JPS635400B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the general formula A novel 2-substituted adenosine-3',5'-cyclic phosphoric acid (hereinafter adenosine-
(3',5'-cyclic phosphoric acid is abbreviated as CAMP).

The compound of the formula has excellent pharmacological effects such as antiallergic action, central nervous system stimulant action, platelet aggregation action, diuretic action, action on the circulatory system (e.g. hypotensive action, myocardial depressant action, etc.), and anticancer action. and is useful as a medicine.

A new compound of formula A halogenating agent is applied to 2-substituted-1,N ⁶ -etheno-adenosine-3',5'-cyclic phosphoric acid represented by (in the formula, R' means a cyano group, a carbamoyl group, or a carboxyl group) It can be manufactured by A compound of the formula in which R is a carboxyl group can also be produced by alkaline hydrolysis of a compound of the formula in which R is a carbamoyl group.

A compound of the general formula (In the formula, X means a halogen atom) is cyanated, and the obtained 2-cyano-
1,N ⁶ -etheno-adenosine-3′,5′-cyclic phosphoric acid (hereinafter referred to as 1,N ⁶ -etheno-adenosine-3′,5′-
It is obtained by alkaline hydrolysis of cyclic phosphoric acid (abbreviated as εCAMP).

A compound of formula may be, for example, 2-mercapto-
It is obtained by halogenating εCAMP (see JP-A-53-12891), and for example, 2-iodo-εCAMP, 2-chloro-εCAMP or preferably 2-bromo-εCAMP is used.

The cyanation reaction of compounds of the formula is carried out using an alkali cyanide such as potassium cyanide, sodium cyanide, etc., preferably in the presence of a polar organic solvent such as N,N'-dimethylformamide, etc., generally at a temperature of from room temperature to 60°C. It lasts from several hours to several days.

The alkaline hydrolysis of 2-cyano-εCAMP is usually carried out using basic substances such as sodium hydroxide, potassium hydroxide, etc. in a hydrophilic solvent such as water or alcohol.
This is carried out using ammonia, organic amines, etc.

2-Cyano-
From εCAMP, 2-carbamoyl-εCAMP, then 2-carboxy-εCAMP, or directly 2-carboxy-εCAMP can be obtained. For example 2
-When cyano-εCAMP is alkaline hydrolyzed at pH 10 to 13, generally 2-
Carbamoyl-εCAMP is obtained. If this reaction mixture is allowed to react for a longer time or by adding an additional amount of basic substance to reach a pH of 12-13.5,
-carboxy-εCAMP is obtained. When 2-cyano-εCAMP, 2-carbamoyl-εCAMP or a mixture of both is subjected to alkaline hydrolysis at pH 12 to 13.5,
Generally, 2-carboxy-εCAMP can be obtained in several minutes to several hours at 0 to 40°C.

The compound () of the present invention can be obtained by deethenoizing the starting material () by allowing a halogenating agent to act thereon. The deetheno reaction is carried out by dissolving or suspending the starting material () preferably in a solvent, adding a halogenating agent, and generally increasing the temperature to 0°C or higher, preferably 0 to 0°C.
It is carried out at 40° C. for up to 10 hours, preferably for 1 to 60 minutes, preferably under stirring. After the reaction is complete,
By adding a basic substance, preferably sodium hydroxide, potassium hydroxide, etc., to make the reaction mixture alkaline, the yield of the target substance () can be significantly increased. At that time, generally the liquid
It is particularly advantageous to maintain the pH at 10 or higher, preferably between 10 and 13, and to stir for several minutes to several hours.

Examples of halogenating agents include bromine, chlorine, N
-Chlorosuccinimide, N-bromosuccinimide, etc. are used, and N-
Bromosuccinimide is preferred. The amount of the halogenating agent to be used is generally at least equimolar to the compound of the formula, preferably 1 to 6 times the molar amount.

Any solvent can be used as long as it dissolves or suspends the starting material () and the halogenating agent and does not chemically react with these reaction participants, such as water, acidic solutions, buffers, organic solvents, etc. is used. As an acidic solution, a solution with a pH of 4.0 or less, preferably between PH0.1 and 1.0, and as a buffer solution,
The pH is 0.1 to 8, preferably PH 0.1 to 4.0. Examples of the organic solvent include any solvent miscible with water, such as alcohol, dimethylformamide, dimethylacetamide, acetone, and dioxane. These solvents can be used alone or in combination of two or more.

