JPH07633B2 - 1- (β-D-erythro-pentofuran-2-urosyl) pyrimidine derivative - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel compound useful as a synthetic intermediate in the production of 2'-alkylidene pyrimidine nucleoside derivatives.

[0002]

2. Description of the Related Art In recent years, as researches on pathogenic viruses of various viral infectious diseases have progressed, attention has been paid to the development of preventive and therapeutic agents therefor. Conventionally, as antiviral agents in chemotherapy, idoxuridine, cytarabine, vidarabine, and acyclovir have been clinically used (for example, Yu Mizushima, Akimasa Miyamoto, 1986 edition, today's therapeutic agents, commentary and handbook, No. 1). 47-50, March 1, 1986
Other than this, various antiviral active nucleosides are being developed as pharmaceuticals.

[0003]

However, the above drugs are not suitable for clinical use because of their antiviral activity spectrum, low absorption, low solubility, easy degradability, appearance of drug resistant virus strains, various side effects, etc. Many have problems such as restrictions. Therefore, there has been a strong demand for the development of new antiviral agents. Under such circumstances, the present inventors have found that the 2'-alkylidene pyrimidine nucleoside derivative has excellent antiviral activity. Therefore, the present invention has a 2'having excellent antiviral activity.
-An object of the present invention is to provide a novel compound useful as a synthetic intermediate in the production of an alkylidene pyrimidine nucleoside derivative.

[0004]

As a result of various studies on the synthetic route of 2'-alkylidene pyrimidine nucleoside derivatives, the present inventors have found that 1- (β-D-erythro-
Pentofuran-2-urosyl) pyrimidine derivative is 2 '
-It was found to be useful in the production of alkylidene pyrimidine nucleoside derivatives and has completed the present invention. Therefore, the present invention provides 1- (β represented by the following formula [II].
The present invention relates to a -D-erythro-pentofuran-2-urosyl) pyrimidine derivative (hereinafter sometimes referred to as a synthetic intermediate of the present invention).

[0005]

[Chemical 2]

(Wherein R ² is a hydrogen atom, a halogen atom or a lower alkyl group, R ⁵ is an alkoxyl group, an amino group or a hydroxyl group, and Z is a protecting group). The present invention will be described in detail below.

1- (β-D- which is a synthetic intermediate of the present invention
The erythro-pentofuran-2-urosyl) pyrimidine derivative is represented by the above formula [II], and R ² and R ⁵ in the formula are as defined above. The lower alkyl group represented by R ² is alkyl having 1 to 3 carbon atoms, and specific examples thereof include methyl, ethyl, propyl and isopropyl. As the halogen atom of R ² , chlorine, fluorine,
Examples include bromine and iodine. The alkoxyl group represented by R ⁵ is a lower alkoxyl having 1 to 3 carbon atoms, and specific examples thereof include methoxy, ethoxy and propoxy. The protecting group for Z may be any group used as a protecting group for a hydroxyl group of a usual nucleoside,
Examples thereof include acyl groups such as acetyl, propionyl, butyryl and benzoyl, alkylidene groups such as benzylidene, arylalkyl groups such as trityl, silyl groups such as tetraisopropyldisiloxyl (TIPDS) and t-butyldimethylsilyl.

The synthetic intermediate of the present invention can be synthesized by applying a known method. For example, a compound in which R ⁵ is an alkoxyl group can be prepared by the following reaction route.

[0009]

[Chemical 3]

(In the formula, R ⁵ 'is an alkoxyl group, and R ² and Z have the same meanings as above.) That is, after protecting the sugar group hydroxyl group of the uridine represented by the formula (A), the 4th position of the base portion is protected. It is halogenated with a halogenating agent and then reacted with an alkoxide to introduce an alkoxyl group to obtain a compound of the general formula (B). N ⁴ − represented by the general formula (B)
The desired compound can be obtained by protecting the sugar moieties 3 ′ and 5 ′ of the alkoxy compound and then oxidizing the sugar moiety 2 ′ hydroxyl group.

The hydroxyl-protecting group in the halogenation reaction is not particularly limited as long as it does not hinder the halogenating reaction, and a usual hydroxyl-protecting group such as an acyl group, an alkylidene group or an arylalkyl group is applied. However, a protecting group which is not eliminated particularly in the presence of an acid, for example, an acyl group is preferable. For example, the acyl protection reaction may be carried out by a conventional method, and a reaction solvent (for example, pyridine, picoline, diethylaniline, dimethylaminopyridine, dimethylformamide, acenitrile, tetrabutylamine, triethylamine or the like alone or a mixed solvent) for the compound of the general formula (A) In the reaction, an acylating agent (for example, an acid anhydride such as acetic acid, propionic acid, butyric acid, benzoic acid, substituted benzoic acid or an acid chloride thereof) is reacted at 3 to 10 times mol at a reaction temperature of 0 to 50 ° C. It can be carried out.

The halogenation reaction can be carried out by a method of reacting a halogenating agent in an inert solvent (eg chloroform, methylene chloride, etc.). As the halogenating agent, thionyl chloride, thionyl bromide, phosphorus oxychloride and the like can be applied, and if necessary, they may be used as an organic solvent solution of dimethyl sulfoxide and the like. The amount used is 1 to 5 with respect to 1 mol of the compound of the general formula (A).
It is about molar. The reaction may be carried out under heating under reflux.

The introduction reaction of an alkoxyl group is carried out by introducing an N ⁴ -halogeno compound of the general formula (A) having a protective group into an alkoxide (eg sodium methoxide) in a reaction solvent (eg alcohols such as methanol, ethanol and propanol). , Potassium methoxide, sodium ethoxide, potassium ethoxide, sodium propoxide, etc.) by heating at about 1 to 5 times mol. The protecting groups at the 3'-position and the 5'-position may be the same as those used in the above halogenation reaction, preferably a silyl protecting group, and particularly TIP.
The DS group is preferred. Explaining the silylation protection as an example, the amount of the silylating agent used can be appropriately selected from the range of 1 to 3 mol with respect to 1 mol of the compound of the general formula (B), and the reaction conditions are the same as in the acylation reaction described above. The conditions of can be adopted.

