JP2783531B2 - Anticancer drug - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to an anticancer drug.

[0002]

2. Description of the Related Art Many nucleic acid-related substances having an anticancer effect are known, and some of them are clinically used as useful pharmaceuticals, such as 5-fluorouracil and cytosine arabinoside. It has been known.

[0003]

However, the above-mentioned anticancer drugs have many problems in their application range, side effects and the like.

[0004]

According to the present invention, there is provided a compound represented by the general formula (I)
V)

[0005]

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[Wherein B is a purine base and R ⁴ is a hydrogen atom or a protecting group] (1R, 2R, 3
S) An anticancer drug comprising a cyclobutane derivative as an active ingredient.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION In the general formula (IV), as the purine base of B, for example,

[0008]

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Compounds represented by the formula: (Where Y ² represents a hydrogen atom, a halogen atom, or an amino group, Y ³ represents a hydrogen atom or an amino group, and Y ⁴ represents
Represents a hydrogen atom or an amino group)

Specific examples of the compound represented by the general formula (IV) are shown below. (B) 9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine (b) 9-[(1R, 2R, 3S) -2,3-bis (Hydroxymethyl) cyclobutan-1-yl] -guanine (c) 2-amino-9-[(1R, 2R, 3S) -2,
3-bis (hydroxymethyl) cyclobutan-1-yl] -purine (d) 2-amino-9-[(1R, 2R, 3S) -2,
3-bis (hydroxymethyl) cyclobutan-1-yl] -6-chloropurine (e) 2,6-diamino-9-[(1R, 2R, 3S)
-2,3-bis (hydroxymethyl) cyclobutane-1
-Yl] -purine (f) 9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -hypoxanthine (g) 2-amino-9-[(1R, 2R, 3S) -2,
3-bis (acetoxymethyl) cyclobutan-1-yl] -purine

The compound represented by the general formula (IV) is

[0012]

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Wherein R ⁴ represents hydrogen or a protecting group;
Represents a leaving group] and a compound represented by the general formula (VI)

[0014]

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Wherein R ⁴ represents hydrogen or a protecting group;
B ¹ represents a purine base], and when a protecting group is present in this compound, the compound is produced by removing the protecting group with an appropriate deprotecting reagent, if desired.

The protective group (R ⁴ ) in the general formulas (V) to (VI) is not particularly limited as long as it is generally used as a protective group, and includes ester-type protective groups such as acetyl and benzoyl. acyl group, dimethylcarbamoyl group, or a carbamoyl group and di-phenylcarbamoyl group, ether type protecting groups such as, t- butyl dimethyl silyl group, or a silyl group such as t-butyl diphenyl silyl group, such as methoxymethyl group (C ₁ - C ₄ ) alkoxy-
(C ₁ -C ₄ ) alkyl group, tetrahydropyranyl group,
Examples include a methyl group substituted with one or more phenyl groups such as a benzyl group, a 4-methoxybenzyl group, and a trityl group.

The leaving group (X) in the general formula (V) includes, for example, sulfonyloxy groups such as methanesulfonyloxy group, p-toluenesulfonyloxy group and trifluoromethanesulfonyloxy group, chlorine, bromine and iodine. And the like.

Examples of the purine base include adenine, hypoxanthine, guanine, 2-amino-6-chloropurine, 2-aminopurine, and 2,6-diaminopurine. These compounds may have a protecting group. For example, general formulas (XII) to (XV)

[0019]

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Wherein R ⁴ is the same as above, and R ⁵ is a C ₁ -C ₅ lower alkyl group such as a benzyl group or a butyl group, or a (C ₁ -C ₅ alkoxy) C ₁ -C ₅ group such as a methoxyethyl group. _{5 represents} an alkyl group or R ⁴ , R ⁶ represents a hydrogen atom, a halogen atom, or NHR ⁴ , and R ⁷ represents a hydrogen atom or NHR ⁴ ].

In the reaction between the compound represented by the general formula (V) and a purine base, for example, the compounds represented by the general formulas (XII) to (XV), the compound represented by the general formula (V) and the compound represented by the general formula (X
The use ratio of the compounds of II) to (XV) is about 0.5 to 10 equivalents, preferably about 1 to 5 equivalents, per equivalent of the former. The reaction between the two is carried out in the presence or absence of a base catalyst.

As the base catalyst, potassium carbonate, lithium hydride, sodium hydride or the like is used, and N, N-dimethylformamide (DMF), dimethyl sulfoxide (D
(MSO), 1,3-dimethyl-2-imidazolinone, hexamethylphosphoric triamide (HMPA) or the like, at a temperature from 0 ° C. to the reflux temperature of the solvent, preferably from about room temperature to about 170 ° C. The amount of the base catalyst used is, for example, 0 equivalent to 2 equivalents, preferably 0.5 to 1.5 equivalents to the compounds of the general formulas (XII) to (XV).
The equivalent is about double equivalent, more preferably about 0.8 to 1.2 equivalent.

The removal of the protecting group (including the alkyl group) of the compound represented by the general formula (VI) can be achieved by using an appropriate deprotecting reagent or a deprotecting method depending on the difference of the protecting group. You. Examples thereof include alkalis such as sodium hydroxide, sodium methylate, and ammonia, acids such as hydrochloric acid and sulfuric acid, fluorine reagents such as tetrabutylammonium fluoride, hydrogenolysis, and the like.

In the compound of the present invention represented by the general formula (IV), B is 2,6-diaminopurine,
The compound which is -aminopurine or 2-amino-6-halopurine can also be produced from the compound represented by the general formula (IVa). For example, the reaction formula (1)

[0025]

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In the formula, R ⁴ is the same as above, X represents a leaving group, and Y ² represents a hydrogen atom, a halogen atom or an amino group. A hydroxyl group of a guanine derivative is protected by the general formula (XX)
And then reacting with a halogenating agent such as phosphorus oxychloride or a sulfonylating agent such as 1,3,5-trimethylbenzenesulfonyl chloride to synthesize a compound represented by the general formula (XXI). .
By heating the compound represented by the general formula (XXI) with ammonia-methanol in a sealed tube, a compound in which Y ² is an amino group in the general formula (IVb) is obtained. After catalytic reduction using the compound, deprotection gives Y ^{2 in the} general formula (IVb).
Is a hydrogen atom. For example, the compound represented by the general formula (XXI) in which X is a halogen atom is deprotected to give Y in the general formula (IVb).
A compound in which ² is a halogen atom is obtained.

The compound of the present invention represented by the general formula (IV) wherein B is hypoxanthine can also be produced from the compound represented by the general formula (IVc). For example, the reaction formula (2)

[0028]

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As shown in the above, for example, the adenine derivative represented by the general formula (IVc) is hydrolyzed after diazotization by treating with nitrous acid, or by treating with a hydrolase such as adenosine deaminase. , The general formula (IV
The compound represented by d) can be obtained.

The compound of the present invention represented by the general formula (IV) can be obtained by reacting the compound of the reaction formula (3)

[0031]

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Wherein R ⁴ represents hydrogen or a protecting group;
X represents a leaving group, B represents a purine base, and B ² represents a substituent capable of leading to B in one step or in multiple steps], for example, when a compound represented by the general formula (V) is sodium azide An amine derivative represented by the general formula (XXII) is synthesized by an ordinary method such as treatment after treatment with an azide ion or the like, and the compound represented by the general formula (XXII) is synthesized by a known method (R. Vince et al. , J. Med.Chem., 2
7,1358 (1984), R. Vince et al., J. Med.Chem., 30,202
6 (1987). YF Shealy and CAO'Dell, J. Heterocyc
lic Chem., 13,1015 (1976), YFShealy et al., J. Het
The compound represented by the general formula (IV) can be obtained via the intermediate represented by the general formula (XXIII) according to erocyclic Chem., 18, 383 (1981).

