WO2005021571A1

WO2005021571A1 - Method for producing n4-benzoylcytidine derivative

Info

Publication number: WO2005021571A1
Application number: PCT/JP2004/012161
Authority: WO
Inventors: Hideki Umetani; Katsutoshi Tsuchiya; Kiyoshi Takahashi
Original assignee: Mitsui Chemicals, Inc.
Priority date: 2003-08-29
Filing date: 2004-08-25
Publication date: 2005-03-10

Abstract

Disclosed is a method for producing a compound represented by the general formula (5) below wherein a compound represented by the general formula (1) below or a salt thereof is reacted with a dimethoxytrityl halide for isolating a compound represented by the general formula (3) below, and then the isolated compound represented by the general formula (3) below is reacted with a benzoic anhydride or a benzoic acid halide in an organic solvent.

Description

Specification

Method for producing N ⁴ -benzoyl cytidine derivative

Technical field

The present invention relates to a method for producing an N ⁴ -benzoylcytidine derivative.

Background art

[0002] In recent years, with the progress of genomic drug discovery, antisense DNA drugs have been rapidly developed. Therefore, the demand for DNA oligomers as raw materials and nucleoside derivatives as raw materials for the oligomers is increasing.

For the production of oligomers, high-purity nucleoside derivatives are required as raw materials. In particular, it is necessary to avoid the incorporation of compounds very similar to the chemical structure of the starting nucleoside derivative into the starting material.

[0003] As nucleoside derivatives, for example,? ^ ⁴ _ base Nzoiru _5, _〇_ (4, 4, over dimethoxy tri chill) _2 '- Dokishi _5- N ^4, such as methyl cytidine - base Nzoirushichijin derivatives important production intermediate for the production of antisense DNA One.

[0004] A conventional method for producing N ⁴ _benzoyl_5'_0_ (4,4'_dimethoxytrityl) _2'-deoxy_5-methylcytidine includes, for example, N ⁴ -benzoyl 5-methyl_2 'as a starting material. A method is known in which _ deoxycytidine is used and 4,4'-dimethoxytrityl chloride is reacted therewith (Non-Patent Document 1, Non-Patent Document 2, and Non-Patent Document 3).

[0005] Specifically describing these methods, the target product is separated and purified by using pyridine as a reaction solvent and subjecting the obtained reaction mixture to column chromatography (Non-Patent Document 2, Non-Patent Document 2). Patent Document 3).

Non-Patent Document 1: Nucleic Acid Research, Vol. 15, No. 1, 219-232 (published in 1987) (Nucleic Acids Research, Vol. 15, No. 1, pp. 219-232 1987) Non-Patent Document 2: Photochemistry And Photobiology Vol. 45, No. 5, pp. 571-574, published in 1987 (Photochemistry and Photobiology Vol. 45, No. 5, pp. 571-574 1987)

Non-Patent Document 3: Chemical Pharmaceutical Bulletin Vol. 34 No. 1 51- 60 pages Published in 1986 (Chemical Pharmaceutical Bulletin, Vol. 34, No. 1, pp. 51—60 1986)

Disclosure of the invention

Problems to be solved by the invention

The present inventors produced N ⁴ -benzoyl-5′-0- (4,4, -dimethoxytrityl) -2, -deoxy-5-methylcytidine according to the method described in Non-Patent Document 1. As a result, in the target product obtained by this method, N ⁴ —benzoyl 3 ′, 5, -〇-bis (4,4, -dimethoxytrityl)-in which an excessive protecting group was introduced at the 3′-position was found. 2 Deoxy-5-methylcytidine was found to be present as an impurity. Furthermore, it was difficult to separate these impurities from the target substance, and purification by column chromatography according to the methods described in Non-Patent Documents 2 and 3 was necessary. When aiming at an industrial production method, purification by column chromatography is not preferable because a large amount of solvent and its removal are required.

[0007] Accordingly, the present invention relates to a method for producing _benzoyl_5, _〇_ (4,4, dimethoxytrityl) -1_5-substituted cytidine derivatives by introducing impurities having an excessively protective group introduced at the 3'-position. It is an object of the present invention to provide a novel production method for reducing the amount.

Means for solving the problem

[0008] The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, unlike the above-mentioned conventional method, the cytidine derivative having a trityl group introduced into the 5'-hydroxyl group has The method of introducing a benzoyl group into an amino group is effective in solving the above-mentioned problem.In addition, a crystal of a cytidine derivative in which a trityl group is introduced into a hydroxyl group at the 5′-position, and the derivative and a specific solvent are solvated. The present inventors have found that the crystals are useful for producing a cytidine derivative in which a trityl group has been introduced into the 5′-hydroxyl group and a benzyl group has been introduced into the 4-amino group, and have completed the present invention.

