SU717037A1 - Method of preparing n-acetyl-d-(-)-alpha-aminophenylacetic acid - Google Patents

Description

The invention relates to an improved state of the art method for the preparation of N-acetyl-O-C-) -c) -aminophenylacetic acid used for the preparation of drugs.

In the patent and technical literature, methods are known for producing H-acetyl-C- (-) - (- aminophenylacetic acid, for example, by selective crystallization of the ammonium salt of M-acetyl-P, L-ck-aminophenylacetic acid) 1. The method is difficult to carry out under conditions of large-scale production due to the crystallization of the racemate and the optical antipode and also does not ensure the optical purity of the final product sufficiently high for the pharmaceutical industry.

There is also known a method of separating D, L-CI-aminophenylacetic acid as a salt with D (-) - camphorsulfonic acid 2.

The disadvantage of this method is the need for chemical cleavage of the diastereoisomeric salt, the need to regenerate the compound used to produce the salt, and the difficulty of isolating the desired product. .

The closest in technical significance is the method according to which D, L-α-aminophenylacetic acid is separated by fermentation with a fungus from the group of the imperfect genus Fusarium nivale Fusarium, which metabolizes the L-isomer P, L-ch-amyiophenylacetic acid. According to this method, the specified microorganism is preliminarily cultured at a temperature of 24 hours in a medium containing,%: D, L-ok-aminrphenylacetic acid 0.1; glucose 5; ammonium nitrate 0.05; yeast extract 0.1; magnesium sulfate semi-sodium 0.05; potassium chloride 0.05. The prepared culture is inoculated into a basic medium containing,%: D, L-dL-aminophenylacetic acid 1,2; glucose 5; potassium phosphate disubstituted 0,1; magnesium sulfate semi-sodium 0.05; calcium chloride dihydrate 0.002; ferric trichloride 0.00l. Aeration at a temperature of 35 hours followed by treatment with amberlite 120-in the H-form gives a yield of 5.3 g of D (-) - rfs-aminophenylacetic acid from 1 liter of fermentation

Liquids that make up 44.2% of the amount taken to separate D, L-1 4-aminophenylacetic acid. Specific rotation of the product in 1.5 and. Salt of nois-acid 3.

This method has the following disadvantages.

j 1. Use for stereospecific enzymatic separation of D, L-d.-aminophenylacetic acid | Fusarlum Microorganism. nivale, which completely metabolizes the L-isomer, i.e. half of the raw material, as a result of which the yield of the target product is only 44.2% of that taken for the separation of D, L-d-acotnophenylacetic acid.

; . 2. Use FOR INSTALLATION. ; the target product of the ion-exchange mode, which is very difficult, expensive and time-consuming for large-scale production.

3. Multi-component and relative complexity of the fermentation substrate.

4. The final product is

D - (-). - " -aminophenylacetic acid, which for the production of K-acetyl-P - (-) - 5K-aminophenylacetic acid, is subject to acylation, which is associated with an additional loss of UI of the desired product. - X;

The aim of the invention is to eliminate the above mentioned points and increase the yield and increase the optical purity of the target product.

This goal is achieved by separating L-acetyl-B, L-C, -aminophenyl-acetic acid in the presence of a nitrogen-containing acidic compound or its salt, for example, sodium diethyldithiocarbamate, nitrilotriucetic acid, iminodiacetic acid or ethylene glycinate; The separation of the racemate is carried out, as a rule, by a complex of enzymes of water extract of pork kidneys, separated by L - (+) -.- ausphenylacetic acid and returned to the process by acylation, preferably XyuMy anhydride, with Teipo; at 85-100 ° C, and a target target. It was acidified by the reaction mass.

The nitrogen-containing acidic compound or its salt is preferably in an amount of 0.005-0.01 mol per 1 kg of base L-acetyl 0.1, -c-aminophenylacetic acid. :

Separation of N-acetyl-D, Lc, -aminophenylacetic acid in the form of a nitrogen-containing acidic compound or its salt deactivates the production of N-acetyl-B- {-) -aminophenylacetic acid of a high degree of optical purity, reduces the consumption of enzyme raw materials, shortens the time; stabilizes the process due to the binding of trivalent metal ions inhibiting complex

enzymes water extract pork kidneys.

The return of the isolated L- (+) - “-aminophenylacetic acid to the process by acylation at a temperature of 85 Doo C reduces losses and increases the yield to 93% of the theoretically possible, and eliminates the need for racemization of the L - (+) 1 isomer. At a temperature of 85-1CO C proteins associated. L - (+) - isomer, denatured and can easily be separated by filtration or center | have hyiovania.

isolation of the desired product by bleeding the reaction mixture to pH 1 below.

Example 1 . :.

