JPH064656B2 - Method for producing phosphoenolpyruvate monosodium salt monohydrate - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to phosphoenolpyruvate (hereinafter referred to as “PE”).
Abbreviated as “P”), more specifically, a method for producing a monosodium salt monohydrate, which is capable of obtaining the above-mentioned compound of high purity in a high yield in a shorter time. Concerning the method.

[Prior Art] PEP is a high-energy phosphate compound, and is an important biochemical and enzymatic chemistry compound that regenerates adenosine triphosphate (ATP). Recently, it has been used in enzyme-catalyzed organic synthesis reactions [eg, Journal of American Chemical Society (J. Amer. Chem.S.
oc.,) 107, 7008 and 7019, 1985.
See the year], and other uses as blood preservatives have also been found.

Chemically synthesizing PEP has been performed for a long time, and particularly, a method using halopyruvic acid and trialkyl phosphite has been carried out. However, the PEP produced by such a method is its esterified product, and is a non-esterified PE that is conventionally present in nature.
No method has been found that allows the synthesis of P to be carried out on an industrial scale.

As a method for producing a non-esterified PEP proposed up to the present, for example, (1) a method using a pyruvic acid and a phosphoric acid derivative protected by a trimethylsilyl group (JP-A-56-113, (2) Dialkylphosphoenolpyruvate is hydrolyzed for a long time (15 hours), and then potassium hydroxide is used to obtain a monopotassium salt (JP-A-59-50).
No. 0,515); (3) Method for hydrogenolysis of monobenzylphosphoenolpyruvate [see Chemist Ber. Vol. 92, p. 952, 1959]; (4) Dialkyl phospho PEP cyclohexylamine salt obtained by reacting enolpyruvic acid with cyclohexylamine for a long time (3 days) was made into PEP by using an ion exchange resin, and then PE was obtained by using alkali hydroxide.
Method for obtaining an aqueous solution of an alkali metal salt of P [Biochemical Preparation (Biochem.Prep.) Vol. 11, 101
Page, 1966].

However, each of the manufacturing methods (1) to (4) above is
Each has the following problems.

The method (1) is not industrially advantageous because it is necessary to protect pyruvic acid and the like with a trimethylsilyl group, resulting in a very high production cost and complicated operations. Also, the PEP obtained is the trisodium salt and this method cannot produce the monosodium salt. Although the method (2) takes too long a production time and finally a high-purity monopotassium salt is obtained, the yield of the monopotassium salt is 50% with respect to the weight of pyruvic acid used as a starting material. It is very low and not industrially advantageous. The method (3) is not suitable because the yield is low and a large amount of by-products hydrogenated to the double bond portion of PEP are produced. The method (4) is not suitable for practical use because it takes an extremely long time to obtain a PEP cyclohexylamine salt from a dialkylphosphoenolpyruvate.

As described above, in order to carry out any of the methods on an industrial scale, it takes too long a manufacturing time, the manufacturing method is complicated, and further, both the purity and the yield of the obtained PEP salt and the like are required. It is not enough because there is no satisfactory method.

[Problems to be Solved by the Invention] In the conventional method for producing a PEP salt, the present invention takes a too long production time and cannot obtain a PEP salt with high purity and high yield. It is an object of the present invention to solve the problems and provide a production method capable of obtaining highly pure PEP monosodium salt monohydrate in a high yield in a short time.

[Means for Solving Problems] In the method for producing a PEP monosodium salt monohydrate according to the present invention, is the dialkylphosphoenolpyruvate hydrolyzed in the presence or absence of a strongly acidic ion exchange resin? Or a step of hydrolyzing dialkylphosphoenolpyruvate and then treating the hydrolyzate with a strongly acidic ion exchange resin; a step of adjusting the pH of the hydrolyzate to 1.8 to 2.5 with sodium hydroxide; a step of adding a water-affinitive organic solvent to the hydrolysis solution of the pH adjusting solution to precipitate crystals;
And a step of separating the precipitated crystals.

Hereinafter, the present invention will be described in detail.

Dialkylphosphoenolpyruvate (hereinafter, referred to as "DAPE" as a starting material for PEP monosodium salt monohydrate.
(Abbreviated as “P”) is represented by the following general formula.

