HU198069B - Process for producing 6-(phenylacetamido)- and 6-(phenoxyacetamido)-2,2-dimethylpenam-3-carboxylic acid-potassium salt from extracts of ferment juices - Google Patents

Abstract

Prepn. of potassium salts of 6-(phenyl-acetamido)- or 6-(phenoxy-acetamido)-, -2,2'di:methyl-pename-3-carboxylic-acids from acetic acid-ester extracts of fermentation brews by precipitating penicillinic acids with potassium-acetate or with a soln. of potassium acetate in a 1-4C aliphatic alcohol and/or an aq. son. of the alcohol. - The precipitation is achieved by adding acetic acid to the extract directly, or following a purification of the extract by washing it with a mixt. of 1-4C aliphatic alcohol and water separating the phases, and treating the solvent contg. phase with active charcoal.

Description

The present invention relates to the preparation of 6- (phenylacetamido) - and 6- (phenoxyacetamido) -2,2-dimethyl-penam-3-carboxylic acid potassium salt (hereinafter G-penicillin potassium salt and V-penicillin potassium salt). , or acetic acid ester extracts of non-harvested fermentation broths for preparation of pharmaceutical preparations.

There are several possibilities for the preparation of G-penlcillin potassium salt and V-penicillin potassium salt of the above purity.

The classic extraction penicillin production technology removes mycelium and other water-insoluble impurities from the fermentation broth by filtration. The resulting filtered fermentation broth at low temperature (5-10 ° C) with organic solvents, especially acetic acid esters (British Patent Nos. 760,351 and U.S. Patent 2,448,878), using appropriate extractors (2,509,010, 2,758,783). and U.S. Patent Nos. 2,758,784), while extracting penicillinic acid with dilute mineral acid.

Proteins from the filtered fermentation broth at the pH of the extraction form a stable emulsion, and suitable surfactants are also used to improve separation (U.S. Patent No. 886,527; U.S. Patent 2,563,779; and British Patent No. 598,597).

From the organic solvent extract thus obtained, the G- or V-penicillinic acid is isolated in the form of its potassium, sodium, calcium or ammonium salts (U.S. Patent Nos. 2,719,149, French Patent 1,270,010 and Hungarian Patent No. 173,447).

Due to their contamination, the penicillin salts produced in the above manner are expensive to process! can be used in the preparation of pharmaceutical compositions.

The low solubility of V-penicillinic acid in water allows direct separation of the filtered fermentation broth with dilute mineral acid (U.S. Patent No. 4,354,971). Thus, the extraction step can be omitted, however, due to the high degree of contamination of the recovered V-penicillinic acid, several reworking is required.

After purification from acetic acid ester extract of the filtered fermentation broth, a purer G-penicillin potassium salt or V-penicillin potassium salt can be obtained after purification with activated carbon according to the procedure described in the -, or phenoxyacetic acid, penicillin degradation products, proteins) potassium salts are more soluble in water than penicillin potassium salts. The presence of water-soluble or partially soluble organic solvents, such as aliphatic alcohols or ketones, facilitates the release of ballast materials into aqueous solution.

Addition of deionized water, an aliphatic alcohol or a saturated aqueous solution of ketone and potassium acetate to the clarified extract precipitates the potassium salt of penicillin, while the potassium salts of the above-mentioned ballast substances are dissolved in the aqueous phase.

The disadvantage of this process is that the high content of phenoxyl or phenylacetic acid in the fermentation broth requires the addition of large amounts of water, which increases product losses.

The process for producing pure V-penylcylcarbamate salt is described in Czechoslovak Patent Application No. 208,049, which is based on the discovery that the precursor phenoxyacetic acid is more soluble in certain organic solvents (e.g., aliphatic ethers, alcohols and ketones) than V-penicillinic acid. however, the potassium salts of these compounds differ only slightly in solubility. Therefore, the crude V-penicillyl potassium salt of the phenoxyacetic acid potassium salt from a fermentation broth containing a high yield precursor is washed with a mixture of aliphatic alcohols and / or ketones containing water adjusted to pH 1.0 with mineral acid. The disadvantage of this method is that it requires a large amount of organic solvent mixture, which can be regenerated only by complex distillation.

It is known that direct extraction of penicillin-containing fermentation broths without filtration can be achieved using a three-phase extraction decanter (German Patent No. 2,701,763). This eliminates the loss of filtration, but the extract thus obtained is significantly more impure than in the case of the filtered fermentation broth. The impurities are mainly derived from vegetable oils (sunflower, soya, palm oil) used for foaming during the fermentation of penicillin, which are hydrolysed enzymatically to free fatty acids depending on the metabolic pathway. While in the classical penicillin processing technology the free fatty acids are removed from the fermentation broth along with mycelium, by direct extraction these compounds remain in the fermentation broth and are dissolved in the organic solvent together with the penicillin during the extraction. Substances that are excreted from the mycelium during extraction further contaminate the extract.

