SE501370C2 - New cyclobutane derivative and process for preparation thereof - Google Patents

Abstract

This invention relates to cyclobutane derivatives represented by general formula (IV), wherein B represents a nucleic acid base and R<4> represents hydrogen atom or a protecting group, which is expectedly useful as an antiviral agent or a carcinostatic agent.

Description

DETAILED DESCRIPTION OF THE INVENTION Examples of B in formula (IV) comprise the compounds, which correspond to the following formulas: NH '2 O adenine 5 ïN (å, 6 NH guanine 4 2 ß a * 12 ßö Among the compounds corresponding to the general formula (IV), those are suitable whose substituents have such a steric configuration that substituent B and its adjacent hydroxymethyl group are in a trans ratio and the hydroxymethyl group adjacent to B and and the other hydroxymethyl group is in a trans ratio, those with a steric configuration of (1R, 2R, 3S) are more suitable.

Concrete examples of the compound, which correspond to the general formula (IV), are given below. In this description, the relative steric configuration of compounds is expressed in the following manner. Thus, when the cyclobutane ring is intended as a plane, a sub- (one side) is denoted by the symbol a, and is denoted by the substituent located below the plane a substituent located above the plane (the other side) by the symbol ß. 1. (1) -9- [1α, 20, 3β) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] adenine 2. 9 - [(1S, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl ] -adenine (prepared in Example 17, outside formula IV) 3. (1) -9 - [(1a, 2a, 36) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine 9 - [( 1S, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (prepared in Example 18, outside formula IV) 5. (1) -9 - [(1ß, 2a, 3ß) - 2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine 6. 9 - [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine 7. (1) -9 - [(1β, 2α, 3β) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine 8. 9 - [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutane -1-yl] -guanine Among the compounds of the present invention, the compounds corresponding to the general formula (IV) can be prepared by bringing a compound corresponding to of the following general formula (V): 4 R OCH 2 X (V) 4 CHZOR 10 15 20 25 30 35 40 3 501 370 (R 4 represents a hydrogen atom or a protecting group and X represents a cleavable group) to react with a nucleic acid base to to give a compound corresponding to the following general formula (VI): R4ocH2 B1 '(VI) 4 CHZOR (R4 represents a hydrogen atom or a protecting group and B1 represents a nucleic acid base) and, when the latter compound has a protecting group, optionally eliminates the protecting group with a suitable protecting group eliminating reagent.

As the protecting group R 4 in the general formula (V) and (VI), any group may be used without limitation as long as it is suitably used as a protecting group. Examples of the protecting group R 4 include ester-type protecting groups such as acyl groups (eg acetyl, benzoyl and the like) and carbamoyl protecting groups such as silyl groups (eg t-butyldimethylsilyl, (eg dimethylcarbamoyl, diphenylcarbamoyl and the like), ether type groups, t -butyldiphenyl-silyl and the like), (C1-C4) alkoxy- (C1-C4) alkyl groups (eg methoxymethyl and the like), benzyl group and the like. Examples of the leaving group X in the general formula (V) include solphonyloxy groups such as methanesulfonyloxy, p-toluenesulfonyloxy, trifluoromethanesulfonyloxy and the like, and halogen atoms such as chlorine, iodine and the like.

Examples of the nucleic acid base include bromine, purine bases such as adenine and guanine.

In the reaction between a compound corresponding to the general formula (V) and a nucleic acid base (eg adenine or guanine), the ratio of the compound with the general to about 10 equivalents of the latter precursor (V ) and the nucleic acid base about 0.5 valents and more suitably about 1 to about 5 per 1 equivalent of the former compound. This reaction is carried out either in the presence of a basic catalyst or in the absence of catalyst. As the basic catalyst, potassium carbonate, lithium hydride, sodium hydride and the like can be used. The reaction is carried out in a solvent, dimethyl sulfoxide (DMSO), formic acid triamide (HMPA) and the like, at a temperature such as N, N-dimethylformamide (DMF), 1,3-dimethyl-2-imidazolinone, hexamethylphosphate ranging from 0 ° C to the reflux temperature of the solvent, preferably from ambient temperature to about 170 ° C. Based on the amount of the nucleic acid base, the basic catalyst is preferably used in an amount of from 0 to 2 equivalents, preferably 0.5 to 1.5 equivalents, and more suitably about 0.8 to 1.2 equivalents.

Elimination of the protecting group (including alkyl group) from the compound of general formula (VI) can be achieved by using suitable reagent, as protecting protecting group, and method, which eliminates protecting group, according to the kind of protecting group. As reagents which eliminate protecting group, reference may be made, for example, to alkalis such as sodium hydroxide, sodium methylate, ammonia and the like, acids such as hydrochloric acid, sulfuric acid and the like, fluorinated reagents such as tetrabutylammonium fluoride and the like. As a method which eliminates protecting group, reference may be made, for example, to hydrogenolysis and the like.

Furthermore, the compounds of general formula (IV) can be synthesized in the following manner. As shown in the following reaction scheme (1): 4 e 4 R ocnz x ___, R ocnz NH2 4 4 CHZQR CHZOR (v) (XXII) 3400112 H2 _ "> š Hoc fl z B CH 0124 CH OH 2 2 (xxxn) (IV) (in this reaction scheme, R 4 represents a hydrogen atom or a protecting group, X represents a cleavable group, B represents a nucleic acid base and B2 represents a substituent convertible to B in one step or several steps). Thus, a compound having the general formula (V) with an azide ion compound, such as sodium azide or the like, and then reduced in the usual manner to synthesize an amine derivative corresponding to the general formula (XXII), and the latter (XXII) is converted into a compound of the general formula (IV) via an intermediate corresponding to the general formula (XXIII), according to known method (R. Vince et al., J. Med. Chem., 27, 1358 (1984), R. Vince et al., J. Med Chem., 30, 2026 (1987), YF Shealy and CA O'Dell, J. Heterocyclic Chem., 13,101S (1976), YF Shealy 18,383 (1981), et al, J. Heterocyclic Ch. em., etc). Furthermore, it was found that a compound corresponding to the general formula (III) is very useful for the preparation of a compound of the general formula (V) or a compound of the present invention , which corresponds to the general formula (IV). Since the compounds of the general formula (III) can be obtained in an optically active form, the compounds of the general formula (IV) are also obtainable in an optically active form.