As the method for oxidizing the 2'-hydroxyl group, chromic acid oxidation using a chromic acid-pyridine-acetic anhydride complex or the like (method A) or activated dimethyl sulfoxide generated by oxalyl chloride-dimethyl sulfoxide or the like is used. Dimethyl sulphoxide oxidation (method B) and the like can be adopted. The reaction is -10 ° C for Method A
It can be carried out at room temperature in the case of Method B at -70 to -10 ° C in the presence of 1 to 10 equivalents of an oxidizing agent.

A compound in which R ⁵ is a hydroxyl group or an amino group can also be prepared by the following reaction route.

[0016]

[Chemical 4]

(In the formula, R ⁵ ″ is a hydroxyl group or an amino group,
R ² and Z are as defined above. That is, the target compound can be obtained by protecting the sugar moieties 3 ′ and 5 ′ hydroxyl groups of the uridine represented by the general formula (A) and then oxidizing the sugar moieties 2 ′ hydroxyl group. The protection reaction of the 3'- and 5'-position hydroxyl groups and the oxidation reaction of the 2'-position hydroxyl group can be carried out in the same manner as the above-mentioned reaction.

The thus-obtained synthetic intermediate of the present invention can be isolated and purified by the usual means for separating and purifying nucleosides. For example, the solvent is distilled off, and then it is subjected to column chromatography to obtain n. -Can be crystallized in a suitable organic solvent such as hexane.

The 2'-alkylidene pyrimidine nucleoside derivative useful as an antiviral agent can be prepared from the synthetic intermediate of the present invention by the method comprising the following first to third steps. The first step; the sugar moiety 2'position of the compound represented by the following general formula [II] is converted to an alkylidene by Wittig reagent,
Step of obtaining a compound represented by the following general formula [III] (wherein R ² is hydrogen atom, halogen atom or lower alkyl group, R ³ is hydrogen atom or lower alkyl group, R ⁵ is alkoxyl group, hydroxyl group or amino group) , Z represents a protecting group)

[0020]

[Chemical 5]

Second step: The sugar moiety-protecting group of the compound represented by the following general formula [III] is eliminated, and the following general formula [I
V] to obtain a compound (in the formula, R ² , R ³ ,
(R ⁵ and Z are as defined above)

[0022]

[Chemical 6]

Third step: When R ⁵ is an alkoxyl group, the compound represented by the following general formula [IV] is hydrolyzed or aminated at the 4-position of the base moiety, and then, if desired, a phosphorus moiety at the 5′-position of the sugar moiety is further added. A step of obtaining a compound represented by the following general formula [I] by oxidation (wherein R ¹ is an amino group or a hydroxyl group, R ⁴ is a hydrogen atom or a phosphoric acid residue, R ² or R
⁽³ and R ⁵ have the same meaning as above)

[0024]

[Chemical 7]

The Wittig reagent used in the alkylidene formation reaction is an alkylidenephosphorane represented by the formula (C ₆ H ₅ ) ₃ P═CH—R ³ (wherein R ³ represents a hydrogen atom or a lower alkyl group). And specifically, triphenylphosphine methylene, triphenylphosethylene, triphenylphosphine propylene, etc. are used. The Wittig reagent used in the reaction has the formula [(C ₆
H ₅ ) ₃ P ⁺ —CH ₂ —R ³ ] X ⁻ (wherein R ³ has the same meaning as described above, X ⁻ represents a halogen ion such as Br ⁻ , I ^−, etc.), and a triphenylphosphonium compound ( For example, methyltriphenylphosphonium bromide, methyltriphenylphosphonyl iodide, ethyltriphenylphosphonium bromide, etc. and a strong alkali (eg, potassium hydride, sodium hydride, n-butyllithium, sodium methoxide, potassium-t). -Butoxide, sodium amide, etc.) prepared by a conventional method is preferably used. The amount of the Wittig reagent used can be appropriately selected from 1 to 3 mol per 1 mol of the synthetic intermediate of the present invention. The alkylideneation reaction can be carried out by reacting the synthetic intermediate of the present invention with a Wittig reagent at room temperature in a solvent (for example, tetrahydrofuran, dioxane, ether, benzene, dimethyl sulfoxide, etc. alone or as a mixed solvent). .

The deprotection reaction may be carried out by appropriately selecting an ordinary treatment such as acidic hydrolysis, alkaline hydrolysis, ammonium fluoride treatment, catalytic reduction or the like depending on the protective group used. When R ⁵ is an alkoxyl group, the amination reaction may be carried out by a conventional method, for example, by reacting methanolic ammonia in a sealed tube. The reaction temperature is 50 to 150 ° C. Further, the hydrolysis reaction may be carried out according to a conventional method, and acidic hydrolysis is particularly preferable. Further, when the compound having a phosphate residue at the 5′-position of the sugar moiety is intended, after completion of the above-mentioned amination reaction or hydrolysis reaction, a normal nucleoside such as phosphorus oxychloride or tetrachloropyrophosphate can be prepared. It can be converted into a free acid form or a salt form by a conventional method by reacting with a phosphorylating agent used for selective phosphorylation at the 5'position.