For the above-mentioned reasons, if a potassium salt is obtained from the extract of the non-ferrous fermentation juice by known methods, a product contaminated with a poorly water-soluble precipitate is obtained, since the potassium salts of fatty acids are precipitated along with the penicillin potassium salt. The purification of such a product is costly, only by multiple extraction refinements or by washing with a large amount of organic solvent, since the potassium salts of oily consistency can be poorly dispersed in the washing mixture.

SUMMARY OF THE INVENTION It is an object of the present invention to provide an economical processing method for producing, in a single step, G or V penicillin potassium salt from extracts of filtered or non-filtered penicillin-containing fermentation broths in a purity and yield not known to those skilled in the art.

Surprisingly, our experiments have shown that when the penicillin salts are separated from the acetic acid ester extracts of the fermentation broths by adding a sufficient amount of concentrated exoic acid to the activated carbon extract before addition of potassium acetate, the purity of the excellent crude purity of the crude product produced, but the product is easy to filter and requires little solvent to wash. The purifying effect of acetic acid can be enhanced by washing the acetic acid ester extract with a water-aliphatic alcohol mixture prior to the operations of clarifying, adding acetic acid and separating the liver, the resulting extract is clarified with activated carbon and then the acetic acid is added. potassium acetate.

Thus, according to the invention, acetic acid is added directly or after washing of the extract with activated carbon, or after washing the extract with a mixture of water-aliphatic alcohol, separation of the phases and the activated carbon of the solvent phase, followed by potassium acetate or its alcoholic and / or aqueous solution of the corresponding penicillin potassium salt.

It has been found that in the presence of acetic acid, the rate of crystal nucleation is reduced, resulting in the formation of crystals of larger particle size. According to our measurements, the crystal size increase can be 5-20 times, depending on the acetic acid content. The larger crystals thus formed have a smaller specific surface area and therefore adsorb less impurities on their surface than smaller ones. However, slow crystal growth also prevents solvent inclusion.

However, increasing the amount of acetic acid not only affects the rate of crystalline nucleation but also affects the conversion rate of the penicillin potassium salt of penicillinic acid. In our experiments, 1.5 tons of acetic acid per volume of extract were found to be optimal in terms of yield and product quality.

The product of the present invention is easily and quickly filtered and requires little solvent washing. At the same time, the amount of contaminants in the product is minimized and the color of the product improves. The presence of acetic acid in the separation of the extracts from the non-harvested fermentation broths is particularly advantageous, since the dissolution of the ballast materials in the aqueous phase also shifts the fatty acid-fatty acid potassium neutral reaction in the opposite direction to the salt formation. In this way, the fatty acids remain in the solvent phase and do not contaminate the product.

According to the present invention, G- or V-penicillin-containing fermentation broth without or after filtration of the mycelium, in the presence of a suitable emulsion breaker, on an appropriate machine with an aliphatic acetic acid ester, e.g. butyl acetate, ethyl acetate, methyl isobutyl ketone and the like. extracted. There are two ways to separate the product from the extract:

the. The extract was clarified with 0.3% charcoal. The clarified extract is mixed with 0.1 to 5.0 wt% acetic acid, preferably 1.5 wt%, followed by 2 wt% C14 aliphatic alcohol, e.g. ethanol, 1-butanol, etc., and 5% v / v aqueous potassium acetate solution or 20 g / l solid potassium acetate solution in one or two portions.

b. The extract is first mixed with 1 to 4 aliphatic alcohols containing 10 to 30% (preferably 20% by volume) of water, preferably 20 to 80% by volume of water, such as ctanol, butanol, etc. After separation of the phases, with 0.3% v / v activated carbon based on the solvent phase.

1.5% acetic acid was added followed by 4% v / v aqueous potassium acetate solution containing 75% v / v solids or twice stoichiometric solid potassium acetate based on the total penicillin content of the extract. After 15-45 minutes, the precipitated product is filtered off through a suitable filtration apparatus. The filtered G- or V-penicillin potassium salt crystals are mixed in a blending device with 0.5 to 5 volumes of C14 alpha alcohol, e.g. ethanol, isobutanol, etc., or ketone e.g. acetone or acetone. the mixture is slurried and the crystals are filtered off. The suspension is repeated one or more times depending on the purity of the product and the product is dried.

The invention is illustrated by the following examples, without limiting our claims.

Example 1

To a 500 cm ³ extract obtained by direct extraction of G-penicillin-containing fermentation broth with isobutyl acetate with an HPLC content of 44,600 IU / cm ³ , 1.5 g of activated carbon was added and, after stirring for 5 minutes, the carbon was filtered off.

To the clarified extract was added 7.5 cm ^{3 of} concentrated acetic acid, followed by 35 cm ^{3 of a} precipitating mixture consisting of 2 volumes of ethanol and 5 volumes of 65% aqueous potassium acetate. After stirring for 30 minutes, the white product precipitated was filtered off with suction. 23.2 g wet product first

30.2 cm ³ and then 27.8 cm ^{3 of} anhydrous alcohols [washed in suspension and dried under vacuum at 45 ° C.