As shown in the following reaction scheme (1): Rs 2 / z Å Å SRI SRI RZCH = CH N z + R cnv = c -> R il \ Aj S121 SR (I) (II) Co (III) 1 Y = Zl / Reaction Scheme (4) (in this reaction scheme, R 1, taken individually, represents an alkyl group having 1 to 5 carbon atoms or an aralkyl group or taken together, two R 1 groups represent a cyclic alkylene group having 2 to 3 carbon atoms, R 2 represents hydrogen atom, alkyl group with 1 to 5 carbon atoms, protected hydroxyalkyl group or protected carboxyl group, R 3 represents hydrogen atom, lower alkyl group having 1 to 5 carbon atoms, lower alkoxy group having 1 to 5 carbon atoms or aralkyloxy group, A represents a straight or branched chain alkylene group having 2 to 5 carbon atoms, Y represents oxygen atom or sulfur atom and Z represents substituted or unsubstituted methyl group, oxygen atom or sulfur atom) is thus caused to react a compound of general formula (I) and a compound of general formula (II) in the presence of a condensation catalyst, wherein a cyclobutane compound ing, which corresponds to the general formula (III), is obtained in a high yield and this product is racemically or optically active according to the kind of catalyst. As the condensation catalyst useful in this reaction, reference may be made, for example, to Lewis acids or combinations of a Lewis acid and an equivalent or excess amount of a ligand. Examples of Lewis acid include titanium compounds such as titanium tetrachloride, dichlorodiisopropoxy titanium and the like, tin compounds such as stannous chloride, stannous chloride, stannous trifluromethanesulfonates and the like and aluminum compounds. , such as dimethylaluminum chloride, diethylaluminum chloride and the like. As a ligand, sterically complicated diols are suitable. Examples of the ligand include compounds having a ring not less than 5-membered (preferably 5- to 8-membered ring) in the molecule and two hydroxy-containing groups on both sides of the ring, such as (2S, 3S) -2, 3-O- - (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol (2R, 3R) -2,3-O- (1-phenylethylidene) -1, 1,4,4-tetraphenyl-1,2,3,4-butanetetraol (Compound B), (2S, 3S) -2,3-O-benzylidene-1,1,4,4-tetraphenyl- 1,2,3,4-butanetetraol (Compound * C), (2R, 3R) -2,3-O-benzylidene-1,1,4,4-tetraphenyl-1,2,3,4-butane- ( 2S, 3S) -2,3-O- (1-phenylethylidene) - (compound A), -tetraol (compound D), -1,1,4,4-tetrakis (4-methoxyphenyl) -1,2,3 , 4-butanetetraol (Compound E), (ZR, 3R) -2,3-O- (1-phenylethylidene) -1,1,4,4-tetrakis (4-methoxyphenyl) -1,2,3 , 4-butanetetraol (Compound F), racemic forms of the above compounds and the like. The compound of general formula (I) and the compound of general formula (II) are used in such a ratio that the amount of compound (II) is 0.1 to 5 equivalents, preferably 0.5 to 2 equivalents, per equivalent of compound (I). The condensation catalyst is used in an amount of from 0.001 to 2 equivalents, preferably 0.01 to 1.2 equivalents, per one equivalent of the compound of general formula (I). This reaction can sometimes be proceeded more efficiently by adding a dehydrating agent, such as molecular sieves 4A and the like, to the reaction system. Examples of the solvent useful in this reaction include hydrocarbon solvents such as pentane, hexane, heptane, petroleum ether , benzene, toluene, ethylbenzene, trimethylbenzene, triisopropylbenzene and the like; halogenated hydrocarbon solvents such as flakes and the like; ethereal solvents such as ether, tetrahydrofuran and the like; acetonitrile and mixtures of these solvents. The reaction temperature is recommended in the range from the freezing point of the reaction solvent to its boiling point and suitably in the range from -50 ° C to about 30 ° C. By, for example, reacting an equivalent of a compound of general formula (I), wherein R 2 represents methoxycarbonyl group, A represents CH 2 CH 2, Y represents oxygen and Z represents oxygen, to react with 1, Equivalents of a compound of general formula (II), wherein R 1 represents methyl group and R 3 represents hydrogen atom, in the presence of a condensation catalyst (combination of 0.05 equivalents of dichlorodiisopropoxy titanium and 8.055 equivalents (2S, 3S) -2,3-O- 1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol (Compound A)) and molecular sieves 4A in a solution mixture consisting of hexane and toluene at a reaction temperature of At 0 ° C, (2S, 3S) -3- -methoxycarbonyl-1,1-bis (methylthio) -2- -carbonylcyclobutane is obtained in a high chemical yield and a high optical yield, if this reaction is carried out in the presence of a combined catalyst consisting of dichlorodiisopropoxy titanium and (2R, 3R) -2,3-O- (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol (compound B), <2R, 3R) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-on-3-yl) -carbonylcyclobutane is obtained. Furthermore, if this reaction is carried out using racemic 2,3-O- (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol, (1) - (2 The 3-methoxycarbonyl-1,1-bis compound of the present invention comprises examples of alkyl group having 1 to 5 carbon atoms, which are represented by R 1, R 2 and R 3, alkyl groups such as methyl, ethyl, butyl and the like; examples of the aralkyl group include alkyl groups substituted with an aromatic ring such as benzyl group, 4-methoxybenzyl group and the like; examples of the protected hydroxyalkyl group include benzyloxymethyl group, acetyloxymethyl group, t-butyldiphenylsilyloxymethyl group and the like; examples of the protected carboxyl group include alkoxycarbonyl groups such as methoxycarbonyl, ethoxycarbonyl and the like and aralkyloxycarbonyl groups such as benzyloxycarbonyl and the like; examples of alkoxy groups having 1 to 5 carbon atoms include methoxy group, allyloxy group and the like; and examples of the aralkyloxy group include benzyloxy group, 4-methoxybenzyloxy group, t-butyldiphenylsilyloxy group and the like. In the present invention, examples of the compound corresponding to the general formula (III) comprise the compounds of the general formula (III), wherein R 1, R 2, as shown in Table 1: R 3, A, Y and z are Table 1 No. Rl 122 123 A yz 13 Me cooMe H cnzcnz oo 14 Me coosn H c fl zcnz oo 15 Me HH cnzc fl z oo 1 s Me cooMe H c fl zc fl z ss 17 Me n oBn cuzcnz oo provided that the two R1 groups in formula (III) may be identical to others. The compound corresponding to the general formula (V) can be prepared, for example, from a compound corresponding to the general formula (III) (herein, R 2 represents a protected carboxyl group and R 3 represents hydrogen atom). according to a procedure corresponding to the following reaction scheme (3): 10 15 20 25 30 35 _ 40 501 370. sR1 SRI lloóc -> - Hoc fl 1 R (II) -_> SR1 2 SRI cooRl ° cnzon (VII) (VIII) 4 _ sR1 4 -f> R ocnz 1 -s »R oc fl z 0 sR CH QR4 CH OR4 - 2 2 (IX) (X) 4 -> - R4ocH2 on -e» R ocnz X 4 R4 cH2oR CH20 (XI) (V) Reaction Scheme (3) (in this reaction scheme, R 1 is the same as in the general formula (III), R 10 and R 11 each represent hydrogen atom, alkyl group having 1 to 5 carbon atoms or aralkyl group, R 4 represents hydrogen atom or a protecting group and X represents an eliminable group).

Thus, in the first step, a compound corresponding to the general formula (III) (herein R 2 represents a protected carboxyl group and R 3 represents hydrogen atom) is reacted in an alcoholic solvent, such as methanol, ethanol or the like, in the presence of a corresponding metallic alkoxide, such as magnesium methoxide, sodium ethoxide or the like, at a temperature ranging from -78 ° C to the boiling point of the solvent (preferably at a temperature not exceeding room temperature), or solvated in an alcoholic solvent, as methanol, ethanol or the like, in the presence of an acid, such as hydrochloric acid, p-tulpenosulfonic acid and the like, or a base, corresponding ester, such as triethylamine, sodium hydroxide and the like, wherein corresponding to the general formula (VII), wherein R 10 and R 11 represent hydrogen atom, alkyl group having 1 to 5 Alternatively, carbon atoms or aralkyl group can be obtained. it is hydrolyzed in the presence of an acid such as hydrochloric acid, p-toluenesulfonic acid and the like, or a base such as sodium hydroxide, potassium carbonate and the like, wherein a carboxylic acid of the general formula (VII) wherein RIG and R11 represent By reducing the compound with the hydrogen atom, can be obtained. The general formula (VII) obtained herein with a metal hydride such as lithium aluminum hydride, di (isobutyl) aluminum hydride, alcohol, sodium borohydride, diborane and the like is obtained one corresponding to the general formula (VIII). Next, the hydroxyl group of the compound of general formula (VIII) is protected to obtain a compound of general formula (IX), and its dithioketal moiety is hydrolyzed in an aqueous solvent with a halogenating agent such as iodine, N-bromosuccinimide. N-chlorosuccinimide, sulfuryl chloride and the like, or a heavy metal compound such as silver nitrate, silver oxide, silver perchlorate, mercury chloride, copper chloride, copper oxide and the like or a combination of these compounds to form a ketone compound corresponding to the general formula (X).

A compound corresponding to the general formula (X) is reduced with a complex metal-hydrogen compound, such as lithium aluminum tri (t-butoxy) aluminum hydride, minium, sodium borohydride, lithium tri (s-butyl) - borohydride, lithium borohydride and the like or a metal hydride such as diisobutylaluminum hydride, diborane and the like as a reduced agent in a solvent such as hydrocarbon type solvent (e.g. pentane, hexane, heptane, petroleum ether, benzene, toluene, ethylbenzene and the like). de), chloride, halogenated hydrocarbon type solvent (eg methylene chloroform and the like), ethereal solvent (eg ether, tetrahydrofuran and the like), alcoholic solvent (eg methanol, ethanol and the like), water or mixture thereof , at a reaction temperature of from -100 ° C to 50 ° C and suitably -80 ° C to 30 ° C to give a compound corresponding to the general formula (XI). In this reaction, one of the stereoisomers can preferably be obtained by selecting the type of reductant and the reaction conditions. If, for example, a 2,3-trans-cyclobutanone corresponding to the general formula ( Xa) as shown in the following reaction scheme (&): 4 ----- e »R ocH2 - <::> == o 4 CHZQR (Xa) 4 4, R oc fl z -« <::> «woH + R ocnzw - <:: >> ~ OH ~ š 4 š 4 c fl oR c fl zoa 2 (xïa) (xïb) UJ! Reaction Scheme (4) (in this scheme, R 4 represents hydrogen atom or a protecting group) is reduced with a sterically relatively simple reducing agent, a 1,2-trans-alcohol is selectively formed, which corresponds to the general formula (Xlb). However, if the reduction is carried out with a highly steric hindered reducing agent, a 1,2-cis-alcohol is suitably formed, which corresponds to the general formula (XIa). For example, if (25,35) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cyclobutanone is reduced with lithium tri (t-butoxy) aluminum hydride or sodium borohydride (both of these reducing agents are not highly sterically hindered), is formed selectively (IR, 28,38) -2,3-bis (t-butyldiphenylsilyloxymethylJ- -cyclobutanol the same starting compound as above is reduced with lithium tri (s- -butyl) borohydride or these reducing agents are highly sterically hindered) is preferably formed ( 1S, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -cyclobutanol (isolation yield 75% and 82%, respectively). Results obtained are shown in Table 2. 10 15 20 25 30 35 12 501 370 Table 2 Conditions Uüwte (%) Reagent Solution medium] Temp. , ° C XIa XIb LiA1 (OtBU) 3H THF -78 ~ RT 9 88 NQBH4 THF-H20 RT 19 80 Li (SBU) 3BH THF -78 75 23 (iBU) 2AlH CHZCIZ -78 _ 74 22 (íBu) 2AlH Toluene1 - 78 82 17 124 = t-supn si (in xa, xra and xrb) 2 The stereoisomers obtained herein, i.e. the compound of the general formula (Xla) and the compound of the general formula (XIb), are both readily convertible from the one after isolation to the other. For example, a compound corresponding to the general formula (Xla) or a compound corresponding to the general formula (XIb) is oxidized again to a ketone corresponding to the general formula (Xa), by means of conventional oxidizing agent or oxidation method (eg metal oxidizing agents such as chromic acid / acetic acid, chromic acid / pyridine and the like, or dimethyl sulfoxide (DMSO) oxidation method such as DMSO-acetic anhydride / acetic acid, DMSO-oxalyl chloride-triethylamine / methylene oxidized On other to the product (Xa) is subjected to a selective reduction. In this way, steric configuration of the 1-carbon atom to which the hydroxyl group is attached is inverted to a compound of general formula (Xla) or a compound of general formula (XIb) using, for example, the Mitsunobu reaction.