Isolation and purification of the 2'-alkylidenepyrimidine nucleoside derivative can be carried out by appropriately combining the methods used for usual isolation and purification of nucleosides or nucleotides. For example, in the case of a nucleoside compound, the solvent may be distilled off and then crystallized from a suitable solvent such as ethanol, or the nucleoside compound may be obtained in a salt form if necessary. In the case of the nucleotide form, it can be purified by ion exchange column chromatography such as ion exchange resin, adsorption column chromatography such as activated carbon, etc., and the free acid form can be obtained by lyophilization or crystallization, and if necessary salt form. Can also be obtained as

The 2'-alkylidene pyrimidine nucleoside derivative thus prepared has the following general formula [I]:
And R ¹ and R ² in the general formula
R ³ and R ⁴ are as defined below. R ² and R
Specific examples of the lower alkyl group of ³ include a lower alkyne group having 1 to 3 carbon atoms, more specifically, methyl, ethyl, propyl, inpropyl and the like. In addition, specific examples of the halogen atom of R ² include chlorine, fluorine, bromine, iodine and the like.

[0029]

[Chemical 8]

(In the formula, R ¹ is an amino group or a hydroxyl group, and R ^{2 is}
Is a hydrogen atom, a halogen atom or a lower alkyl group, R ³
Represents a hydrogen atom or a lower alkyl group, R ⁴ represents a hydrogen atom or a phosphoric acid residue). Typical examples of such a compound include, for example, 2′-methylidene-2′-deoxyuridine, 2′-methylidene thymidine, 2'-ethylidene thymidine, 2'-methylidene-2'-deoxycytidine,
2'-methylidene-2'-deoxy-5-fluorouridine, 2'-methylidene-2'-deoxy-5-chlorourilidine, 2'-methylidene-2'-deoxy-5-
Bromouridine, 2'-methylidene-2'-deoxy-
Included are nucleosides such as 5-iodouridine and their 5'-phosphate esters.

The compound also includes a salt form, and examples of such a salt include R of the general formula [I].
^{When 4} is a hydrogen atom, an acid addition salt such as hydrochloride or sulfate, and when R ⁴ is a phosphoric acid residue, an alkali metal salt such as sodium salt, potassium salt or lithium salt, calcium salt, etc. Examples thereof include any pharmaceutically acceptable salt such as an alkaline earth metal salt or an ammonium salt.

The 2'-alkylidene pyrimidine nucleoside derivative has antiviral activity against viruses, particularly viruses belonging to the herpesvirus family such as herpes simplex virus and cytomegalovirus, and prevents infections caused by these viruses. , Useful for treatment. Among them, a compound group in which R ² in the general formula [I] is a hydrogen atom, a halogen atom or a methyl group, and R ³ is a hydrogen atom has a strong antiviral activity against herpes simplex virus (HSV). ing.

The dose of the 2'-alkylidene pyrimidine nucleoside derivative as an active ingredient of the drug varies depending on the severity of the patient, the tolerability of the drug, etc., and should not be finally decided by the judgment of the doctor. There is usually 0.1 to 10 g per adult per day, preferably 0.2 to 5
g, which is administered once or in divided doses. The administration method can take any form suitable for the administration route.

The pharmaceutical composition containing the 2'-alkylidene pyrimidine nucleoside derivative is provided for administration in a conventional manner by using any necessary pharmaceutical carrier or auxiliary agent. For example, in the case of a composition preparation for oral administration, it is provided in a form suitable for absorption from the digestive tract, and tablets, capsules,
It may be prepared as a solid form such as powder, sugar-coated tablet and granule, or as a liquid preparation such as syrup, suspension and elixir. In the case of fixatives, binders such as syrup, gum arabic, gelatin, sorbit, tragacanth, polyvinylpyrrolidone, lactose, sugar, corn starch, calcium phosphate, sorbit, glycine, and other bandage agents, magnesium stearate, talc, polyethylene. A lubricant such as glycol or silica, a disintegrating agent such as potato starch, a wetting agent, a stabilizer, and an auxiliary agent such as a flavoring agent can be selectively used for formulation according to pharmaceutical considerations. In the case of liquid preparations, sorbit syrup, methylcellulose, glucose / sugar syrup, gelatin, and
A suspending agent such as hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel, hydrogenated edible fat and the like, an emulsifier, a preservative such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate and sorbic acid can be used. When preparing a composition formulation for injection administration, if necessary, a pH adjusting agent, a buffering agent, a stabilizing agent, a preservative, a solubilizing agent, etc. are added to the 2'-alkylidene pyrimidine nucleoside derivative, and then, by a conventional method, Subcutaneous, intramuscular, and intravenous injections.

The test methods and results for the anti-HSV action and anti-CMV action of the 2'-alkylidene pyrimidine nucleoside derivative will be described below. Test method (HSV) A. Human fetal lung-derived cells were transferred to Eagle MEM medium (10%
Subculture with quasi-fetal serum). B. The subcultured above was used as a parental culture, and the cell suspension obtained by diluting this two-fold was 96 μl at a rate of 150 μl / well.
Seed in wells of a well and incubate at 37 ° C. for 4 to 5 days in a carbon dioxide incubator. C. Discard the culture medium and infect 100% to 50% of the tissue culture infectious dose.
20 times (100 to 320 TCID ₅₀ ) HSV type 1
(HSV-1) VR-3 strain or HSV type 2 (HS
V-2) Inoculate MS strain. After incubating at 37 ° C for 1 hour, the virus solution was discarded and Eagle MEM medium containing the test compound at an appropriate concentration (2.5% quasi-fetal serum was added)
Add 150 μl and incubate at 37 ° C. The test compound is usually subjected to 0.5-log 10-fold serial dilution in the range of 100 to 1 μg / ml for the test. D. After culturing for 2 to 3 days, the degree of cytopathic effect (CPE) in each well is observed under a microscope when the cells containing the test compound are completely denatured due to virus infection, and scores 0 to 4 are assigned. E. The minimum concentration that inhibits CPE by 50% or more (CPE score of 2 or less) is defined as the minimum effective concentration (MIC) of the test compound.

