JPH0931092A - Production of 2'-3'-di-o-acyl atp - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject compound useful in pharmaceutical field as a purinergic ATP receptor antagonist, etc., on an industrial scale in high yield at a low cost by acylating an ATP (salt) in the presence of a specific phosphonic acid ester derivative. SOLUTION: The objective 2',3'-di-O-acyl ATP useful in pharmaceutical field as a purinergic ATP receptor antagonist, etc., is produced in high yield by dissolving adenosine-5'-triphosphate(ATP) or its salt in water, bringing the solution into contact with a pyridinium-type cation exchange resin at <=4 deg.C to form ATP pyridinium salt, dissolving the salt in a mixture of methanol and tri-(n-butyl)amine, distilling out the solvent under reduced pressure, adding dimethylformamide, adding an acylation agent (e.g. isonicotinic acid) and tri-(n- butyl)amine and reacting in the presence of a phosphonic acid ester derivative expressed by formula [R<1> is a lower alkyl, a (substituted) aryl or an aralkyl; R<2> is cyano or azido].

Description

Detailed Description of the Invention

[0001]

The present invention relates to 2'-3'-di-O-
The present invention relates to a novel method for producing acyladenosine-5'-triphosphate (hereinafter abbreviated as 2'-3'-di-O-acyl ATP) or a salt thereof.

[0002]

2. Description of the Prior Art 2'-3'-di-O-acyl ATP is
It is useful in the field of medicine as an antagonist of a plinazic ATP receptor. Conventionally, as a method for producing 2′-3′-di-O-acyl ATP, for example, 1,1′-carbonyldiimidazole is used as a condensing agent, and a sodium salt of ATP and a carboxylic acid are mixed in an aqueous solution at room temperature at a temperature of 1 To the reaction for 2 days [Am. J. Physiol. , 257
(6) 2834 (1982)], 2-fluoro-1-methylpyridinium p-toluenesulfonate as a condensing agent, and a method of reacting a tributylammonium salt of ATP with a carboxylic acid in dichloromethane at room temperature for 1 to 2 days [New J ． Chem. , 17, 325 (19
93)] and the like are known. However, since all of these methods had a yield of 30% or less and were obtained as a mixture with a 2'- or 3'-monoacyl compound, they cannot be industrially applied and the production with a good yield is possible. Development of law was desired. Further, these acylation reactions are low in reactivity, which is not preferable as a general method applicable to many acylating agents.

[0003]

Therefore, an object of the present invention is to provide an industrially useful method capable of producing 2'-3'-di-O-acyl ATP in high yield, and to provide 2'-
It is an object of the present invention to provide a general method for producing 3′-di-O-acyl ATP.

[0004]

Under such circumstances, the inventors of the present invention have conducted extensive studies and as a result, as a result, 2'-3'-di-O, which has an extremely high yield and generality as compared with the known method, has been obtained. −
The present invention has been completed by finding out a method capable of producing acyl ATP. In the present invention, ATP or a salt thereof is acylated to obtain 2 '.
3'-di -O- acyl ATP or a process for the preparation of a salt thereof, the following equation the _{^{reaction, (R 1 O) 2 P}} (O) R 2 ( wherein, ^{R 1} represents a lower alkyl group or a substituted group R ² represents a cyano group or an azido group), and the aryl group or aralkyl group which may have 2′-3′-di- The present invention relates to a method for producing O-acyl ATP or a salt thereof. In addition, the present invention also provides, as a condensing agent, a compound represented by the following formula: (R ¹ O) ₂ P (O) R ² (wherein R ¹ represents a lower alkyl group or an aryl group or aralkyl group which may have a substituent, R ² represents a cyano group or an azido group), and ATP or a salt thereof is reacted with an acylating agent to produce 2′-3′-di-O-acyl ATP. Is provided. The method of the present invention can be preferably carried out in the presence of a base. That is, the present invention acylates ATP or a salt thereof to produce 2'-3'-di-
In the method for producing O-acyl ATP or a salt thereof,
The reaction is represented by the following formula: (R ¹ O) ₂ P (O) R ² (wherein, R ¹ is a lower alkyl group or an aryl group or aralkyl group which may have a substituent, and R ² is a cyano group or 2'-, which is carried out in the presence of a phosphonate derivative represented by azido group) and a base.
The present invention relates to a method for producing 3'-di-O-acyl ATP or a salt thereof.