Now, the Mitsunobu reaction in the latter method will be explained below. Thus, a compound corresponding to the general formula (Xla) or (XIb) is reacted with a three-woven phosphorus compound, such as triphenylphosphine, trimethylphosphite, triethylphosphite or the like, a carboxylic acid such as acetic acid, benzoic acid or the like, and an azodicarboxylic acid. such as diethyl azodicarboxylate or the like, in a solvent, such as hydrocarbon type solvents (for example, pentane, hexane, heptane, petroleum ether, benzene, toluene, ethylbenzene and the like), halogenated hydrocarbon solvents (e.g. methylene chloride, chloroform and the like), ethereal solvent (for example, ether, tetrahydrofuran and the like) or a mixture thereof at a reaction temperature of -100 ° C to 5000 (preferably -50 ° C to 30 ° C) to form an ester . Then the ester is hydrolyzed with an alkali such as potassium carbonate, sodium hydroxide, sodium methylate, ammonia and the like or an acid such as hydrochloric acid, sulfuric acid and the like, or reduced with a complex metal-hydrogen compound such as lithium aluminum hydride, lithium tri -butyl) -borohydride, metal hydride, lithium borohydride and the like or one such as diisobutylaluminum hydride, diborane and the like. By this treatment, a compound corresponding to the general formula (Xlb) or a compound corresponding to the general formula (Xla) can be obtained. For example, if (1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol is reacted with triphenylphosphine benzoic acid-diethyl azodicarboxylate in benzene to form (1S, 2S, 3S) -1-benzoyl-2,3 - -bis (t-butyldiphenylsilyloxymethyl) cyclobutane and the latter is reduced with diisobutane / aluminum hydride in toluene, the benzoyl group can be eliminated, and (IS, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol is obtained. By converting the secondary hydroxyl group of the compound thus obtained, which corresponds to the general formula (XI), to a cleavable group, a compound corresponding to the general formula (V) can be prepared.

The experimental examples presented below will show that the compounds of the present invention exhibit extensive and strong antiviral and carcinostatic effects.

Experimental Example 1 Antiviral effects against Herpes Simplex 1-type virus (HSV-1) and Herpes Simplex 2-type virus (HSV-II) viruses, both of which belong to DNA viruses, were examined by the following methods. 10 15 20 25 30 35 14 501 370 (Method 1) A multiplate with 6 wells and with a single layer of Vero cell (derived from the kidney cell of African green monkey) was infected with 100 to 150 PFU (plaque forming units) virus. After adsorption at 37 ° C for 1 hour, a layer of agar medium ("Eagle MEM medium" containing 1.52 agar) was placed on top, culturing was performed at 37 ° C for 48 hours at 5% (v / v) carbon dioxide. contained a varied sample concentration thereon and an incubator. Plaque formation was measured, from which 50% inhibitory value (IC50) was determined.

Experimental Example 2 Antiviral activity against human cytomegalovirus (HCMVQ, which belongs to the DNA virus, was examined by the following method.

(Method 2) The anti-HCMV effect was determined as follows. Thus, a 35 mm shell with a single layer of human embryonic fibroblast was infected with 100 PFU HCMV (A0169 strain). After adsorption for 1 hour, a layer of a medium (0.5% agarose, 2% fetal calf serum) containing a varied concentration of test compound was placed on top and culture was performed at 37 ° C for 10 days in a 5% (v / v) carbon dioxide incubator. Then plaque formation was measured from which 50% inhibitory value (IC50) was determined.

Experimental Example 3 Antiviral activity against hepatitis B virus (HBV), which belongs to the DNA virus, was examined by the following method.

(Method 3) According to the procedure of Dulbecco (Prcc. Natl. Acad.

Sci. USA, 84, 444 (1987)), cultured liver cell strain HB611, which produces and releases active hepatitis B virus, was cultured in a modified "Eagle medium" (GIBCO) at 37 ° C at a CO 2 concentration of 5% in the presence of 10% fetal calf serum, 200 micrograms / ml G418, 100 u / ml penicillin and 100 u / ml streptomycin The culture fluid was inoculated on a 6-well plate at a ratio of 5 x 104 cells / well (35 mm). One or two days later 50% confluence was achieved when a predetermined quantity of test compound was added and cultivation was continued, then cultivation was continued for 15 days while filling the medium with a fresh medium, which contained the same concentrations of the chemicals at 3 day intervals, then the medium was removed and the cell bodies were treated with 0.5 ml of Lysis buffer (10 mM Tris HCl, pH 7.8 / 5 mM Na 2 EDTA, 1% SDS / 0.1 mg / ml Pronase K) at 37 ° C for 1 hour and dissolved therein, The DNA thus obtained was purified by RNase treatment, phenol-chloroform treatment and ethanol precipitation.

Then, 5 micrograms of DNA was subjected to Hind III treatment and the DNA sample was analyzed by Southern method using 32 L-labeled hepatitis B virus DNA as a sample.

Experimental Example 4 Antiviral activity against human immunodeficiency virus (HIV), which belongs to DNA virus, was examined by the following method.

(Method 4) MT-4 cell (about 50,000 cells / ml) was introduced into a 24-well tray to which was added 100 micrograms of a solution containing a predetermined quantity of test compound. Culture was performed at 37 ° C for 2 hours in a 5% (v / v) carbon dioxide incubator.

Then 103 to 104 units of HIV infection were added and cultured for 4 days, after which part of the culture fluid was applied to a slide and immobilized with acetone and the development of viral antigen was tested by fluorescence antibody method.

As the primary antibody of the fluorescent antibody method, serum from the AIDS patient was used. Like the secondary antibody, FITC-labeled human IgG was used.

Experimental Example 5 Carcinostatic effects on human cervical cancer HeLa S3 cell, mouse leukemia L1210 cell and human leukemia P388 cell were examined by the following methods. (Method 5) HeLa S3 cell was made into a suspension (7.5 x 103 cells / ml). The suspension was spread on a flat plate with 96 wells at a rate of 0.2 ml / well. After culturing it at 37 ° C for 24 hours in a 5% carbon dioxide incubator, 10 microliters of a sample was added and culturing was performed for 72 hours. Then the culture fluid was taken from each well and immobilized with methanol, after which 0.1 ml / well of 0.05 % methylene blue / 10 mM Tris-HCl (pH 8.5) solution was added and staining was performed at room temperature for 30 minutes. The colored fluid in each well was removed using an aspirator and washed three times with clean water. Then 3% HCl was added at a rate of 0.2 ml / well and sealed and allowed to stand at room temperature for about 10 hours to extract pigment from the cells. Optical absorbance of each well at 660 nM was measured using the Dynatic Microplate Reader, from which the growth inhibition value (X) at varying concentrations was calculated according to the following equation, the results were plotted on a logarithmic probability paper, from which 50% inhibitory concentration (IC50, micrograms / ml) was determined.

Growth inhibition value (X) = (1 ~ A1 / A0) x 100 where A1 = absorbance in the treated group, A0 absorbance in the control group.

(Method 6) Cells of P388 and L1210 were spread on a 24-well plate. After addition of a sample, it was grown at 37 ° C for 48 hours in a 5% CO 2 incubator. After culturing, the number of cells was counted using a coal tar counter. The growth inhibition value in the treated group, calculated on that in the control group, was calculated from the following equation, from which 50% inhibitory concentration (C150, micrograms / ml) was determined: The results obtained are as shown in Table 3.

Growth inhibition value (Z) = (1- (NT-NO) / NC-No] x 100 cell number in treated group, where NT NO cell number at the beginning of culture, NC cell number in control group_ 10 15 20 25 17 501 370 Table 3 Pre- IC50 (pg / ml) ingar nr Hsv-1 Hsv-2 HIV Hcmv Hav HeLas3 9388 L121o 6 7,3 8,9 0,03 12 o, o24 14 0,58 1,1, 8 0,03 0,13 0, 3 0.4 o, s6 19 1.6 8.1 Experimental Example 6 The activity of the sample (Compound No. 11) against Herpes Simplex virus type 2 (HSV-2) infection was examined in mice.

Balb / c mice (6 weeks old, males) were inoculated intraperitoneally (l.p_) with HSV-2 strain 188 at 4.8 x 105 PFO / 0.2 ml per mouse.

The sample was prepared in saline and applied either orally (p.o.) or i.p. at the indicated doses once daily for 10 days after 6 hours after virus infection. Mortality was recorded for 20 days after inoculation. The effectiveness of the test was stated as an extension of survival time and the number of survivors. Acicloriv (ACV: commercialized antiviral agent) was used as a reference sample.

Results: Results obtained are as shown in Table 4. 501 370 18 Table 4 Before010gf 'Method' D05; Mean Survivor ~ Survival total (mg / kg / day) ti0_ (0ag) v Control 7.7 0/10 nrle 1.p. 20 20.0 10/10 10 20.0 10/10 5 20.0 10/10 2.5 18.9 9/10 1.25 19.8 7/10 po 20 19.7 9/10 10 19, 8 9/10 5 18.9 8/10 2.5 14.7 4/10 1.25 12.4 2/10 ACV 1.p. 100 5.4 2/10 50 16.1 5/10 25 12.7 1/10 12.5 11.1 1/10 p.o. 100 14/3 2/10 50 14.8 4/10 25 11.1 0/10 12 / S 9.3 0/10 10 15 20 25 30 35 40 B 501 370 To evaluate the in vivo potency of compound no. 8 compared with ACVt Compound No. 8 was highly effective in reducing the mortality value of mice infected with HSV-2 regardless of the route of administration (Table 4). The mean lifespan of mice in a control group was 7.7 days, while a number of survivors were observed through an observation period of 20 days in a wide dosage range of compound No. 8 from 1.25 to 20 mg / kg per day. Survivors were also observed by treatment with ACV. However, the number of survivors and an effective dose range were much greater with compound No. 3 than with ACV.