Test Method (CMV) A. Human fetal lung-derived cells were transferred to Eagle MEM medium (10%
Subculture with quasi-fetal serum). B. The subcultured above was used as a parental culture, and a cell suspension obtained by diluting this two-fold was prepared at a rate of 150 μl / well.
Seed the wells in a semi-microwell and incubate at 37 ° C for 4-5 days in a carbon dioxide incubator. C. Discard the culture,
About 50 Plaque Homing Unit CMV AD169
Inoculate the strain. After incubating at 37 ° C for 1 hour,
Eagle MEM medium containing test compound at an appropriate concentration (2.
Add 400 μl of 5% quasi-fetal serum) and incubate at 37 ° C. The test compound is usually diluted in a range of 100 to 1 μg / l in 0.5 log 10-fold steps and used for the test. D. After culturing for 4 to 6 days, the infected cells are stained with 0.5% crystal violet staining solution, and the number of plaques formed is calculated under a microscope. E. The minimum effective concentration (MIC) of the test compound is defined as the minimum concentration that inhibits the number of plaques formed by 50% or more with respect to the number of plaques in the control group containing no test compound.

[0037]

[Table 1]

[0038]

EXAMPLES The present invention will be described in detail below with reference to examples and application examples of the present invention. Example 1: 1- (3,5-O-TIPDS-β-D-erythropentofuran-2-urosyl) -4-ethoxy-
2-pyrimidinone (formula [II], R ² = H, R ⁵ = OC _{2
H 5, Z (3 ')} - Z- (5') = TIPDS) Synthesis 2 of ', 3', 5'-tri -O- acetyl uridine 3.3
5 g was dissolved in 50 ml of chloroform, 8.1 ml of thionyl chloride and 0.5 ml of dimethylformamide were added, and the mixture was refluxed for 6 hours and 30 minutes, and then dried under reduced pressure. The residue was dissolved in 20 ml of ethanol, 30 ml of 1N sodium ethoxide was added, the mixture was refluxed for 2 hours, neutralized with 1N hydrochloric acid, the precipitated salt was filtered off, and the solution was concentrated to dryness. This was adsorbed on a silica gel column (4 × 31 cm), the fraction containing the target compound was eluted with 16% ethanol / chloroform, and the solvent was distilled off to obtain crude crystals of the target product. This was recrystallized from ethanol to obtain 2.08 g of 4-O-ethyluridine (yield 84.2%).

Melting point: 136 to 137.5 ° C. Elemental analysis value: C ₁₁ H ₁₆ N ₂ O ₆ 1/3 H ₂ 0 calculated value C: 46.97%, H: 6.09%, N: 9 .9
6%, O: 36.98%, measured value C: 46.91%, H: 6.02%, N: 9.9.
8%, O: 37.09%

7.04 g of 4-O-ethyluridine was dissolved in 80 ml of pyridine, ice-cooled, and then 1,1,3,3.
-Dichlorotetraisopropyldisiloxane 9.57g
Was added and the reaction was stirred at room temperature for 4 hours and 30 minutes. Ice water was added, the solvent was evaporated, the residue was partitioned with chloroform-water,
After drying the chloroform layer, the solvent was distilled off, the residue was adsorbed on a silica gel column (10 × 130 cm), and the portion eluted with 40% ethyl acetate-hexane was collected and concentrated.
12.3 g of 3 ', 5'-O-TIPDS body was obtained. Next, 2.7 ml of oxalyl chloride was dissolved in 40 ml of methylene chloride and cooled to -70 ° C. Under argon flow,
4.8 ml of dimethylsulfoxide dissolved in 20 ml of methylene chloride was added dropwise over 20 minutes, and then stirred for 30 minutes. The above 3 ', 5'-O-TIPDS compound (12.3 g) dissolved in 50 ml of methylene chloride was added dropwise thereto, and the mixture was stirred at -70 ° C for 2 hours, and then triethylamine 2 was added.
0 ml was added and the mixture was further stirred for 1 hour. The reaction solution was returned to room temperature, partitioned by adding water, the methylene chloride layer was separated, the solvent was distilled off, the residue was dissolved in ethyl acetate, and partitioned with water. The ethyl acetate layer was concentrated to dryness, adsorbed on a silica gel column (5 × 28 cm), and 20% ethyl acetate-n-
Fractions containing the target substance eluted with hexane were collected, and the solvent was distilled off, followed by crystallization from n-hexane to obtain 10.2 g of the target substance.
(Yield 72.1%) was obtained.

Melting point: 157.5-159 ° C. Elemental analysis: Calculated as C ₂₀ H ₄₀ N ₂ O ₇ Si ₂ C: 53.87%, H = 7.86%, N = 5
46% measured value: C: 53.73%, H: 7.87%, N: 5.
57%

Application Example 1 Preparation of Hydrochloride of 2'-Methylidene-2'-deoxycytidine 232 mg of potassium hydride was added to 2.4 ml of dimethyl sulfoxide under an argon stream, and the mixture was stirred at room temperature for 40 minutes. Methyltriphenylphosphonium bromide (2.2 g) was dissolved in dimethylsulfoxide (8 ml), and the above potassium hydride-dimethylsulfoxide mixture was added dropwise thereto in an argon stream under ice cooling, followed by stirring for 10 minutes. To the above 1
1.02 g of crystals of-(3,5-O-TIPDS-β-D-erythropentofuran-2-urosyl) -4-ethoxy-2-pyrimidinone were added to 10 ml of dimethyl sulfoxide.
What was dissolved in was added dropwise under an argon stream, and cooled under ice 2
Stir for hours. To this, 10 ml of 1N ammonium chloride aqueous solution was added, and further 50 ml of ethyl acetate and 40 ml of water.
Added and distributed. The organic layer was concentrated under reduced pressure, adsorbed on a silica gel column (2.4 × 30 cm), and eluted with a mixed solvent of n-hexane-ethyl acetate to obtain a 2′-methylideneated compound. 320 mg of the compound obtained above was dissolved in 10 ml of tetrahydrofuran, 1 ml of tri-n-butylammonium fluoride was added, and the mixture was stirred at room temperature for 10 minutes. Neutralize with acetic acid After neutralize with acid, use silica gel column (2.4 x
(12 cm) and eluted with chloroform-ethanol, and the elution fraction containing the target compound was collected and deprotected 2 '.
-Methylidene-4-O-ethyl compound 155 mg (yield 30
%) Was obtained.