As the acylating agent used in the method of the present invention, a saturated or unsaturated aliphatic carboxylic acid, a saturated or unsaturated cycloaliphatic carboxylic acid, an aromatic carboxylic acid, an araliphatic carboxylic acid, or a heterocyclic Carboxylic acids such as cyclic carboxylic acids or their reactive derivatives can be used. Further, these carboxylic acids may be substituted with a substituent that does not inhibit the reaction of the method of the present invention. Examples of these carboxylic acids include (1) an azido group in the molecule,
A carboxylic acid having a functional group such as a nitro group, a cyano group, an acetylamino group, an O-silyl group or a carboxylic acid having an unstable functional group which cannot be easily converted into an acid chloride, (2) Generally reactive Poor long-chain carboxylic acid,
(3) sterically bulky carboxylic acid, (4) sparingly soluble carboxylic acid, (5) conventional esterification method, for example, acid anhydride method (Schotten method), WSC method (method using water-containing condensing agent) , CDI method (method using non-aqueous condensing agent), Mukaiyama method (2-fluoro-1-methylpyridinium as condensing agent)
Non-aqueous method using p-toluenesulfonic acid), DC
Examples thereof include a carboxylic acid that does not react with Method C (method using a non-aqueous condensing agent).

The preferred acylating agent used in the method of the present invention has 1 to 3 carbon atoms which may have a substituent.
0, more preferably an alkanoic acid or alkenoic acid having 1 to 20 carbon atoms, 7 to 15 carbon atoms which may have a substituent,
More preferably, an aromatic carboxylic acid having 7 to 10 carbon atoms,
An arylalkanecarboxylic acid or an arylalkenecarboxylic acid having 8 to 16 carbon atoms, more preferably 8 to 10 carbon atoms, which may have a substituent, and optionally having 1
Examples thereof include 4- to 7-membered heterocyclic carboxylic acids containing one or more nitrogen atoms, oxygen atoms or sulfur atoms as different atoms. Substituents of these carboxylic acids include a lower alkyl group having 1 to 10 carbon atoms, preferably 1 to 5 carbon atoms, a hydroxyl group, a lower alkoxy group having 1 to 10 carbon atoms, preferably a lower alkoxy group having 1 to 5 carbon atoms, 1 to 1
Examples thereof include an amino group and an azido group which may be substituted with 0, preferably a lower alkyl group having 1 to 5 carbon atoms.

The following formula used in the method of the present invention: (R ¹ O) ₂ P (O) R ² (wherein R ¹ represents a lower alkyl group or an aryl group or aralkyl group which may have a substituent). , R ² represents a cyano group or an azido group), and the lower alkyl group of the substituent R ¹ of the phosphonate derivative represented by the formula ( ^{1) is} a lower alkyl group having 1 to 10 carbon atoms, preferably 1 to 5 carbon atoms, Specific examples thereof include a methyl group, an ethyl group, an isopropyl group and the like. As the aryl group or aralkyl group which may have a substituent of the substituent R ^1, an aryl group or an aralkyl group having 6 to 12 carbon atoms, preferably 6 to 10 carbon atoms, specifically, a phenyl group, Examples thereof include naphthyl group and benzyl group. These substituents are not particularly limited as long as they do not inhibit the reaction, and examples thereof include a lower alkyl group having 1 to 5 carbon atoms. The substituent ^{R 2,} a cyano group (-CN) or azido group (-N ₃₎ is preferable. Specific examples of the phosphonate derivative of the present invention include, for example, dimethylphosphorocyanidate [(MeO) ₂ P (O) C.
N], diethylphosphorocyanidate [(EtO) ₂ P
(O) CN], diisopropylphosphorocyanidate [(i-PrO) ₂ P (O) CN], diphenylphosphorocyanidate [(PhO) ₂ P (O) CN], diphenylphosphoroazide [(PhO)] ₂ P (O) N ₃ ] and the like. A more preferable example is diethylphosphorusocyanidate [(EtO) ₂ P (O) CN]. The phosphonate derivative used in the present invention can be produced by a known method or according to the method. For example, diethylphosphorocyanidate can be prepared according to the method of Shioiri [J. Am. Chem. Soc. , 9
4, 603 (1975)].