That is, the present invention provides

1.General formula (1) [0010]

(Wherein, R 1 is an alkyl group having 1 to 4 carbon atoms, an alkenyl group having 2 to 4 carbon atoms, an alkynyl group having 2 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, or A halogen atom, R 2 represents a hydrogen atom, an alkoxyl group having 1 to 4 carbon atoms, an alkoxyxylene group having 1 to 4 carbon atoms having a substituent, or a halogen atom.) Or a salt thereof; General formula (2)

[0012] [Formula 2]

[0013] (X represents a halogen atom.) The compound represented by the general formula (3)

[0014] [Formula 3]

(Wherein R1 and R2 are the same as described above),

Further, the compound represented by the general formula (3) and the compound represented by the general formula (4)

[0016] [Formula 4] (4)

(Wherein X represents a benzoyloxy group or a halogen atom). A compound represented by the following general formula (5):

[0018] [Formula 5]

(Wherein R 1 and R 2 are as defined above),

2. General formula (1) [0020] [Formula 6]

(Wherein, R 1 is an alkyl group having 1 to 4 carbon atoms, an alkenyl group having 24 carbon atoms, an alkynyl group having 2 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, or halogen R 2 represents a hydrogen atom, a C 1-4 alkoxyl group, a C 1-4 alkoxylinole group having a substituent, or a halogen atom), and a salt thereof, Equation (2)

[0022] [Formula 7]

(X represents a halogen atom), a compound represented by the general formula (3)

[0024] [Formula 8]

(Wherein R1 and R2 are the same as described above),

3. General formula (1)

[0026] [Formula 9]

(Wherein, R 1 is an alkyl group having 1 to 4 carbon atoms, an alkenyl group having 24 carbon atoms, an alkynyl group having 2 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, or halogen R 2 represents a hydrogen atom, a C 1-4 alkoxyl group, a C 1-4 alkoxylinole group having a substituent, or a halogen atom.) Equation (2) [0028] [Formula 10]

[0029] A compound represented by the formula (3) is reacted with a compound represented by (X represents a halogen atom).

(Wherein R 1 and R 2 are the same as above :).

4.General formula (3)

[0032] [Formula 12]

(Wherein R1 and R2 are the same as described above) and a compound represented by the general formula (4) [0034] [Formula 13]

(Wherein X represents a benzoyloxy group or a halogen atom), a compound represented by the general formula (5)

[0036] [Formula 14]

(Wherein R 1 and R 2 are as defined above),

5. General formula (3)

[0038] [Formula 15]

(Wherein R 1 and R 2 are as defined above) and a compound represented by the general formula (4) [0040] [Formula 16]

(Wherein X represents a benzoyloxy group or a halogen atom). The general formula (6) by-produced in the reaction of the compound represented by

[0042] [Formula 17]

(Wherein R 1 and R 2 are as defined above), and the compound represented by the general formula (5)

[0044]

[0045] The production method according to the above 1 or 4, comprising a step of converting the compound into a compound represented by the formula (wherein, R1 and R2 are as defined above).

6.The method according to any one of the above items 1, 4, and 5, wherein the reaction is carried out using an organic solvent immiscible with water as an organic solvent, and performing the reaction in the presence of a basic compound of an alkali metal or an alkaline earth metal. Described manufacturing method,

7.The method according to any one of the above items 1, 4, and 5, wherein in the reaction using benzoic anhydride as the compound represented by the general formula (4), the reaction is performed in the presence of an organic amine.

8. The method according to the above item 7, wherein a salt formed by benzoic acid and the organic amine is precipitated in an organic solvent used for the reaction, as the organic amine.

9. The production method according to the above 8, wherein the organic amine is dicyclohexynoleamine,

10. The method according to any one of items 1 to 9, wherein R1 in the general formulas (1), (3), (5), and (6) is a methyl group and R2 is hydrogen. Production method,

11. General formula (3) [0046]

(Wherein R 1 and R 2 are as defined above),

12. General formula (7)

[0048]

[In the formula, m and n are each an integer up to 110, Rl and R2 are as described above, and Q is a general formula (8)

[0050] [Formula 21]

0

R3 ( ₈ )

Wherein R 3 represents an alkyl group having 13 to 13 carbon atoms, and R 4 represents an alkyl group having 2 to 5 carbon atoms, and a compound represented by the general formula (9) [0052] [Formula 22]

(Wherein R5 and R6 each represent an alkyl group having 13 to 13 carbon atoms), or a compound represented by the general formula (10)

[0054] [Formula 23]

R7- CN (10)

(Wherein, R 7 represents an alkyl group having 13 to 13 carbon atoms)

. A crystal of the compound represented by

13. The crystal of the compound according to claim 12, wherein R5 and R6 in the general formula (9) are both methyl groups.

14. A crystal of the compound according to any one of 1-11 above, wherein R1 in the general formula (3) or (7) is a methyl group, and R2 is hydrogen,

It is.

The invention's effect

According to the present invention, a novel compound represented by the general formula (5) such as N ⁴ _benzoyl_5, _0_ (4,4, -dimethoxytrityl) _2, _doxy-5-methylcytidine is provided. The ability to provide manufacturing methods. According to one embodiment of the present invention, a compound represented by the general formula (3) such as 5′-0- (4,4′-dimethoxytrityl) -12′-deoxy-5-methylcytidine is Crystallization is possible by forming a solvate with an ester solvent, a ketone solvent or a nitrile solvent. The production route provided by the present invention via this compound as an intermediate is that N ⁴ -benzoyl-3,5,5-〇_bis (4,4, -dimethoxytrityl) _2, -de 3,5, __- bis (4,4, -dimethoxytrityl) -2'-deoxy-5-methylcytidine, a precursor of oxy-5-methylcytidine, can be easily removed by recrystallization or the like. it can. Furthermore, in the next step, 5′_0_ (4,4′-dimethoxytrityl) -3, N ⁴ —dibenzoyl 2′-dexoxy 5—by-produced when the compound represented by the general formula (5) is produced. It is possible to convert a compound represented by the general formula (6) such as methylcytidine into a compound represented by the general formula (5). it can. Therefore, it is possible to manufacture high-quality products while effectively using by-products.

[0057] As is clear from the above, the present invention is suitable for an industrial production method because a high-purity product can be efficiently produced with a simple operation.