In preparing the compound of the general formula (V) and the compound of the present invention represented by the general formula (IV),
The compound represented by the general formula (III) is very useful. Further, since the compound represented by the general formula (III) can be produced optically active, the compound represented by the general formula (IV) can also be produced optically active. That is, the reaction formula (4)

[0034]

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[0035] [wherein R ¹ is 2 carbon atoms attached an alkyl group, an aralkyl group or two R ¹ having 1 to 5 carbon atoms
To 3 are cyclic alkylene groups, R ² is hydrogen, an alkyl group having 1 to 5 carbon atoms, a protected hydroxyalkyl group or a protected carboxyl group, and R ³ is hydrogen, C ₁ -C ₅ lower. It represents an alkyl group, a C ₁ -C ₅ lower alkoxy group, or an aralkyloxy group. A represents a linear or branched alkylene group having 2 to 5 carbon atoms;
Represents an oxygen or sulfur atom, and Z represents a substituted or unsubstituted methylene group or an oxygen or sulfur atom.] And a compound represented by the general formula (I) and a compound represented by the general formula (II) When the reaction is carried out using a condensation catalyst, a racemic or optically active compound of the general formula (II)
The cyclobutane compound represented by I) is obtained in a high yield.

As the condensation catalyst in this reaction, for example, a Lewis acid or a combination of a Lewis acid with an equivalent or excess of a ligand can be mentioned. Examples of the Lewis acid include titanium compounds such as titanium tetrachloride and dichlorodiisopropoxy titanium, tin compounds such as tin dichloride, tin tetrachloride and divalent tin triflate, and aluminum compounds such as dimethyl aluminum chloride and diethyl aluminum chloride. Can be As the ligand, diols which are sterically hindered are preferable, for example, (2S, 3S)-
2,3-O- (1-phenylethylidene) -1,1,1
4,4-tetraphenyl-1,2,3,4-butanetetraol (compound A), (2S, 3S) -2,3-O-
Benzylidene-1,1,4,4-tetraphenyl-1,
2,3,4-butanetetraol (compound C), (2
(S, 3S) -2,3-O- (1-phenylethylidene)
In the molecule of -1,1,4,4-tetrakis (4-methoxyphenyl) -1,2,3,4-butanetetraol (compound E) or the like, a 5- or more-membered ring, preferably 5-8
And those having a membered ring and a group having a hydroxyl group bonded on both sides of the ring.

The ratio of the compound represented by the general formula (I) to the compound represented by the general formula (II) is 0.1 to 5 equivalents, preferably 0.5 to 2 equivalents, per equivalent of the former. The amount of the condensation catalyst to be used is 0.001 to 2 equivalents, preferably 0.01 to 1.2 equivalents, per 1 equivalent of the compound of the formula (I). In this reaction, the reaction may proceed more efficiently by adding a dehydrating agent such as Molecular Sieves 4A to the reaction system. Examples of the solvent for the reaction include hydrocarbon solvents such as pentane, hexane, heptane, petroleum ether, benzene, toluene, ethylbenzene, trimethylbenzene, and triisopropylbenzene, halogenated carbon solvents such as chlorofluorocarbons, ethers, and tetrahydrofuran. Examples of the solvent include ether solvents and solvents such as acetonitrile and mixed solvents thereof. The reaction temperature is preferably about the boiling point to the freezing point of the reaction solvent, and more preferably about -50 ° C to about 30 ° C.

For example, when R ² is a methoxycarbonyl group, A
Is CH ₂ CH ₂ , Y is oxygen, and Z is oxygen (1 equivalent), R ¹ is a methyl group, R ³
Is a compound of the general formula (II) wherein R is hydrogen (1.25 equivalents)
With dichlorodiisopropoxytitanium (0.05 equivalent) and (2S, 3S) -2,3-O-
Using a combination of (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,2,3,4-butanetetraol (Compound A) (0.055 equivalent), molecular sieves 4A In addition, by reacting at 0 ° C. in a hexane-toluene mixed catalyst, (2S, 3S)
-3-Methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-one-3-yl) carbonylcyclobutane can be obtained with high chemical yield and high optical yield.

Examples of the alkyl group having 1 to 5 carbon atoms represented by R ¹ , R ² and R ³ include an alkyl group such as a methyl group, an ethyl group and a butyl group, and an aralkyl group includes a benzyl group. And an alkyl group substituted with an aromatic ring such as 4-methoxybenzyl group.
Further, as the protected hydroxyalkyl group, for example, benzyloxymethyl group, acetyloxymethyl group,
and a t-butyldiphenylsilyloxymethyl group. Examples of the protected carboxyl group include an alkoxycarbonyl group such as a methoxycarbonyl group and an ethoxycarbonyl group, and an aralkyloxycarbonyl group of a benzyloxycarbonyl group. Examples of the alkoxy group having 1 to 5 carbon atoms include a methoxy group,
Examples include an allyloxy group and the like, and examples of the aralkyloxy group include a benzyloxy group, a 4-methoxybenzyloxy group, and a t-butyldiphenylsilyloxy group.

As the compound represented by the general formula (III), for example, the compound represented by the general formula (III)

[0041]

[Table 1] R ¹ R ² R ³ AYZ ────────────────────────────── Me COOMe H CH ₂ CH ₂ 00 Me COOBn H CH ₂ CH ₂ 00 Me COOMe H CH ₂ CH ₂ S S

And the like. However,
Two R ¹ of the compound represented by the general formula (III) may be the same or different. The compound represented by the general formula (V) can be obtained, for example, by using the reaction formula (5)

[0043]

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[Wherein R ¹ is the same as in the general formula (III), R ¹⁰ and R ¹¹ represent hydrogen or an alkyl group or an aralkyl group having 1 to 5 carbon atoms, and R ⁴ represents hydrogen or a protecting group. And X represents a leaving group).
It can be prepared from a compound represented by the general formula (III) (where R ² is a protected carboxyl group and R ³ is hydrogen).

First, a compound represented by the general formula (III) (where R ² is a protected carboxyl group and R ³ is hydrogen) is treated with magnesium methoxide in an alcoholic solvent such as methanol or ethanol. -78 ° C with the corresponding metal alkoxide such as sodium ethoxide
Or near the boiling point of the solvent, preferably at room temperature or lower, or in an alcoholic solvent such as methanol or ethanol, hydrochloric acid, an acid such as p-toluenesulfonic acid, or triethylamine, sodium hydroxide or the like. By solvolysis in the presence of a base, the corresponding ester in which R ¹⁰ and R ¹¹ are groups other than hydrogen in the general formula (VII) is also converted to an acid such as hydrochloric acid or p-toluenesulfonic acid. Alternatively, by hydrolysis in the presence of a base such as sodium hydroxide or potassium carbonate, the compound represented by the general formula (VII)
^A carboxylic acid is obtained wherein ¹⁰ and R ¹¹ are hydrogen.