(In the formula, R represents an alkyl group having 1 to 4 carbon atoms.) Such DAPEP is, for example, chemistry review, 6
Volume 1, p. 607, 1962, obtained by reacting halopyruvic acid with a trialkylphosphite represented by the following formula: P (OR) ₃ (wherein R has the same meaning as above). be able to.

Using such DAPEP as a starting material, PEP monosodium salt monohydrate is produced.

First, DAPEP is hydrolyzed. The amount of water used in this case is preferably 1.5 to 10 times the amount of DAPEP. The reaction temperature is 5 to 50 ° C, preferably 20
The reaction time varies depending on the reaction temperature, but is usually 5 to 10 hours. After completion of this hydrolysis,
Although it is possible to immediately shift to the next treatment step, in order to obtain a high-purity PEP monosodium salt monohydrate with high yield, hydrolysis is carried out in the presence of a strongly acidic ion exchange resin in this treatment step. Alternatively, treatment with the resin can be performed after hydrolysis. This ion exchange resin is
The particle size and the like are not particularly limited, and commercially available products such as Dowex (manufactured by Dow Chemical Co.), Amberlite (manufactured by Rohm and Haas Co.), and Diaion (manufactured by Mitsubishi Kasei) can be used. The resin is used as an H-type ion exchange resin when used. The amount of such resin used is 1/5 to 10 times the amount of DAPEP.

The method of hydrolysis in the presence of such a resin is not particularly limited, and examples thereof include a method of stirring DAPEP, water and a strongly acidic ion exchange resin in a suitable container at room temperature for a certain period (batch method). The method of treating the hydrolyzed liquid with a strongly acidic ion exchange resin is not particularly limited, either, the same batch method as described above, or a method of passing the hydrolyzed liquid through a column filled with the resin (column type). ) Can be mentioned.

Then, after the treatment step is completed, the pH of the hydrolyzed solution is 1.8 to 2.5, preferably 1.8 to 2.2, and more preferably 1.8 to 2.0 using an aqueous sodium hydroxide solution. Adjust to. If the pH value in this case is too high or too low, both the purity and yield of PEP monosodium salt monohydrate cannot be satisfied at the same time. Moreover, as the sodium hydroxide aqueous solution used in this step, one having a concentration of 0.1 to 5 N can be usually used.

After adjusting the pH of the hydrolyzed solution in this manner, crystals of PEP monosodium salt monohydrate can be precipitated by adding a water-affinitive organic solvent.

Examples of the water-affinitive organic solvent used in this step include acetone, ethanol, isopropanol, acetonitrile and dioxane, which can be dissolved in water at a desired ratio. The amount of the organic solvent used is appropriately 2 to 20 times the amount of the hydrolysis liquid of the pH adjusting liquid. It should be noted that in this case, if the amount of the hydrolyzed liquid is too large, that is, if the ratio of the dissolved PEP monosodium salt monohydrate to the amount of the liquid is small, it is said that crystal precipitation is difficult. In some cases, it is necessary to previously concentrate the above-mentioned hydrolyzed liquid to about 1 to 5 times the amount of DAPEP which is the starting material.

The crystals thus precipitated can be separated by a method such as filtration or centrifugation.

EXAMPLES OF THE INVENTION The present invention will be described in more detail with reference to examples.
The analysis of the purity was carried out by NMR spectrum (solvent: heavy water, internal standard substance: hydroquinone) and high performance fluid chromatograph (column: TSK gel QAE-2SW, carrier liquid: 0.15M KCl aqueous solution, detector: UV2).
20 mm, internal standard substance: maleic acid).
Both measurement results agree within the error range. Further, the sodium was analyzed by an atomic absorption method, and the yield was determined by the yield (%) of the produced PEP monosodium salt monohydrate with respect to bromopyruvic acid as a starting material.

Synthetic Example Synthesis of DAPEP A solution of 8.4 g of brompyruvic acid in 20 ml of absolute ethanol was added dropwise to a solution of 6.8 g of trimethylphosphite in 40 ml of anhydrous ether at a rate such that ether slowly refluxed while stirring. After completion of dropping, the reaction solution was further reacted for 1 hour, and then the reaction solution was concentrated under reduced pressure to obtain 10.5 g of a viscous liquid. This viscous liquid was confirmed by NMR (proton, phosphorus 31) analysis to contain 90% or more of dimethylphosphoenolpyruvate.