With a strong antiviral action, the compounds of the present invention, corresponding to the general formula (IV), are expected to be effectively useful in controlling a number of viral diseases, such as herpes labialis, herpes zoster herpes zoster, simple infection from herpes II). , the plant organs, viruses 1 and 2 (HSV-I, varicell zoster virus (VZV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV), viral hepatitis, viral diseases of the respiratory tract, viral diseases of the digestive organs, AIDS, ATL, etc. Furthermore, since they have a carcinostatic effect, they are expected to be useful as carcinostatic agents.

When using the compounds of the present invention, obtained in the manner indicated above as an antiviral drug or a calcinostatic drug, they can be administered orally, intravenously or percutaneously to warm-blooded animals. Although the dose may vary depending on the symptoms and age of the warm-blooded animal and the method of administration, it is usually 0.1 to 500 mg / kg / day. The compounds of the present invention are administered in the form of a composition which is prepared by mixing them with suitable excipients. granule, As a form of composition, tablet, powder, capsule, injection, cream, suppository and the like may be used.

Next, the preparation of the compounds of the present invention will be concretely illustrated by means of the following examples.

Example 1: Preparation of (-) - (2S, 3S) -3-methoxycarbonyl-1,1- -bis (methylthio) -2- (oxazolidin-2-one-3-yl) -carbonyl-cyclobutane Example 40 1-1 In an atmosphere of argon gas, 80 ml of 1,3,5-trimethylbenzene (TMB) was added to dichloroisopropoxy titanium (905 mg, 3.8 mmol) and (2S, 35) -2 , 3-O- (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,2,4,4-butanetetraol (Compound A), (2.22 g, 4.2 mmol), and the mixture stirred at room temperature for 30 minutes. Then powdered molecular sieves 4A (3.2 g) were added to the resulting solution and stirred for a while, then 3 - [(E) -3- - (methoxycarbonyl) -propenoyl] -oxazolidin-2-one (3.98 g, 20 mmol) was added and the resulting suspension was cooled to -15 ° C. Then slowly 1,1-bis (methylthio) ethylene (3.61 g, 30 mmol) dissolved in 20 ml of TMB solution was added and the temperature of the whole mixture was raised from this temperature to 0 ° C for a period of 3 + h stirring. A saturated aqueous solution of sodium hydrogencarbonate was added to the reaction mixture, the inorganic material was filtered off with celite and the organic material was extracted with ethyl acetate. The extract solution was washed with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate hexane = 1: 2, v / v) to obtain (-) - (ZS, 3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidine -2-on-3-yl) -carbonylcyclobutane (3.94 g, 62%). This compound has an optical purity of 86% ee (cf. Example 8).

NMR (500 MHz, CDCl 3) δ 2.01 (3H, s), 2.10 (3H, s), 2.54 (1H, dd, J = 9.9, 12.3Hz), 2.7o (1H , aa, J = 7.9, 12.3Hz), 3.86 (1H, dt, Jd = 9.9Hz, Jt = 7 / 9Hz), 3.71 (3H, s), 4.02 (1H, aaa, J = s, 4, 8.9, 11, oHz), _; 4712 (1H, dada, J = 7.6, 9.3, 11.0Hz), - "4.39-4.47 (2n, m), s, o1 (1H, d, J = 7.9Hz) .

IR (~ fen) cm -1; 1690. 1780, 1730, 10 15 20 25 30 35 40 21 501 370 In such a manner as in Examples 1-1, 3 - [(E) -3- - (methoxycarbonyl) -propenoyl1-oxazolidin-2-one (3 98 g, 30 mmol), dichlorodiiso-mmol), 1,1-bis (methylthio) ethylene (3.61 g, propoxytitanium (474 mg, 2.0 mmol), (2S, 3S) -2,3- O- (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol (Compound A) (1.16 g, 2.2 mmol) and powdered molecular sieves 4Å (4 g) reacting in a solvent mixture consisting of toluene (160 ml) and hexane (120 ml), da (-) - (25,3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2- ( oxa- at 0 ° C for 40 minutes to give bilzolidin-2-one-3-yl) -carbonylcyclobutane (6.13 g, 96%) ((010 = 1010.4 ° (C 1.34, CH 2 Cl 2) It was recrystallized from methylene chloride-isopropyl ether system to give a compound ((030 = -11.10 (c 1.15, CH 2 Cl 2))) in a yield of 5.30 g (83%).

Optical purity of this compound was 98% ee or more as determined by the method set forth in Example 8.

Examples 1-3 In a manner such as in Examples 1-1, 3 - ((E) -3- - (methoxycarbonyl) -propenoyl] -oxazolidin-2-one (19.9 g, 100 1,1-bis ( methylthio) ethylene (15.03 g, (2S, 3S) -2,3-O- (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol (compound A) mmol), 125 mmol), dichlorodiisopropoxy titanium (1.18 g, 5.0 mmol), (2.91 g, 5.5 mmol) and powdered molecular sieves 4A (20 g) to react at 0 ° C in In a solvent mixture consisting of toluene (800 ml) and hexane (600 ml), (-) - (2S, 3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidine-2 - on-3-yl) carbonylcyclobutane (26.69 g, 84%) 1.00, CH 2 Cl 2)). to obtain (UMD = -1o, s ° << = Example Gel 2: Preparation of (+) - (2R, 3R) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-one -3-yl) -carbonylcyclobutane In an atmosphere of argon gas, dichloroisopropoxy titanium (125 mg, 0.53 mmol), (2R, 3R) -2,3-O- (1-phenylethylidene) -1,1,4,4- -tetraphenyl-1,2,3,4-butanetetraol (Compound B) (305 mg, 0.58 mmol) and toluene (5 mL) together and stir at room temperature for 1 h. One portion (0.5 mL, 0.053 mmol) of the resulting solution is taken out and added to powdered molecular sieves 4Å (100 mg), and toluene (1.5 ml) was added thereto.To the obtained solution 10 15 20 25 30 22 501 370 sattesp3-C (E) ~ 3- (methoxycarbonyl] -propenoyl-oxazolidin-2-one (107 mg, 0.534 mmol) and petroleum ether (b.p. about 80 ° C) (PE) (2 ml) and the resulting mixture was cooled to 0 ° C. To this suspension was slowly added a solution of 1,1-bis (methylthio) -ethylene in PE (1.5 ml) and the resulting mixture (115 mg, 0.956 mmol) was stirred at 0 ° C for 3 hours. Then 0.2 M phosphate buffer (pH 7) was added to the reaction mixture o and the inorganic material was filtered off with celite, after which the organic material was extracted with ethyl acetate. The extract solution was washed with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate, after which the solvent was distilled off under reduced pressure. The residue was purified by fractional silica gel thin layer chromatography I + (ethyl acetate: hexane = 1: 1, v / v) to give (+) - (2R, 3R) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-on-3-yl) -carbonylcyclobutane (162 mg, 95%). This product had an optical purity of 98% ee (cf. Example 9).

NMR and IR of this product were identical to those of the compound of Example 1.

Example 3: Preparation of (+) - (ZR, 3R) -3-methoxycarbonyl--1,1-bis (methylthio) -2- (oxazolidin-2-one-3-yl) -carbonylcyclobutane The following table illustrates the results of a study of the conditions of preparation of this compound (including Example 2). The other conditions and treatment methods were the same as in Example 1 or 2. ob) MeO a) SMe Catalyst- sme Qxz + <sme 0 ° C SMG O MS4A O = C-Oxz 10 15 20 25 30 ': 3s 40 23 501 370 .No M Solution of Catalyst Time _ Yield Optical purityc) agent (eq) “(h) <° fi> <2 ee) Ö) l TMB 1.0 075 89 93.2 2 TMB 0.1 0.5 99 94,0 _ 6) 3 T PE lf0 3 78 98f3 4 T-PE 0,1 3 95 97f7 S Et2O 1,0 18 45 - a) Oxz denotes oxazolidin-2-on-3-yl group 4 * b) A mixture (1.0: 1.1) of TiCl 2 (i-PrO) 2 and compound B c) Optical purity was determined from NMR of bis-MTPA ester of compound (VII) d) TMB means 1,3,5- trimethylbenzene e) T-PE means toluene-petroleum ether.

Example Gel 4: Preparation of (+) - 1,1-bis (methylthio) -2- (oxazolidin-2-one-3-yl) -carbonylcyclobutane The reaction of Example 2 was repeated except that 3-E (E) -3 - (methoxycarbonyl) propenoyl-1-oxazolidin-2-one was replaced with 3-propenoyl-oxazolidin-2-one to give (+) - 1,1-bis (methylthio) -2- (oxazolidin-2-one-3 -yl) -carbonylcyclobutane (82%). This product has an optical purity of 88% ee (cf. Example 10).