The compound represented by the general formula (VII) thus obtained is prepared by using a metal hydride such as lithium aluminum hydride, di (isobutyl) aluminum hydride, sodium borohydride, lithium borohydride, diborane or the like. To obtain an alcohol represented by the general formula (VIII). Further, the dithioketal moiety of the compound represented by the general formula (IX) obtained by protecting the hydroxyl group of the compound represented by the general formula (VIII) can be converted to, for example, iodine, N-bromosuccinimide, N- Hydrolysis using halogenating agents such as chlorosuccinimide, sulfuryl chloride, or heavy metal compounds such as silver nitrate, silver oxide, silver perchlorate, mercury chloride, copper chloride, copper oxide, or a combination of these compounds To the ketone represented by the general formula (X).

The compound represented by the general formula (X) is used as a reducing agent such as lithium aluminum hydride, lithium tri (t-butoxy) aluminum, sodium borohydride, lithium tri (s-butyl) hydride.
Using a metal hydride complex compound such as boron or lithium borohydride or a metal hydride such as diisobutylaluminum hydride or diborane, and a solvent such as pentane, hexane, heptane, petroleum ether, benzene, toluene, ethylbenzene, etc. A hydrogen-based solvent, a carbon-based solvent such as methylene chloride and chloroform, an ether-based solvent such as ether and tetrahydrofuran, an alcohol-based solvent such as methanol and ethanol, water, and a mixed solvent thereof are used. ℃ to 5
Reduction at 0 ° C., preferably at about −80 ° C. to about 30 ° C. gives the compound represented by the general formula (XI). During this reduction reaction, by selecting the type of reducing agent used and the reaction conditions,
One of the stereoisomers can be preferentially obtained. For example, the reaction formula (6)

[0048]

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Wherein R ⁴ represents hydrogen or a protecting group.
When cyclobutanone, which is 2,3-trans represented by the general formula (Xa) as shown in the following formula, is reduced with, for example, a reducing agent which is relatively sterically intricate, the cycloformanone represented by the general formula (XIb) which is 1,2-trans The alcohol represented by the formula (XIa), which is 1,2-cis, is preferentially obtained by, for example, treating with a reducing agent having a large steric hindrance.

That is, (2S, 3S) -2,3-bis (t
-Butyldiphenylsilyloxymethyl) -1-cyclobutanone was converted to lithium hydride (t
Reduction with (-butoxy) aluminum or sodium borohydride selectively gives (1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (88% isolated yield each). , 80
%) And lithium hydride (s
-Butyl) or di (isobutyl) aluminum borohydride to give (1S, 2S, 3S) -2,3
-Bis (t-butyldiphenylsilyloxymethyl) cyclobutanol is preferential (75% isolated yield, 8% isolated yield, respectively).
2%). The results are shown in the following table.

[0051]

[Table 2] ──────────────────────────────────── condition yield (%) ───── ─────────────────────────────── reagent solvent temp.XIa XIb ───────────── ─────────────────────── LiAI (OtBu) ₃ H THF −78−RT 988 NaBH ₄ THF-H ₂ O RT 19 80 Li (sBu) ₃ BH THF −78 75 23 (iBu) ₂ AIH CH ₂ Cl ₂ −78 74 22 (iBu) ₂ AIH Toluene −78 82 17 ──────────────────── ──────────────── R ⁴ = t-BuPh ₂ Si (XIa, XIb)

The stereoisomers obtained here, ie, the compound represented by the general formula (XIa) and the compound represented by the general formula (XI
After isolation, the compounds of the formula b) can easily be converted into other isomers of one another. For example, the general formula (XI
a) a compound represented by formula (XIb) or a metal oxidizing agent such as chromic acid / acetic acid, chromic acid / pyridine, dimethyl sulfoxide (DMSO)-
Reoxidation to the ketone represented by the general formula (Xa) by a usual oxidation reagent or oxidation method such as acetic anhydride / acetic acid, DMSO oxidation of DMSO-oxalyl chloride triethylamine / methylene chloride, etc., and further selective reduction. Method,
Alternatively, the configuration of the 1-position carbon having a hydroxyl group of the compound represented by the general formula (XIa) and the compound represented by the general formula (XIb) may be determined, for example, by the Mitsunobu reaction (Mitsumobu Re
action) to reverse it.

For example, a compound represented by the general formula (XIa) or a compound represented by the general formula (XIb) is converted to a trivalent phosphorus compound such as triphenylphosphine, trimethyl phosphite, triethyl phosphite, acetic acid, benzoic acid and the like. Carboxylic acids such as acids, and azodicarboxylic esters such as diethyl azodicarboxylate as solvents, for example, pentane, hexane, heptane, petroleum ether, benzene,
Using a hydrocarbon solvent such as toluene and ethyl benzene, a carbon halide solvent such as methylene chloride and chloroform, an ether solvent such as ether and tetrahydrofuran, and a mixed solvent thereof, the reaction temperature is from -100 ° C to 50 ° C. Preferably, the ester formed by reacting at about -50 ° C to about 30 ° C is hydrolyzed with an acid such as an alkali such as potassium carbonate, sodium hydroxide, sodium methylate and ammonia, hydrochloric acid, sulfuric acid, or hydrogen. Lithium aluminum hydride, lithium trihydride (s-
Butyl) boron or a metal hydride complex such as lithium borohydride or a metal hydride such as diisobutylaluminum hydride or diborane to obtain a compound represented by the general formula (XIb)
Or the compound represented by the general formula (XIa) can be obtained.

For example, (1R, 2S, 3S) -2,3-
Bis (t-butyldiphenylsilyloxymethyl) cyclobutanol is reacted with triphenylphosphine / benzoic acid / diethylazodicarboxylate in benzene to give (1S, 2S, 3S) -1-benzoyl-2,3-
After leading to bis (t-butyldiphenylsilyloxymethyl) cyclobutane, the mixture was reduced with diisobutylaluminum hydride in toluene to remove the benzoyl group, thereby obtaining (1S, 2S, 3S) -2,3-bis (t-). (Butyldiphenylsilyloxymethyl) cyclobutanol is obtained. The compound represented by the general formula (V) is produced by introducing the secondary hydroxyl group of the compound represented by the general formula (XI) thus obtained into a leaving group.

The following experimental examples show that the compounds of the present invention exert a broad and strong anticancer effect.

