KR800001478B1 - Process for 1,2,4 triazole nucleosides - Google Patents

Abstract

Glycosylhydroxyl blocking triazole nucleoside (I; R1 =alkyl carboxylate or cyano group) was prepared by the reaction of 3-substituted 1,2,4-triazole (II; G = a part of pentofranosyl) with pentofranose of 1,2,3,5-o-acyl blocking sugar(III; AC =CH3CO-; Bz = C6H5CO-; R2 and R3 are H, 5'-phosphate, 3,5-phosphate or hydroxyl) under the pressure and in the presence of acid catalyst at 35≰C, and formed 3-substituted nucleoside of 3-thiocarboxamide or carboxamide(IV) by aminolysis and silylation.

Description

Preparation of 1,2,4-triazole nucleoside

The present invention provides a 1,2,4 triazole nucleoside derivative using a 3-substituted 1,2,4 triazole represented by the following general structural formula as a famous compound which is an antiviral agent as a starting material. It is related to the general formula as follows.

Wherein G is a pentofuranosyl moiety, R ₁ is cyano, carbox amido, thiocarbox amido, methyl carboxylate, carbox amidoxyl carbox, amidine or the physiologically acceptable latter Means acid addition salts.

The present invention also contains similar 0-acylated compounds and 5'-phosphates and 3 ', 5'- cyclic phosphates, as well as ammonium and alkali metal salts of phosphates.

The structural formula of these compounds is as follows.

In the formula, R ₁ has the meaning as described above, either R ₂ or R ₃ means hydrogen, and the other means 5′-phosphate and 3 ′, 5′-cyclic phosphate or hydroxyl group.

The compounds according to the invention exhibit antiviral activity and the compounds of the bisylated or deblocked structure exhibit greater water solubility than conventional nucleoside antiviral agents.

The 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carbox amide nucleoside of the present invention exhibits a large spectrum of antiviral as well as antitumoral properties.

Methyl 1,2,4-triazole-3-carboxylate and 3-cyano-1,2,4-triazole are used as starting materials for the synthesis of 1,2,4-triazole nucleosides.

The former compound is Latuija PSR Zinatnu Akad. Vestis. According to the process of Cipens and Grinsteins ′ introduced in Kim Ser (1965) [2] 204-08 (see CA 63, 1243, 1965) 3-cyano-1,2,4- It can be easily prepared by oxidizing the triazole and then esterifying the resulting acid.

3-cyano-1,2,4-triazole is described by Cipens et al. It can be prepared by the synthesis of many stages introduced in Supra, more conveniently, by addition reaction of cyanogen and hydrazine to give 1-cyanoform amidine hydrazide, followed by closure of triatyl orthoformate and acid catalyst ring To prepare a 3-cyano compound.

Methyl 1,2,4-triazole-3-carboxylate is produced by reaction with hexamethyl sirazane in the presence of an acid catalyst at 35 ° C. under reduced pressure, whereby 0-acylated haloche reacts with GX to produce 0-benzoylate. To prepare a mixture of 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester and 5-carboxylic acid methyl ester, the reaction scheme of these processes is as follows. .

In the above scheme, B _Z means benzoyl.

3-carboxylic acid methyl ester is separated by column chromatography on fractional crystals or silica gel.

Dibenzolation and amination result in 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide.

A good method of synthesizing the present invention is an acid catalyst fusion of substituted triazoles with 0-acylblocked sugars (1-0-acetyl-2,3,5-tri-0-benzoyl-β-D-ribopranose). Following deblocking and aminolysis, the reaction schemes are as follows.

AC means CH ₃ CO- and B _x means C ₆ H ₅ CO—. Fusion is optionally with 3-cyano-1,2,4-triazole and 3-cyano-1- (2,3,5-tri-0-acetyl-β-D-tibofuranosyl) -1 , 2,4-triazole is converted to active 3-carboxamido nucleoside by reaction with hydrogen peroxide in the presence of aqueous ammonia and fused with methyl 1,2,4-triazole-3-carboxylate Yield is smaller than when this is done.

3-cyano-1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole may be obtained by silylation or fusion process, but the present invention Active 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-thiocarboxamide by reaction with a hydrogen sulfide and diacylation in the presence of triethylamine useful as an intermediate of In a similar manner to generating nucleosides, nucleoside 1,2,4-triazole-3-carbox amidine is 3-carboxes from 3-cyano-1,2,4-triazole nucleosides. Regardless of whether or not intermediates of amidoxime are produced, their reaction schemes are as follows.