2-carbamoyl-CAMP is 2-cyano-
It can also be obtained by de-ethenoizing εCAMP and then alkaline hydrolysis. 2-carboxy-
CAMP is either deethenolyzed from 2-cyano-εCAMP and then subjected to alkaline hydrolysis in a higher PH range, or
Alternatively, it can also be obtained by alkaline hydrolysis of 2-carbamoyl-CAMP.

The de-etheno reaction of 2-cyano-εCAMP is carried out in the same manner as described above. The subsequent alkaline hydrolysis or alkaline hydrolysis of 2-carbamoyl-CAMP is usually carried out using basic substances such as sodium hydroxide, potassium hydroxide, ammonia, organic amines, etc. in a hydrophilic solvent such as water or alcohol. It will be done.

After de-ethenoization of 2-cyano-εCAMP,
A basic substance is added to the reaction mixture to adjust the pH to 10 or higher, preferably 10 to 13, and the alkaline hydrolysis is generally carried out at a temperature of 0 to 40°C for several minutes to several hours.
-Carbamoyl-CAMP is obtained. The deethenoization reaction mixture of 2-cyano-εCAMP was heated to PH12~
13.5, 2-carboxy-CAMP is obtained by alkaline hydrolysis under the same conditions as above.
Furthermore, when 2-carbamoyl-CAMP is subjected to alkaline hydrolysis at pH 12 to 13.5, 2-carboxy-CAMP is generally obtained in several minutes to several hours at 0 to 40°C. If the final alkaline hydrolysis is carried out at a lower pH, longer reaction times are required due to the formation of 2-carboxy-CAMP.

Note that in the starting materials used in the preparation of the compounds of the formula (for example, the compounds of the formula and the formula), the cyclic phosphoric acid moiety may be in the form of a free acid or a salt. As the salt, for example, an alkali metal salt, an ammonium salt or an organic ammonium salt is used.
In the case of a starting material that also has a carboxyl group, it can be similarly used as a free carboxylic acid or a salt thereof.

Isolation and purification of the target substance of the formula is carried out by conventional methods, and depending on the operating conditions, the compound of the formula can be obtained in the form of a free acid or a salt thereof in the cyclic phosphoric acid moiety and/or carboxyl group. For example, column chromatography using activated carbon, ion exchange resin, alumina, silica gel, etc., organic solvent precipitation using alcohols, acetone, dioxane, etc., precipitation using pH adjustment, etc. can be used alone or in combination. .

Conventional methods are used to convert a free acid type target substance into a salt, or to convert a salt into a free acid. Examples of salts include alkali metal salts such as sodium salts and potassium salts, and ammonium salts or organic ammonium salts such as triethyl salts. Examples include ammonium salts and tributylammonium salts.

Preparation of starting materials: Example A 2-cyano-εCAMP: 2.0 g of 2-bromo-εCAMP was suspended in 500 ml of dimethylformamide, and 1.0 g of potassium cyanide was added to the suspension.
After stirring thoroughly, the mixture was reacted at room temperature for 16 hours with stirring.

After the reaction is complete, add Dowex 50-
After neutralizing by adding X8 (H ⁺ type),
Evaporate the liquid to dryness. The residue was dissolved in a small amount of water, passed through a column (1.4φ x 30cm) packed with Dowex 50-X8 (H ⁺ type, 100-200 mesh), eluted with water, and then the target compound was extracted. When the eluted fraction containing the fraction is concentrated and left at 5°C, 2-cyano-εCAMP crystallizes, and when this is separated and dried, 1.11 g of crystals of 2-cyano-εCAMP are obtained.

