition E, mg / capsule: a) Active ingredient 250 b) Microcrystalline cellulose 125 c) Lactose B.F. 125 d) Ethyl cellulose 13
___
513
PRI me R 52. Ophthalmic solution. Active ingredient, g 0.5 Sodium chloride of analytical grade, g 0.9 Thiomersal, g 0.001 Purified water, ml Up to 100 pH is ensured at the level of 7.5
PRI me R 53. Composition for injection. Active ingredient, g 0,200 Sterile, pyrogen-free, phosphate buffer (pH 9.0), ml Up to 10
The active ingredient is dissolved in most of the phosphate buffer (35-40 ° ^C), then made up to volume and filtered through a sterile micropore filter into a 10 ml glass amber vial (type I), which was closed with sterile stopper and hermetic lid.

PRI me R 54. Intramuscular injection. Active ingredient, g 0.20 Benzyl alcohol, g 0.10 Glycofurol 75, g 1.45 Water for injection, ml Up to 3.00
The active ingredient was dissolved in glycofurol. Then benzyl alcohol was added and dissolution was carried out, then water was added to 3 ml. The mixture was then filtered through a sterile microporous filter and hermetically sealed in sterile 3 ml amber glass vials (type I).

PRI me R 55. Syrup suspension Active ingredient, g 0.25 Sorbitol solution, g 1.50 Glycerin, g 2.00 Dispersible cellulose, g 0.075 Sodium benzoate, g 0.005 Flavoring substance, Peach 17.42.3169, ml 0 , 0125 Purified water to a volume of 5.00 ml
Sodium benzoate was dissolved in a portion of purified water and a sorbitol solution was added. The active ingredient was added and dispersed. A thickening agent (dispersible cellulose) was dispersed in glycerin. Both of these dispersions were mixed and brought to the required volume using purified water. More thickening was provided, if necessary, by an increase in the slice of the suspension.

PRI me R 56. Suppository, mg / suspension: Active ingredient (63 m) 250 Refractory fat. B.F. (Witepsol X15-Dynamite Nobel) 1700
____
1950
The active ingredient was used in the form of a powder in which at least 90% of the particles have a diameter of 63 m or less.

One-fifth of Uitepsola X15 melted in a cup equipped with a jacket through which steam was passed, at ⁴⁵ ° C maximum. The active ingredient was sieved through a sieve with a hole diameter of 200 m and added to the molten base with stirring using a silverson equipped with a cutting head until a uniform dispersion was obtained.

Maintaining the mixture temperature at 45 ^C., was added Uitepsol X15 remaining in suspension and by means of mixing achieved a homogeneous mixture. The suspension was passed through completely with a diameter stainless steel sieve openings of 200 m and under continuous stirring, allowing the mixture to cool to 40 ^C. At a temperature in the range from 38 to 40 ^C. 2.02 g of the mixture is filled into suitable plastic molds. Further, the suppositories were allowed to cool to room temperature.

PRI me R 57. Pessaries, mg / pessary: Active ingredient 63 m 250 Anhydrous dextrose 380 Potato starch 363 Magnesium stearate 7
____
1000
These ingredients were directly mixed and pessaries were obtained by direct compression of the resulting mixture.

 PRI me R 58. Antiviral toxicity and bioavailability test.

 Determination of antiviral activity against the Baricella-Zoster virus.

 The inhibitory effect of the compounds on replication of VVZ (Oka strain) was studied using the ELISA procedure (Berkovits, F.E. and Levin, I, J. 1985), Antimicrob. Agents and Chemother.), Which was modified as follows. Infection was started in the presence of the drug, and not before it was added. At the end of the third day of incubation of the drug and the virus with uninfected cells (human diploid fibroblasts, strain MRG-5), 96-well plates were centrifuged for 5 min at a speed of 200 g to precipitate the separated cells before glutaraldehyde was fixed. In the modified ELISA, alkaline phosphatase-conjugated anti-human IgG was used as the second antibody. The rate of breakdown of para-nitrophenyl phosphate by dissecting alkaline phosphatase was determined using the procedure described in (Tadepally, S.M. Quinie, R.P. and Averett, D.R. 1986, Antimicrob, Agents and Chemother, 29, p. 93- 98), Uninfected cells were used to obtain “pure” reaction rates, which were subtracted from the rates obtained in the presence of the virus. This assay is suitable for detecting viral progeny in cultures that were initially infected with 15 to 3600 infectious particles per well.

 A study of growth inhibition of uninfected mammalian cells.

 The ability of the proposed compounds to inhibit the growth of D98 cells (human) and L-cells (mice) was measured by determining the number of cells with different concentrations of the compound (Rideut, J.L. et al. 1982, J. Med. Chem, 25, p. 1040 -1044). Then this number of cells was compared with the number obtained in the experiment without compound. Cell counting was carried out either by direct particle counting after trypsinization of the monolayer, or by spectrophotometric determination of the amount of living strain made up of cells.

Comparable results were obtained using both procedures. Data analysis. The concentration of the compound, resulting in 50% of the control values (IC ₅₀ ), was calculated either by direct interpolation from the log graphs of the concentration of the compounds relative to the percentage of the control value, or using a computer program that analyzes in accordance with the same algorithm. In these calculations, data in the range from 20 to 80% of the control were used (see Table 2).

 Analysis and data on bioavailability.

 Two Long Evans rats received a dose of the compound using a tube inserted into the stomach of Example 3 10 mg / kg or molar equivalents of the compounds of Examples 16, 24 and 32.

 Animals were placed in metabolic cells and urine was collected in periods from 0 to 24 hours and from 24 to 48 hours after dosing. Urine was analyzed by reverse phase high performance liquid chromatography. Results of a dose excreted from urine within 48 hours. Example Dose, 3 4.9 / 2.5 16 11 24 7.9 32 15.9

Claims (1)

The method of obtaining substituted purine arabinosides of General formula I

where R ₁ halogen, methoxy, ethoxy, propoxy, amino group, monosubstituted with methyl or C ₃ C ₆ -cycloalkyl, or disubstituted with C ₁ C ₅ -alkyl, or a nitrogen-containing heterocyclic ring containing 4 7 carbon atoms, linked to the purine radical through a nitrogen atom ;
R _{2 is} hydrogen, a halogen or an amino group,
or its pharmaceutically acceptable derivatives other than 2 ′, 3 ′, 5 ′ triacetate and 2 ′, 3 ′, 5 ′ tribenzyl derivatives, where R _{1 is} chloro or fluoro, R _{2 is} chloro, fluoro, hydrogen or amino, provided that when the target product is not a pharmaceutically acceptable derivative, as noted above, and R _{2 is} hydrogen, R ₁ cannot mean methoxy, methylamino, dimethylamino, piperidino or pyrrolidino, and when R _{2 is an} amino group, R ₁ cannot mean methylamino,
characterized in that the compound of General formula II

where R ₁ and R ₂ have the indicated meanings,
reacted with a compound of general formula III

where X is a pyrimidine or purine base,
with the exception of the bases of the specified general formula II and, if necessary, if the obtained compound is a compound of the general formula I, it is converted into a pharmaceutically acceptable derivative or, if the obtained compound is a pharmaceutically acceptable derivative, it is converted to another pharmaceutically acceptable derivative.

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