In the above scheme, G means glycosyl powder as described above.

Carbox amidine is obtained in free form from a synthesis comprising a carbox amidoxyl intermediate.

In contrast, a good preparation produces carboxamidine in the form of its hydrochloride salt directly from the 3-cyano compound in the presence of ammonium chloride and other physiologically soluble addition salts (e.g., hydrogen sulfide, hydrogen iodide, citric acid, conversion, Phosphoric acid addition salts) can be obtained by ion exchange or by neutralizing the hydrogen chloride with sodium bicarbonate and then reacting the resulting free carbox amidine with a suitable acid.

The 3-carbox amido, 3-thiocarbox amido and 3-carbox amidine nucleosides of the present invention can be converted to the ammonium or alkali metal salts of the corresponding 5'-phosphates or phosphates and treated. have.

In an analogous manner, active nucleosides are formed by forming 5-phosphates of the intermediates of the invention, 3-methyl carboxylate, 3-cyano and 3-carboxamidoxyl compounds, before acting on three sites of triazole aglycone. .

One skilled in the art knows enough how to phosphorylate nucleosides.

Specifically, the 5 'phosphate of 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide is 1- (β-D-ribofuranosyl)- It is formed by reacting 1,2,4-triazole-3-carboxylic acid methyl ester with POCl ₃ in trimethyl phosphate followed by hydrolysis in ice water to produce free 5-phosphate.

The formation of ammonium salts and the amination of methyl esters occur violently in treatment with aqueous ammonia.

Free 5-phosphate of alkali metal salts or 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide nucleotides can be obtained by ion exchange resins from the ammonium salt of phosphate have.

The 3,5-cyclphosphate of the active carbox amido, thiocarbox amido and carbox amidine compounds of the present invention for the strengthening of cell tissue transfer can be converted to the corresponding 5-phosphate dicyclohexyl carbodiimide (DCC). It is hardened by). Optionally, cyano, methyl carboxylate and carbox amidooxyl 5′-phosphate intermediates form the active cycle nucleotides by cyclization before they are acted on three sites of aglycone.

The cycle phosphate can be applied as in the case of 5'-phosphate in free form or in the form of ammonia or alkali metal salts.

While the present invention specifically describes acetyl or benzoyl which blocks 2 ', 3' and 5 'hydroxy fractions of glycosyls, any acyl protects these hydroxy from side reactions such as dehydrogenation in the synthesis of the active agents of the present invention. It can be seen that the use of acyl group to do.

As reported by Sutherland et al. Of Biochim et Biophys Acta 148, 106 (1967), acylation of cycle nucleotides enhances cell transfer.

In a similar manner, the nucleotides and nucleoside freeglycosyl hydroxyls of the invention can be acylated for enhanced solubility, e.g. alkyl or alkaline sulphates such as dosyl, mesyl, brosyl, nisyl, etc. The same result is obtained using a polyyl group.

It can be seen that the 1-degassing of the sugar reagent described above is used for the purpose of separating by-products from any degassing while characterized by acetyl or halo in the silylation process.

Although β-D-ribofuranosyl and kifuranosyl powder are mentioned in the examples of the above reference, other glycosyl powders are used in the present invention, that is, 2′-deoxy-β-D-ribofuranosyl-2′-deoxy- It can be seen that α-D-ribofuranosyl and β-D-arabino furanosyl can be used.

When using 2-dioxytriazole nucleoside sugar, 2-dioxy-1,3,5-tri-0-acetyl-3-ribofuranos, 2-dioxy-3,5-di-0 And compounds produced by treating-(paratoluoyl) -D-ribofuranosyl chloride with mercury acetate in tetrahydrofuran.

Arabinosyl triazole nucleosides can be prepared by a silylation process using 2,3,5-tri-0-benzyl-D-arabino-furanosyl chloride.

The biological activity of the ribofuranosyl triazole of the present invention and the activity in the substituted arabino furanosyl triazole are shown as follows.

In view of the antiviral properties of “Ara-A” and “Ara-C”, the biological activity of the corresponding 2-dioxy-β-D-ribofuranosyl triazoles so far known is 5-uredo-2′-di. It is implied by the antivirality known to date of oxyuridine.

Ribofuranosyl triazoles of the present invention are particularly effective against DNA bacteria such as helpers types 1 and 2.