Experimental Example 1 Human cervical cancer HeLa S ₃ ,
The anticancer effect on mouse leukemia L1210 and P388 cells was tested by the following method. (Method 1) HeLa S ₃ cells were 7.5 × 10 ³ cells
It was adjusted to ls / ml and seeded at 0.2 ml / well on a 96-well flat bottom plate. 37 ° C, 5% CO ₂
After culturing for 24 hours in an incubator, 1
0 μl was added and the cells were cultured for 72 hours. After the culture, the culture solution of each well is extracted, fixed with methanol, and then 0.05%
Methylene blue / 10 mM-Tris-HCI (pH
8.5) The solution was added at a rate of 0.1 ml / well and stained at room temperature for 30 minutes. Each well was stained with an aspirator and then washed three times with pure water. 3%
After adding HCl at a rate of 0.2 ml / well, the cells were sealed with a seal and left at room temperature for about 24 hours to extract the dye from the cells. The absorbance at 660 nm of each well was measured with a Dynatech microplate reader, and the growth inhibition rate (%) at each concentration was calculated by the formula shown below, and plotted on a log-probability paper to show a 50% inhibitory concentration (IC ₅₀ value). , Μg / ml). Growth inhibition rate (%) = (1−A ₁ / A ₀ ) × 100 where A ₁ = absorbance of drug-treated group A ₀ = absorbance of control group

(Method 2) Cells of P388 and L1210 were seeded on a 24-well plate, added with a drug, and cultured at 37 ° C., 5% CO ₂ for 48 hours. The number of cells after completion of the culture was calculated using a Coulter counter. The growth inhibition rate of the treated group with respect to the control group was calculated from the following formula, and L
50% inhibitory concentration (IC
₅₀ value, μg / ml). Growth inhibition rate (%) = _{[1- (N T -N 0) /} (N c -N
₀ ] × 100 where _NT = number of cells in the drug-treated group N ₀ = number of cells at the start of culture N _c = number of cells in the control group

[0058]

[Table 3] ────────────────────────────────────IC ₅₀ (μg / ml) compound── ───────────────────────────── No. HeLaS ₃ P388 L1210 ─────────────── ──────────────────── (a) 14 0.58 1.1 (b) 19 1.6 8.1 ────────────────

The compound of the present invention represented by the general formula (IV) is expected to be used as an anticancer agent because it has an anticancer effect. When the compound of the present invention obtained as described above is used as an anticancer agent, it can be administered orally, intravenously, or transdermally. The dose varies depending on the patient's condition, age, and administration method, but is usually 0.1 to 500 mg / kg.
/ Day. The compound of the present invention is administered in the form of a preparation prepared by mixing with an appropriate preparation carrier. As the form of the preparation, tablets, granules, fine granules, powders, capsules, injections, creams, suppositories and the like are used.

[0060]

The production of the compound of the present invention will be specifically described below with reference to examples.

Embodiment 1 (-)-(2S, 3S) -3-methoxycarbonyl-
1,1-bis (methylthio) -2- (oxazolidine-
Production of 2-one-3-yl) carbonylcyclobutane

Embodiment 1-1. In an argon atmosphere, dichlorodiisopropoxy titanium (905 mg, 3.8 mmol)
1) and (2S, 3S) -2,3-O- (1-phenylethylidene) -1,1,4,4-tetraphenyl-1,
2,3,4-butanetetraol (compound A) (2.2
(2 g, 4.2 mmol), 1,3,5-trimethylbenzene (TMB) (80 ml) is added, and the mixture is stirred at room temperature for 30 minutes. To this solution, add powdered molecular sieves 4A.
(3.2 g) and stirred for a while.
[(E) -3- (methoxycarbonyl) propenoyl]
-Oxazolidine-2-one (3.98 g, 20 mmol
1) and cool to -15 ° C. 1,1-
Bis (methylthio) ethylene (3.61 g, 30 mmo
1) TMB solution (20 ml) was gradually added, and the temperature was raised to 0 ° C. over 3 hours from the same temperature with stirring. A saturated aqueous solution of sodium hydrogen carbonate is added to the reaction solution, the inorganic substance is removed by filtration using celite, and the organic substance is extracted with ethyl acetate. The extract is washed with saturated saline, dried over anhydrous sodium sulfate, and the solvent is distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: hexane = 1: 1).
2,-(v / v)) and purified by (-)-(2S, 3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2-
(Oxazolidin-2-one-3-yl) carbonylcyclobutane (3.94 g, 62%) is obtained. The optical purity of this compound is 86% ee (see Example 3).

NMR (500 MHz FT, CDCl ₃ )
δ: 2.01 (3H, s), 2.10 (3H, s) 2.54 (1H, dd, J = 9.9, 12.3 Hz),
2.70 (1H, dd, J = 7.9, 12.3 Hz),
3.86 (1H, dt, J _d = 9.9 Hz, J _t = 7.
9Hz), 3.71 (3H, s), 4.02 (1H, d
dd, J = 6.4, 8.9, 11.0 Hz), 4.12
(1H, ddd, J = 7.6, 9.3, 11.0H
z), 4.39-4.47 (2H, m) 5.01 (1H, d, J = 7.9 Hz). IR (neat) cm ^-1 : 1780, 1730, 169
0.

Embodiment 1-2. Almost in the same manner as in Example 1-1, 3-[(E) -3- (methoxycarbonyl) propenoyl] -oxazolidin-2-one (3.98 g, 2
0 mmol), 1,1-bis (methylthio) ethylene (3.61 g, 30 mmol), dichlorodiisopropoxytitanium (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 m
mol), powdered molecular sieves 4A (4 g) and toluene (160 ml) and hexane (120 m
1) By reacting at 0 ° C. for 40 minutes in a mixed solvent,
(-)-(2S, 3S) -3-methoxycarbonyl-
1,1-bis (methylthio) -2- (oxazolidine-
2-on-3-yl) carbonylcyclobutane (6.1
3g, 96%) ([α] _D = -10.4 ° (c 1.3
4, CH ₂ Cl ₂ )). This was recrystallized from methylene chloride-isopropyl ether to give 5.30 g.
(83%) ([α] _D = -11.1 ° (c 1.15,
CH ₂ Cl ₂ )) is obtained. The optical purity of this compound is 98% ee or more (determined by the method described in Example 3).

Embodiment 1-3. Almost in the same manner as in Example 1-1, 3-[(E) -3- (methoxycarbonyl) propenoyl] -oxazolidin-2-one (19.9 g, 1
00 mmol), 1,1-bis (methylthio) ethylene (15.03 g, 125 mmol), dichlorodiisopropoxytitanium (1.18 g, 5.0 mmol), (2
(S, 3S) -2,3-O- (1-phenylethylidene)
-1,1,4,4-tetraphenyl-1,2,3,4-
Butanetetraol (Compound A) (2.91 g, 5.5)
mmol), powdered molecular sieves 4A (20 g)
Using toluene (800 ml) and hexane (60 ml).
0 ml) By reacting in a mixed solvent at 0 ° C. for 5 hours, (−)-(2S, 3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-one-3- Yl) carbonylcyclobutane (2
6.69 g, 84%) ([α] _D = -10.5 ° (c
1.00, CH ₂ Cl ₂ )).

Embodiment 2 FIG. Production of (-)-(2S, 3S) -2,3-bis (methoxycarbonyl) -1,1-bis (methylthio) cyclobutane Example 2-1. (-)-(2S, 3S) -3-methoxycarbonyl-1,1-bis (methylthio) -2- (oxazolidin-2-one-3-yl) carbonyl prepared in Example 1-1 in an argon atmosphere. Cyclobutane (3.
94 g, 12.3 mmol) in methanol (25 m
l) is 1M-dimethoxymagnesium / methanol (2
5 ml, 25 mmol) and stir at room temperature for 1 hour.
After the reaction solution is concentrated under reduced pressure, a saturated aqueous ammonium chloride solution is added, and the mixture is extracted with ether. After the ether extract is washed with a saturated saline solution, the solvent is distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: hexane = 1: 9, v / v) to give (-)-(2S, 3
S) -2,3-Bis (methoxycarbonyl) -1,1-
Bis (methylthio) cyclobutane (2.17 g, 67
%). The optical purity of this compound is 86% ee (see Example 3).