Brief Description of Drawings

FIG. 1 is a chart showing a chart obtained by measuring an object obtained in Example 1 by XRD.

FIG. 2 is a view showing a chart obtained by measuring an object obtained in Example 2 by XRD.

BEST MODE FOR CARRYING OUT THE INVENTION

In the compounds represented by the general formula (1), the general formula (3), the general formula (5), the general formula (6), and the general formula (7), R1 is an alkyl having 1 to 4 carbon atoms. Represents an alkynyl group having 2 to 4 carbon atoms, an alkenyl group having 2 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, or a halogen atom, and R2 represents a hydrogen atom, 1 to 4 carbon atoms. Represents an alkoxyl group, an alkoxyl group having 1 to 4 carbon atoms having a substituent, or a halogen atom.

In the general formula (1), the general formula (3), the general formula (5), the general formula (6), and the general formula (7), with respect to the alkoxyl group having a substituent of R2 and having 14 carbon atoms, It represents an alkoxyl group belonging to the range in which the number of carbon atoms falls within the applicable range and represents a group having a plurality of substituents at arbitrary positions. Examples of the substituent include an alkoxy group, an aryl group and the like. Specific examples include methoxymethoxy group, butoxymethoxyl group, pentyloxymethoxyl group, trichloroethoxymethoxyl group, methoxyethoxymethoxyl group, methoxyethoxyl group, benzyloxyl group, benzyloxymethyl group, methoxybenzyloxymethoxyl group. And the like.

The configuration of R2 in the compounds represented by the general formulas (1), (3), (5), (6) and (7) is not particularly limited. Or you can take the S configuration.

The compound represented by the general formula (1) or a salt thereof is reacted with the compound represented by the general formula (2) to isolate the compound represented by the general formula (3). The compound represented by the general formula (5) is reacted with the compound represented by the general formula (4) in an organic solvent. ) Can be produced.

As the compound represented by the general formula (1), a compound in which R1 in the general formula (1) is a methyl group and R2 is a hydrogen atom can be preferably applied to the production method of the present invention.

[0060] As the compound represented by the general formula (1), a commercially available product can be usually used.

As the salt of the compound represented by the general formula (1), usually, a salt composed of the compound represented by the general formula (1) and an acid is used.

The acid may be an organic acid or an inorganic acid.Specifically, inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, etc., methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, trifluoromethanesulfonic acid, dichloroacetic acid And the like, among which hydrochloric acid is preferred.

The salt of the compound represented by the general formula (1) can be produced by reacting the compound represented by the general formula (1) with the above-mentioned acid.

X in the general formula (2) represents a halogen atom.

The compound represented by the general formula (2) is not particularly limited as long as it is at least 1 equivalent to the compound represented by the general formula (1) or a salt thereof. Use equivalents.

In the reaction for obtaining the general formula (3) from the compound represented by the general formula (1) or a salt thereof and the compound represented by the general formula (2), a solvent is usually used.

[0061] The solvent is not limited as long as it does not react with the compound represented by the general formula (2), but may be an aliphatic solvent such as hexane, pentane, heptane, or cyclohexane. Hydrocarbon solvents, aromatic solvents such as benzene, toluene, xylene, tamene, cymene, and anisol; halogen solvents such as chloroform, dichloromethane and dichloroethane; ester solvents such as ethyl acetate, isopropyl acetate and butyl acetate; Ketone solvents such as acetone, methyl ethyl ketone and methyl isobutyl ketone; nitrile solvents such as acetonitrile and propionitrile; amide solvents such as dimethylformamide; urea solvents such as dimethyl imidazoline and dimethyl propylene urea; pyridine , Collidine and other nitrogen-containing aromatic solvents, t-butyl methyl ether, dioxa And ether-based solvents such as tetrahydrofuran, among which pyridine, dimethinolephonoremamide, dimethylimidazoline, tetrahydrofuran, and dioxane are preferred. More preferably, it is pyridine. [0062] The amount of the solvent to be used is not limited as long as it does not hinder the reaction, but usually it is used in an amount of 3 to 20 times the amount of the substrate.

The reaction between the compound represented by the general formula (1) or a salt thereof and the compound represented by the general formula (2) is usually performed in the presence of a base in order to trap hydrogen halide generated in the reaction. Examples of the base include organic bases such as pyridine, lutidine, collidine, triethylamine and tributylamine. Of these, pyridine is preferred.

[0063] The amount of the base used is not particularly limited as long as it is at least 1 equivalent to the compound represented by the general formula (1) or a salt thereof. I do.

[0064] The reaction temperature may be any temperature that is sufficient for the reaction to proceed, and may be any temperature as long as it does not cause decomposition, but is preferably from -10 ° C to 40 ° C, more preferably -5 ° C. More than 2

5 ° C.

[0065] The reaction pressure is not particularly limited, but the reaction is usually performed under atmospheric pressure.

The compound represented by the general formula (3) or the compound represented by the general formula (3) is obtained from a reaction solution obtained by reacting the compound represented by the general formula (1) or a salt thereof with the compound represented by the general formula (2). Each of the compounds represented by the formula (7) can be isolated as crystals.

When each of the compound represented by the general formula (3) or the compound represented by the general formula (7) is isolated as a crystal, an excess of the general formula ( It is possible to decompose the compound represented by 2), reduce impurities or inorganic salts by filtration or liquid separation, and distill off the reaction solvent.