Melting point: 157.5-159 ° C. Elemental analysis value: Calculated as C ₁₂ H ₁₆ N ₂ O ₅ : C: 53.72%, H: 6.01%, N: 1
0.44% measured value: C: 53.80%, H: 5.99%, N: 1
0.37%

2'-methylidene-4-O-ethyl derivative 15
0 mg of ammonia saturated methanol solution under ice-cooling 10 ml
Was dissolved in the mixture, placed in a sealed tube, and heated at 100 ° C. for 2 days. After allowing to cool, add 2 ml of 2N hydrochloric acid, concentrate and concentrate with ethanol-
Crystallized from water, 125 mg of the title compound (yield 81.
7%). Melting point: 148 to 155 ° C. (decomposition point) Elemental decomposition: C ₁₀ H ₁₃ N ₃ O ₄ .HCl.1 / 2H ₂ O Calculated value: C: 42.14%, H: 5.31%, N: 1
4.74% Found: C: 42.55%, H: 5.26%, N: 1
4.69%

Example 2: 1- (3,5-O-TIPDS
-Β-D-erythropentofuran-2-urosyl) thymine (formula [II], R ² = CH ₃ , R ⁵ = hydroxyl group, Z
Synthesis of (3 ′)-Z (5 ′) = TIPDS 4.13 g of 5-methyluridine was dissolved in 50 ml of pyridine, and 5.57 g of 1,1,3,3-dichlorotetraisopropylzine siloxane was added under ice cooling. Stir at room temperature for 6 hours. A small amount of water was added to this, and the mixture was stirred for 30 minutes and then concentrated to dryness under reduced pressure. The residue was partitioned with chloroform-water, the organic layer was concentrated and then adsorbed on a silica gel column (5 x 21 cm) to give a 3 ', 5'-protected form from the elution fraction of 2% ethanol-chloroform. On the other hand, oxalyl chloride 1.
7 ml was dissolved in 40 ml of methylene chloride, cooled to -70 ° C., a mixed solution of 3 ml of dimethyl sulfoxide and 20 ml of methylene chloride was added dropwise under an argon stream, and the mixture was further stirred for 30 minutes. This solution was added to the above 3 ', 5'-protected body 8.0.
A solution prepared by dissolving 4 g in 50 ml of methylene chloride was added dropwise and further stirred at -70 ° C for 2 hours. Triethylamine (6.6 ml) was added dropwise to this, and the mixture was stirred for one and a half hours, then returned to room temperature, chloroform and water were added for partitioning, the organic layer was concentrated to dryness under reduced pressure, and developed on a silica gel column (4 × 28 cm), The fraction eluted with 40% ethyl acetate-n-hexane was concentrated and crystallized from n-hexane to obtain 6.68 g of the target compound (yield 83.2%).

Melting point: 168 to 170 ° C. Elemental analysis: C ₂₂ H ₃₈ N ₂ O ₇ Si ₂ calculated value: C: 52.98%, H: 7.68%, N: 5.
62% Actual value: C: 52.93%, H: 7.71%, N: 5.
61%

Application Example 2: Preparation of 2'-methylidene thymidine 455 mg of potassium hydride was added to 5 ml of dimethyl sulfoxide under an argon stream, and the mixture was stirred at room temperature for 50 minutes. On the other hand, 4.28 g of methyltriphenylphosphonium bromide was dissolved in 10 ml of dimethylsulfoxide, and the solution containing the above calcium hydride was added dropwise to this solution under ice cooling, followed by stirring for 20 minutes. 1- (3,5-
O-TIPDS-β-D-erythropentofuran-2-
Urosyl) thymine 1.5 g tetrahydrofuran 5 ml
What was melt | dissolved in the mixed solvent of 5 ml of dimethyl sulfoxide and was added dropwise, and stirred at room temperature for 10 hours. Next, the reaction solution was neutralized with 1N ammonium chloride, 120 ml of ethyl acetate and 120 ml of water were added to the mixture, and the organic layer was concentrated to dryness. The residue was purified by a silica gel column (2.4 x 24 cm).
Then, the fractions eluted with 20% ethyl acetate-n-hexane were collected to obtain a 2'-methylidene compound. This is dissolved in 10 ml of tetrahydrofuran, and tetra-n-fluoride is added.
Butyl ammonium / tetrahydrofuran 1 molar solution 1
ml was added and the mixture was stirred at room temperature for 10 minutes for deprotection. Next, after neutralization with acetic acid, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (2.4 × 14 cm), and the fractions eluted with a 7% ethanol-chloroform solution were collected and concentrated to 2′-.
257 mg of crystalline powder of methylidene thymidine (yield 85
%) Was obtained.