These compounds are combined with the phosphorus free nucleosides lacking the trans 2-oxygen atom of the glycosyl fraction, since no antivirality is apparent when testing the virus in Example 16 infra and apparently no enzyme can be present in tissue culture. Thus these nucleosides combine with phosphorus to form active 5′-phosphate prior to treatment and optionally 5′-phosphate can be cyclized prior to treatment and generate active 5′-phosphate metabolites in VIVO cleavage.

Phosphorylation process is the same as the above-described ribolide manufacturing process.

The invention is explained in more detail according to the following examples.

In this example, all parts and percentages are by weight and all temperatures mean degrees Celsius unless otherwise inferred, and all processes are performed at 35 ° C. under reduced pressure in a rotary evaporator.

Example 1

[1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester and 1- (2,3,5 -Tri-0-benzoyl-β-D-ribofuranosyl) -1,2,4-triazole-5-carboxylic acid methyl ester (silication step)]

(A) Preparation of methyl N- (trimethylsilyl) -1,2,4-triazole-3-carboxylate

A suspension of methyl 1,2,4-triazole-3-carboxylate (14.0 g, 1.0 m mole) and hexamethyldisirazane (100 ml) are refluxed with stirring until ammonia production is complete (Ca 2 h).

Excess hexamethyl disyrazane is removed under reduced pressure to give 20.6 g (100%) of N-trimethyl silyl derivative of methyl 1,2,4-triazole-3-carboxylate.

(B) Halo Sugawaribosilization

Methyl N- (trimethylsilyl) -1,2,4-triazole-3-carboxylate (20.6 g, 110 m mole) and 2,3,5-tri-0-benzoyl-D-ribofuranosyl bromide (52.2 g, 100 m mole) was left in anhydrous acetonitrile (300 ml) for 3 days at 25 ° C.

The solution is removed and the residue is recrystallized from ethanol.

Recrystallization from ethyl acetate-ethanol and column chromatography of the chloroform and the filtrate on silica gel resulted in pure 1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl having a melting point of 137-139 ° C. ) -1,2,4-triazole-3-carboxylic acid methyl ester (25.1 g, 44.0%) is obtained.

The fast moving component from the silica gel column is crystallized from ethanol to give 1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl) -1,2,4-tria having a melting point of 112-124 ° C. Sol-5-carboxylic acid methyl ester (13.2 g, 23.1% is obtained).

Example 2

[1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester (fusion step).]

1,2,4-triazole-3-carboxylate (12.7 g, 100 m mole) and 1-0-acetyl-2,3,5-tri-0-benzoyl-β-D-ribofuranose (55,4 g , 110m mole) is heated in an oil bath while maintaining 160-165 ℃.

After the sugar is dissolved, bis (P-nitrophenyl) phosphate (400 mg) is added to the mixture with stirring to heat the mixture to 160-165 ° C. for 15-20 minutes under reduced pressure.

The residue is crystallized from ethyl acetate ethanol to give 42.5 g (74.5%) of product having a melting point of 137-139 ° C.

Example 3

[1- (2,3,5-Tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester]

1,2,4-triazole-3-carboxylate (12.7 g, 0.10 mole) and 1,2,3,5-tetra-0-acetyl-β-D-ribofuranose (31.8 g, 0.10 mol) Heat at 160-165 ° C in an oil bath until the sugar melts.

Bis (P-nitrophenyl) phosphate (250 mg) was added stock and heated to 160-165 ° C. and stirring continued for 15-20 minutes under reduced pressure.

The residue is dissolved in hot benzene and the solution is filtered and then cyclohexane is added to the filtrate to give a crystal product (30.0 g, 77.8%) with a melting point of 107-109 ° C.

Example 4

[1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester]

1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester (25.0 g, 43.8 m mol) and sodium The solution of methoxide (400 mg) dissolved in methanol (200 ml) was refluxed for 45 minutes.

The solution is neutralized by treatment with BiO-Red AG 50-X2 (H), filtered and the filtrate is concentrated to a syrup. The syrup is determined from methanol-methyl-acetate to give 8.9 g (70.5%) of product having a melting point of 117-119 ° C.

Example 5

(A) 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carbox amide.

[Method 1]

1- (2,3,5-tri-0-benzoyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester (16.0 g, 28.0 m mol) is methanol The solution contained in (300 ml, pre-saturated with anhydrous ammonia at 0 ° C.) is kept at 25 ° C. for 3 days in a sealing pressure flask.