1.1. To a solution of 1.6 kg (8.29 M) K-acetyl-O, L-ck-aminophenium acetic acid, 0.32 kg (8.0 M) sodium hydroxide, 2.9 g (0.13 M) of diethyldithiocarbamate trihydrate sodium in 13 liters of water: at a temperature of the reaction medium of 40-42 0 and pH 7, B-7.5, 0.2 l of aqueous dilution of blue pe pods prepared; stirring 60 g of rough, non-fatty kidneys with 0.25, liter of oil for 1 hour. Fermentation is carried out at; temperature 40 + 2C. . for .16 h / after which the mixture is cooled; to 10-15 ° C, filtered off, L - (+) -o1-aminophenyl acetic acid in the amount of 0.61 kg (4.0 M), and the filtrate is acidified with hydrochloric acid to a pH below 1. Viped N- acetyl-b - (-) - A-anethnophenylacetic acid is washed and dried at 50-60 ° C. Obtain 0.76 kg (3.94 M product,. Is 95% of theoretically possible. The specific rotation of the product ct. -185, which corresponds to 98% optical purity of 1} -acetyl-P - (-) - D -Amino-filament SU suny; ki slots.

Claims (3)

1.2. The isolated L - (+) - o (α-aminophenylacetic acid) vol. Amount of 0.51 kg (4.0 M), 160 g (4 M) of sodium hydroxide and 2.0 L of water is heated to and 510 g (5 , 6 M) of acetic anhydride at such a rate that the temperature of the reaction mass was maintained at 95100 C. At the temperature of the reaction mass is stirred for 1 h, cooled to 70-80 ° C, 10 g of activated carbon is added and filtered. 30-40 ° C, acidified with hydrochloric acid to a pH below 1 and cooled to 5-8 ° C. The precipitated M-acetyl-0, L-precipitate produced from h-aminophenylacetic acid filtered, washed and dried, followed by return to the process. The yield is 0.75 kg (3.88 M), which is 97% of the theoretically possible. The total yield of H-acetyl-O - (-) - n; -am | butphenylacetic acid, 89.4% of theoretically possible, Pr and m and p 2. 2.1. To a solution of 1.6 kg (8.29 M) C-acetyl-0, L-c. -Aminophenylacetic acid, 0.32 kg ( 8.0 M) sodium hydroxide, 1.5 g (0.008 M) nit nitrate of the ylotrilotse acid in 13 l of water at a reaction medium temperature of 4042 ° C and a pH of 7.3-7.5 add 0.2 l of aqueous extract of eubic acid} kidneys, prigovovlennoy mixing 60 g of roughly ground non-fat kidneys with 0 , 25 l of water for 1 h at a temperature of 2-5C. The fermentation is carried out at a tekteratura for 112 hours, after which the mixture is cooled with another lb-15 ° C, L - (+) aminophenylacetic acid is filtered in the amount of 0.62 kg (4.1 M), and the filtrate is acidified to T) acid to a pH of 1 and se 1. The precipitated N-acetyl-p- (-) - d-aminophenyl acetic acid is washed and dried at 50-60 s. 0.77 kg (4.03 M) of product is obtained, which is 96.3% of the theoretically possible. The specific rotation of the product is 3, 186, which corresponds to the 98.4% optical purity of N-auetil-L- (-) - d-aminophenylacetic acid. . 2.2. The isolated precipitate of L - (-) -A.-aminophenylacetic acid in the amount of 0.62 kg. (4.1 M), 1b4 g, (4., 1M sodium HYDROXIDE and 2.0 p of water. Load up to and adding 5-10 g (5.0 M) of acetic anhydride at such a rate that the temperature of the reaction mass is maintained at 95-100 ° C. At this temperature, the reaction mass is stirred for 1 h, cooled to 70-BOC, added. 10 g of activated carbon and filtered. The filtrate is cooled to 30-40 0, acidified with hydrochloric acid to pH below 1 and cooled to 5-8 ° C. The precipitated s-acetyl-O, 1 s -shnofeni y-cs acid filters washed, dried and then returned to the process, 0.7.7 K (3.99 M), which is 97% of the theoretically possible. The overall yield of N-acetyl-O - (-) - ck-aminophenylacetic acid 92, 8% of theoretically possible Example 3. 3.1. To a solution of 1.6 kg (8.29 M) N-acetyl-B, L-ck -aminophenylacetic acid 0.32 kg (8.0 M) sodium hydroxide, 2, 13 g (0.016 M) of iminodiuXUS acid in 13 l of water at a TiS temperature of the reaction medium 40-42 ° C and pH 7.3-7.5 were added 0.2 l of aqueous stretching of porcine, kidney, prepared by mixing 60 g of coarsely ground non-fatted kidneys with 0.25 L of water JB for 1 hours at a temperature of 2-5 ° C. Fermentation is carried out at a temperature of 14 hours, after which the mixture is cooled | up to 10-15. c, filter L- (+) - "-aminophenyl acetic acid in an amount of 0.61 kg (4.0 M): and the filtrate is acidified with hydrochloric acid to a pH below 1. The precipitated