KR20020071637A - A purification method of pravastatin using methanol - Google Patents

Abstract

PURPOSE: A method for purifying pravastatin containing impurities using methanol is provided, by just controlling the concentration of methanol of the adsorption solution and the elution solution. CONSTITUTION: The method comprises the step of carrying out the adsorption chromatography of the solution containing pravastatin and other impurities with a methanol solution and a column charged with a HP20 resin or a HP20ss resin, wherein the concentration of methanol of the adsorption solution is maintained to be 10% or less during adsorption and the concentration of methanol of the elution solution is maintained to be 20% or more during elution.

Description

Purification of pravastatin using methanol {A purification method of pravastatin using methanol}

The present invention relates to a method for purifying pravastatin, and more particularly, purastatin is purified by adsorption chromatography using HP20 resin or HP20ss and methanol.

Pravastatin precursor compactin is disclosed in US Pat. Nos. 3,983,140 and 1,453,425 as substances capable of lowering cholesterol concentrations. It is a substance that is effective in heart disease such as myocardial infarction or arteriosclerosis caused by hyperlipidemia, especially hyperlipidemia by lowering cholesterol concentration.

Strains used in the preparation of such pravastatin precursors include Penicillium brevicompactin, Penicillmyces sp., Trichoderma longibraiatum, Tricorderma pseudo Trichoderma psudokoning, Hyphomyces chrisopomus, and Penicillium citrinum (Japanese Patent Publication No. 4-349034). Pravastatin precursors can be prepared by such strains or chemical transformations. In addition, Korean Patent No. 0186758 discloses a method of preparing pravastatin precursor compactin and its derivatives by aerobic culturing of Gliocladium sp. YJ-9515 (KCTC 02528BP).

Korean Patent No. 100212482 discloses a method for preparing pravastatin sodium using Streptomyces exfoliatus YJ-118, and No. 0186706 to Streptomyces roseochromogenes subsp. A method of preparing pravastatin sodium by adding and culturing pravastatin precursor to a culture medium of KTCC 12385 is described.

Pravastatin is produced by fermenting and converting microorganisms to pravastatin precursor compactin and its derivatives. At this time, in the fermentation broth sticking to fermentation, pravastatin precursor compactin and its derivatives remain, which makes it difficult to purify.

The present invention provides a method for purifying only pravastatin by column chromatography of pravastatin produced from pravastatin precursor using methanol and HP20 or HP20ss, and adjusting the methanol concentration at the time of adsorption and elution. In general, pravastatin is produced by fermenting a precursor with a strain, which is difficult to purify. According to the method of the present invention, such pravastatin precursor and pravastatin can be efficiently separated.

The present invention uses the adsorption chromatography using HP20 resin or HP20ss as a method for purifying pravastatin,

At the time of adsorption, the concentration of methanol in the adsorption solution

The present invention relates to a method for purifying pravastatin by removing impurities by eluting at a concentration of 20% or more of the methanol in the eluting solution during elution.

In the present invention, the production strain of pravastatin precursor is prepared using Streptomyces sp.

That is, pravastatin is prepared from the precursor compactin by adding a culture solution containing the pravastatin precursor compactin and derivatives thereof to a strain of the genus Streptomyces and co-culturing the same together. Thereafter, HP20 resin and HP20ss are developed in packed column chromatography to adsorb pravastatin in this fermentation product to the resin. In this case, a solution containing methanol at a concentration of 20% or less is used as the adsorption solution. Pravastatin is obtained by eluting the adsorbed pravastatin with an eluent containing at least 20% methanol.

As the fermentation filtrate and the mixed solution used, 1 g / l of pravastatin, 0.2 g / l of ML236B and its derivatives were used in the adsorption liquid.

Example 1 Effect of Methanol (IPA) on Adsorption of Adsorbents on HP20 Resin

HP20 resin capacity: 200ml, adsorption liquid volume: 1000ml (pravastatin 1000mg / l, Compectin and its derivatives 200mg / l), the chromaticity was measured by OD660nm, the color of the adsorption stock solution was converted to 100.

Methanol concentration in total sediment (%) 0 10 20 30 40 50 Content in Permeate (mg) Pravastatin (mg) 561 723 803 843 870 891 yield(%) 56 72 80 84 87 89 Compectin and its derivatives (mg) 0 0 25 130 168 181 Chromaticity (%) 22 27 35 45 56 54

result:

The adsorption filtrate which adsorbed the alcohol content below 10% contained only pravastatin and the yield was 56-72%. The pravastatin yield was 80% in the passage of the adsorbent containing 20% alcohol. The adsorbent containing 30% or more of alcohol contained the same ratio as that of the original adsorbed by pravastatin, compactin and its derivatives.

From this result, it can be seen that it is possible to improve the purification yield while removing impurities by adsorbing the alcohol concentration in the adsorption liquid at 20% or less.

Example 2 Influence of Methanol Concentration in Eluent When Elution Resin

1000 ml of alcohol-free adsorbent was filled in 200 ml of HP20 resin and adsorbed, and eluted by passing 1000 ml of eluent with different methanol concentration in each eluent.

Concentration of Methanol in Eluent (%) 0 10 20 30 40 50 Eluent content after elution (mg) Pravastatin (mg) 265 298 405 410 411 413 Compectin and its derivatives (mg) 0 0 40 145 146 167 Chromaticity (%) 11 21 32 35 35 36

In case of methanol content of eluent less than 10%, only pravastatin eluted. In 20%, only pravastatin 405 mg and compactin and its derivatives were eluted; Derivative content increased.

Therefore, it is possible to improve the purification yield while removing impurities by eluting with a methanol concentration of 20% or less in the eluent.

When purvastatin is purified using the purification method of the present invention, pravastatin is purified by simply passing a fermentation broth containing pravastatin and other impurities through a column packed with HP20 resin, and adjusting only the methanol concentration of the adsorption solution and the elution solution without further processing. It can be obtained in high yield.

Claims (2)

In a method for purifying pravastatin by adsorption chromatography using methanol and HP20 resin, A method of adsorbing with a concentration of methanol of not more than 10% at the time of adsorption and eluting with a concentration of methanol of at least 20% at the time of elution. In a method for purifying pravastatin by adsorption chromatography using methanol and HP20ss resin, A method of adsorbing with a concentration of methanol of not more than 10% at the time of adsorption and eluting with a concentration of methanol of at least 20% at the time of elution.