The solvent is removed and the product is determined from ethanol to give 6.7 g (98%) of material having a melting point of 174-176 ° C.

(B) 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide

[Method 2]

1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester (10.0 g, 26.0 m mol) is methanol (70 ml) in anhydrous ammonia at 0 ° C. and a saturated solution at 0 ° C. are left at 25 ° C. for 18 hours in a closed pressure flask.

The product is determined from ethanol to give 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide (5.70 g, 90.0%).

(C) 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide

[Method 3]

3-cyano-1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole (705 mg, 2.0 mmol) 28% aqueous ammonia solution (20 ml ) And 30% hydrogen peroxide (3.0 ml) are stirred at 25 ° C. for 6 hours. 3.0 ml parts of 30% hydrogen peroxide were added to the stock and stirring was continued at 25 ° C. for 12 hours.

Excess hydrogen peroxide is added to destroy the excess platinum black, the solution is filtered and the filtrate is evaporated to dryness. The residue is dissolved in methanol and silica gel (5.0 g) is added to the solution.

The solvent is built up and removed and the silica gel mixture is applied on a silica gel column.

Treatment with ethyl acetate-methanol (1: 1) affords 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide (250 mg, 51.2%).

Example 6

[1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide 3′-5′-cyclephosphate ammonium salt]

1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide 5′-phosphate ammonium salt (5.7 g, 15.9 m mol) in pyridine solvent The resulting solution by adding, N′-dicyclohexylcarboxamidine (4.65 g, 15.9 m mol) is vacuum evaporated several times with pyridine to make anhydrous syrup.

The syrup is dissolved in 1 L of pyridine and added dropwise through reflux condensation to reflux anhydrous solution of dicyclo hexylcarbodiimide (16.4 g, 76.6 mmol) contained in 3 L of pyridine for at least one hour.

The solution is further refluxed for 2 hours, 200 ml of water is slowly added and the solution is vacuum evaporated after 12 hours, and 200 ml of water and 50 ml of ether are added to the residue. The suspension is stirred vigorously, filtered to separate the liquid layer and extracted with 2 x 100 ml of ether.

Pass the liquid layer through Dowex 50 (NH ₄ ⁺ type, 100-200 mesh) and evaporate to syrup.

100 ml of ethanol is added to the syrup and the resulting mixture is left at room temperature for 12 hours to filter the residue precipitate and the filtrate is left at room temperature for 2 weeks.

The resulting precipitate (1.2 g) is dissolved in 5 ml of hot water and then 40 ml of ethanol is added.

The resulting crystals were filtered off and dried at 78 ° C. under vacuum for 12 hours on P ₂ O ₅ to obtain 1.05 g of 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide. Obtain 3 ', 5'-cycle phosphate ammonium salt.

α ₂₅ ^D = 67 ° (C = 1 H = 0) IR (KBr)

Example 7

[1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide 5-phosphate ammonium salt]

1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester (259 mg, 1.00 m mol) of trimethylphosphate (3.0 ml) and phosphoryl chloride (0.20 ml) The mixed solution is stirred at 0 ° C. for 1.5 hours, added ice water and neutralized by treating the solution with sodium bicarbonate.

The solution is extracted with chloroform and the aqueous layer is cooled to 0 ° C. and saturated with ammonia.

The solution is left at 25 ° C. for 16 hours, then filtered and the filtrate is concentrated to a small amount.

Ethanol was added to form a precipitate, which was dissolved in water, passed through a Bio-Rad AG 50W-X ₂ (NH ₄ ) Column (20ml), and the fraction containing nucleotides was concentrated. Yarn) -1,2,4-triazole-3-carboxamide 5-phosphate (190 mg, 53.0 mmol) of ammonium salt.

Example 8

[1- (β-D-ribofuranosyl) -1,2,4-triazole-3-thiocarboxamide]

3-cyano-1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole (8.0 g, 22.7 m mol) triethylamine (14.0 ml) and ethanol (200 ml) are stirred at 25 ° C. while emulsified hydrogen gas is passed through the solution for 2 hours.

The solvent is removed and the residue is treated with a solution containing sodium methoxide (600 mg) in methanol (150 ml) at 25 ° C. for 3 hours. Treat the solution with Bio-Rad AG 50W-X ₂ (H) to make it neutral, then filter and remove the solvent.