Melting point: 191-193℃ (decomposition) Elemental analysis: C ₁₃ H ₁₁ N ₆ O ₆ P・0.5H ₂ O C H N Theoretical value (%) 40.37 3.03 21.66 Actual value (%) 40.31 3.10 21.71 Ultraviolet absorption spectrum : λ ^0.1N-HCl _nax 242.5, 249.0 (shoulder) 273.0 (shoulder), 28
1.0,
294.0, 333.5nm Example B 2-carbamoyl-εCAMP: Dissolve 1.0g of 2-cyano-εCAMP in 10ml of 0.7N aqueous sodium hydroxide solution (PH12.9),
Allow to react under stirring for a minute. Adjust the pH of the reaction mixture to 1.0 using concentrated hydrochloric acid, collect the precipitated crystals,
After washing with a small amount of water and drying, 793 mg of 2-carbamoyl-εCAMP crystals are obtained.

Melting point: 196-199℃ (decomposition) Elemental analysis: C ₁₃ H ₁₃ N ₆ O ₇ P・H ₂ O C H N Theoretical value (%) 37.65 3.34 20.12 Actual value (%) 37.68 3.62 20.29 Ultraviolet absorption spectrum: λ ^0.1N-HCl _nax 239.5, 278.0, 308.5nm Example C 2-carboxy-εCAMP: Dissolve 1.0g of 2-carbamoyl-εCAMP in 10.0ml of 1.3N-sodium hydroxide aqueous solution (PH13.2),
After reacting at room temperature for 20 minutes with stirring, the pH of the reaction mixture is adjusted to 1.0 using concentrated hydrochloric acid. The precipitated crystals were separated and dried to obtain 843 mg of 2-carboxy-εCAMP crystals.

Melting point: 200-202℃ (decomposition) Elemental analysis: C ₁₃ H ₁₂ N ₅ O ₈ P・1.5H ₂ O C H N Theoretical value (%) 36.57 3.47 16.61 Actual value (%) 36.79 3.54 16.51 Ultraviolet absorption spectrum : λ ^0.1N-HCl _nax 237.5, 276.5, 284.0 (shoulder), 306.0
(shoulder)
nm λ ^0.1N-NaOH _nax 237.5, 270.0 (shoulder), 322.0 nm By operating in the same manner as above but using 2-cyano-εCAMP instead of 2-carbamoyl-εCAMP, 2-carboxy-εCAMP is obtained.

Example 1 2-carbamoyl-CAMP: (a) 500mg of 2-carbamoyl-εCAMP in 0.1N-
Dissolve in 500 ml of hydrochloric acid, add 250 mg of N-bromosuccinimide, and react at room temperature for 5 minutes with stirring.

Then, the reaction mixture was passed through a column (1.6φ x 30cm)
After washing with water, elution is performed using ethanol/water/28% ammonium hydroxide (volume ratio 10:10:1), and the eluted fraction containing the target compound is concentrated. The concentrate obtained is passed through a column packed with Dowex 50-X8 (H ⁺ form, 100-200 mesh) and eluted with water. The elution fraction containing the target compound is concentrated and cooled to 5°C, resulting in 2-carbamoyl-
CAMP crystallizes and is separated and dried to obtain 10 mg of 2-carbamoyl-CAMP crystals.

Melting point: 213-216℃ (decomposition) Elemental analysis: C _{H N 6} O ₇ P・H ₂ O C H N Theoretical value (%) 33.85 3.85 21.54 Actual value (%) 33.82 3.65 21.50 Ultraviolet absorption spectrum: λ ^0.1N-HCl _nax 258.5 (shoulder) 265.0, 295.0nm λ ^0.1N-NaOH _nax 258.5 (shoulder), 265.0, 293.0nm (b) 2-carbamoyl-εCAMP500mg to 0.1N-
Dissolved in 500 ml of hydrochloric acid, added 250 mg of N-bromosuccinimide, reacted at room temperature for 5 minutes with stirring, then added 6N aqueous sodium hydroxide solution to adjust the pH of the reaction mixture to 12.0, and reacted for 1 hour. Stir.

Next, the pH of the mixture was adjusted to 2.0 using concentrated hydrochloric acid, adsorbed onto activated carbon packed in a column (1.6φ x 30cm), washed with water, and mixed with ethanol/water/28
% ammonium hydroxide (volume ratio 10:10:1) and concentrate the eluted fraction containing the target compound. Purification is performed in the same manner as in (a) below to obtain 140 mg of 2-carbamoyl-CAMP crystals.

This product has the same physicochemical properties as the compound obtained in (a).