The crystal of the compound represented by the general formula (3) or the crystal of the compound represented by the general formula (7)

Of these, a crystal of a compound in which R1 in the general formulas (3) and (7) is a methyl group and R2 is a hydrogen atom is preferable.

M and n in the general formula (7) are each an integer of up to 110, and R1 and R2 in the general formula (7) are as described above.

[0069] In Formula (8), R3 represents an alkyl group having 13 to 13 carbon atoms, and R4 represents an alkyl group having 25 carbon atoms.

[0070] Among the compounds represented by the general formula (8), R3 in the general formula (8) is a methyl group; Is preferably an ethyl or butyl group.

R5 and R6 in the general formula (9) each represent an alkyl group having 13 to 13 carbon atoms.

[0071] Among the compounds represented by the general formula (9), a compound in which R5 and R6 in the general formula (9) are methyl groups is preferable.

[0072] In the general formula (10), R7 represents an alkyl group having 13 to 13 carbon atoms.

[0073] Among the compounds represented by the general formula (10), a compound in which R7 in the general formula (10) is a methyl group is preferable.

[0074] The solvent used for producing the crystal of the compound represented by the general formula (3) or the crystal of the compound represented by the general formula (7) includes hexane, isohexane, pentane, heptane, and cyclohexane. Aliphatic hydrocarbon solvents such as xane, aromatic hydrogen solvents such as benzene, toluene, xylene, tamene, cymene, and anisol; halogen solvents such as chloroform, dichloromethane, and dichloroethane; ethyl acetate, isopropyl acetate, and butyl acetate Ester solvents such as acetone, methyl ethyl ketone, methyl isobutyl ketone, etc., nitrile solvents such as acetonitrile, propionitrile, alcohol solvents such as methanol, ethanol, isopropyl alcohol, etc. Amide solvents such as dimethylformamide, dimethylimidazoline, dimethylpropyl Examples include urea solvents such as urea, nitrogen-containing aromatic solvents such as pyridine and collidine, ether solvents such as t-butyl methyl ether, dioxane, and tetrahydrofuran, and water. In particular, when an ester solvent, a nitrile solvent, or a ketone solvent is used, the compound represented by the general formula (3) forms a solvate to form a crystal of the compound represented by the general formula (7). can get. Regarding the solvent used for crystal production, a single solvent or a mixed solvent may be used in any ratio as long as the solubility is sufficient for crystallization.

When preparing a crystal of the compound represented by the general formula (7), ethyl ethyl acetate, butyl acetate, acetonitrile, and methyl isobutyl ketone are preferred, and methyl isobutyl ketone is more preferred.

[0076] The temperature for producing the crystal of the compound represented by the general formula (3) or the crystal of the compound represented by the general formula (7) is arbitrary as long as it is a temperature sufficient for crystallization. However, a range of -10 ° C to 50 ° C is desirable, more preferably -10 ° C to 40 ° C. [0077] In producing crystals of the compound represented by the general formula (3) or crystals of the compound represented by the general formula (7), the compound represented by the general formula (3) is dissolved. By adding a new solvent to the solution, the solubility of the compound represented by the general formula (3) is reduced, thereby making it possible to manufacture the compound. Further, in order to reduce the solubility, the compound can be produced by removing the solvent from a solution in which the compound represented by the general formula (3) is dissolved.

[0078] Production of crystals of the compound represented by the general formula (3) or crystals of the compound represented by the general formula (7)

The amount of the solvent at this time may be arbitrary as long as it is an amount sufficient for crystallization, but is preferably between 3 times and 100 times the amount of the compound of the general formula (3), more preferably 5 times. The double power is also 50 times.

[0079] Crystals of the compound represented by the general formula (7) can be converted into crystals of the compound represented by the general formula (3) by solvent replacement or drying under reduced pressure.

The compound represented by the general formula (3) or (7) obtained as described above is obtained by removing the impurity represented by the general formula (5). Can be used for manufacturing.

The compound represented by the general formula (5) can be produced by reacting the compound represented by the general formula (3) and the compound represented by the general formula (4) in an organic solvent.

[0080] X in the general formula (4) represents a benzoyloxy group or a halogen atom.

Of the compounds represented by the general formula (4), benzoic anhydride and benzoyl chloride are preferred because they are easily available.

The compound represented by the general formula (4) is not particularly limited as long as it is at least 1 equivalent to the compound represented by the general formula (3), but from the viewpoint of economy, 115 equivalents are used.

The organic solvent is not particularly limited as long as the reaction proceeds, but examples thereof include ketone solvents such as acetate, methyl ethyl ketone, and methyl isobutyl ketone, t-butyl methyl ether, tetrahydrofuran, and the like. Ether solvents, aromatic solvents such as benzene, toluene, xylene, cumene, cymene, and anisol; alcohol solvents such as isopropyl alcohol and butanol; halogen solvents such as methylene chloride, chloroform, and dichloroethane; and ethyl acetate , Isopropyl acetate, butyl acetate and other ester solvents, acetonitrile Nitrile solvents and the like can be mentioned.

[0082] The amount of the organic solvent to be used is not limited as long as it does not hinder the reaction, but is usually 3 to 20 times the amount of the reaction substrate.

The reaction temperature may be set to an arbitrary temperature if the reaction between the compound represented by the general formula (3) and the compound represented by the general formula (4) proceeds and the decomposition of the general formula (3) does not occur. Can be set. Usually, it is 30 ° C. or higher and the boiling point of the solvent or lower.

[0083] The reaction pressure is not particularly limited, but the reaction is usually performed under atmospheric pressure.

