JPH0764845B2 - Purification method of dideoxyadenosine - Google Patents

Description

TECHNICAL FIELD The present invention relates to a novel purification method of 2 ′, 3′-dideoxyadenosine (hereinafter abbreviated as DDA) produced by a chemical synthesis method.

[Conventional technology]

As a method for synthesizing DDA, RKRobins using a 2'-deoxyadenosine (hereinafter abbreviated as 2'-DA) derivative as a raw material
Et al. (Biochemistry, 5 , 224 (1966)), JGMoffat et al.'S method (J. Am. Chem, Soc, 95 , 40
25 (1973)), and the method of MJ Robins et al. Using adenosine (hereinafter abbreviated as Ado) as a raw material (Tetrahedron Letters,
25 , 367 (1984)).

However, in these DDA reaction solutions, for example, JGMoffat
In these methods, 3'-DA is contained as an by-product impurity.

Similarly, according to the method of MJ Moffat et al.
In the production method of reacting with -acetoxyisobutyryl bromide and then reducing with palladium-carbon, 2'-DA and 3'-DA are by-produced in addition to DDA. 2'-DA, 3'-DA and DD
A is only the difference in the presence or absence of OH groups structurally, and mutual separation of these is possible at the laboratory level by TLC (thin layer chromatography), liquid chromatography, etc. There is a big problem in producing profitable products.

Under acidic conditions, DDA easily cleaves the N-glycoside bond to produce deoxyribose and adenine (hereinafter abbreviated as Ade). Therefore, in the DDA synthesis reaction solution or DDA
Ade is easily generated during recrystallization of. Since Ade has low solubility, it is difficult to separate it from DDA once it is produced.

As described above, the DDA reaction solution obtained by the chemical synthesis method contains 2'-DA, 3'-DA, adenosine, Ade, etc. derived from raw materials or by-products, depending on the synthetic raw material. These mutual separations have not yet been established on an industrial level.

[Problems to be solved by the invention]

It is desired to develop an industrially excellent purification method for DDA derived from a synthetic method that solves the above-mentioned drawbacks.

[Means for solving problems]

The present inventors have conducted extensive studies to solve the above-mentioned problems, and as a result, by treating a DDA-containing solution with a non-polar porous adsorbent resin, DDA of 2'-DA, 3'-DA, adenosine Ade, etc. It was found that high-purity DDA can be separated from impurities.

That is, the present invention relates to DDA produced by a chemical synthesis method.
A method for purifying DDA, which comprises adsorbing DDA to a non-polar porous resin in purifying the DDA.

The starting material of the present invention may be obtained by a chemical synthesis method or may be unpurified DDA derived therefrom, and the degree of purity does not matter. Therefore, DDA synthesis reaction solution and D obtained by recrystallization were used.
It may be DA coarse x'al.

An operation method for feeding the DDA-containing solution described in the present invention to the nonpolar porous resin will be described.

The DDA-containing solution may contain impurities such as 2'-DA, 3'-DA adenosine and Ade, and the concentration of DDA is not limited as long as it is below the solubility.

Next, the non-polar porous resin used here, for example, the matrix is a styrene-divinylbenzene-based copolymer or,
Any derivative thereof, for example, a substance which is a polymer obtained by halogenating the derivative and increasing the specific gravity thereof can be used. For example, Diaion HP series, SP series (above, Mitsubishi Kasei), XAD-4 (Rohm and Haas), OC10
31 (Bayer Co., Ltd.) and the like can be used, but any other nonpolar porous resin may be used as long as it has equivalent properties. In particular, SP207 (Mitsubishi Kasei Co., Ltd.), which has a high specific gravity, is suitable because the resin does not float when the DDA-containing solution is fed and the operability is good.

The liquid contacting method of the non-polar porous resin and the DDA-containing solution includes a batch method and a column method, but the column method is preferable because of its simple operation.

There is no particular restriction on the flow rate through the column, and usually SV = 0.5
˜4.0, preferably about SV = 1˜2.