(c) 500mg of 2-cyano-εCAMP in 500mg of 0.1N-hydrochloric acid
250 mg of N-bromosuccinimide dissolved in ml
After reacting at room temperature for 5 minutes with stirring, the pH of the reaction mixture was adjusted to 12.0 by adding 6N aqueous sodium hydroxide solution, and the mixture was stirred for 3 hours.

Next, the pH of the mixture was adjusted to 2.0 using concentrated hydrochloric acid, adsorbed onto activated carbon packed in a column (1.6φ x 30cm), washed with water, and mixed with ethanol/water/28
% ammonium hydroxide (volume ratio 10:10:1) and concentrate the eluted fraction containing the target compound. Purification is performed in the same manner as in (a) below to obtain 103 mg of 2-carbamoyl-CAMP crystals.

The physicochemical properties of the obtained compound (a) were consistent with the product.

Example 2 2-carboxy-CAMP: (a) 500 mg of 2-carboxy-εCAMP was dissolved in 500 ml of 0.1N hydrochloric acid, 250 mg of N-bromosuccinimide was added thereto, and the mixture was reacted with stirring at room temperature for 5 minutes. Afterwards, the pH of the reaction mixture was adjusted to 12.0 using 6N-sodium hydroxide aqueous solution.
Stir for 1 hour.

Next, the pH of the mixture was adjusted to 2.0 using concentrated hydrochloric acid, adsorbed onto activated carbon packed in a column (1.6φ x 30cm), washed with water, and mixed with ethanol/water/28
% ammonium hydroxide (volume ratio 10:10:1) and concentrate the eluted fraction containing the target compound. Dowex 50
Pass through a column packed with -X8 (H ⁺ form, 100-200 meshes) and elute with water. The eluted fraction containing the target compound is concentrated and cooled to 5°C to crystallize 2-carboxy-CAMP.
When the crystals are separated and dried, 108 mg of 2-carboxy-CAMP crystals are obtained.

Melting point: 190-193℃ (decomposed) Elemental analysis: C ₁₁ H ₁₂ N ₅ O ₈ P・1.5H ₂ O C H N Theoretical value (%) 33.00 3.75 17.50 Actual value (%) 33.02 3.75 17.50 Ultraviolet absorption spectrum : λ ^0.1N-HCl _nax 258.0 (shoulder), 264.5, 289.5 nm λ ^0.1N-NaOH _nax 258.0 (shoulder), 263.5, 290.0 (shoulder) nm (b) 1.3N-hydroxylation of 1.0 g of 2-carbamoyl-CAMP Dissolved in 10ml of sodium aqueous solution (PH13.2),
After reacting at room temperature for 20 minutes with stirring, the pH of the reaction mixture is adjusted to 1.0 using concentrated hydrochloric acid.
The precipitated crystals were separated and dried to obtain 450 mg of 2-carboxy-CAMP crystals.

This product has the same physicochemical properties as the compound obtained in (a).

Claims (1)

[Claims] 1. General formula 2-substituted-adenosine-3',5'-cyclic phosphoric acid represented by the formula (wherein R means a carbamoyl group or a carboxyl group). 2 General formula A halogenating agent is applied to 2-substituted-1,N ⁶ -etheno-adenosine-3',5'-cyclic phosphoric acid represented by (in the formula, R' means a cyano group, a carbamoyl group, or a carboxyl group) A general formula characterized by A method for producing 2-substituted adenosine-3',5'-cyclic phosphoric acid represented by the formula (wherein R means a carbamoyl group or a carboxyl group). 3. The method according to claim 2, wherein the reaction mixture is made alkaline after the action of the halogenating agent. 4 2-cyano-1,N ⁶ -etheno-adenosine-3',5'-cyclic phosphoric acid is treated with a halogenating agent and then subjected to alkaline hydrolysis,
The manufacturing method according to claim 2. 5. The method according to claim 4, wherein the alkaline hydrolysis is carried out at a pH of 10 or higher. 6. Characterized by alkaline hydrolysis of 2-carbamoyl-adenosine-3',5'-cyclic phosphoric acid,
Method for producing 2-carboxy-adenosine-3',5'-cyclic phosphoric acid. 7. The production method according to claim 6, wherein the alkaline hydrolysis is carried out at a pH in the range of 12 to 13.5.