[0084] The reaction between the compound represented by the general formula (3) and the compound represented by the general formula (4) is performed using an organic solvent that is immiscible with water as an organic solvent, and The reaction can be carried out in the presence of a basic compound of an alkali metal or an alkaline earth metal.

Examples of the water-immiscible organic solvent include ketone solvents such as methyl ethyl ketone and methyl isobutyl ketone, and ether solvents such as t-butyl methyl ether and tetrahydrofuran.

Aromatic solvents such as benzene, toluene, xylene, tamene, cymene, and anisol; halogen solvents such as methylene chloride, chloroform, dichloroethane; and ester solvents such as ethyl acetate, isopropyl acetate, and butyl acetate. No.

[0086] The amount of the organic solvent to be used is not limited as long as it does not hinder the reaction, but usually it is 3 to 20 times the amount of the reaction substrate.

Examples of the basic compound of an alkali metal or an alkaline earth metal include sodium hydrogen carbonate, sodium carbonate, sodium hydroxide, potassium hydrogen carbonate, potassium carbonate, potassium hydroxide and the like.

The basic form of alkaline metal or alkaline earth metal compound is to be used after dissolving in water.

The

The amount of the alkali metal or alkaline earth metal basic compound used is not particularly limited as long as it is set so as to maintain alkalinity at the end of the reaction.

[0087] In the reaction between the compound represented by the general formula (3) and the compound represented by the general formula (4), benzoic anhydride was used as the compound represented by the general formula (4). In this case, the reaction is preferably performed in the presence of an organic amine. As the organic amine, those having low reactivity with benzoic anhydride are preferred.For example, diisopropylamine, triethylamine, trippropinoleamine, tributylamine, triamylamine, trihexylamine, triheptinolamine, trioctylamine, Tertiary amines such as diisopropylethylamine, pyridine, collidine and dimethylaniline, and secondary amines such as diisopropylamine and dicyclohexylamine.

Among the organic amines, when a salt formed by reacting benzoic acid by-produced with the organic amine precipitates in the organic solvent used in the above reaction, the yield of the target compound tends to be improved. This is particularly preferred because

Such organic amines include, for example, symmetric amines, but are not particularly limited. Specific examples include dicyclohexynoleamine.

The amount of the organic amine is not particularly limited, but is usually 1 equivalent or more and 3 equivalents or less from an economic viewpoint.

By reacting the compound represented by the general formula (3) with the compound represented by the general formula (4), the compound represented by the general formula (5) is mainly produced, and is represented by the general formula (6). Compounds are by-produced.

A method for producing a compound represented by the general formula (5) by reacting a compound represented by the general formula (3) with a compound represented by the general formula (4) in an organic solvent, General formula by-produced in the reaction

By including the step of treating the compound represented by (6) with a base, the compound represented by the general formula (6) can be selectively converted to the compound represented by the general formula (5).

The base used for the conversion of the compound represented by the general formula (6) to the compound represented by the general formula (5) includes a benzoyl group at the N ⁴ position which is not hydrolyzed and a benzoyl group at the 3′-position. There is no particular limitation as long as the group can be hydrolyzed. Examples of such a base include alkali metal hydroxides and alkali metal hydroxides and alkoxides.

The amount of the base used is not particularly limited as long as it is set so that the hydrolysis reaction proceeds, but is from 1 equivalent to 10 equivalents from an economic viewpoint.

The reaction temperature is not particularly limited as long as it is set so that the benzoyl group at the N ⁴ position is not hydrolyzed, and is usually preferably −20 ° C. or more and 20 ° C. or less.

The method for recovering the compound represented by the general formula (3) is described in Examples and Non-Patent Document 1. This can be done with reference to the method described above.

[0088] In this way, it has become possible to industrially produce high-purity N ⁴ -benzoyl-5, _0_ (4,4, dimethoxytrityl) _5_-substituted cytidine derivatives. Example

Hereinafter, the present invention will be described in more detail by way of examples, but the present invention is not limited thereto. In the following, 5, _〇_ (4,4, dimethoxytrityl) _2, -deoxy-5-methylcytidine is referred to as (I) and 3,, 5, -0-bis (4,4, -dimethoxytrityl) -2 Deoxy-5-methyl cytidine (II), N ⁴ -benzoyl-5 '_'_ (4,4'-dimethoxytrityl) _2'-Doxy-5-methyl cytidine (III), 5 , — O— (4,4, -Dimethoxytritinol) -3, N ⁴ -dibenzoyl-2′-doxy-5_methylcytidine (IV) and 3 ′, 5′_bis (4,4′_dimet (Xithrityl) -14_ (N-benzoyl) _5-methyl-2'-deoxycytidine is abbreviated as (V), and methyl isobutyl ketone is abbreviated as MIBK.

[0090] Regarding the HPLC conditions of (1) and (II), a column of YMC_Pack CN A512 (size 6.0 X 150 mm manufactured by YMC) was used, the temperature of the column oven was 40 ° C, and the eluate was acetonitrile. : LOOmM aqueous solution of triethylamine acetate = 42:58, the flow rate was set to 1.0m1Z, and the observation wavelength was set to I = 254nm.

[0091] Regarding the HPLC conditions of (111), (IV) and (V), Develosil 〇DS_MG_5 (size 4.6

X 250mm column (manufactured by Nomura Chemical Co., Ltd.), the column oven temperature is 40 ° C, the eluent is acetonitrile: lOOmM aqueous solution of triethylamine acetate = 90:10, the flow rate is 1.0 OmlZ, and the observation wavelength is I = The measurement was performed at 254 nm.