NMR (500 MHz FT, CDCl ₃ )
δ: 2.01 (3H, s), 2.12 (3H, s),
2.47 (1H, dd, J = 9.0, 12.2 Hz),
2.50 (1H, dd, J = 9.4, 12.2 Hz),
3.63-3.75 (2H, m), 3.69 (3H,
s), 3.72 (3H, s), IR (neat) cm ^-1 : 1730.

Embodiment 2-2. 1M- in argon atmosphere
Dimethoxymagnesium / methanol (400 ml, 4
(00 mmol) at 0 ° C. and (−)-(2S, 3S) -3
-Methoxycarbonyl-1,1-bis (methylthio)-
2- (Oxazolidin-2-one-3-yl) carbonylcyclobutane (26.0 g, 81.4 mmol) is added, and the mixture is stirred at the same temperature for 15 minutes. A saturated aqueous ammonium chloride solution is added to the reaction solution, and the mixture is extracted with ether. After the ether extract is washed with a saturated saline solution, the solvent is distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate: hexane = 1: 9, v / v) (-)-.
(2S, 3S) -2,3-bis (methoxycarbonyl)
-1,1-bis (methylthio) cyclobutane (20.7
3 g, 96%).

Embodiment 3 FIG. Production of (−)-(2S, 3S) -2,3-bis (hydroxymethyl) -1,1-bis (methylthio) cyclobutane Lithium aluminum hydride (56
2 mg, 14.8 mmol) in an ether suspension at 0 ° C.
(-)-(2S, 3S)-manufactured in Example 2-1
2,3-bis (methoxycarbonyl) -1,1-bis (methylthio) cyclobutane (1.96 g, 7.4 mm
ol) (10 ml) was slowly added thereto, followed by stirring at 0 ° C. for 2 hours. After adding an aqueous saturated sodium sulfate solution to the reaction solution to decompose excess reducing agent, anhydrous sodium sulfate is added and the mixture is stirred for a while. After the inorganic substance is removed by filtration and further washed with hot isopropyl alcohol, the filtrate and the washing solution are combined and the solvent is distilled off under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate; hexane: methanol = 15: 20: 1, v / v / v) to give (-)-(2
S, 3S) -2,3-bis (hydroxymethyl) -1,
1-bis (methylthio) cyclobutane (1.48 g, 9
6%).

NMR (270 MHz FT, CDCl ₃ )
δ: 2.02 (1H, dd, J = 9.1, 12.1H
z), 2.05 (3H, s), 2.06 (3H, s),
2.28 (1H, dd, J = 8.1, 12.1 Hz),
2.47-2.68 (2H, m), 3.26 (2H, b
rs), 3.54 (1H, dd, J = 8.8, 10.3)
Hz), 3.67-3.77 (2H, m), 3.83
(1H, dd, J = 5.0, 10.4 Hz). IR (neat) cm ^-1 : 3350.

A part of the compound was obtained by a conventional method ((R) -α-
Methoxy-α-trifluoromethyl-phenylacetyl chloride ((R) -MTPACl), dimethylaminopyridine (DMAP) / pyridine (pyr) led to bis (R) -MTPA ester. 500MHzNM
In R, the signal of the methyl group of this compound derived from the racemate is 1.852, 1.856, 1.912, 1.9.
A total of four signals at 70 ppm were observed. The signals of the methyl group of the substance were 1.852 and 1.912 ppm, which were larger than those of the others.
ee.

The title compound (-)-(2S, 3S)
-2,3-Bis (hydroxymethyl) -1,1-bis (methylthio) cyclobutane can be recrystallized from ethyl acetate-hexane to obtain a crystal having an optical purity of 100%. ([Α] _D = -32.0 ° (c
_{1.03, CH 2 Cl 2))} .

Embodiment 4 FIG. Production of (+)-(2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1,1-bis (methylthio) cyclobutane (-)-(2S, 3S) -2,3- Bis (hydroxymethyl) -1,1-bis (methylthio) cyclobutane (1.37 g, 6.58 mmol), triethylamine (2.8 ml, 20 mmol), 4-dimethylaminomethylpyridine (catalytic amount) and DMF (1 ml) To a solution dissolved in methylene chloride (25 ml) is added t-butyldiphenylsilyl chloride (4.52 g, 25 mmol), and the mixture is stirred at room temperature overnight. The reaction solution is concentrated under reduced pressure, dissolved in ether, washed with water, washed with saturated saline, and dried over anhydrous sodium sulfate. After distilling off the solvent under reduced pressure, the ether solution is purified by silica gel column chromatography (ether: hexane = 1: 20, v / v) (+).
-(2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1,1-bis (methylthio)
Cyclobutane (4.50 g, 100%) is obtained.

NMR (200 MHz FT, CDCl ₃ )
δ: 0.99 (9H, s), 1.02 (9H, s),
2.06 (6H, s), 2.09-2.25 (2H,
m), 2.85 (1H, m), 3.52-3.73 (3
H, m), 3.90 (1H, dd, J = 9.0, 10.
8 Hz), 7.26-7.47 (12H, m), 7.5
5-7.74 (8H, m).

Example 5 Preparation of (+)-(2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cyclobutanone N-chlorosuccinimide (1.60 g, 12 mmol)
l) and silver nitrate (2.29 g, 13.5 mmol) in 8
Dissolve in 0% acetonitrile aqueous solution (45 ml) and add 25%
C. (+)-(2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1,1-bis (methylthio) cyclobutane (2.06 g, 3 mmol) in acetonitrile (6 ml) and methylene chloride (1 ml) is added quickly and stirred for 10 minutes. A saturated aqueous sodium sulfite solution (3 ml) was added to the reaction solution, and the mixture was stirred for 1 minute. Then, a saturated aqueous sodium hydrogen carbonate solution (3 ml) was added.
, And the mixture is stirred for 1 minute. Saturated saline (3 ml) is further added, and the mixture is stirred for 1 minute. After adding a methylene chloride-hexane mixed solution (1: 1, v / v) (60 ml) to this solution, the insoluble matter is removed by filtration using Hyflo. After the filtrate was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (ether: hexane =
1:10, v / v) (+)-(2S, 3S)-
This gives 2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cyclobutanone (1.49 g, 82%).

NMR (270 MHz FT, CDCl ₃ )
δ: 1.02 (9H, s), 1.04 (9H, s),
2.74-3.05 (2H, m), 3.29 (1H,
m), 3.69 (1H, dd, J = 3.7, 10.6H
z), 3.82 (1H, dd, J = 4.8, 10.3H
z), 3.88 (1H, dd, J = 4.8, 10.3H
z), 3.97 (1H, dd, J = 4.0, 10.6H
z), 7.32-7.44 (12H, m), 7.62-
7.68 (8H, m), IR (neat) cm ^-1 : 1785.

