JPS59152399A - Preparation of nucleic acid derivative - Google Patents

Abstract

PURPOSE:To prepare the titled compound useful as a starting substance for the synthesis of nucleic acid in genetic engineering, easily, by reacting a nucleoside with a bifunctional silicon compound in the presence of a base, reacting the product with an organic carboxylic acid halide, and hydrolyzing the product. CONSTITUTION:A nucleoside is made to react the a bifunctional silicon compound of formula (R<1>-R<4> are alkyl or phenyl; X is halogen; n is 0 or 1) in the presence of a base (preferably pyridine) to protect two hydroxyl groups at the deoxyribose segment of said nucleoside. The obtained reaction product is made to react with an organic carboxylic acid halide (e.g. benzyl chloride, anisoyl chloride, etc.) and hydrolyzed to obtain the objective N-acylated nucleoside.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a method for producing a nucleic acid derivative, more specifically,
A new compound is produced as an intermediate from a difunctional silicon compound and a nucleoside, and then reacted with an organic carboxylic acid halide and then hydrolyzed to easily produce N-
The present invention relates to a method for producing acylated nucleosides.

In recent years, nucleic acids and their derivatives have rapidly come into the spotlight with the development of genetic engineering, and have also attracted considerable attention in the pharmaceutical field, and research on them has been actively conducted.

By the way, the nucleoside that is the starting material for this nucleic acid derivative is a polyfunctional compound containing primary and secondary sugar hydroxyl groups, and an amine group in the base part. Conventionally, the sugar hydroxyl group and amino group in the nucleoside are treated with excess acid chloride or acid anhydride, and then only the sugar hydroxyl group is selectively hydrolyzed with an alkali [Journal. Of the American Chemical Society (
J. Am. Ohem, Soc, Vol. 85, p. 3821 (1963), etc.).

However, in this method, it is necessary to select the alkali treatment time for each compound.
Furthermore, since nucleic acids are easily degraded by acids, there are many operational problems, such as the need for careful neutralization, and methods for synthesizing only some derivatives have been established.

The present inventors have conducted intensive research to avoid such complicated operations and provide a method for easily producing the desired N-acylated nucleonds. It was discovered that the object could be achieved by temporarily protecting the compound using a silicon compound, reacting it with an organic carboxylic acid halide, and then hydrolyzing it. Based on this knowledge, the present invention was completed. Ta.

That is, the present invention provides for a nucleoside in the presence of a base,
A difunctional silicon compound represented by the general formula % formula % (1) (in the formula, R1, R2, R3 and R4 are an alkyl group or a phenyl group, X is a halogen atom, and n is 0 or 1) is reacted. The present invention provides a method for producing an N-arylated nucleondo, which comprises protecting two hydroxyl groups of the deoxyribose moiety of the nucleoside, reacting with an organic carboxylic acid halide, and then hydrolyzing the nucleoside. be.

The nucleoside used in the method of the present invention is a deoxynucleoside whose sugar moiety is deoxyribose, and the reaction between this nucleoside and the bifunctional silicon compound represented by the general formula (1) is carried out in the presence of a base, preferably anhydrous. It is carried out in the state of Although this reaction proceeds at room temperature, the reaction can be accelerated if necessary by heating. By this reaction,
As shown in the following formula, the bifunctional silicon compound acts on the two hydroxyl groups of the deoxyribose moiety of the nucleoside, 2 and an intramolecular cyclic silicon compound represented by the general formula (11) is formed. The cyclic silicon derivative of deoxynucleoside represented by the above general formula (11) is a new compound that has not been described in any literature.

The base used in this reaction is preferably a methyl group or a phenyl group as R in the solvent, and a chlorine atom as X, such as dimethyldichlorosilane, diphenyldichlorosilane, methylphenyldichloroh7rane, 1,3-dichlorotetramethyl di/loxane, 1,3-dichloro-
Examples include 1,3-dimethyl-1,3-diphenyldisiloxane.