NMR (500 Mzrr, CDCl 3) δ: 2.00 (3H, s), 2.12 (3H, s), 2.26-2.33 (ln, m), 2.34-2.38 (ln, m). 2.39-2.48 (ln, m). 2.52-2.61 (ln, m), δ 4.00-4 13 (ZH, m), 4.41 (za, t, J = s, oHz), 4.63 (1H Example 5: Preparation of (-) - (25.3S) -2,3-bis (methoxycarbonyl) -1,1-bis (methylthio) -cyclobutane Example 5 In an atmosphere of argon gas, 1M-dimethoxymagnesium / methanol (25 mL, 25 mmol) was added to a methanolic solution (25 mL) of (-) - (2S, 3S) -3-methoxycarbonyl-1,1-bis ( methylthio) -2- (oxazolidin-2-one-3-yl) carbonylcyclobutane (3.94 g, 12.3 mmol) obtained in Example 1-1 and stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, saturated aqueous solution of ammonium chloride was added to the concentrate and it was extracted with ether.The ether extract was washed with saturated aqueous solution of sodium chloride and then the solvent was distilled off under reduced pressure.The residue was purified by silica gel column chromatography (ethylene chloride); / volume) to give C -) - (2S, 3S) -2,3-bis (methoxycarbonyl> -1,1-bis (methylthio) cyclobutane <2.17 g, 67 %). This compound has an optical purity of 86% ee (cf. Example 8).

NM (500 MHzFT, CDCl 3) δ: 201 (3H, s), δ 2.12 (3H, s), 2.47 (in, da, J = 9.0, 12.2Hz), 2.50 ( ln, da, J = 9.4, 12.2Hz), 3.63-3.75 (za, m), 3.69 (su, S), 3.72 (3H, S), 1 in (raw ) cm *: 1730.

Example 5-2 In an atmosphere of argon gas was added (~) ~ (2S, 3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-on-3-yl) carbonyl cyclobutane (26.0 g, 81.4 mmol) to 1M-dimethoxymagnesium / methanol (400 mL, 400 mmol) at 0 ° C and the mixture was stirred at this temperature for 15 minutes. Saturated aqueous solution of ammonium chloride was added to the reaction mixture, and it was extracted with ether. After washing the extract solution with saturated aqueous sodium chloride solution, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate-hexane = 1: 9, v / v) to give (-) - (2S, 3S) -2,3-bis (methoxycarbonyl) -1,1-bis- (methylthio) cyclobutane (20 , 73 g, 96%).

Example Gel 6: Preparation of (+) - (ZR, 3R) -2,3-bis (methoxycarbonyl) -1,1-bis (methylthio) -cyclobutane The procedure of Example 5-2 was repeated except that (+) - (2R, 3R) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-one-3-yl) carbonylcyclobutane, prepared in Example 2, was used as the starting compound. Thus, (+) - (2R, 3R) -2,3-bis (methoxycarbonyl) -1,1-bis (methylthio) cyclobutane was obtained in a yield of 95%. This compound had an optical purity of 98% ee (cf. Example 9).

The NMR and IR of this compound were identical to those of the compound of Example 5.

Example Gel 7: Preparation of (+) - 2-methoxycarbonyl-1,1-bis (methylthio} cyclobutane The procedure of Example 6 was repeated except that (+) - 1,1-bis (methylthio) -2- (oxazolidine 2-On-3-yl) -carbonylcyclobutane, prepared in Example 4, was used as the starting compound to give (+) - 2-methoxycarbonyl-1,1-bis cyclobutane in 83% yield. had an optical purity of 88% ee (cf. Example 10).

NMR (500 MHzFT, CDCl 3) δ: 1.97 (3H, s), 2.07 (3H, s), 2.18-2.30 (3H, m), 2.49 (1H, m), 3 , 37 (1H, m), 3.67 (an, s). - Gas Zd 501 370 Example Gel 8: Preparation of (-) - (28,35) -2,3-bis (hydroxymethyl) -1,1-bis (methylthio) cyclobutane In an atmosphere of argon gas was slowly added an ethereal solution (10 ml) of (-) - (2S, 3S) -2,3-bis (methoxycarbonyl) -1,1- prepared in -bis (methylthio) cyclobutane (1.96 g, 7.4 mmol), Example 5-1, to an ethereal suspension of lithium aluminum hydride (562 mg, stirred at 0 DEG C. for 2 hours. 14.8 mmol) at 0 DEG C. and the mixture was obtained after adding saturated aqueous solution. of sodium sulfate to the reaction mixture to decompose the excess reducing agent, anhydrous sodium sulfate was added and stirred for a while. Then inorganic material was filtered off and washed with hot isopropyl alcohol. The filtrate and washings were combined and the solvent was distilled off therefrom under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate-hexane-methanol = 15: 20: 1, v / v / v) to obtain (-) - (2S, 3S) -2,3-bis (hydroxymethyl> -1,1-bis (methylthio ) - cyclobutane (1.48 g, 96%).

NMR (270 MZFT, CDCl 3) δ: 2.02 (ln, ad, J = 9.1, 12.1Hz), 2.05 (3H, S), 2.06 (3H, s), 2.28 ( 1H, dd, J = 8, l, 12 / l fl z), 2.47-2.68 (za, m), 3/26 (2H, brs), 3.54 (ln, da, J = a, a , 10, 3Hz), 3.67-3.77 (za, m), 3.83 (1H, dd, J = 5.0, 10.4Hz), IR (ken) cm -1: 3350.

A portion of this compound was taken out and converted to bis (R) -TMPA ester in the usual manner using (R) -Q- -methoxy-G-trifluoromethylphenylacetyl chloride ((R) -MTPACI), dimethylaminopyridine (DMAP) / pyridine (pyr). 501 370 In 500 MHz NMR, four methyl group signals derived from racemic mixture were observed at 1.852, 1.856, 1.912 and 1.970 ppm. Among them, the methyl group signals at 1,852 and 1,912 ppm were larger than others, from which optical purity was determined as 86% ee.

The title compound (-) - (2S, 3S) -2,3-bis (hydroxymethyl) -1,1- -bis (methylthio) cyclobutane can be made into a crystal of 100% optical purity by recrystallization from ethyl acetate-hexane system (EGJD = -32.00 (c 1.03, CH 2 Cl 2)).

Example 9: Preparation of (+) - (2R, 3R) -2,3-bis (hydroxymethyl) -1,1-bis (methylthio) -cyclobutane The procedure of Example 8 was repeated except that (+> - (2R, 3R) -2,3-bis (methoxycarbonyl) -1,1-bis (methylthio) cyclobutane, prepared in Example 8, was thus obtained (+) - (2R, 3R) -2,3-bis ( hydroxymethyl) -1.1-bis (methylthio) cyclobutane in a yield of 70%.

The NMR and IR of this compound were identical to those of the compound of Example 8.

A portion of this compound was taken out and converted to (R) -TMPA ester according to standard method (CR) -MTPAC1, DMAP / pyr).

NMR of the product was studied according to Example 8. Thus, it was found that signals of 1.856 and 1.970 ppm were larger than the others and optical purity was 98% ee.

Apart from the above, part of the title compound was recrystallized from ethyl acetate-hexane system to give a colorless crystal. By X-ray analysis of its single crystal, its absolute steric configuration was determined as (2R, 3R), as shown in the title.

Example 10: Preparation of (+) - 2-hydroxymethyl-1,1-bis- (methylthio) cyclobutane The procedure of Example 8 was repeated except that (+) - 2-methoxycarbonyl-1,1-bis (methylthio) cyclobutane, obtained in Example 7 was used. Thus, (+) - 2-hydroxymethyl-1,1-bis (methylthio) cyclobutane was obtained in a yield of 54%. Δ las 28 501 370 NM (500 MHzFT, CDCl 3) δ: 1.92 (1H, m), 2.05 (3H, s), 2.07 (3H, m). 2.12-2.32 (4H, m), 2.68 (1H, m), 3.73 (ln, dd, J = 4.8, 11, 8Hz), 3.84 da, J = 7ISI ll ; 8Hz) ~ A portion of this compound was taken and converted to (R) -MTPA ester according to standard method (CR) -MTPAC1, DMAP / pyr).

In 500 MHz NMR, four methyl group signals for this were shown at 1.696, compound derived from racemic mixture, 1.927, 1.930 and 1.955 ppm. Among them, the signals of 1.927 and 1.955 ppm were larger than the others, from which optical purity was determined as 88% ee.

Example Gel 11: Preparation of (+) - (2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1,1-bis (methylthio) cyclobutane In methylene chloride (25 ml) was dissolved (-) - (2S , 3S) -2,3-bis- (hydroxymethyl) -1,1-bis (methylthio) cyclobutane (1.37 g, 20 mmol), 6.58 triethylamine (2.8 ml, 4-dimethylaminomethylpyridene) -butyldi- mmol), din (catalytic amount) and DMF (1 ml). phenylsilyl chloride (4.52 g, 25 mmol) to the solution and stirred at room temperature overnight. After concentrating the reaction mixture under reduced pressure, the concentrate was dissolved in ether, washed with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. After distilling off the solvent from the ethereal solution, the residue was purified by silica gel column chromatography (ether: hexane = 1:20, v / v) to give (+) - (2S, 3S) -2,3-bis (t- -butyldiphenylsilyloxymethyl ) -1,1-bis (methylthio) cycloobutane <4.0 SO, 100%). NMR (200 MHz, CDCl 3) δ: 0.99 (9H, s), 1102 (9H, s), 2.06 (6H, s), 2.09-2 / 25 (2H, m), 2.85 (1n, m), 3.52-373 (an, m), I 3.90 (1H, dd, J = 9, 0.1q, sHz), 7/26 -7.47 (12H, m), 0. 9 7.55-7.74 (one, m), Example 12: Preparation of (-) - (ZR, 3R) -bis (t-butyldiphenylsilyloxymethyl) - 1,1-Bis (methylthio) cyclobutane t-butyldiphenylsilyl chloride (292 mg, 1.06 mol) was added to a solution of <+) - (2R, 3R) -2,3-bis (hydroxymethyl) -1,1-bis (methyl 1,5 1.55 in DMF thio) cyclobutane (82 mg, 0.39 mmol), imidazole (106 mg, mmol) and 4-dimethylaminomethylpyridine (catalytic amount) (4 mL) and the resulting mixture were stirred at room temperature overnight. addition of ZM phosphate buffer (pH 7.0) to the reaction mixture was extracted with ethyl acetate. The extract solution was washed with water and then with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate, after which the solvent was distilled off under reduced pressure. The residue was purified by preparing silica gel TLC Cetylacetatzhexane = 1:10, v / v) to give (-) - (2R, 3R) -2,3-bis (t-butyldiphenylsilyloxymethyl) ~ 1,1-bis- (methylthio) cyclobutane (266 mg, 100%).