N-acetyl precipitate O - (-) - oC-aminophenylacetic acid is washed and dried at 50-60 ° C. 0.76 g (3.94 m) of product is obtained, amounting to em 95% of theoretically possible. The specific rotation of the product is ek -l86, which corresponds to 98.4% optical purity of N-ace | TIL-B- (-) -c-aminophenylacetic acid: Lots. 3.2. The isolated precipitate of L - (+) - L-aminophenylacetic acid in the amount of 0.61 kg (4.0 M), 160 g (4.0 M) of sodium hydroxide and 2.0 L of water is heated to and 510 g (5 , 0 M) acetic anhydride at such a rate that the temperature of the reaction mass was maintained 95-100s. In this temperature, the reaction mass is stirred for 1 hour, b cooled to 70-8., LO of activated carbon is added and filtered. The filtrate is cooled down to 30-40 ° C, acidified with hydrochloric acid to a pH below 1 and cooled to. The precipitated L-acetyl-0, L- "S-aminophenylacetic acid is filtered off, washed and dried, followed by return to the process. O yield, .75 kg (3.88 M), which is 97% of the theoretically possible. The total yield of N-acetyl-O - (-) - d-aminophenylacetic acid is 89.4% of the theoretically possible. Example 4. 4.1. To a solution of 1.6 kg (8.29 M) M-acetyl-P, L- (K-aminophenyl acetic acid, 0.32 kg (8.0 M). Sodium hydroxide, 3.0 g (0.01 M) ethylenediaminetetraacetic acid in 13 l of water at a temperature of the reaction medium of 40-42 ° C and a pH of 7.3-7.5, 0.2 l of aqueous extract of pig kidneys is added, prepared by stirring 60 g of coarsely ground non-degreased buds with 0.25 l of water for 1 hour at -Teure 2-5, C. Fermentation is carried out at a temperature of 40 + 2 ° C for 18 hours, after which the mixture is cooled to 10-15 seconds, filtered L- (+) - (. -aminophenylacetic acid in the amount of 0.62 kg (4.1 M), and the filtrate podki hydrochloric acid / until the pH is below 1. The precipitated N-aiietil-O- (-) - -aminophenylacetic acid precipitate is washed and dried at 50-60 C. The product obtained is 0.77 kg (4.03 M), which is em 96.3% of theoretically possible. The specific rotation of the product ek JJ -ISff, which corresponds to the 99% optical purity of N-acetyl-D- (-) - ois-aminophenylacetic acid 4. 2. The isolated precipitate of L- (+) - in α-aminophenyl acetic acid in coli-7 ./. 0.62 kg (4.1 M), 164 g (4.1 sodium hydroxide, and 2.0 l of water are heated until 510 g (5.0 M) are added at the same time as before). speed so that the temperature of the reaction mass is maintained at EB-HW C At this temperature, the reaction mixture is stirred for 1 hour, cooled to 70-80 seconds, 10 g of activated carbon is added and filtered. The filtrate is cooled to 30-40 ° C with hydrochloric acid to the pH is below 1 and cooled to 5-8 ° C. The precipitated H-acetyl-T precipitate, .-. L-aminophenylacetic acid is filtered off, washed and returned to the process. Approximately 0.77 kg {3.99 M} Than 97% of theoretically possible. The total yield of N-acetyl-E - (-) -aminophenylacetic acid is 92.8% bt of the theoretically possible. The invention claims 1. A method for producing N-acetyl-b- (-) - "k-aminophenylacetic acid by stereospecific enzymatic separation of the racemate, from - l and h: a y i and the fact that, with the aim of increasing yields and increasing optical CCST target product : i5 .., tri ,. .37 N-acetyl-O, L- (; -aminophenyl acetic acid) is separated in the presence of a nitrogen-containing acidic compound or its salt, released during the process L - (+) - dk -apdanofonyl acetic acid acylylite and borax at the beginning of the process. 2 Method POP.1, distinguishing and and, besides, the acid-desiccant acid compound is taken in an amount of 0.005-0.01 mol per 1 kg of initial H-acetyl-P ,.-aminophenyl succinic acid occ., 3. Method POP.1, which differs in that, as the nitrogen-containing acid compound, EXTERNAL sodium dis tithyldithiocarbamate, nitrilotriacetate acid, iminodiuacetic acid or ethylenediaminetetraacetic acid. 4. Method according to claim 1, characterized in that the acylation of L - (+) -. -amyophenylacetic acid is yrbid t at 85-100 C. The information points received by the expert during examination 1. U.S. Patent B3660477, CL 260-518, published 1972. 2. Postnikov, GS, et al., Chemicals., .., 1 & 3, 1, 39-44. 3. Japan patent number 7015433, .kl. 36 25, published. 1970 (prototype) ..