[Example 1]

5_Methyl_2'_doxycytidine hydrochloride l.lg (40 mmol) was subjected to azeotropic dehydration twice with pyridine (200 ml), and then dissolved in 160 ml of pyridine. After charging 14.22 g of 4,4'-dimethoxytrityl chloride at room temperature, the mixture was stirred at room temperature for 3 hours. When the reaction mixture was analyzed by HP LC, the largest impurity was (II), and (I) / (II) = 92.0 / 8.0 (area ratio). After adding 8.74 g of sodium hydrogen carbonate and stirring at room temperature for 2 hours, pyridine was concentrated under reduced pressure. After adding 300 ml of ethyl acetate and 200 ml of water to the concentrated residue and separating the mixture, the organic layer is mixed once with a mixed solvent of 50 ml of saturated aqueous sodium bicarbonate and 150 ml of water, and mixed with 100 ml of saturated saline and 100 ml of water. Washed once with solvent. After concentrating the organic layer, 200 ml of MIBK was added, and the obtained crystals were filtered and washed. After drying at room temperature and observing the crystals by ¹ H-NMR, (I) was obtained as a solvate of (I) having a composition of (I) / MIB K = 3/2. Further, when dried at 40 ° C., the composition was (I): MIBK = 1: 6, and a yield of 19.3 g (purity conversion yield: 85./.) Was obtained. When this product was analyzed by HPLC, the product area was 99.5% (area), the maximum impurity was (II), and the purity was very high, 0.3% (area).

[0092] The analysis values of the crystal of (I): MIBK = 1: 6 are shown below.

[0093] 'H-NMR (DMSO-d6) δ ppm: 0.85 (6H, d: MIBK), 1.49 (3H, s), 2.00 (1H, m: MIBK), 2.06 (3H, s: MIBK), 2.0-2.2 (2H, m),

2.29 (2H, d: MIBK), 3.19-3.22 (2H, m), 3.73 (6H, s), 3.82_3.89 (lH, m), 4.25—4.29 (lH, m), 5.29 (lH , d),

6.23 (1H, t) 6.7-6.8 (lH, br), 6.90 (4H, d), 7.2_7.45 (10H, m), 7.49 (lH, s). Obtained by measuring the target substance by XRD The chart is shown in FIG.

[Example 2]

0.55 g of 5-methyl-2'-deoxycytidine hydrochloride was subjected to azeotropic dehydration twice with 10 ml of pyridine, and then dissolved in 5 ml of pyridine. After charging 0.74 g of 4,4'-dimethoxytrityl chloride at room temperature, the mixture was stirred at room temperature for 4.5 hours. After adding sodium hydrogen carbonate and stirring at room temperature for 2 hours, pyridine was concentrated under reduced pressure. After adding 10 ml of ethyl acetate and 10 ml of water to the concentrated residue and separating the mixture, the organic layer was washed once with a mixed solvent of 1 Oml of saturated saline and 100 ml of water. After the organic layer was dried over sodium sulfate, the solvent was distilled off. Acetonitrile (10 ml) was added to the obtained residue, and the mixture was crystallized at room temperature, filtered, and dried. Observation of the obtained crystals by ¹ HN MR revealed that (I) had a composition of (1) / acetonitrile = 3Zl as an acetonitrile solvate. The yield was 0.55 g. When this product was analyzed by HPLC, the area of the product was 99.6% (area), and the maximum impurity was (II), 0.2% (area). The chart obtained by measuring the target product by XRD is shown in FIG.

[0094]

[Example 3]

2, -Deoxy-5-methylcytidine hydrochloride 8. After azeotroping Og with 100 ml of pyridine, 120 ml of lysine and 10.2 g of 4,4'-dimethoxytrityl chloride were added and reacted at room temperature for 4 hours. When this reaction solution was analyzed by HPCL, it was (I) / (II) = 93.7 / 6.3 (area ratio). To this was added 6. lg of sodium bicarbonate, and the mixture was stirred for 1 hour, and then concentrated under reduced pressure to 80 g. Subsequently, MIBK100 ml was added, and the mixture was washed three times with 130 ml of water. The obtained MIBK layer was concentrated under reduced pressure, the precipitate was collected by filtration, and recrystallized from MIBK100 ml and pyridine 50 ml to obtain 12.7 g of (I). When the obtained (I) was analyzed by HPLC, (Ι) / (Π) = 100/0, and (II) was removed.

[Example 4]

582 mg (1.07 mmol) of (I) synthesized in Example 1 was azeotropically dehydrated twice with 1 Oml of dehydrated pyridine. After dissolving in 4 ml of pyridine, 292 mg of benzoic anhydride was added at room temperature, and the mixture was reacted at room temperature for 6 hours. Then, 30 mg of benzoic anhydride was added, and the mixture was allowed to stand overnight. After stirring at 40 ° C for 1 hour, the mixture was cooled and added with 50 zL of water under ice cooling. Further, 0.8 ml of a 30% aqueous solution of sodium hydroxide was added at 15 ° C, and 1 ml of pyridine, 2 ml of ethanol and 7 100 / iL were stirred at 5 ° C to 0 ° C for 3 hours. . After neutralization by adding a 7% aqueous hydrochloric acid solution, 1 Oml of water and 1 Oml of ethyl acetate were added and separated. The organic layer was washed once with 10 ml of a saturated aqueous sodium hydrogen carbonate solution and 10 ml of a saturated saline solution, and then the organic layer was concentrated. To the organic layer after concentration, 2.5 g of MIBK was added, and the mixture was added dropwise to 40 ml of hexane at room temperature. The precipitated solid was filtered, washed with hexane, and dried under reduced pressure to obtain 0.48 g of (III) as a light brown solid. According to the HPLC analysis, (III) was 99.4% (area), and the largest impurity was (V) 0.2% (area), indicating that it was very high purity.