Embodiment 6 FIG. Production of 2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol Example 6-1. Under an argon atmosphere, lithium tri (t-butoxy) aluminum (1.27 g, 5.0 m
mol) in tetrahydrofuran (THF) (10 ml)
And cool to -78 ° C. Add (+)-
(2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cyclobutanone (1.21
g, 2.0 mmol) in THF, and slowly warm to room temperature over several hours with stirring. 0.2M in reaction solution
-After decomposing excess reducing agent by adding a phosphate buffer, methylene chloride is added and inorganic substances are removed by filtration. The filtrate is extracted with methylene chloride, dried over anhydrous sodium sulfate, and the solvent is distilled off. The residue was separated and purified by silica gel column chromatography (ethyl acetate: hexane = 1: 9-1: 4, v / v) to give (+)-(1S, 2S, 3S) -2,3-
Bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (0.104 g, 9%)

NMR (200 MHz FT, CDCl ₃ )
δ: 1.00 (9H, s), 1.07 (9H, s),
1.97-2.25 (2H, m), 2.32 (1H,
m), 2.48 (1H, m), 3.17 (1H, d, J
= 7.3 Hz), 3.56 (2H, d, J = 5.8H)
z), 3.88 (1H, dd, J = 5.6, 11.4H)
z), 3.98 (1H, dd, J = 4.0, 11.4H
z), 4.46 (1H, m), 7.26-7.48 (1
2H, m), 7.54-7.72 (8H, m),

And (+)-(1R, 2S, 3S) -2,
3-bis (t-butyldiphenylsilyloxymethyl)
Cyclobutanol (1.068 g, 88%) is obtained.

NMR (200 MHz FT, CDCl ₃ )
δ: 1.03 (9H, s), 1.04 (9H, s),
1.60-1.73 (2H, m), 1.92 (1H,
m), 2.10-2.37 (2H, m), 3.52-
3.70 (3H, m), 3.77 (1H, dd, J =
4.5, 10.4 Hz), 4.03 (1H, m), 7.
27-7.46 (12H, m), 7.56-7.69
(8H, m).

Embodiment 6-2. Under argon atmosphere, (+)
-(2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-cyclobutanone (4.1
To a solution of 2 g (6.8 mmol) in toluene (70 ml) was slowly added 1 M diisobutylaluminum hydride / toluene (8.2 ml, 8.2 mmol) at -78 ° C, and the mixture was stirred at the same temperature for 10 minutes. A 0.2 M phosphate buffer (pH 7) is added to the reaction solution, and after stirring for a while, an excess of methylene chloride is added and inorganic substances are removed by filtration. The filtrate was extracted with methylene chloride, dried over anhydrous sodium sulfate,
The solvent is distilled off. The residue was subjected to silica gel column chromatography (ethyl acetate: hexane = 1: 9-1: 4, v /
v) to separate and purify, (+)-(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%) is obtained.

Embodiment 6-3. From (+)-(1R, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol, (+)-(1S, 2S, 3S) -2,3-bis (t
-Butyldiphenylsilyloxymethyl) cyclobutanol Step 1. In an argon stream, (+)-(1R, 2S, 3
S) -2,3-Bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (700 mg, 1.15 m
mol), benzoic acid (167 mg, 1.37 mmol)
And triphenylphosphine (362 mg, 1.38
mmol) in benzene (10 ml) was added to diethylazodicarboxylate (217 μl, 1.38 mmol).
l) and stir at room temperature overnight. After evaporating the volatile substances under reduced pressure, the reaction solution 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%) is obtained.

NMR (200 MHz FT, CDCl ₃ )
δ: 0.97 (9H, s), 1.06 (9H, s),
2.23-2.40 (2H, m), 2.45 (1H,
m), 2.85 (1H, m), 3.71 (2H, d, J
= 5.4 Hz), 3.83 (1H, dd, J = 6.1,
10.5 Hz), 3.99 (1H, dd, J = 7.3,
10.5Hz), 5.48 (1H, aperent)
q. J = 6.5 Hz), 7.21-7.45 (14H,
m), 7.49-7.72 (9 Hz), 7.98 (2
H, d, J = 7.1 Hz).

Step 2. (+)-(1S, 2S, 3S) -1-benzoyl- obtained in step 1 in an argon stream.
To a solution of 2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutane (790 mg, 1.1 mmol) in toluene (10 ml) at −78 ° C., 1M diisobutylaluminum hydride / toluene (2.6 ml, 2.
6 mmol) is gradually added, and the mixture is stirred at the same temperature for 30 minutes.
A 0.2 M phosphate buffer (pH 7) is added to the reaction solution, and after stirring for a while, an excess of methylene chloride is added and inorganic substances are removed by filtration. The filtrate is extracted with methylene chloride, dried over anhydrous sodium sulfate, and the solvent is distilled off. The residue was subjected to silica gel column chromatography (ether: hexane =
1: 5, v / v) and purified by (+)-(1S, 2
(S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (644 mg, 95
%).

Embodiment 7 FIG. Production of (+)-(1S, 2S, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (+)-(1S, 2S, 3S) -2,3 To a solution of -bis (t-butyldiphenylsilyloxymethyl) cyclobutanol (911 mg, 1.5 mmol) and triethylamine (0.6 ml, 4.3 mmol) in methylene chloride at 0 ° C was added methanesulfonyl chloride (0.17 ml,
2.2 mmol) and stir at 0 ° C. for 15 minutes. A 0.2 M phosphate buffer is added to the reaction solution, and the mixture is extracted with ether. After the ether extract is dried over anhydrous sodium sulfate, the solvent is distilled off. The residue was subjected to silica gel column chromatography (acetate: hexane = 1: 6, v /
v) and purified by (+)-(1S, 2S, 3S) -2,3-
Bis (t-butyldiphenylsilyloxymethyl) -1
Obtaining methanesulfonyloxycyclobutane (quantitative).

NMR (200 MHz FT, CDCl ₃ )
δ: 1.04 (9H, s), 1.06 (9H, s),
2.25-2.53 (3H, m), 2.78 (1H,
m), 2.87 (3H, s), 3.65 (2H, d, J
= 4.0 Hz), 3.85 (1H, dd, J = 6.3,
10.5 Hz), 3.93 (1H, dd, J = 6.5,
10.5Hz), 5.25 (1H, m), 7.32-
7.50 (12H, m), 7.60-7.75 (8H,
m).

This compound was recrystallized from ether-hexane to give a crystal ([α] _D = + 12.0 ° (c
1.01, CH ₂ Cl ₂ )) (100% optical purity).

Example 8 Production of (-)-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine (Compound A) Step 1 (+)- Preparation of 9-[(1R, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -adenine DMF of adenine (100 mg, 0.74 mmol)
(4 ml) 60% sodium hydride (30 m
g, 0.75 mmol) and stir for 1 hour. (+)-(1S, 2S, 3S) -2,3-bis (t
-Butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (254 mg, 0.37
mmol) in DMF (1.5 ml).
Stir at C for 6 hours. After cooling, a 0.2 M phosphate buffer is added, and the mixture is extracted with ethyl acetate. After the extract is dried over anhydrous sodium sulfate, the solvent is distilled off. The residue was purified by silica gel column chromatography (methylene chloride: methanol = 30: 1, v / v) to give (+)-9-[(1
(R, 2R, 3S) -2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -adenine (126 mg, 47%) is obtained.

NMR (200 MHz FT, CDCl ₃ )
δ: 0.98 (9H, s), 1.06 (9H, s),
2.30-2.65 (3H, m), 3.07 (1H,
m), 3.62-3.84 (4H, m), 4.81 (1
H, m), 5.61 (2H, 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-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine (Compound A) ) -9-[(1R, 2R, 3S)
-2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -adenine (118 m
g, 0.16 mmol) in 4 ml of methanol solution (2 ml) in 4N hydrochloric acid / dioxane (0.17 ml, 0.65 mm).
ol) and stirred overnight at room temperature. After evaporating the solvent under reduced pressure, the reaction solution was added with water, and the ether-soluble portion was removed.
Neutralize with 0.1N sodium hydroxide solution and evaporate the solvent. The crude product was subjected to Sephadex HP-20 column chromatography (water: methanol = 1: 0-1: 1, v / m
v), silica gel column chromatography (methylene chloride: ethanol = 5: 1, v / v), and further separation and purification by Sephadex LH-20 column chromatography (methanol) to give (-)-9-[(1R, 2R). , 3
S) -2,3-Bis (hydroxymethyl) cyclobutan-1-yl] -adenine (compound A) (37 mg, 91
%).