Melting point: 161-162 ° C. Elemental analysis: Calculated as C ₁₁ H ₁₄ N ₂ O ₅ : C: 49.58%, H: 5.83%, N: 1
1.57% Found: C: 49.46%, H: 5.91%, N: 1
1.48%

Application Example 3: Preparation of 2'-ethylidene thymidine 455 mg of potassium hydride was added to 5 ml of dimethyl sulfoxide under a stream of argon, and the mixture was stirred at room temperature for 50 minutes. On the other hand, 4.44 g of ethyltriphenylphosphonium bromide was dissolved in 10 ml of dimethylsulfoxy, and the solution containing potassium hydride was added dropwise to this solution under ice cooling, followed by stirring for 20 minutes. 1-obtained in Example 2 in this solution
A solution prepared by dissolving 1.5 g of (3,5-O-TIPDS-β-D-erythropentofuran-2-urosyl) thymine in a mixed solvent of 5 ml of tetrahydrofuran and 5 ml of dimethyl sulfoxide was added dropwise and stirred at room temperature for 12 hours. . Next, the reaction solution was neutralized with 1N ammonium chloride, 140 ml of ethyl acetate and 140 ml of water were added to the mixture, and the organic layer was concentrated and dried.
cm), and the fractions eluted with 10% ethyl acetate-n-hexane were collected to obtain a 2'-ethylidene compound. This was dissolved in 10 ml of tetrahydrofuran, 1 ml of a tetra-n-butylammonium fluoride / tetrahydrofuran 1 mol solution was added, and the mixture was stirred at room temperature for 30 minutes for deprotection.
Next, after neutralization with acetic acid, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (2 × 10 cm), and the fractions eluted with a 7% ethanol-chloroform solution were collected and concentrated to 2′-.
190 mg of non-crystalline powder of ethylidene thymidine was obtained.

Elemental analysis: Calculated as C ₁₂ H ₁₆ N ₂ O ₅ : C: 53.72%, H: 6.01%, N: 1
0.44% Found: C: 53.68%, H: 6.15%, N: 1
0.39%

Example 3: 1- (3,5-O-TIPDS
-Β-D-erythropentofuran-2-urosyl) -5
- fluorouracil (formula ^{[II], R 2 = F} , R 5 = hydroxyl, Z (3 ') - Z (5') = TIPDS) Synthesis of 5-fluorouridine 2.42g was dissolved in pyridine 30ml of ice-cold 3.3 g of lower 1,1,3,3-dichlorotetraisopropyldisiloxane was added and stirred for 2 hours,
It returned to room temperature and stirred for 1 hour and 30 minutes. A small amount of water was added to this, and after stirring, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (2.4 × 23 cm), and 25% ethyl acetate-n was added.
-The fractions eluted with hexane were collected and 3 ', 5'-O-
A TIPDS body was obtained. 3 ', the 5'-O-TIPDS body 3.91g was dissolved in methylene chloride 10 ml, 4 equivalents of chromic acid complex (chromium trioxide (CrO ₃₎
3 g, pyridine 5 ml, and acetic anhydride 3 ml added to and mixed with 80 ml of methylene chloride), and the mixture was added at room temperature for 1 hour.
After stirring at -4 ° C for 14 hours, 4 equivalents of chromic acid complex was further added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was added dropwise to 600 ml of ethyl acetate, and silica gel (6 × 15c
m), the filtrate was concentrated to dryness under reduced pressure, the residue was developed on a silica gel column (2.4 × 21 cm), and 20
Fractions eluted with% ethyl acetate-n-hexane are collected,
2.8 g (yield 71.6%) of the target compound was obtained.

Melting point: 183-186 ° C. Elemental analysis: Calculated as C ₂₁ H ₃₅ N ₂ O ₇ FSi ₂ C: 50.15%, H: 7.01%, N: 5.
57% measured value: C: 50.01%, H: 7.22%, N: 5.
49%

Application Example 4: Synthesis of 2'-methylidene-2'-deoxy-5-fluorouridine 1.1 g of potassium hydride was added to 12 ml of dimethyl sulfoxide under an argon stream, and the mixture was stirred at room temperature for 1 hour. On the other hand, 11 g of methyltriphenylphosphonium bromide was dissolved in 25 ml of dimethylsulfoxide, and the solution containing potassium hydride was added dropwise to this solution under ice cooling.
Stir for 0 minutes. 1- (3,5-OT)
A solution prepared by dissolving 1.4 g of IPDS-β-D-erythropentofuran-2-urosyl) -5-fluorouracil in 25 ml of dimethyl sulfoxide was added dropwise and stirred at room temperature for 10 hours. Next, the reaction solution was neutralized with 1N ammonium chloride, 200 ml of ethyl acetate and 200 ml of water were added for partitioning, the organic layer was concentrated to dryness, and the residue was developed on a silica gel column (2.4 × 22 cm). % Ethyl acetate-n
The fractions eluted with -hexane were collected to obtain a 2'-methylidene derivative. This was dissolved in 5 ml of tetrahydrofuran, 4 ml of a tetra-n-butylammonium fluoride / tetrahydrofuran 1 mol solution was added, and the mixture was stirred at room temperature for 30 minutes for deprotection. Next, after neutralizing with acetic acid, the mixture was concentrated to dryness under reduced pressure and developed on a silica gel column (2.4 × 17 cm),
Fractions eluted with 7% ethanol-chloroform solution were collected and concentrated to give 2'-methylidene-2'-deoxy-5.
-0.37 g (54% yield) of fluorouridine was obtained.

Melting point: 154-156 ° C. Elemental analysis: Calculated as C ₁₀ H ₁₁ N ₂ O ₅ F: C: 46.55%, H: 4.30%, N: 1
0.86% Found: C: 46.49%, H: 4.41%, N: 1
0.78%

Example 4: 1- (3,5-O-TIPDS
-Β-D-erythropentofuran-2-urosyl) -5
-Synthesis of iodouracil (formula [II], R ² = I, R ⁵ = hydroxyl group, Z (3 ′)-Z (5 ′) = TIPDS) 10.0 g of 5-iodouridine was dissolved in 100 ml of pyridine and iced. 8.94 g of 1,1,3,3-dichlorotetraisopropyldisiloxane was added under cooling, and the mixture was stirred for 1 hour and 30 minutes, returned to room temperature and further stirred for 3 hours. A small amount of water was added to this, and after stirring, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (3 × 30 cm), and the fractions eluted with 25% ethyl acetate-n-hexane were collected and 3 ′, 5. '-
An O-TIPDS body was obtained. 3 ', 5'-O-TIPDS
Dissolve 13.65 g of the body in 30 ml of methylene chloride,
Equivalent amount of chromic acid complex (chromium trioxide (CrO
₃ ) 9 g, 15 ml of pyridine, and 9 ml of acetic anhydride added to 230 ml of methylene chloride and mixed) were added, and the mixture was stirred at room temperature for 2 hours, then 2 equivalents of chromic acid complex was added, and the mixture was stirred at room temperature for 2 hours. After stirring, the reaction solution was added dropwise to 1.5 ml of ethyl acetate, and the mixture was washed with silica gel (10 x 20
cm), the filtrate is concentrated to dryness under reduced pressure, the residue is developed on a silica gel column (3.0 × 32 cm), and 2
Fractions eluted with 0% ethyl acetate-n-hexane were collected to obtain 4.4 g of the target compound.