[Example 5]

2.0 g of (I) and 0.92 g of benzoic anhydride were added to 20 ml of MIBK and reacted at 70 ° C for 2 hours.

When observed by HPLC, the yield of (III) was 85% and the yield of (IV) was 1%. Next, the organic layer was washed with a 5% by weight sodium carbonate solution, and the separated organic layer was ice-cooled. To this was added dropwise 10 ml of an 80% ethanol aqueous solution in which 0.44 g of sodium hydroxide was dissolved, and the mixture was reacted at the same temperature for 4 hours to convert (IV) to (III). After neutralization with 2N aqueous hydrochloric acid, (IV) was not detected by HP LC observation, and (III) was obtained in a yield of 86%.

[Example 6]

Synthesized in Example 3 in MIBK100 ml and 100 g of a 5% by weight aqueous sodium hydrogen carbonate solution (I) 10.0 g and 5.Og of benzoic anhydride were added and reacted at 70 ° C. for 4 hours. After the completion of the reaction, the resultant was cooled to room temperature. Observation by HPLC revealed that the yield of (II) was 97% and the yield of (IV) was 2%. The separated organic layer was ice-cooled, and 80% aqueous ethanol in which 2.21 g of sodium hydroxide was dissolved was added dropwise at 5 ° C or lower. After reacting for 4 hours under ice cooling, 10 ml of water in which 3.34 g of acetic acid was dissolved was added dropwise at 5 ° C or lower. Observation by HPLC revealed that (IV) had disappeared and (I II) power was about 8%. The organic layer was separated, washed with a saturated sodium hydrogen carbonate solution, concentrated under reduced pressure, and added with 25 ml of MIBK. This MIBK solution was slowly dropped into 650 ml of hexane. The precipitate was collected by filtration and dried under reduced pressure to obtain 11.3 g of (III) (yield 94%). Observation of the obtained compound by HPLC revealed that it was 99.4% (area).

[Example 7]

The procedure was performed in the same manner as in Example 6 except that sodium hydrogen carbonate was changed to sodium carbonate. After the reaction was completed, the yield of (ΠΙ) was 75%, and the yield of (IV) was 21%. After completion of the hydrolysis, (IV) was converted to (III) without being observed, and the yield was 96%. The obtained compound was 10.8 g (yield 90%). Observation of the obtained compound by HPLC revealed that it was 99.2% (area).

[Example 8]

To MIBKlOml, 10 g of a 5% by weight aqueous solution of sodium hydrogencarbonate, (1) 1.Og, and 289 μl of benzoinole chloride were added and reacted at 70 ° C. for 3 hours. Further, benzoyl chloride (96 μl) was added and reacted for 2 hours, cooled to room temperature, and separated. Observation by HPLC revealed that the yield of (III) was 95% and the yield of (IV) was 1%. The separated organic layer was ice-cooled, and 5 ml of an 80% aqueous ethanol solution containing 0.22 g of sodium hydroxide was added dropwise and reacted for 4 hours. The reaction mixture was separated into two layers, and when the organic layer was observed by HPLC, (IV) had disappeared and (III) was 96%. The organic layer was washed with a saturated sodium hydrogen carbonate solution, concentrated under reduced pressure, and added with 3 ml of MIBK. This MIBK solution was slowly added dropwise to 70 ml of hexane. The precipitate was collected by filtration and dried under reduced pressure to obtain (III) in l.Og (yield 90%). Observation of the obtained compound by HPLC revealed that it was 99.1% (area).

[Example 9]

2.0 g of (I), l.lml of tri-n-butylamine and 0.92 g of benzoic anhydride were added to 20 ml of MIBK. And reacted at 70 ° C for 2 hours. Observation by HPLC revealed that the yield of (III) was 87% and that of (IV) was 3%. Next, the organic layer was washed with a 5% by weight sodium carbonate solution and a saturated aqueous solution of ammonium chloride in this order, and the separated organic layer was ice-cooled. To this, 10 ml of an 80% aqueous ethanol solution in which 0.44 g of sodium hydroxide was dissolved was added dropwise, and the mixture was reacted at the same temperature for 4 hours to convert (IV) into (III). After neutralization with 2 N aqueous hydrochloric acid, HPLC observation showed that (III) was obtained in a yield of 89%.

[Example 10]

10.6 g of (I), 4.45 g of dicyclohexylamine, and 5.28 g of benzoic anhydride were added to MIBKlOOml, and reacted at 70 ° C. for 4 hours. At the end of the reaction, the dicyclohexynoleamine salt of benzoic acid was in a precipitated state. When the reaction mass was observed by HPLC, the yield of (III) was 93%, and the yield of (IV) was 3%. Next, the precipitated dicyclohexynoleamine salt of benzoic acid was filtered, and the filtrate was cooled to 3 ° C. To this, 50 ml of 80% ethanol in which 2.34 g of sodium hydroxide was dissolved was added dropwise while maintaining the temperature at 5 ° C or lower, and the mixture was stirred under ice cooling for 4 hours. Neutralized with 2 ml of 2N hydrochloric acid, and the organic layer was observed by HPLC. As a result, (IV) had completely disappeared and (III) was obtained in a yield of 94%.