NMR (200 MHz FT, CD ₃ OD)
δ: 2.24 (1H, m), 2.37 (1H, appa)
rent q, J = 9.5 Hz), 2.62 (1H,
m), 2.88 (1H, m), 3.65-3.74 (4
H, m), 4.71 (1H, applicable q, J
= 5 Hz), 8.20 (1H, s), 8.26 (1H,
s). UV λ max (H ₂ O) nm: pH 1,259; pH
7,259; pH 13,259. HRMS (FAB) Calcd for [C ₁₁ H ₁₅ N ₅ O ₂ + H] ⁺ ; 2
50.1304 Found; 250.1305.

The compound ([α] _D = -44.7 °) was recrystallized from ether-methanol.
(C 0.98, pyridine)) (optical purity> 98%).

Embodiment 9 FIG. Production of (+)-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (Compound B) (+)-2-Amino-9-[(1R, 2R, 3S)-
Preparation of 2,3-bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -6- (2-methoxyethoxy) purine 2-amino-6- (2-methoxyethoxy) purine (1
Lithium hydride (6 mg, 0.75 mmol) is added to a suspension of 55 mg (0.74 mmol) in DMF (4 ml) and stirred for 1 hour. (+)-(1S, 2S,
3S) -2,3-Bis (t-butyldiphenylsilyloxymethyl) -1-methanesulfonyloxycyclobutane (450 mg, 0.65 mmol) in DMF (1.5
ml) and add the solution and stir at 145 ° C. for 6 hours. After cooling, a 0.2 M phosphate buffer and ethyl acetate are added, the insolubles are filtered off, and the mixture is extracted with ethyl acetate. After the extract is dried over anhydrous sodium sulfate, the solvent is distilled off. The residue is
Silica gel column chromatography (methylene chloride:
(Ethyl acetate = 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% Get)

NMR (200 MHz FT, CDCl ₃ )
δ: 1.01 (9H, s), 1.05 (9H, s),
2.25-2.60 (3H, m), 2.92 (1H,
m), 3.44 (3H, s), 3.60-3.94 (6
H, m), 4.50-4.90 (5H, m), 7.15
−7.55 (12H, m), 7.55-7.80 (8
H, m), 7.67 (1H, s).

As the solvent for this reaction, DMSO and HMP
When A is used, the reaction proceeds at 75 ° C. and 100 ° C., respectively, and the same results as when DMF is used are obtained.

Step 2. Production of (+)-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (compound b) (+)-2-amino produced in Step 1 −9-[(1R,
2R, 3S) -2,3-Bis (t-butyldiphenylsilyloxymethyl) cyclobutan-1-yl] -6
(2-Methoxyethoxy) -purine (79 mg, 0.1
4N-hydrochloric acid / dioxane (0.1 ml, 0.4 mmol) was added to a methanol solution (2 ml) of 0 mmol), and the mixture was stirred at room temperature overnight. After distilling off the solvent under reduced pressure,
Water was added to remove the ether-soluble portion, the solvent was distilled off, and 2N-
An aqueous hydrochloric acid solution (2 ml) is added, and the mixture is heated under reflux for 1 hour. After the reaction solution was neutralized with a 1 M sodium hydroxide solution, Sephadex HP-20 column chromatography (water:
Methanol = 1: 0-1: 4, v / v),
(+)-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (compound b) (23 mg, 88%) is obtained.

NMR (200 MHz FT, CD ₃ OD)
δ: 2.18 (1H, m), 2.32 (1H, appa)
rent q, J = 10.0 Hz), 2.50 (1H,
m), 2.79 (1H, m), 3.62-3.74 (4
H, m), 4.54 (1H, applicable q, J
= 8.8 Hz), 7.89 (1H, s). UV λmax (H ₂ O) nm: pH 1,253,27
7 (sh); pH 7, 253, 268 (sh); pH 1
3,256 (sh), 267 HRMS (FAB) Calcd for [C ₁₁ H ₁₅ N ₅ O ₃ + H] ⁺ : 2
66.1253 Found; 266.1125.

This compound was recrystallized from water to give a crystal ([α] _D = + 26.5 ° (c 0.99,
0.1 N-NaOH)) (optical purity> 98%). Compounds (c), (d), (e) and (f) are obtained in the same manner as in this example using the corresponding base.

Embodiment 10 FIG. 2-amino-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -6
-Production of chloropurine (compound d) 2-amino-9-[(1R, 2R, 3S) -2,3-bis (acetoxymethyl) cyclobutan-1-yl] -6
Preparation of -chloropurine 9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine (compound b) (265 mg, 1.0 mmol) in anhydrous dimethylformamide (1 0.5 ml) suspension to pyridine (0.5 m
l) and acetic anhydride (1 ml) are added and stirred at 75 ° C. for 30 minutes. The solvent was distilled off from the reaction solution under reduced pressure, azeotroped several times with toluene, and then dried to obtain crude 9-[(1R, 2R, 3S).
-2,3-bis (acetoxymethyl) cyclobutane-1
-Yl] -guanine (250 mg) is obtained. The obtained crude 9-[(1R, 2R, 3S) -2,3-bis (acetoxymethyl) cyclobutan-1-yl] -guanine (350 mg) and tetraethylammonium chloride (331 mg, 2.0 mmol) were anhydrous. It was dissolved in acetonitrile (2 ml) and N, N-dimethylaniline (13
0 μl, 1.0 mmol) and phosphorus oxychloride (0.5
7 ml, 6.0 mmol) and heat to reflux at 100 ° C. for 10 minutes. The reaction solution was distilled under reduced pressure to remove volatile substances,
Crushed ice and methylene chloride were added to the residue, and the mixture was stirred at 0 ° C. for a while, then neutralized with a saturated aqueous solution of sodium hydrogen carbonate, and concentrated.
After stirring at ℃ for a while, extract with methylene chloride. After the methylene chloride extract is dried over anhydrous sodium sulfate, the solvent is distilled off. The residue was subjected to silica gel column chromatography (methylene chloride: ethyl acetate = 1: 1, v / v
v) and purified by 2-amino-9-[(1R, 2R, 3S)
-2,3-bis (acetoxymethyl) cyclobutane-1
-Yl) -6-chloropurine (335 mg, 91%) is obtained.

NMR (200 MHz FT, CDCl ₃ )
δ: 2.02 (3H, s), 2.11 (3H, s),
2.18-2.73 (3H, m), 3.09 (1H,
m), 4.25 (4H, d, J = 5.9 Hz), 4.5
9 (1H, applicable q, J = 8.4 Hz),
4.65 (2H, br), 7.88 (1H, s).

