Resolution method of beta-amino acid

Abstract

The beta-amino acid resolving process includes the first N acylation of amino acid; hydrolyzing the acyl radical of the N acylated beta-amino acid in some one configuration in high selectivity with properly selected enzyme while not affecting its antimer and extracting according to the difference of different matters in water solubility and fat solubility to obtain one kind of corresponding optically pure beta-amino acid; and hydrolyzing the acyl radical of the N acylated beta-amino acid in one other configuration with one other kind of selected enzyme to obtain one other kind of optically pure beta-amino acid similarly. The said process is stable, environment friendly, safe, low in cost and with mild production conditions, and the product reach optical purity of 99.9 wt%. The present invention provides one new way for producing optically pure beta-amino acid.

Description

technical field

[0001] The invention relates to a method for splitting beta-amino acids, specifically a method for splitting and preparing single-configuration beta-amino acids by using two acylases (acyltransferases), and belongs to the field of organic synthesis. Background technique

[0002] Optically active β-amino acids exist in natural substances such as alkaloids and antibiotics, free optically active unnatural amino acids and their derivatives exhibit interesting pharmacological effects, and are also used in the synthesis of modified peptides, which serve as With the increasing importance of important intermediates for the preparation of pharmaceuticals, their isolation has attracted increasing interest. Optically active β-amino acids are important intermediates of many drugs that have been marketed or are about to be marketed abroad. Such as: antibacterial drug Deoxynegamycin (2-[2-[3(R), 6-Diaminohexanoyl]-1-methylhydrazino]acetic acid) developed by Vicuron Pharma...

Examples

Embodiment 1

[0063] Embodiment one (the resolution of comparative example 1 3-phenyl-3-amino-propionic acid ethyl ester)

[0064] a, Synthesis of the substrate: 10g of 3-phenyl-3-amino-propionic acid (purchased) was put into a 100mL reaction bottle, 50mL of ethanol was added, 5g of thionyl chloride was added dropwise at 0°C, and the reaction was carried out at room temperature for about 2-8 hours. complete. The solvent was evaporated to dryness to obtain 13 g of hydrochloride of ethyl 3-phenyl-3-amino-propionate, with a yield of 95%;

[0065] b, split reaction and detection:

[0066] 10mmol (mmol) racemic compound 3-phenyl-3-amino-propionic acid ethyl ester hydrochloride was dissolved in 50mL water, and the pH was adjusted to 7.5-8.5. 10 mg Amano lipase PS was added at a reaction temperature of 30° C., and after a reaction time of 3 hours, part of the reaction liquid was taken out for processing. Add 50 mL of ethyl acetate to extract (R) ethyl 3-phenyl-3-amino-propionate, evaporate to d...

Embodiment 2

[0067] Embodiment two (the resolution of comparative example 2 3-phenyl-3-phenylacetamido-propionic acid)

[0068] a. Synthesis of racemic substrate: 10g of 3-phenyl-3-amino-propionic acid (purchased) was put into a 100mL reaction bottle, 50mL of water was added, 12.5g of potassium carbonate was added, and 14.5g of phenylacetyl chloride was added dropwise at -20°C , low temperature (below 0°C) for 2 hours. The pH was adjusted to 1, and 16 g of white solid was precipitated, with a yield of 93%.

[0069] b. Split reaction and detection:

[0070] 10 mmol of the racemate compound 3-phenyl-3-phenylacetamido-propionic acid was dissolved in 50 mL of NaOH solution (1 mol / L), and the pH was adjusted to 7.5-8.5. 10 mg of penicillin G acylase was added at a reaction temperature of 30° C., and after a reaction time of 30 minutes, part of the reaction solution was taken to separate the product. Use 1M HCl to adjust the pH to 1, extract unreacted S-3-phenyl-3-phenylacetamido-propionic ac...

Embodiment 3

[0073] Embodiment three splits 3-phenyl-3-acetamido-propionic acid with two kinds of enzymes (small test)

[0074] a. Synthesis of racemate substrate: 3-phenyl-3-amino-propionic acid can be purchased conveniently from the market. Dissolve 10 mmol of the racemic compound 3-phenyl-3-amino-propionic acid in 10 mL of NaOH solution (2 mol / L). 12 mmoles of acetic anhydride was added dropwise at 0°C, and the reaction was complete after 5 minutes. Adjust the pH to 1, extract the product with MTBE (methyl tert-butyl ether), and evaporate to dryness on a rotary evaporator to obtain 3-phenyl-3-acetamido-propionic acid.