Application Example 5: Synthesis of 2'-methylidene-2'-deoxy-5-iodouridine 22.0 g of methyltriphenylphosphonium bromide was added to 100 parts of tetrahydrofuran.
3 ml of n-butyllithium was added dropwise under argon flow, and the mixture was stirred for 1 hour. To this solution was added 1- (3,5-O-TIPDS-β-D-erythropentofuran-2-urosyl) -5-iodouracil.
A solution prepared by dissolving 0 g in tetrahydrofuran (20 ml) was added dropwise at -10 ° C, and the mixture was stirred for 30 minutes, and further stirred at room temperature for 1 hour and 30 minutes. Next, the reaction solution is neutralized with 1N ammonium chloride, then 200 ml of ethyl acetate and 200 ml of water are added.
Was added and distributed, the organic layer was concentrated to dryness, the residue was developed on a silica gel column (3 x 23 cm), and 20% ethyl acetate-
Fractions eluted with n-hexane were collected, 300 mg of the 2'-methylidene compound was dissolved in 5 ml of tetrahydrofuran, 1.1 ml of tetra-n-butylammonium fluoride / tetrahydrofuran 1 molar solution was added, and the mixture was stirred at room temperature for 30 minutes. Deprotected. Next, after neutralizing with acetic acid, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (2.4 × 17 cm), and the fractions eluted with a 7% ethanol-chloroform solution were collected and concentrated to 2′-. 118 mg of methylidene-2'-deoxy-5-iodouridine were obtained.

Melting point: 169-172 ° C. Elemental analysis: Calculated as C ₁₀ H ₁₁ N ₂ O ₅ I: C: 32.82%, H: 3.03%, N: 7.
65% measured value: C: 32.76%, H: 3.15%, N: 7.
60%

Example 5: 1- (3,5-O-TIPDS
-Β-D-erythropentofuran-2-urosyl) -5
- bromouracil (formula ^{[II], R 2 = Br} , R 5 = hydroxyl, Z (3 ') - Z (5') = TIPDS) dissolved synthesis of 5-bromo uridine 3.32g of pyridine 30 ml, ice 3.3 g of 1,1,3,3-dichlorotetraisopropyldisiloxane was added under cooling, and the mixture was stirred for 2 hours, returned to room temperature, and further stirred for 1 hour and 40 minutes. A small amount of water was added to this, and after stirring, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (2.4 × 25 cm), and the fractions eluted with 25% ethyl acetate-n-hexane were collected and 3 ′. , 5'-
An O-TIPDS body was obtained. 3 ', 5'-O-TIPDS
4.30 g of the body was dissolved in 10 ml of methylene chloride, and 4 equivalents of chromic acid complex (chromium trioxide (CrO 3
₃ ) 3 g, 5 ml of pyridine, and 3 ml of acetic anhydride added to 80 ml of methylene chloride and mixed) were added, and the mixture was stirred at room temperature for 2 hours. After stirring, the reaction solution was added dropwise to 300 ml of ethyl acetate and filtered through silica gel (6 × 10 cm),
The filtrate was concentrated to dryness under reduced pressure, the residue was developed on a silica gel column (2.4 × 32 cm), and 20% ethyl acetate-n was added.
-The fractions eluted with hexane were collected and the title compound 2.4
g was obtained.

Application Example 6: Synthesis of 2'-methylidene-2'-deoxy-5-bromouridine 3.3 g of methyltriphenylphosphonium bromide was dissolved in 20 ml of tetrahydrofuran, and n was added under a stream of argon.
-Butyllithium (6.6 ml) was added dropwise under ice cooling, and the mixture was added to 5
Stir for 0 minutes. 1- (3,5-O-
After 650 mg of TIPDS-β-D-erythropentofuran-2-urosyl) -5-bromouracil dissolved in 10 ml of tetrahydrofuran was added dropwise at -10 ° C,
The mixture was stirred for 1 hour and further at room temperature for 4 hours. Next, the reaction solution is neutralized with 1N ammonium chloride, and then ethyl acetate 1
00 ml and 100 ml of water were added for partitioning, the organic layer was concentrated to dryness, the residue was developed on a silica gel column (2.4 × 18 cm), and the fractions eluted with 20% ethyl acetate-n-hexane were collected and collected. '-A methylidene body was obtained. This was dissolved in 5 ml of tetrahydrofuran and tetra-n-butylammonium fluoride / tetrahydrofuran 1 molar solution 1.4
ml was added, and the mixture was stirred at room temperature for 30 minutes for deprotection. Next, after neutralization with acetic acid, the mixture was concentrated to dryness under reduced pressure, developed on a silica gel column (2.4 × 12 cm), and the fractions eluted with a 7% ethanol-chloroform solution were collected and concentrated to 2 ′.
-Methylidene-2'-deoxy-5-bromouridine was obtained as an amorphous powder.