The volume loading of the DDA-containing solution fed to the column depends on the concentration of the DDA-containing solution, and at the same time, the resin loading of DDA (g / lR) is 5-40 g / lR, preferably 10-30 g / l-
R is suitable in terms of separability and economy.

The liquid contact temperature to the column is not particularly limited as long as it is 10 to 60 ° C. At this temperature DDA and impurities 2'in solution
There is almost no difference in separability from -DA, 3'-DAAdo, Ade.

Next, the method for eluting DDA from the column will be described. As the liquid separating agent, a lower aliphatic alcohol aqueous solution is suitable. For example, it is an aqueous solution of methyl alcohol, ethyl alcohol, isopropyl alcohol, or the like. The elution rate is
SV = 1-2 is good.

The actual refining operation using a non-polar porous resin may be as follows. That is, the column packed with the resin is used for DDA.
After feeding a fixed amount of the contained solution, it is pressed with water to elute hydrophilic impurities. Then, using an aqueous alcohol solution, 2 '
-DA, 3'-DA, Ado and Ade are eluted, and DDA is eluted by further increasing the alcohol concentration. Then, this DDA fraction was concentrated, crystallized, and then cooled to obtain highly pure DDA.
Can be collected.

〔Example〕

 Hereinafter, the present invention will be described in detail with reference to examples.

Example 1 Using Ado as a raw material, JGMoffat (J. Am. Chem. Soc., 95 , 4025
(1973)) and concentrated the DDA reaction solution, and DDA4.4g / dl, 3′-DA3.2g / dl, Ado0.1g / dl, Ade, 0.1g /
10 ml of dl solution was prepared. This solution is a non-polar porous resin
SP207 (manufactured by Mitsubishi Kasei) 50 ml (column φ × L = 20 mm × 1
After feeding SV = 2 to 60mm), press water (2RV),
Next, it was eluted with 900 ml of 5% isopropyl alcohol (designated as fraction-1). Finally 20% isopropyl alcohol 20
Elution with 0 ml (designated fraction-2).

By liquid chromatography analysis of each fraction, fraction-1
Contains 3'-DA, Ade, Ado (each recovery rate, 95
%, 98%), Fraction-2 contains 3'-DA (recovery rate 5%) and D
DA (recovery rate 98%) was included. Fraction-2 was concentrated,
After precipitating DDA, 350 mg of high-purity DDA crystal was obtained (yield 80%).

Example 2 A DDA reaction solution prepared in the same manner as in Example 1 was concentrated to give DDA (2.2 g).
/ dl 3'-DA1.1g / dl, 2'-DA1.0g / dl, Ado0.1g / dlAde, 0.
100 ml of a 1 g / dl solution was obtained.

This solution is a non-polar porous resin SP207 (manufactured by Mitsubishi Kasei) 2
After feeding SV = 2 to 00 ml (column φ × L = 30 mm × 300 mm), water pushing (2 RV) was performed, and then elution was performed with 500 ml of 10% ethyl alcohol (fraction-1). Finally, it was eluted with 1200 ml of 20% ethyl alcohol (designated as fraction-2).

Fraction-1 was 2'by liquid chromatography analysis
-DA, 3'-DAAdo, Ade are included (recovery rate: 98%, 9
4%, 99%), Fraction-2 has 3'-DA (recovery rate 3%), DDA
(Recovery rate 95%) was included. Fraction-2 was concentrated and DD
After depositing A, 1700 mg of high-purity DDA crystal was obtained (yield 77%).

〔The invention's effect〕

As described above, the present invention can efficiently separate and purify DDA by treatment with a non-polar resin, and is expected to have a great potential for industrialization.

Claims (3)

[Claims] 1. A 2 ', 3'-produced by a chemical synthesis method.
A method for purifying 2 ', 3'-dideoxyadenosine, which comprises adsorbing 2', 3'-dideoxyadenosine on a non-polar porous resin when purifying dideoxyadenosine. 2. The method according to claim 1, wherein the substance to be purified contains at least one of 2'-deoxyadenosine, 3'-deoxyadenosine, adenosine and adenine as impurities. 3. The method according to claim 1, wherein the non-polar porous resin contains a styrene-divinylbenzene copolymer or a derivative thereof.