By using these difunctional compounds, a ring is formed between the nucleoside and this is more stable than the product produced when a monofunctional compound such as trimethylchlorosilane is used. The formation of by-products can be reduced by effectively protecting sugar hydroxyl groups. Moreover, this cyclic body is easily hydrolyzed in pyridine water.

In the method of the present invention, the cyclic silicon compound of deoxy/nucleoside thus obtained is treated with an organic carboxylic acid...ride, and then water is added to decompose the cyclic silicon compound, and then impurities are removed. The target product is isolated by extraction and removal.

By using benzoyl chloride, anisoyl chloride, acetyl chloride, etc. as the organic carboxylic acid halide, corresponding N-acylated nucleosides such as N-pensoyl nucleoside, N-acylated nucleoside, and N-acetyl nucleoside can be obtained. It will be done.

As described above, according to the method of the present invention, N-acylated nucleosides can be obtained very easily, and these are useful as starting materials for nucleic acid synthesis in genetic engineering or as intermediates for pharmaceuticals.

Next, the present invention will be explained in more detail with reference to Examples.

Example 1 Anhydrous conditions F1 Deoxyazonone (], dissolved in 2 mmotwoviridi/2 me, the space of the reaction vessel was filled with nitrogen
, exchanged. Then, 1.1 times the mole of dimethyldichloro7rane was added using a syringe and allowed to react at 30°C for 30 minutes, and then 1.1 times mole of benzoyl chloride was added using a syringe and the mixture was heated at 30°C. After 2 hours of reaction, 2 liters of water was added and the mixture was decomposed for over 4 hours. When the obtained product was separated and quantified using high performance liquid chromatography, it was found that N-
Benzoyldeoxyadenosine was obtained with a yield of 60%.

Example 2 Example 1 except that 5 times the mole of benzoyl chloride was used instead of 1.1 times the mole of benzoyl chloride in Example 1.
When treated in the same manner as above, N,N-dibenzoyldeoxyadenosine was obtained in a yield of 72%.

Example 3 The treatment was carried out in exactly the same manner as in Example 1, except that 1,3-dichlorobutramethyldisiloxane was used in place of dimethyldichlorosilane as the siliconizing agent in Example 1. N-benzoyldeoxyadenosine is 63
% yield.

Example 4 2 mmot of deoxycytidine hydrochloride was suspended in 5 me of pyridine, and 1 mole of triethylamine was added to make 3.
Stir for 0 minutes. Then 1.02 times molar ty)+,
:+-Cyclorotetramethylzineroxanxane After being allowed to react for 30 minutes, 1.1 times the mole of anisoyl chloride
After treatment for several hours, add the same amount of water and hydrolyze overnight.7.
Then, after performing ether extraction twice, the aqueous layer] ml
! When the mixture was added with triethylamine and concentrated, N-anisoyldeoxycintidine was separated in a yield of 9:3%.

Note that anhydrous conditions were maintained in the same manner as in Example 1 until the treatment of anisoyl chloride was completed.

Example 5 5 ml of 72 mmot of deoxyazonone under anhydrous conditions
! of p-nitrobenzoyl chloride was suspended in pyridine for 7 hours, and 1.1 times the mole of 1,3-dichlorobutramethyldisiloxane was reacted with it for 30 minutes.
Allowed time to react. Next, the same amount of water was added and hydrolyzed overnight, followed by ether extraction, and 11ne triethylamine was added to the aqueous layer and concentrated.7), N-, p-
Nitrobenzoyldeoxy/adenosine was isolated in 64% yield.

The elemental analysis values of the obtained N-p-nitrobenzoyldeoxyadenosine are shown below.

Claims (1)

[Claims] 1. Presence of a base - in the nucleoside, the general formula % (in the formula, Hl, R2, R3 and H4f are an alkyl group or a phenyl group, and X is a halogen atom, n1dO or 1) A difunctional silicon compound represented by is reacted to protect the two hydroxyl groups of the deoxyribose moiety of the nucleoside, and then an organic carboxylic acid halide is reacted, followed by hydrolysis). Method for producing acylated nucleosides. 2. The method according to claim 1, wherein the base is pyridine.