The product is crystallized from aqueous ethanol to give 4.5 g (76.1%) of thiocarboxamide having a melting point of 173-175 ° C.

Example 9

[3-Cyano-1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole]

3-cyano-1,2,4-triazole (9.41 g, 0.10 mol) and 1,2,3,5-tetra-0-acetyl-β-D-ribofuranose (31.8 g, 0.10 mol) The mixture is heated while maintaining 150 ° C. in the oil weight.

Bis (P-nitrophenyl) phosphate (100 mg) was added with stirring and heated to 150 ° C. for 15 minutes under reduced pressure.

The residue is dissolved in chloroform, the solution is filtered, the solvent is removed and the residue is determined from ether to give 28.2 g (80%) of product having a melting point of 96-97 ° C.

Example 10

[1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamidine hydrochloride]

3-cyano-1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole (7.04 g, 20.0 mmol), ammonium chloride ( 1.07 g, 20.0 mmol) and anhydrous ammonia (150 ml) are heated to 85 ° C. for 18 hours in a bomb.

After excess ammonia is removed, the residue is crystallized from acetonitrile-ethanol to give 5.30 (95%) of the product having a melting point of 177-179 ° C.

Example 11

3-cyano-1- (2,3,5-tri-0 in 100 ml in 1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamidoxin ethanol A solution of acetyl-β-D-ribofuranosyl) -1,2,4-triazole (3.0 g, 8.52 mmol) and excess hydroxylamine is refluxed for 2 hours with stirring.

The solvent is removed and the product is determined from aqueous ethanol to give 2.0 g (90.7%) of carboxamidoxin having a melting point of 212-214 ° C.

Example 12

1- (β-D-chishirofuranosyl) -1,2,4-triazole-3-carboxamide tetra-0-acetyl-D-chishirofuranosyl (12.7 g, 40.0 m mol) methyl 1, A mixture of 2,4-triazole-3-carboxylate (5.08 g, 40.0 mmol) and bis- (P-nitrophenyl) phosphate (50 mg) was maintained at 160-165 ° C. in an oil bath for 20 minutes under reduced pressure. Heat.

The residue is cooled to dissolve in chloroform and the residue product is purified by chromatography with chloroform acetone (20: 1) solution on silica gel.

2.0 g of purified 1- (2,3,5-tri-0-acetyl-β-D-kishirofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester with saturated methanol Treat with ammonia at 25 ° C. for 16 hours.

The solvent is removed and the residue is crystallized from liquid methanol to give 0.7 g of product having a melting point of 194 ° -196.5 ° C.

Example 13

[1- (2,3,5-Tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide]

1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide (488 mg, 2.00 m mol) in pyridine (in 10.0 ml solvent) containing acetic anhydride solution (1.0 ml) ) Is left at 0-5 ° C. for 22 hours.

Ethanol (1.0 ml) was added thereto and the solution was stirred at room temperature for 2 hours. The solvent is removed, water is added to the residue, and the mixture is extracted with methylene chloride (treated three times with 20 ml) and the organic layer is extracted with aqueous sodium hydrogen carbonate and water.

The organic layer was dried over magnesium sulfate, filtered, evaporated and applied to a silica gel layer to make a syrup and treated with chloroform-methanol (19: 1) to give a pure product (560 mg, 75.6%) as an amorphous solid.

1- (2,3,5-tri-0-acetyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide in mice infected with Infurenza A ₂ (Jap 305) The dose of 125-250mg / kg / Day was orally administered twice a day for 9 days, which not only increased surviving rats but also prolonged survival time.

Example 14

[1- (β-D-arabinofranosyl) -1,2,4-triazole-3-carboxamide]

In anhydrous methylene chloride (100 ml), methyl-N- (trimethylsilyl) -1,2,4-triazole-3-carboxylate (4.12 g, 22.0 mmol) and 2,3,5-tri-0-benzoyl- The solution containing D-arabinofuranosyl chloride (8.78 g, 20.0 m mol) is left at 25 ° C. for 48 hours.

The solution is evaporated to dryness and the solution in which the residue is dissolved in methylene chloride is washed with sodium bicarbonate and water.

The methylene chloride solution is dried over magnesium sulfate, concentrated, applied to a layer of silica gel in benzene to make a syrup, and the column is treated with benzene-ether (7: 3) to give 1.9 g of syrup product.

This is treated with methanol (50 ml) saturated at 0 ° C. and anhydrous ammonia for 48 hours in a pressure flask of 25 ° C.