The NHR and IR of this compound were identical to those of the compound of Example 11.

Example Gel 13: Preparation of (+) - <2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cycloobutanone N-chlorosuccinimide (1.60 g, 12 mmol) and silver nitrate (2.29 g , tonitrile (45 ml), 13.5 mmol) was dissolved in 80% aqueous solution of ace- to which was rapidly added at 25 ° C a solution, - 10 15 20 25 30 35 30 501 370 solution of (+) - (2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1,1-bis (methylthio) cyclobutane (2.06 g, 3 mmol) in a mixture consisting of acetonitrile (6 ml) and methylene chloride (1 ml). The resulting mixture was stirred for 10 minutes. After adding 3 ml of saturated aqueous solution of sodium sulfide to the reaction mixture and stirring it for 1 minute, saturated aqueous solution of sodium hydrogencarbonate (3 ml) was added and stirring for 1 minute, and then saturated aqueous solution of sodium chloride (3 ml) was added. and stirred for 1 minute.

Then 60 ml of a mixture of methylene chloride and hexane (1: 1, v / v) was added to this solution and insoluble matter was filtered off using Hyfldš (manufactured by Johns-Manville Co.). After drying the filtrate over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (ether hexane = 1:10, v / v) to obtain (+) - (2S, 3S) - -2,3-bis (t butyldiphenylsilyloxymethyl) -1-cyclobutanone (1.49 g, 82%).

NMR (270 MHz, CDCl 3) δ: 1.02 (9H, S), 1.04 (9H, s), δ 2.74-sfos (zu, m). 3.29 (ln, m). 3.69 (1H, da, J = 3.7, 10.6Hz), 3.82 (ln, ad, J = 4, ß, 10.3Hz), zfae (ln, da, J = 4, a, 10, 3Hz), 3.97 (ln, ad, J = 4.0, 10.6Hz), 7.32-7.44 (12H, m), A7 / 62-7.68 (BH, m), IR (δ) cm -1: 1785. Example 14: Preparation of (-) - (2R, 3R) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cyclobutanone The procedure of Example 13 was repeated except that (-) - (2R, 3R) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1,1-bis (methylthio) cyclobutane was used. There was thus obtained (-) - (2R, 3R) -2,3-bis- (t-butyldiphenylsilyloxymethyl) -1-cyclobutane in a yield of 99%.

The NMR and IR of this compound were identical to those of the compound of Example 13.

Example 15: Preparation of 2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol Example 15-1 In an atmosphere of argon gas, lithium tri (t-butoxy) - 5.0 mmol) was added to tetrahydrofuran (THF) aluminum hydride ( 1.27 g, (10 ml) and cooled to -78 ° C. Solution of (+) - (2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-2.0 mmol) To this suspension a -cyclobutanone (1.21 mg, in THF) was added and the temperature of the mixture was slowly raised to room temperature with stirring for several hours. After adding 0.2 M phosphate buffer to the reaction mixture to decompose the excess reducing agent, methylene chloride was added and inorganic The filtrate was extracted with methylene chloride and the methylene chloride layer was dried over anhydrous sodium sulfate, purified and purified by silica gel column chromatography (ethyl, after which the solvent was distilled off. The residue sepacetate hexane = 4: v / v to 1: 9 to 1: 1). (+) - (IS, 25.35) -2,3-bis (t-butyldiphenylsilyloxime yl) cyclobutanol (0.104 g, 9%): NM (zoo MHZFT, CDCl 3) δ: I 1.00 (QH, s), 1.07 (9H, s), 1.97-2.25 (zu , m), 2.32 (1H, m), 2.48 (1H, m), 10 15 20 25 30 535 32 501 370: (17 (1H, d, J = 7.3Hz), 3.56 < 2H. d, J = s, etc.>, 3} 88 (1H. dd, J = 5.6, 11 / 4Hz), 3.98 (1H. dd, J = 4f0, 11.4Hz), 4 46, (1H , m), 7.26-7.48 (lzn, m), 7554-772 (BH, m), and (+) - (IR, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl cyclobutanol (1,068 g, 881): NMR (zoomnzir, coc13) δ: 1/03 (9H, s), 1.04 (9H; s), 1.60-1.73 (za, m ), 1.92 (1H, m), 2.10-2.37 (za, m), 3, §2 ~ 3f7O (BH, m), 3 77 (ln, da, J = 4, s, 1o , 4Hz), I 4.03 (1n, m), 7.27-7.46 (12H, m), 7.56-7769 (an, m).

Example Gel 15-2 In an atmosphere of argon gas, 1M-diisobutylaluminum hydride / toluene (8.2 mL, 8.2 mmol) was slowly added at -78 ° C to a solution of (+) - (2S, 3S) -2, The 3-bis (t-butyldiphenylsilyloxymethyl) -1- in toluene (70 mL) and mixture After addition of cyclobutanone (4.12 g, 6.8 mmol) the mixture was stirred at this temperature for 10 minutes. batch of 0.2 M phosphate buffer (pH 7) to the reaction mixture and stirring it for a while, an excess of methylene chloride was added and inorganic material was filtered off. The filtrate was extracted with methylene chloride and dried over anhydrous sodium sulfate, and then the solvent was distilled off. The residue was purified by silica gel column chromatography hexane = 1: 9 to 1: 4, v / v) to give (+) - (1S, 2S, 3S) - -2.3 ~ bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (3.38 g, 82%) and (+) - (1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (0.69 g, 17%).

Example Gel 15-3: Conversion from (+) - (1R, 2S, 3S) -2,3-bis- (t-butyldiphenylsilyloxymethyl) cyclobutanol to (+) - - (15.2S, 3S) -2.3 -bis (t-butyldiphenylsilyloxymethyl) -cyclobutanol Step 1 In an atmosphere of argon gas, diethyl azodiarboxylate 1.38 mmol) was added to a solution of (+) - (1R, 2S, 3S) - (700 mg, (217 microliters, -2, 3-Bis (t-butyldiphenylsilyloxymethyl) cyclobutanol 1.37 mmol) and triphenylphosphine 1.15 mmol), benzoic acid (167 mg, (362 mg, 1.38 mmol) in benzene (10 ml) and the resulting mixture were stirred at room temperature overnight After distilling off volatiles from the reaction mixture, the residue was purified by silica gel column chromatography (ether hexane = 1:10, v / v) to give (+) - (1S, 2S, 3S) -1-benzoyl-2,3-bis ( t--butyldiphenylsilyloxymethyl) cyclobutane (791 mg, 97%).

NMR (200 MzFT, CDCl 3) δ: 0.97 (sn, s), 1.06 (9H, s), 2.23-2.40 (2H, m), 2.45 (1H, m), 2 85 (1H, m), 71- (za, d, J = s 4Hz), I (1H, dd, J = 6.1, 10, 5Hz), (1H} da, J = 7.3, 10o, snz), '48 (1H, 1mpa fi mrt q, J = 6.5Hz), 7.21-7.45 (14H, m), 7.49-7.72 (9H, m), 7.98 (zu, a, J = j¿1Hz). Step 30 In an atmosphere of argon gas, 1M-diisobutylaluminum hydride / toluene (2.6 mL, 2.6 mmol) was slowly added at -78 ° C to a solution of (+) - (1S, 2S, 3S) -1-Benzoyl-2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutane (790 mg, 1.1 mmol) in toluene (10 mL) and the mixture was stirred at this temperature for 30 minutes. 0.2 M phosphate buffer (pH 7) was added to the reaction mixture and stirred for a while, after which an excess amount of methylene chloride was added and inorganic material was filtered off. The filtrate was extracted with methylene chloride, the methylene chloride layer was dried over anhydrous sodium sulfate and the solvent was distilled off therefrom. The residue was separated and purified by silica gel column chromatography. Ceterzhexane = 1: 5, v / v %).

Example Gel 16: Preparation of (+) - (1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane Methanesulfonyl chloride (0.17 mL, 2.2 mmol) was added at 0 ° C to a solution of (+) - (1R, 2S, 3S) -2,2-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (911 mg, 1.5 mmol) and triethylamine (0.6 mL, 4.3 mmol ) in methylene chloride and the mixture was stirred at 0 ° C for 15 minutes. After adding 0.2 M phosphate buffer to the reaction mixture and extracting it with ether, the extract solution was dried over anhydrous sodium sulfate and the solvent was distilled off. The residue: was obtained by silica column column chromatography (ethyl acetate-hexane = 1: 6, v / v) to give (+) - (1R, 2S, 35) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane ( 1.034 g, 100%).