13.2 g of G-penicillin potassium salt are obtained.

total pnicnicin potassium salt content (iodometry): 99.8% G-penicillin potassium salt content (HPl.C): 86.3%

Color (Ph.HG.VI): Zj [a] ³ p = * 290 ”(c = 0.5, water)

Yield: 91.1% based on extract (penicillin) extract.

Example 2

A 5 liter extract was obtained by direct extraction of G-penicillin-containing fermentation broth with isobutyl acetate having a HPLC content of 39.250 IU / cm ³ with 1 liter of ethanol-water 1: 1 and the phases were separated. To the solvent phase is added 16.5 g of activated carbon and after 5 minutes of stirring the carbon is filtered off.

To the washed and clarified extract was added 75 cm ^{3 of} concentrated acetic acid and 200 cm ^{3 of} 75% potassium acetate in water. After stirring for 30 minutes, the precipitated white crystals were filtered off. THE

201.5 g of moist product is washed with 262 cm ³ of anhydrous ethanol and 242 ml of anhydrous ethanol the washing procedure was repeated. The product was dried in a vacuum oven at 45 ° C.

117.45 g of G-penicillin potassium salt are obtained.

total penicillin potassium salt content (iodometry): 99.6% G-penicillin potassium salt content (HPLC): 96.2%

Color (Ph.Hg.VL) Z, [a] p ² = + 289 ° (c = 0.5, water)

Yield: 92.0% based on the content of extract G-penicillin.

Example 3

A 1000 cm ³ extract was obtained by direct extraction of G-penicillin-containing fermentation broth with isobutyl acetate having a HPLC content of 49.450 IU / cm ³ of concentrated acetic acid from Example 1, followed by addition of 16.3 g of solid potassium acetate and stirring for 30 minutes. The precipitated product is filtered off with suction.

198 069

56.3 g of a wet product are obtained, which are first washed with 73.2 cm ³ of anhydrous alcohol and then 67 m ⁶ cm ^{3 of} anhydrous alcohol, dried under vacuum at 45 ° C.

29.8 g of solid potassium salt of G-penicillin were obtained, total penicillin potassium salt content (iodometry): 99.9% of penicillin potassium salt G (HPLC: 96.5%).

Color (Ph.Hg.VI.): Z, [α | ² θ = + 290 ° (c = 0.5, water)

Yield based on G-penicillin content of extract: 92.9%.

Example 4

Penicillin V obtained broth containing 500 cm ³ of isobutyl acetate extract direct extraction with HPLC drug content of 61900 IU / cm ^3, as described in Example 1, treated with charcoal, then 7.5 cm ³ cc. acetic acid and 35 cm <^{3> of} the precipitate of Example 1 were used to precipitate the V-penicillin potassium salt.

After filtration of the crystals, 38.2 g of a wet product are obtained, which are washed as above in Example 1 with 49.6 cm <^3> and then 45.8 cm <^{3> of} anhydrous ethanol. The washed product was dried under vacuum at 45 ° C.

19.3 g of potassium salt of V-penicillin are obtained.

total penicillin potassium salt content (iodometry): 99.1% V-penicillin potassium salt content (HPLC: 96.1% p-hydroxy-V-penicillin potassium salt content (HPLC) 0.9% phenoxyacetic acid potassium salt content (HPLC) 0.05% color (Ph.Hg.VI.) Z, [a] p ² = + 220 ° (c = 0.1, water)

Yield based on V-penicillin content of extract: 91.8%

As an advantage of our process, it can be mentioned that using pure extracts from penicillin-containing fermentation broths without extraction of the mycelium and by direct extraction, the pure G- or V-penicillin intermediate can be obtained in a high yield. washing with large quantities of organic solvents; recrystallization.

The purity of the G- or V-penicillin potassium salt produced is characterized by the fact that its active ingredient content is min. 99% as measured by HPLC min. 96%, color: Ph.Hg.VI. according to min. Zi. The content of para-hydroxy-V-penicillin is less than 1% and that of phenoxyacetic acid is less than 0.1% in the case of potassium V-penicillin.

The yield of the product is between 90-93% based on the content of solvent extract and 85-88% based on the content of the starting ferric compound.

Claims (3)

Claims A process for preparing potassium salts of 6- (phenylacetamido) and 6- (phenoxylacetamido) -2,2-dimethyl-penam-3-carboxylic acid. 20 obtained by the extraction of fermentation broths with acetic acid ester extraction and separation with activated charcoal or a C 1-4 aliphatic alcoholic and / or aqueous solution, characterized in that the extract is added to the extract directly or to the extract. After washing the aliphatic alcohol-water mixture with carbon atoms, separating the phases and clarifying the solvent phase with activated charcoal, acetic acid is added. Process according to claim 1, characterized in that the volume of the extract is added in an amount of 0.1-5.0% by volume, preferably 1.5% by volume, of acetic acid. Process according to claim 1 or 2, characterized in that the extract is present in an amount of 10-30% by volume, preferably 20% by volume, 20-80% by volume, preferably 50% by volume, of aliphatic C 1-4 aliphatic alcohols. alcohol-water-alcohol mixture 35 wash.