[Comparative example]

N ⁴ -Benzoyl 2'-doxy-5-methylcytidine (0.3 g) was azeotropically dehydrated twice with dehydrated pyridine (20 ml) and then dissolved in dehydrated pyridine (30 ml). After adding 328 mg of 4,4'-dimethoxytrityl chloride, the mixture was reacted at room temperature for 10 hours. After stirring methanol for 1 hour, the solvent was distilled off under reduced pressure. To the concentrated residue were added 20 ml of black-mouthed form and 30 ml of a 5% by weight aqueous sodium hydrogen carbonate solution, and the layers were separated. The organic layer was filtered through a membrane filter and then concentrated under reduced pressure. 7 ml of getyl ether was added to the concentrated residue, and the precipitated component was filtered through a membrane filter. The mixture was added dropwise to 100 ml of hexane and further stirred for 2 hours. The precipitate was filtered, washed with hexane, and dried under reduced pressure to obtain 0.42 g of a solid. When this solid was analyzed by HPLC, it was found that (III) was 77.0% (area), the raw material was 21.0% (area), and the impurities 3, 5, 5, 5-bis (4 , 4, - dimethoxytrityl) _N ⁴ _ Benzoiru _2, - Dokishi - 5-methyl cytidine have been included 1 2.6% (area). Industrial applicability

According to the present invention, an N ⁴ -benzoinolecitidine derivative can be synthesized with a high degree of purity by simple means, so that it can be produced industrially advantageously and has high industrial applicability.

Claims

Claims [1] General formula (1)

[Chemical 1]

(In the formula, R1 is an alkyl group having 1 to 4 carbon atoms, an alkenyl group having 24 carbon atoms, an alkynyl group having 2 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, or a halogen atom. R 2 represents a hydrogen atom, an alkoxyl group having 14 to 14 carbon atoms, an alkoxyxylene group having 14 to 14 carbon atoms having a substituent, or a halogen atom), or a salt thereof, and a compound represented by the general formula (2 )

[Formula 2]

(X represents a halogen atom) to react with a compound represented by the general formula (3)

[Formula 3]

(Wherein R1 and R2 are the same as described above).

[Formula 4]

(Wherein X represents a benzoyloxy group or a halogen atom). A compound represented by the general formula (5)

[Formula 5]

(Wherein R1 and R2 are as defined above).

General formula (1)

[Formula 6]

(In the formula, Rl represents an alkyl group having 1 to 4 carbon atoms, an alkenyl group having 2 to 4 carbon atoms, an alkynyl group having 2 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, or a halogen atom. And R 2 represents a hydrogen atom, an alkoxyl group having 14 to 14 carbon atoms, an alkoxyxylene group having 14 to 14 carbon atoms having a substituent, or a halogen atom, or a salt thereof, and a compound represented by the general formula ( 2)

[Formula 7]

[Formula 8]

(Wherein, R1 and R2 are the same as described above). [3] General formula (1)

[Formula 9]

[Formula 10]

[Formula 11]

(Wherein, R1 R2 is the same as described above).

[4] General formula (3)

[Formula 12]

And general formula (4)

[Formula 13]

[Formula 14]

(Wherein, R1 and R2 are as described above).

[5] General formula (3)

[Formula 15]

(Wherein, R1 and R2 are as described above) and a compound represented by the general formula (4)

[Formula 16]

(Wherein, X represents a benzoyloxy group or a halogen atom).

[Formula 17]

(Wherein R1 and R2 are _{t 1 as} described above) by treating the compound represented by the general formula (5)

[Formula 18]

5. The production method according to claim 1, comprising a step of converting into a compound represented by the formula (wherein R1 and R2 are as defined above).

[6] The method according to any one of claims 1, 4, and 5, wherein the organic solvent is an organic solvent that is immiscible with water, and the reaction is performed in the presence of a basic compound of an alkali metal or an alkaline earth metal. The production method according to item 1.

[7] The reaction according to any one of claims 1, 4, and 5, wherein the reaction is carried out in the presence of an organic amine, in a reaction using benzoic anhydride as the compound represented by the general formula (4). The method according to 1.

[8] As the organic amine, a salt formed between benzoic acid and the organic amine is used as an organic solvent for the reaction. 8. The production method according to claim 7, wherein a substance precipitated in a medium is used.

9. The method according to claim 8, wherein the organic amine is dicyclohexynoleamine. 10. The method according to claim 11, wherein R1 in the general formulas (1), (3), (5), and (6) is a methyl group and R2 is hydrogen. .

General formula (3)

[Formula 19]

(Wherein R1 and R2 are as defined above).

[12] General formula (7)

[Formula 20]

Wherein m and n are each an integer from 1 to 10, Rl and R2 are as described above, and Q is a general formula (8)

[Formula 21]

(Wherein R3 represents an alkyl group having 13 to 13 carbon atoms, and R4 represents an alkyl group having 2 to 5 carbon atoms), a general formula (9)

[Formula 22]

(Wherein, R5 and R6 represent an alkyl group having 13 carbon atoms)

Or a compound represented by the general formula (10)

[Formula 23]

R7- CN (10)

(Wherein, R7 represents an alkyl group having 13 to 13 carbon atoms)

Represents the amount of the compound represented by. ] The crystal | crystallization of the compound represented by this.

[13] The crystal of the compound according to claim 12, wherein R5 and R6 in the general formula (9) are both methyl groups.

[14] The crystal of the compound according to any one of [11] to [13], wherein R1 in the general formula (3) or (7) is a methyl group, and R2 is hydrogen.