The solvent was removed and the product was determined from methylene chloride-cyclohexane to give 1- (2,3,5-tri-0-benzyl-β-D-arabinofranosyl) -1,2, having a melting point of 100-102 ° C. 1.3 g (12.6%) of 4-triazole-3-carboxamide are obtained.

A suspension of palladium black and methanol (20 ml) was obtained by reducing palladium chloride (300 mg) with hydrogen and then in 1- (2,3,5-tri-0-benzyl-β-D-arabinofuranosyl in methanol (25 ml). The stock is added to a solution containing 1,2,4-triazole-3-carboxamide (600 mg).

The mixture is mixed at 20 ° C. in 25 ml of hydrogenator for 2 hours, the catalyst is removed by filtration and the solution is passed through a layer of Amberlite IR 45 (OH) (15 ml).

The solvent is removed and the residue is determined from ethanol to give 250 mg of product having a melting point of 189-191 ° C.

Example 15

(A) 1-0-Acetyl-2-dioxy 3,5-di-0-P-toluoyl-D-ribofuranos

2-dioxy-3,5-di-0-P-toluoyl-D-ribofuranosyl chloride (15.6 g, 40.0 m mol) mercury acetate (12.7 g, 40.0 m mol) and tetrahydrofuran (200 ml) The mixture was stirred at 25 ° C. for 10 hours.

The solvent is removed, chloroform is added to the residue, and the mixture is extracted with 30% aqueous potassium iodide (4 treatments of 60 ml each) and water.

The organic layer was dried over magnesium sulfate, filtered and concentrated to a syrup (16.0 g, 97%) and the product was determined from cyclohexane benzene to give a pure compound with a melting point of 88-91 ° C.

(B) 1- (2-dioxy-3,5-di-0-P-toluoyl-α-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester 1- (2-Dioxy-3,5-di-0-P-toluoyl-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxylic acid methyl ester 1-0- Acetyl-2-dioxy 3,5-di-0-P-toluoyl-D-ribofuranose (4,54 g, 11.0 m mol), methyl 1,2,4-triazole-3-carboxylate (1.27 g, 10.0 m mol) and a mixture of bis (P-nitrophenyl) phosphate (10 mg) are heated with stirring while maintaining 150 ° C. in an oil bath for 10-15 minutes under reduced pressure.

The residue is dissolved in methylene chloride and the solution is filtered and washed with aqueous sodium hydrogen carbonate and water. The organic layer is dried over magnesium sulfate, filtered and evaporated to syrup.

This anmeric mixture mixture is column chromatographed with chloroform on silica gel to give α-anomer, which is then crystallized from cyclohexane-methylene chloride to give 780 mg of pure product having a melting point of 94-95 ° C.

As a result, the fraction from the silica gel layer contains a mixture of α and β anomers. These fractions are mixed and the mixture is purified by column chromatography with chloroform on silica gel to give β-anomer as a syrup.

(C) 1- (2-dioxy-β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide

1- (2-dioxy-3,5-di-0-P-toluoyl-β-D-ribofuranosyl) -1,2,4-triazole-3 in saturated ammonia and saturated methanol at 0 ° C The solution containing carboxylic acid methyl ester (650 mg, 1,35 mmol) is left at 25 ° C. for 3 days in a closed compression bottle.

The solvent was removed and the residue was treated in silica gel followed by ethyl acetate-methanol (9: 1) to give 1- (2-dioxy-β-D-ribofuransil) -1,2,4- amorphous solid. Triazole-3-carboxamide (250 mg, 80.8%) is obtained.

(D) 1- (2-dioxy-α-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide

1- (2-dioxy-3,5-di-0-P-toluoyl-α-D-ribofuranosyl) -1,2,4-tria in anhydrous ammonia and saturated methanol (25 ml) at 0 ° C. The solution containing sol-3-carboxylic acid methyl ester (780 mg, 1.63 mmol) is left at 25 ° C. for 3 days in a closed compression flask.

The solvent was removed and the residue product was applied to the silica gel layer and treated with ethyl acetate-methanol (9: 1) to give 1- (2-dioxy-α-D-ribofuranosyl) -1,2,4- as an amorphous solid. Triazole-3-carboxamide (350 mg, 94.4%) is obtained.

By the same process applied to convert the separated methyl ester monomer to its carboxamide, the mixture of the methyl ester monomer obtained in the step (B) was reacted with ammonia to react α and β anomers of the carboxamide. Obtain a mixture.