Step 2. 2-amino-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -6
Preparation of -chloropurine (compound d) 2-amino-9-[(1R, 2R, 3S) -2,3-bis (acetoxymethyl) cyclobutan-1-yl] -6
-Chloropurine (98 mg, 0.27 mmol) was dissolved in methanol (2 ml) and potassium carbonate (90 mg,
(0.65 mmol) and stirred at 0 ° C. for 30 minutes. After neutralizing the reaction solution with 1N-hydrochloric acid, Sephadex HP2
Purified with 0 (water-60% methanol aqueous solution) and 2-amino-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -6-chloropurine (compound d) (65 mg, 86%).

NMR (200 MHz FT, CD ₃ OD)
δ: 2.20 (1H, m), 2.37 (1H, m),
2.57 (1H, m), 2.89 (1H, m), 3.6
9 (4H, d, J = 5.5 Hz), 4.64 (1H, a
parent q, J = 8.8 Hz), 8.19 (1
H, s). UV λmax (H ₂ O) nm: pH 1,244 (s
h), 313; pH 7, 248 (sh), 306; pH
13, 248 (sh), 305.

Example 11 Preparation of 2-amino-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -purine (compound C) Preparation of 2-amino-9-[(1R, 2R, 3S) -2,3-bis (acetoxymethyl) cyclobutan-1-yl] -purine (compound) 2-amino-9-[(1R, 2R, 3S) -2,3-Bis (acetoxymethyl) cyclobutan-1-yl] -6
-Chloropurine (87 mg, 0.24 mmol) was dissolved in methanol (2 ml), and 10% palladium on carbon (1
3 mg) and stirred overnight at room temperature under a hydrogen atmosphere.
A small amount of a saturated aqueous solution of sodium hydrogencarbonate was added to the reaction solution, the catalyst was filtered off, the solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (methylene chloride: methanol = 20: 1, v / v). 2-amino-9-
[(1R, 2R, 3S) -2,3-bis (acetoxymethyl) cyclobutan-1-yl] -purine (56 mg,
71%).

NMR (200 MHz FT, CDCl ₃ )
δ: 2.01 (3H, s), 2.12 (3H, s),
2.28-2.71 (3H, m), 3.11 (1H,
m), 4.17-4.34 (4H, m), 4.60 (1
H, applicable q, J = 8.6 Hz), 5.1
1 (2H, brs), 7.80 (1H, s), 8.70
(1H, s),

Step 2. Preparation of 2-amino-9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -purine (compound C) 2-amino-9-[(1R, 2R, 3S ) -2,3-Bis (acetoxymethyl) cyclobutan-1-yl] -chloropurine (40 mg, 0.12 mmol) was dissolved in methanol (2 ml) and potassium carbonate (40 mg, 0.2
9 mmol) and stirred at 0 ° C. for 30 minutes. After neutralizing the reaction solution with 0.1 N hydrochloric acid, Sephadex HP20
(Water-50% methanol aqueous solution) to give 2-amino-
9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -purine (compound C) (27 mg, 90%) is obtained.

NMR (200 MHz FT, CD ₃ OD)
δ: 2.10-2.66 (3H, m), 2.90 (1
H, m), 3.70 (4H, d, J = 5.6 Hz),
4.65 (1H, applicable q, J = 8.7H
z), 8.20 (1H, s), 8.54 (1H, s), UV λmax (H ₂ O) nm: pH 1,252 (s
h), 312; pH 7,244 (sh), 304; pH
13, 244 (sh), 304.

Embodiment 12 FIG. 2,6-diamino-9-[(1R, 2R, 3S) -2,
Preparation of 3-bis (hydroxymethyl) cyclobutan-1-yl] -purine (compound e) 2-amino-9-[(1R, 2R, 3S) -2,3-bis (acetoxymethyl) cyclobutan-1-yl -6
-Dissolve chloropurine (87 mg, 0.24 mmol) in methanol (2 ml) and cool to -78 ° C. This solution is saturated with liquid ammonia and placed in a sealed tube at 100 ° C for 1 hour.
Heat for 2 hours. After evaporating the volatile substance under reduced pressure, the reaction solution was dissolved in water, neutralized with 0.1 N sodium hydroxide, and purified with Sephadex HP20 (water-40% methanol aqueous solution) to obtain 2,6. -Diamino-9-[(1R,
2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -purine (compound e) (41 m
g, 82%).

NMR (200 MHz FT, CD ₃ OD)
δ: 2.19 (1H, m), 2.31 (1H, m),
2.54 (1H, m), 2.75 (1H, m), 3.6
1-3.74 (4H, m), 4.49 (1H, appa)
rent q, J = 8.6 Hz), 7.90 (1H,
s), UV λ max (H ₂ O) nm: pH 1,253,29
1, pH 7, 255, 280; pH 13, 255, 28
0.

Example 13 Preparation of 9-[(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -hypoxanthine (compound F) 9-[(1R, 2R, 3S) ) -2,3-Bis (hydroxymethyl) cyclobutan-1-yl] -adenine (compound I) (25 mg, 0.10 mmol in water (2.5 m
l) and sodium nitrite (138 mg, 2.0
mmol) and cooled to 0 ° C., followed by acetic acid (0.13 mmol).
ml) and stirred at room temperature for one day. The reaction solution was 1N-
After neutralization with sodium hydroxide, Sephadex HP2
0 (water-40% methanol aqueous solution) to purify 9-[(1
(R, 2R, 3S) -2,3-bis (hydroxymethyl)
Cyclobutan-1-yl] -hypoxanthine (Compound F) (23 mg, 90%) is obtained.

NMR (200 MHz FT, CD ₃ OD)
δ: 2.25 (1H, m), 2.38 (1H, m),
2.58 (1H, m), 2.90 (1H, m), 3, 6
6-3.74 (4H, m), 4.76 (1H, appa
rent q, J = 8.5 Hz), 8.04 (1H,
s), 8.20 (1H, s). UV λ max (H ₂ O) nm: pH 1,250; pH
7,250; pH 13,254.

[0111]

The anticancer agent of the present invention has an excellent anticancer effect.

──────────────────────────────────────────────────続 き Continued on the front page (31) Priority claim number Japanese Patent Application No. 1-158223 (32) Priority date Hei 1 (1989) June 22, (33) Priority claim country Japan (JP) (72) Inventor Kenichi Matsubara 3-18-1-804 Yamadahigashi, Suita-shi, Osaka (72) Inventor Takemitsu Nagahata 2-6-25 Kamishita, Toyonaka-shi, Osaka Senri-zaka Noki Heights 208 (72) Inventor Hiroo Hoshino Gunma 1-14-5 Heiwacho, Maebashi-shi, Japan (72) Inventor Junichi Seki 239, Iwanacho, Takasaki-shi, Gunma (72) Inventor Koichi Narasaka 1-3-17-310, Ecchujima, Koto-ku, Tokyo (72) Inventor Yujiro Hayashi 2-6-2 Minamiyukiya, Ota-ku, Tokyo Examiner Mika Fuji (56) References JP-A-2-6478 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 31/52 C07D 473/18 C07D 473/34

Claims (3)

(57) [Claims] 1. A compound of the general formula (IV) [Wherein B is a purine base and R ⁴ is a hydrogen atom or a protecting group]. (1R, 2R, 3S) cyclobutane derivative represented by the following formula: 2. The compound represented by the general formula (IV) is 9-
The anticancer agent according to claim 1, which is [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -adenine. 3. A compound represented by the general formula (IV):
The anticancer agent according to claim 1, which is [(1R, 2R, 3S) -2,3-bis (hydroxymethyl) cyclobutan-1-yl] -guanine.