Elemental analysis: Calculated as C ₁₀ H ₁₁ N ₂ O ₅ Br: C: 37.65%, H: 3.48%, N: 8.
78% Found: C: 37.49%, H: 3.55%, N: 8.
79%

Example 6: 1- (3,5-O-TIPDS
-Β-D-erythropentofuran-2-urosyl) uracil (formula [II], R ² = hydrogen atom, R ⁵ = hydroxyl group, Z
Synthesis of (3 ′)-Z (5 ′) = TIPDS 3.91 g of uridine was dissolved in 50 ml of pyridine, 5.57 g of 1,1,3,3-dichlorotetraisopropyldisiloxane was added under ice cooling, and the mixture was mixed at room temperature with 6 Stir for hours. A small amount of water was added to this, and the mixture was stirred for 30 minutes and then concentrated to dryness under reduced pressure. The residue was partitioned with chloroform-water, the organic layer was concentrated and then adsorbed on a silica gel column (5 x 21 cm) to give a 3 ', 5'-protected form from the elution fraction of 2% ethanol-chloroform. On the other hand, 1.7 ml of oxalyl chloride was dissolved in 40 ml of methylene chloride, cooled to -70 ° C., and 3 ml of dimethyl sulfoxide and 2 ml of methylene chloride were fed under an argon stream.
The 0 ml mixed solution was added dropwise, and the mixture was further stirred for 30 minutes. A solution prepared by dissolving 7.8 g of the above 3 ', 5'-protected compound in 50 ml of methylene chloride was added dropwise to this solution, and further at -70 ° C.
And stirred for 2 hours. 6.6 ml of triethylamine
Was added dropwise, and the mixture was stirred for 1 hour and a half, then returned to room temperature, chloroform and water were added for partitioning, the organic layer was concentrated to dryness under reduced pressure, developed on a silica gel column (4 × 28 cm), and 40% ethyl acetate-n was added. -Concentrate the fraction eluted with hexane and n-
Crystallization from hexane gave 6.53 g of the title compound.

Application Example 7: Synthesis of 2'-methylidene-2'-deoxyuridine Meanwhile, methyltriphenylphosphonium bromide 22.0 g
Was dissolved in 100 ml of tetrahydrofuran, 37.5 ml of n-butyllithium was added dropwise under an argon stream, and 1
Stir for hours. 1- (3,5-OT)
A solution of 3.2 g of IPDS-β-D-erythropentofuran-2-urocil) uracil dissolved in 20 ml of tetrahydrofuran was added dropwise at -10 ° C, and then stirred for 30 minutes,
Further, the mixture was stirred at room temperature for 1 hour and 30 minutes. Next, add 1
After neutralizing with the specified ammonium chloride, add ethyl acetate 2 to it.
00 ml and 200 ml of water were added for distribution, the organic layer was concentrated to dryness, the residue was developed on a silica gel column (3 × 24 cm), and the fractions eluted with 20% ethyl acetate-n-hexane were collected and 2′- A methylidene body was obtained. 240 mg of this 2'-methylidene compound was dissolved in 5 ml of tetrahydrofuran, and 1 ml of a tetra-n-butylammonium fluoride / tetrahydrofuran 1 mol solution was added, followed by stirring at room temperature for 30 minutes for deprotection. Next, after neutralizing with acetic acid, the mixture was concentrated to dryness under reduced pressure and developed on a silica gel column (2.4 x 14 cm) to give 7%.
Fractions eluted with an ethanol-chloroform solution were collected and concentrated to obtain 77 mg of 2'-methylidene-2'-deoxyuridine crystalline powder.

Elemental analysis: Calculated as C ₁₀ H ₁₂ N ₂ O ₅ : C: 50.00%, H: 5.04%, N: 1
1.66% Found: C: 49.88%, H: 5.13%, N: 1
1.59%

Application Example 8: Preparation of 2'-methylidene-2'-deoxychlorouridine Using 5-chlorouridine instead of uridine in Example 6, 1- (3,5-O-TIPDS-β-D- Erythropentofuran-2-urosyl) -5-chlorouracil (formula [II], R ² = Cl, R ⁵ = hydroxyl group, Z (3 ′)-
Z (5 ′) = TIPDS) was synthesized and reacted with it in the same manner as in Application Example 7 to obtain 2′-deoxy-2′-methylidene-5-chlorouridine. Mp: 149-152 ° C. Elemental _{analysis: C 10 H 11 N 2 O} 5 Cl Calculated: C: 43.68%, H: 4.03%, N: 1
0.19% Found: C: 43.77%, H: 3.98%, N: 1
0.20%

Application Example 9: 2'-methylidene thymidine-
Production of 5'-phosphoric acid 2.54 g of 2'-methylidene thymidine was added to 60 ml of trimethyl phosphoric acid and ice-cooled, 1.53 g of phosphorus oxychloride was added dropwise thereto, and the mixture was further stirred for 1 hour. The reaction solution was poured into 100 g of ice water containing 8 g of sodium hydrogen carbonate and stirred for 1 hour as it was.
l Add and distribute. The aqueous layer was concentrated, adsorbed onto the anion exchange resin Dowex 1 (formic acid type) and eluted with a 1 molar formic acid solution, the fractions containing the target substance were collected, concentrated, lyophilized and 2'-methylidene thymidine. -5'-phosphoric acid is obtained.

Continuation of the front page (56) References JOURNAL OF ORGANIC CHEMISTRY; VOL. 35 N O. 10 P. 3552-3558 (1970) JOURNAL OF AMERICA N CHEMICAL SOCIETY; VOL. 89 NO. 11 P. 2697-2705 (1967)

Claims (1)

[Claims] 1. A 1- (β-D-erythro-pentofuran-2-urosyl) pyrimidine derivative represented by the formula [II]. [Chemical 1] (In the formula, R ² represents a hydrogen atom, a halogen atom or a lower alkyl group, R ⁵ represents an alkoxyl group having 1 to 3 carbon atoms , and Z represents a protecting group.)