Example 16

The present invention was tested for anti-viral agents by bacterial evaluation (V.R) method of Sid well, et al, Appl, Micbrobial 22,787 (1971) against large and small bacteria of DNA and RNA type.

V.R> 1.0 represents the antiviral limit, and if V.R is 0.5-0.9, it shows appropriate antivirality. If V.R is 0.5, it is either less or less antiviral.

The following results report is obtained by testing the monolayers of KB or RK ₁₃ tissues together in a micro test II (Falcon plastics) plastic panel as compared to several conventional antiviral agents. Among the Basler bacteria used in the test, herpes type 1 herpes type is related to labiaris (cold wound), herpes zoster Keratitis and herpes encephalitis. Herpes zoster 2 causes herpes zheniratis and usually infectious sexually transmitted diseases.

Myxoma causes death of livestock and curing rabbits, and involves bronchial disease and severe swelling.

Gastric rabies (pseudorabies) not only causes infectious bulb palsy, but is also known as a pathogen that causes severe mad itch in cattle, sheep, pigs, dogs and minks. Parainfurenza bacteria become a source of bronchial disease in humans, especially young children, and become a hospital for conjunctivitis in livestock.

Vaccinia is non-toxic and sometimes has side effects from the smallpox bacterium used to inoculate smallpox.

Rhino type 13 is one of several bacteria that are usually included in the common cold.

[Test Table 1]

Compound (1) was tested in rabbits and rats to show anti-HSV geratitis clearly at non-toxic doses.

The compound inhibited mouse tail-induced HSV pathology when partially treated at the site of infection.

This compound was also tested in mice infected with Infurenza A ₂ , Infurenza B and Parainfurenza 1 and showed high antiviral properties.

As indicated in Test Table 1 above, the compounds exhibited antiviral spectra and were broader than the effects exhibited by 1 Du, Ara-A or Ara-C.

1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide also inhibits growth of Pseudomonas Aeruginosa bacteria and Candida Albicans bacteria and Cryptococcus diffluents.

The compounds of (1-6) are all less toxic and can be dissolved in an aqueous medium.

1- (β-D-ribofuranosyl) -1,2,4-triazole-3-carboxamide also contains antitumor properties.

Three groups of rats consisting of six rats (C 57 Blk / 6 rats) were treated post-treatment with adeno calcinoma-755, and the rats of group 2 were compounded (100 mg / kg × 7 days and 200 mg / kg). 7 days), while the third treatment group received only Saline.

On the 16th day, the treated animals were compared with the control group. The tumor inhibition was 18% and 63%.

A similar test showed that L-1210 decamia (animal 1 × 10 ⁵ cells and treated DBA / 2 mice extended the survival time over the control group by 31% when treated with this compound.

When treated with Swiss mice at a dose of 250 mg / kg × 14 days, 80% of the mice survived the pain of intraperitoneal intubation of the ascites.

The same effect was seen in ViVO activity against Novikoff Hepatoma.

It was found by the present invention that 1,2,4-triazole-3-carboxamide alone can exhibit sufficient antivirality.

When tested in the above-described method, the VR score report is in herpes type 1. 0,6, 1.0 Laryngeal. 0.8 Faraene Furenza 0.6 and 0.6. The compounds were tested for adeno and reno form 13, and the VR values were 0.0 and 0.3, respectively.

Claims (1)

Reduction of the penofuranose of the 3-substituted 1,2,4-triazole represented by the general formula (IV) and the 1,2,3,5-0-acyl blocking sugar represented by the general formula (III) at 35 ° C Fusing in the presence of an acid catalyst to form a glycosyl hydroxyl blocked triazole nucleoside represented by formula (II), followed by aminoylsis and silylation to form 3-thiocarboxamide or carboxamide, etc. Method for producing 1,2,4 triazole nucleoside represented by the general formula (I), characterized in that to form a 3-substituted nucleoside of the compound.

Wherein R ¹ is an alkylcarboxylate or cyano group, which is a carboxamido, thiocarboxamido, methyl carboxylate, carboxamidoxyl, carboxamidine or physiologically acceptable acid addition. Salt. G means pentofuranosyl moiety. AC is CH ₃ CO- and Bz is C ₆ H ₅ CO- and R ₂ or R ₃ means hydrogen, as well as 5′-phosphate and 3′5 ′,-cyclic phosphate as well as hydroxyl groups.