NMR (zoo mzrr, cDc13) 6: 1/04 (sn, m), 1.05 (sn, m), 2.07-2.23 (za, m), 2.43 (1H, m), , 64 (za, m), 2.91 (BH, m), 3.52-3.65 (sn, m), 3.76 (ln, ad, J = 4, or 11.0Hz). 10 15 20 25 30 šss 4.97 (ln, 7.24-7í4a 7.56-7 / 70 Exemgel 17: 35 m). (12H, m), (BH, m), 501 370 Preparation of (-) - 9-EC1S, 2R, 3S) -2,3-bis- (hydroxymethyl) cyclobutan-1-yl] -adenine (compound 2) Step 1: Preparation of (+) - 9 - {(1S, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -adenine To a suspension of adenine (177 mg, ( 6.5 ml) was added 60% sodium hydride (52 mg, 1.3 mmol) 1.3 mmol) in DMF After stirring the mixture for 1 hour a solution of 9 (+) - (IR, 28.35) - 2,3-bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (the reaction mixture and stirred at 14,500 for 6 hours. 450 mg, 0.65 mmol) in DMF (1.5 mL) to After cooling, 0.2 was added. M phosphate buffer and the mixture was extracted with ethyl acetate.

The extract solution was dried over anhydrous sodium sulfate, and then the solvent was dried over anhydrous sodium sulfate, and then the solvent was distilled off. column chromatography (methylene chloride / methanol volume / volume) The residue was separated and purified by silica gel 5021 to 30: 1, to give unreacted <+) - C (100 mg, phenylsilyloxymethyl) cyclobutan-1-yl1-adenine (203 mg, NMR (400 Mezrr, I 0.75 (sn, 1.03 (sn, 2.42-2.54 2.92-3.08) .53 (ln, 3.68 (la, 3.73-3.85 5.23 (la, 7.20-7.46 7.60-7.72 8.10 (ln, CD3OD) 8: S) , so, (2H. m).

(ZH. M), dd, J = 4] o, ad, J = ß, e, (2H. M). m), (16H, m). (4H, m).

S), 11.0nz>, 11_0Hz), _ / 22% recovery) and (+) - 9 - [(1S, 2R, 3S) -2,3-bis (t-butyldi- 43%). Step 2: Preparation of (-) - 9- [C1S, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine (Compound 2) To a methanolic solution (1 ml) of (+) - 9 - [(1S, 2R, 3S) - -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] adenine (203 mg, 0.28 mmol ), obtained in step 1, was added 4N hydrochloric acid / dioxane (0.15 ml, 0.6 mmol). After stirring the mixture at room temperature overnight, the solvent was distilled off from the reaction mixture under reduced pressure, and water was added. After removal of ether-soluble substances, it was neutralized with 0.1 N sodium hydroxide solution and the solvent was distilled off. The crude product thus obtained was purified by "Sephadex LH-20" column chromatography (methanol) water = 1: 1, v / v) and then washed with acetone to give (-) - 9-C (1S, 2S, 3S) -2 1,3 bis (hydroxymethyl) cyclobutan-1-yl] adenine (Compound 2) (26 mg, 37%).

NM (400 MMZFT, CD 3 OD) δ 2 / 43-2.57 (zu, m), 2 va (1a, m), 1 3.03 (1H, m), 3.41 (1H, ad, J = 6 , 2, 11.4Hz), -3.46 (ln, ad, J = 7.3, 11.4Hz), 3 74 (zu, d, J = s, zHz), / s zs (ln, Uppg fi mgt q , J = s, 1az), I 20 (1H, s), 8.37 (1H, s).

UV Xmax (H 2 O) nm: pH 1.258; pH 7.260; pH 13,260.

Example Gel 18: Preparation of (-) - 9 - [(1S, 2R, 3S) -2,3-bis- (hydroxymethyl) cyclobutan-1-yl] -guanine (Compounds fl) Step 1: Preparation of (-) - 2 -amino-9-E (1S, 2R, 3S) -2,3-bis- (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl1-6- - (2-methoxyethoxy) -purine Lithium hydride (10 mg, 1.3 mmol ) was added to a suspension of 2-amino-6- (2-methoxyethoxy) -purine (274mg, 1.3 mmol) in DMF 10 15 20 25 30 35, 40 37 '501 570 (6.5 ml) Then a solution of (+) - (1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxy-0.65 mmol) in DMF and the mixture was stirred for 1 h. .5 ml) to the reaction mixture and stirred at 145 ° C for 6 hours. After cooling, 0.2 M phosphate buffer was added and ethyl acetate insoluble material was filtered off and then the filtrate was extracted with ethyl acetate. The extract solution was dried over anhydrous sodium sulfate and the solvent. distilled off_ The residue was purified by silica gel column chromatography Cetylacetatzhexane = 5: 1 to 3: 1, v / v) to give unreacted (+) - (1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (157 mg, 35% recovery) and (-) - 2-amino-9- [C1S, 2R, 35) -2,3-bis (t-butyldiphenylsilyloxy methyl) cyclobutan-1-yl- 1- (2-methoxyethoxy) purine (160 mg, 31%).

Nma (400 MzFT, cD 30 OD) 6: 0/79 (9H, S), 10 3 (9H, S), 2 / 38-2.40 (za, m), 2.92 (1H, m), 2, 98 (1H, m), 3.54 (1H, dd, J = 4, 0, 11, oHz), 3.68 (1H, dd, J = 8.8, 11, oHz), 3; 72-3 , 85 (4H, m), 4756-4.67 (zu, m), 5.18 (1H, disclosed ~, fq, J-sßfggz), 7.23-7.47 (len, m), 7, 50-7.60 (4H, m), 8 / .07 (1H, s).

Step 2: Preparation of C -) - 9 - [(1S, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (compound Å) 2 N hydrochloric acid <1 ml) was added (-) - 2-amino-9- - {<1S, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutane-1- 0.17 mmol), γ1] -6- (2-methoxyethoxy) -purine (140 mg obtained in step 1) and the mixture was heated under reflux for 1 hour. After distilling off the solvent from the reaction mixture under reduced pressure, water was added and ether-soluble substances were removed. The solvent was distilled off, then neutralized with the 0.1 N solid, purified by HP-20 column chromatography (aqueous methanol 1: 0 to 1: 3, v / v) and then washed with acetone to give (-) - 9 - [(1S, 2R, 38) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (compound Å) (15 mg, 32%).

NMR (400 MHz, 2 / 36-2.55 / 2.72 (ln, 2.90 3145) 1H, (2H, 3 70 (ZH, I 5 05) 1H, I 7 94 (1H, 7 CD3 (2H, m ), OD) 8: m). M). D, J = 7.0Hz), - Ö, J = 6; 2HZ), obviously q, J = 8; 1Hz), S).

UV kmax (H 2 O) nm: pH 1.2S3.279 (charge); pH 7,252,272 (erisats); pH 13, 257 (approach), 267.

Example 19: Phenylsilyloxymethyl> -1-methanesulfonyloxycyclobutane The treatment of Example 16 was repeated, except that butanol was used. -butyldiphenylsilyloxymethyl) -methanesulfonyloxycyclobutane in a quantitative yield.

NMR (200 MzFT, CDCl 3) δ: 1.04 (9H, 1.06 (9H, - zizs-2.53 2 78 (1H, I 2 87 (3H, f 3.65 (zu, S), S)) .

(BH. M), m), S). d, J = 4, 0Hz), Preparation of (+) - (1S, 2S, 3S) -2,3-bis (t-butyldi- (+) - (1S, 28.3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclo- Thus, (+) - (1S, 2S, 3S) -2,3-bis (t-3 20 25 25 135 V40 39 '501 370 3.85 (ln, da, J = s, 3, 10 5Hz), I 3r93 (1H, then, J = 6.5, 10, 5Hz), 5.25 (ln, m), 7.32-7.50 (lzn, m), 7. so ~ 7] 75 (sn, m) - / By recrystallizing this compound from the ether-hexane system, a crystalline product ([&]) D = + 12.0 ° (c 1.01, CH 2 Cl 2)) ( optical purity 100%) is obtained.

Example Gel 20: Preparation of (-) - 9 - {(1R, 2R, 3S) -2,3-bis- (hydroxymethyl) cyclobutan-1-yl] -adenine 9? (Compound 6) Step 1: Preparation of (+) - 9 - {(1R, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -adenine 0.74 mmol) in DMF 0.75 mmol). To a suspension of adenine (100 mg, (4 ml) was added 60% sodium hydride (30 mg, stirring the mixture for 1 hour was added a solution of (+) - -ClS, 2S, 35) -2,3- bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (254 mg, 0.37 mmol) in DMF (1.5 mL) to the reaction mixture and stirred at 145 ° C for 6 h. After cooling, 0.2 M phosphate buffer was added. and the mixture was extracted with ethyl acetate.The extract solution was dried over anhydrous sodium sulfate and the solvent was distilled off, the residue was purified by silica gel column chromatography (methylene chloride / methanol = 30: 1, v / v) to give (+) - 9- - [( IR, 2R, 38) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yix-aaenin <12s mg, 47%) NMR (zoo m fl zif, CDCl 3) δ 98 (9H, S), 1 , 06 (9H, s). 2.30 - 2.65 (3H, m). 3.07 (1H, m). 3.62 - 3.84 (4H, m). 4.81 (1H, m), δ 15 20 25 30 I35 40 40 501 370 5.61 (ZH, brs), 7.20-7.52 (12H, m), 7.52-7.75 (8H , m), 7.85 (1H, s), 8.33 (1H, s).