Example 1 5.0 g of dimethylphosphoenolpyruvate prepared above was dissolved in 40 ml of water, 10 g of strongly acidic ion exchange resin Dowex 50H type was added, and the mixture was stirred at room temperature for 5 hours. A 3N aqueous NaOH solution was carefully added to the liquid from which the ion exchange resin had been removed until the pH reached 2.0. The liquor was then concentrated to about 15 ml. Then, 50 ml of ethanol was added to the concentrated solution while stirring to precipitate crystals.
Then, the precipitated crystals were separated and dried under reduced pressure at room temperature to obtain 4.1 g of white crystals (yield: 82% based on bromopyruvic acid).

The obtained crystal was confirmed by NMR spectrum, IR spectrum and elemental analysis to be PEP monosodium salt monohydrate represented by the following formula.

The decomposition point of this monosodium salt monohydrate was 120 ° C., and as a result of analysis by high performance liquid chromatography, the purity was 98.8%.

Example 2 A PEP monosodium salt monohydrate was obtained in exactly the same manner as in Example 1 except that the ion exchange resin was not used.
The yield was 3.6 g (yield 72%), and the purity was 97.5.
%Met.

Examples 3 to 5 PEP monosodium salt monohydrate was obtained in the same manner as in Example 1 except that the addition amount of the resin was changed as shown in Table 1.

Example 6 Amberlite IR-22 was used as an ion exchange resin.
g PEP monosodium salt monohydrate was obtained in the same manner as in Example 1 except that g was used. The yield was 3.7 g (74
%) And the purity was 98.5%.

Examples 7 to 9 PEP monosodium salt monohydrate was obtained in the same manner as in Example 1 except that the pH of the hydrolyzed solution was changed as shown in Table 2.

Examples 10 to 12 PEP monosodium salt monohydrate was obtained in the same manner as in Example 2 except that the pH of the hydrolyzed solution was set as shown in Table 3.

Comparative Example P according to the method described in JP-A-59-50015
EP monosodium salt monohydrate was prepared.

First, 5.0 g of dimethylphosphoenolpyruvate was dissolved in 10 ml of water and hydrolyzed at room temperature with stirring for 15 hours. Then, a 3N aqueous sodium hydroxide solution was added to the sewage decomposition solution to adjust the pH of the decomposition solution to 2.8. Thereafter, 50 ml of ethanol (in the example of the publication, ethanol was added in an amount 2.7 times as much as that of dimethylphosphoenolpyruvate, but with this addition, the amount of crystals deposited was very small, and the subsequent separation operation, etc. Since it was difficult, the amount of ethanol was increased) was added with stirring to precipitate a colorless solid. The solid was confirmed by NMR and the like to be PEP monosodium salt monohydrate. However, the yield was 3.4 g (68% yield),
The purity was 79.8%.

[Effects of the Invention] As described above, according to the production method of the present invention, not only the production time can be significantly shortened as compared with the conventional method, but also a higher-purity PEP monosodium salt monohydrate can be obtained. It can be obtained in a higher yield (see Examples and Comparative Examples above).

Such a production method of the present invention can be carried out on an industrial scale, and is extremely effective as a production method of PEP monosodium salt monohydrate useful as a blood preservative.

Claims (2)

[Claims] 1. A dialkylphosphoenolpyruvate is hydrolyzed in the presence or absence of a strongly acidic ion exchange resin, or after dialkylphosphoenolpyruvate is hydrolyzed, the hydrolyzed solution is subjected to a strongly acidic ion exchange. Step of treating with resin; pH of the hydrolyzed solution is 1.8 to 2 with sodium hydroxide.
5, a step of adding a water-affinitive organic solvent to the hydrolyzed solution after pH adjustment to precipitate crystals; and a step of separating the precipitated crystals; Method for producing monosodium salt monohydrate. 2. The pH of the hydrolyzed solution is 1.
The manufacturing method according to claim 1, wherein the manufacturing method is adjusted to 8 to 2.0.