The ATP used in the method of the present invention is preferably used as its salt. Although various salts of ATP can be used, ammonium salts thereof are preferred. The ammonium salt of ATP includes ATP
An ammonium salt which can be produced by a usual method with a general amine, more preferably a tertiary amine can be used. Examples of the tertiary amine include trialkylamines having the same or different C1 to C10, more preferably C1 to C6 alkyl groups, amines having a 5- to 7-membered alicyclic group, pyridine, picoline, etc. Examples thereof include a heterocyclic amine which may have a substituent. More specifically, for example, trimethylamine, triethylamine, tripropylamine, triisopropylamine, tri-n-butylamine, triisobutylamine, tri-
t-butylamine, tri-n-amylamine, tri-n
Examples thereof include tri-lower alkylamines having an alkyl group having 1 to 6 carbon atoms such as hexylamine, dimethylethylamine, and dimethylpropylamine, and pyridine. The ammonium salt of ATP of the present invention can be produced, for example, by reacting ATP, preferably an organic salt of ATP, for example, a pyridine salt, with a tertiary amine in a solvent. As the solvent, alcohols such as methanol, ethanol and propanol are used. The reaction proceeds instantaneously at 0 to 50 ° C., preferably around room temperature.
Although the ammonium salt obtained by this method can be purified by a usual method, it can be directly used for the next reaction without purification.

The reaction of ATP or a salt thereof of the present invention with an acylating agent such as a carboxylic acid is carried out by reacting both with a di-lower alkylphosphorusanidate in an anhydrous solvent, more preferably in the presence of a base, The target 2'-3'-di-O-
Acyl ATP is obtained. As the solvent, for example, anhydrous dimethylformamide, dimethylsulfoxide, tetrahydrofuran, toluene, dichloromethane, chloroform and the like can be used. As the base, for example,
In addition to the tri-lower alkylamines exemplified above, sodium hydride, DBU (1,5-diazabicyclo [5.4.
[0] Undecene-5), potassium carbonate, sodium hydroxide and the like can be used. The reaction proceeds at 0 to 50 ° C., preferably around room temperature, for several hours to several days. Separation / purification of the desired product can be carried out by an ordinary method, for example, a solvent extraction method, a salt exchange method, a chromatography method, a recrystallization method, a lyophilization method, or the like alone or in combination. The target compound can also be isolated in the form of a salt. When this reaction is carried out in the presence of a tertiary amine as a base, a salt of the corresponding amine is usually obtained, but when another salt is desired, it can be converted into a new salt by a usual salt exchange method. .

2'-3 'produced by the method of the present invention
Specific examples of -di-O-acyl ATP include the following. 2'-3'-di-O-acetyl-ATP, 2'-3'-di-O-propionyl-ATP,
2'-3'-di-On-butyryl-ATP, 2'-
3'-di-O-isobrityl-ATP, 2'-3'-di-O-n-valeryl-ATP, 2'-3'-di-O-isovaleryl-ATP, 2'-3'-di-O -Capryloyl-ATP, 2'-3'-di-O-enanthyl-AT
P, 2'-3'-di-O-caprylyl-ATP, 2'-
3'-di-O-pelargonyl-ATP, 2'-3'-di-O-palmitoyl-ATP, 2'-3'-di-O-stearoyl-ATP, 2'-3'-di-O-ole. Oil-ATP, 2'-3'-di-O-benzoyl-ATP,
2'-3'-di-Op-toluoyl-ATP, 2'-
3'-di-O- (4-t-butylbenzoyl) -AT
P, 2'-3'-di-O- (2-aminobenzoyl)-
ATP, 2'-3'-di-O- (4-aminobenzoyl) -ATP, 2'-3'-di-O- (3,4,5-trimethoxybenzoyl) -ATP, 2'-3 ' -The-O
-Sartyloyl-ATP, 2'-3'-di-O-cinnamoyl-ATP, 2'-3'-di-O-naphthoyl-A
TP, 2'-3'-di-O- (5-azidonaphthoyl)
-ATP, 2'-3'-di-O-nicotinoyl-AT
P, 2'-3'-di-O-isonicotinoyl-ATP,
And their salts.