[0075] b. β-Amino acylase resolution reaction and detection

[0076] 10 mmol of the racemic compound 3-phenyl-3-acetylamino-propionic acid was dissolved in 50 ml of NaOH solution, and the pH was adjusted to 7.0-8.0. Add 10 mg of β-Amino acylase at a reaction temperature of 40°C, and after 30 minutes of reaction time, adjust the pH to 1 with 1M HCl, and extract unreact...

technical field

[0001] The invention relates to a method for splitting beta-amino acids, specifically a method for splitting and preparing single-configuration beta-amino acids by using two acylases (acyltransferases), and belongs to the field of organic synthesis. Background technique

[0002] Optically active β-amino acids exist in natural substances such as alkaloids and antibiotics, free optically active unnatural amino acids and their derivatives exhibit interesting pharmacological effects, and are also used in the synthesis of modified peptides, which serve as With the increasing importance of important intermediates for the preparation of pharmaceuticals, their isolation has attracted increasing interest. Optically active β-amino acids are important intermediates of many drugs that have been marketed or are about to be marketed abroad. Such as: antibacterial drug Deoxynegamycin (2-[2-[3(R), 6-Diaminohexanoyl]-1-methylhydrazino]acetic acid) developed by Vicuron Pharma...

Examples

Embodiment 1

[0063] Embodiment one (the resolution of comparative example 1 3-phenyl-3-amino-propionic acid ethyl ester)

[0064] a, Synthesis of the substrate: 10g of 3-phenyl-3-amino-propionic acid (purchased) was put into a 100mL reaction bottle, 50mL of ethanol was added, 5g of thionyl chloride was added dropwise at 0°C, and the reaction was carried out at room temperature for about 2-8 hours. complete. The solvent was evaporated to dryness to obtain 13 g of hydrochloride of ethyl 3-phenyl-3-amino-propionate, with a yield of 95%;

[0065] b, split reaction and detection:

[0066] 10mmol (mmol) racemic compound 3-phenyl-3-amino-propionic acid ethyl ester hydrochloride was dissolved in 50mL water, and the pH was adjusted to 7.5-8.5. 10 mg Amano lipase PS was added at a reaction temperature of 30° C., and after a reaction time of 3 hours, part of the reaction liquid was taken out for processing. Add 50 mL of ethyl acetate to extract (R) ethyl 3-phenyl-3-amino-propionate, evaporate to d...

Embodiment 2

[0067] Embodiment two (the resolution of comparative example 2 3-phenyl-3-phenylacetamido-propionic acid)

[0068] a. Synthesis of racemic substrate: 10g of 3-phenyl-3-amino-propionic acid (purchased) was put into a 100mL reaction bottle, 50mL of water was added, 12.5g of potassium carbonate was added, and 14.5g of phenylacetyl chloride was added dropwise at -20°C , low temperature (below 0°C) for 2 hours. The pH was adjusted to 1, and 16 g of white solid was precipitated, with a yield of 93%.

[0069] b. Split reaction and detection:

[0070] 10 mmol of the racemate compound 3-phenyl-3-phenylacetamido-propionic acid was dissolved in 50 mL of NaOH solution (1 mol / L), and the pH was adjusted to 7.5-8.5. 10 mg of penicillin G acylase was added at a reaction temperature of 30° C., and after a reaction time of 30 minutes, part of the reaction solution was taken to separate the product. Use 1M HCl to adjust the pH to 1, extract unreacted S-3-phenyl-3-phenylacetamido-propionic ac...

Embodiment 3

[0073] Embodiment three splits 3-phenyl-3-acetamido-propionic acid with two kinds of enzymes (small test)

[0074] a. Synthesis of racemate substrate: 3-phenyl-3-amino-propionic acid can be purchased conveniently from the market. Dissolve 10 mmol of the racemic compound 3-phenyl-3-amino-propionic acid in 10 mL of NaOH solution (2 mol / L). 12 mmoles of acetic anhydride was added dropwise at 0°C, and the reaction was complete after 5 minutes. Adjust the pH to 1, extract the product with MTBE (methyl tert-butyl ether), and evaporate to dryness on a rotary evaporator to obtain 3-phenyl-3-acetamido-propionic acid.

[0075] b. β-Amino acylase resolution reaction and detection

[0076] 10 mmol of the racemic compound 3-phenyl-3-acetylamino-propionic acid was dissolved in 50 ml of NaOH solution, and the pH was adjusted to 7.0-8.0. Add 10 mg of β-Amino acylase at a reaction temperature of 40°C, and after 30 minutes of reaction time, adjust the pH to 1 with 1M HCl, and extract unreact...

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