Step 2: Preparation of (-) - 9 - [(IR, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine (Compound 6) To methanolic solution (2 ml) of (+) - 9 - [(1R, 2R, 3S) -2,3-Bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] adenia (118 mg, 0.16 mmol) obtained in step 1 was added 4 N Hydrochloric acid / dioxane (0.17 mL, 0.65 mmol) and the mixture was stirred at room temperature overnight. After distilling off the solvent from the reaction mixture under reduced pressure, water was added and ether-soluble substances were removed. Then it was neutralized with 0.1 N aqueous solution of sodium hydroxide and the solvent was distilled off. The crude product thus obtained was purified by Sephadex HP-20 column chromatography (aqueous methanol = 1: 0 to 1: 1, v / v) and then by silica gel column chromatography (methylene chloride-ethanol = 5: 1, v / v). and further thereafter by Sephadex LH-20 column chromatography (methanol) to give (-) - 9 - [(1R, 2R, 3S) -2,3-bis- (hydroxymethyl) cyclobutan-1-yl] -adenine (compound 8) (37 mg, 91%).

NMR (z 2 MHzFT, cD 3 OD) δ 2 24 (in, m).

I 2737 (1 fl, Lmpamart q, J = 9.5Hz), (1H, m) r 2 88 (1H, m), I 3 ss-3.74 (4H, m).

I 41 a, 20 (1H, s), 8.26 (1H, s).

UV max (H 2 O) nm: pH 1.259; pH 7.259; pH 13.259- \ '11 (m, upiäe fi baff. ~ q. J = a.sHz); - 10 15 20 25 30 35 40 41 501 370 HRMS (FAB) Ber. for [C 11 H 15 N 5 O 2 + H] +; 250, 1304 Found: 250, 1305 By recrystallizing this compound from ether-methanol system, a crystalline product was obtained ([α] D = -44.7 ° (c 0.98, pyridine) (optical purity: 98% or more ).

Example 21; Preparation (+) - 9 - [(1R, 2R, 3S) -bis (hydroxymethyl) -cyclobutan-1-yl] -guanine (Compound 8) Step 1: Preparation of (+) - 2-amino-9 - [( 1R, 2R, 3S) -bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -6- (2-methoxyethoxy) -purine Lithium hydride (6 mg, 0.75 mmol) 2-amino-6- (2- methoxyethoxy) -purine (155 mg, was added to a suspension of 0.74 mmol) in DMF (4 ml) and the mixture was stirred for 1 hour. Then a solution of (+) - (1S, 2S, 3-S) was added. 2,3-bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (450 mg, 0.65 mmol) in DMF (1.5 mL) to the reaction mixture and for 6 h.

After cooling, 0.2 M phosphate buffer was added and ethyl acetate was stirred at 145 ° C and insoluble matter was filtered off and the filtrate was extracted with ethyl acetate. After drying the extract solution over anhydrous sodium sulfate, the solvent was distilled off. The residue was purified by 10: 1, v / v) to give (+) - 2-amino-9 - [(1R, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl ] - 6- (2-methoxyethoxy) -purine (88 mg, 30%). silica gel column chromatography (methylene chloride: ethyl acetate = NMR (200 MHz, CDCl 3) δ 1.01 (9H, S), 1205 (9H, S), ziz-2.60 (3H, m), 2.92 (1H, m ), 3.44 (3H, s), 94 (6H, m), (SH, m), 3.60-3, 4 / SO-4.90 7.15-7.55 (lzn, m), 7.55-7.80 (sn, m), 7/67 (1H, S) When the DMSO and HMPA were used as solution, the reaction proceeded at 75 ° C with similar results in the case of DMF.

Step 2: Preparation of (+) cyclobutan-1-yl] -guanine (compound 8) agent in this reaction and 100 ° C, respectively, to give results -9 - [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) - To methanolic solution (2 ml) of (+) - amino-9 - [(1R, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutane-1-ethoxy) -purine (79 mg, 0.10 mmol), N-hydrochloric acid / dioxane (0.1 ml, obtained in step 1, -yl] -6- (2-methoxy was added 4 0.4 mmol). After stirring the mixture overnight at room temperature, it was distilled off. Then the water was added, ether soluble substances N-hydrochloric acid (2 were removed, the solvent was distilled off, 2 ml) was added and the mixture was heated under reflux for 1 hour. After neutralizing the reaction mixture with 1 M aqueous solution of sodium hydroxide was purified. den = 1: 0 to 1: 4, VOlym / - (+) - 9 - [(1R, 2R, 3S) -2,3-bis- (hydroxymethyl) cyclobu- HP-20 "column chromatography (aqueous methanol volume) for to give tan-1-yl] -guanine (compound 8) (23 mg, 88%) mm (zoo n) s; 2 18,) (1H 2; 32 (1H, m), obviously Q. 2.50 (1H, m), 2/79 (1H, m), 3; 524.74 (m, m), 4 [54 (1H, 7.89 (1H, s), UV kmax (H 2 O) nm: / Upnenbárt q, J = s, sHz), by Sephadex J = 10 OHZ), PH 1, 253.277 (approach); pH 7.253, sea (approach); pH 13.2S6 (approach) δ, 267_: mms (ïAs) + 1iH1sN503 + H] runner: zss, 1251.

Ber. for [C. By recrystallizing this compound from water, a crystalline product ([α] D = + 26.5 ° (c 0.99.0.1 N-NaOH) could be: Example 22: (optically pure - 98% or more) is obtained.

Preparation of (1R, 2R, 3S) -1-amino-2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutane Step 1: Preparation of (1R, 2R, 3S) -1-azido-2,3-bis (t -butyl-diphenylsilyloxymethyl) cyclobutane In an atmosphere of argon gas, sodium azide (975 mg, 15 mmol) was added to a solution of (+) - (1S, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1 -methanesulfonyloxycyclobutane (1.03g, 1.5mmol) in DMF (6ml) and the mixture was stirred at 120 ° C for 2 hours.

After cooling, the reaction mixture was diluted with water and extracted with ether, and the ether extract solution was washed twice with water and then with saturated aqueous sodium chloride solution. After drying the ethereal solution over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (methylene chloride / hexane = 1: 4, v / v) to give (1R, 2R, 3S) -1-azido-2,3 bis (t-butyldiphenylsilyloxymethyl) cyclobutane (992 mg, quantitative yield). 1 IR (neat) cm -1: 2100.

NMR (200 MHz, CDCl 3) δ: 1rO 3 (9H, s), 1/05 (9H, s), 1/91 (1H, m), 2.02-2.29 (2H, m), 2.44 (1n, m), 3.51-3.65 (4H, m), 3.71 (1H, dd, J = 4, 0Hz, 10 / znz), 7.31-7.44 (izn, m) 7.7 S7-7; 67 (8H, m). 441 Preparation of (1R, 2R, 3S) -1-amino-2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutane (1R, 2R, 3S) -1-azido- 2,3-bis (t-butyldiphenylsilyloxymethyl) - dissolved in ethyl acetate Step 2: as prepared in step 1, (100 mg) stirred loose cyclobutane (992 mg), (2 ml). After adding the 10% palladium carbon in an atmosphere of hydrogen at room temperature overnight. After filtering off the catalyst, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography, v / v / v) (Hexane ethyl acetate: triethylamine = to give 67%). cyclobutane (642 mg, NMR (200 MHz, CDCl 3) δ: -1103 (QH, S), 1.05 (9H, 5), 1.42 (ln, m), 1/54 (2H ), 1.86-2.05 (zu, m), 2.24 (1H, m), 3.13 (1H, m), 3.54-3.65 (sn, m), 3.75 ( 1H, dd, H = 4/1, 10, sHz>, 7.27-7.46 (lzn, m), 7.58-7.69 (ß m. M).

INDUSTRIAL APPLICABILITY: The cyclobutane derivatives of the present invention are useful carcinostatic as medicinal drugs, such as antiviral agents, agents and the like. (1R, 2R, 3S) -1-amino-2,3-bis (t-butyldiphenylsilyloxymethyl) -

Claims (10)

10 15 20 25 30 35 40 46 * 501 370 Patent claims Cyclobutane derivatives corresponding to the following general formula (IV): wherein CH 2 B represents B adenine or guanine and R 4 represents a hydrogen atom, an acyl group or a silyl group and wherein the steric configuration of the substituents of formula (IV) is ( 1R, 2R, 3s). The cyclobutane derivative according to claim 1, wherein the protecting group R 4 is an acetyl group. Cyclobutane derivative according to claim 1, which is 9 - [(1R, 2R, 3S) -2,3-bis-hydroxymethyl) cyclobutan-1-yl] guanine or 9 - [(1R, 2R, 3S) -2, 3-bis- (hydroxymethyl) cyclobutan-1-yl] adenine. An antiviral agent comprising as its active ingredient a cyclobutane derivative corresponding to the following general formula (IV): 4 R OCH 2 B (IV) 4 CH 2 OR wherein B represents adenine or guanine and R 4 represents a hydrogen atom, an acyl group or a silyl group, and wherein the steric configuration of the substituents of formula (IV) is (1R, 2R, ss). The antiviral agent according to claim 4, wherein the cyclobutane derivative is 9 - [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] guanine or 9 - [(1R, 2R, 3S) -2, 3-bis (hydroxymethyl) cyclobutan-1-yl] adenine. ' The antiviral agent of claim 4, wherein the virus is Herpes Simplex virus, cytomegalovirus, hepatitis B virus or immunodeficiency virus. 116 501 370 Cyclobutane derivatives according to claim 1 or 3, for pharmaceutical use. 8. 9 - [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine for use in the treatment of diseases caused by DNA viruses. The compound of claim 8, wherein the DNA virus is Herpes Simplex virus, cytomegalovirus or hepatitis B virus. 10. 9 - [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] adenine for use in the treatment of diseases caused by a virus.