[0011]

The present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

Example 1 Preparation of 2'-3'-di-O-isonicotinoyl-ATP: (a) 1.0 g of the disodium salt of ATP (trihydrate) was dissolved in 20 ml of water and the temperature was kept at 4 ° C or lower. 150 ml of water was eluted with respect to 10 ml of cation exchange resin, Dowex 50W × 8 (200-400 mesh. Pyridinium type: manufactured by Dow Corning). The eluate was freeze-dried to give A
1.07 g of powder of TP pyridinium salt was obtained. (B) ATP pyridinium salt 20 mg (0.03 mm
was dissolved in a mixed solution of 1 ml of methanol and 22 mg (0.06 mmol) of tri- (n-butyl) amine. Next, the colorless caramel obtained by distilling off the solvent under reduced pressure was dissolved in 0.5 ml of dimethylformamide. 55 mg (0.3 m of tri- (n-butyl) amine was added to this solution.
l) was added, followed by 110 mg of isonicotinic acid (0.9
mmol) and diethyl phosphorocyanidate 147 m
g (0.9 mmol) was added and the mixture was stirred at room temperature. 91 mg (0.9 mmol) of triethylamine was gradually added dropwise to this reaction solution, and the mixture was stirred at room temperature for 72 hours. After the reaction,
The solvent was distilled off under reduced pressure at a bath temperature of 40 ° C or lower, and water was added to the residue.
The mixture was added with ml and back-extracted, and washed with ethyl acetate. The aqueous layer was separated and the solvent was distilled off under reduced pressure at 40 ° C. or lower to obtain 324 mg of a pale yellow oily substance. 1 ml of this oily substance is ethanol
The extract was dissolved in, and subjected to Sephadex LH-20, and eluted with ethanol. The solvent was distilled off from the fraction containing the target compound under reduced pressure to obtain 27 mg (yield: 90%) of 2-tri-n-butylamine salt of the target compound.

IR value cm ⁻¹ (KBr): 1741 ¹ H-NMR value δ (D ₂ O) ppm: 4.42-4.52 (2H, m, C5′-H × 2) 5.0-5 .1 (1H, brs.C4'-H) 6.2-6.3 (1H, m, C3'-H) 6.3-6.4 (1H, m, C2'-H) 6.88 ( 1H, d, C1'-H) 8.22 (1H, m, Aden-H) 8.42 (3H, d, Py-H) 8.52 (2H, d, Py-H) 8.75 (1H , S, Aden-H) 8.9 (2H, m, Py-H) 9.05 (2H, d, Py-H)

Example 2-14 Various carboxylic acids were used and treated in the same manner as in Example 1 to obtain compounds shown in Tables 1 to 3 below.

[0015]

[Table 1]

[0016]

[Table 2]

[0017]

[Table 3]

[0018]

INDUSTRIAL APPLICABILITY The present invention uses 2'-from ATP or a salt thereof.
Provided is an industrially useful method capable of producing 3'-di-O-acyl ATP in high yield, and 2'-3'-di-O.
-A general method for producing acyl ATP can be provided.

Claims (6)

[Claims] 1. ATP or a salt thereof is acylated to obtain 2'-
In the method for producing 3′-di-O-acyl ATP or a salt thereof, the reaction is performed by the following formula: (R ¹ O) ₂ P (O) R ² (wherein R ¹ represents a lower alkyl group or a substituent). 2′-3′-di-O, which is carried out in the presence of a phosphonate derivative represented by R ² which may have an aryl group or aralkyl group, wherein R ² represents a cyano group or an azido group. -Method for producing acyl ATP or a salt thereof. 2. A method for producing 2′-3′-di-O-acyl ATP or a salt thereof, which comprises performing the method according to claim 1 in the presence of a base. 3. The method according to claim 1, wherein the ATP salt is an ammonium salt. 4. The production method according to claim 1, wherein the salt of ATP is a pyridinium salt. 5. The production method according to claim 2, wherein the base is tri-lower alkylamine. 6. The production method according to claim 1, wherein the acylating agent is an aliphatic carboxylic acid, an aromatic carboxylic acid, or a reactive derivative thereof.