Preparation method of (2S,3S)-2-hydroxy-4-phenylbutane derivative

Abstract

The invention discloses a preparation method of a (2S,3S)-2-hydroxy-4-phenylbutane derivative. The preparation method comprises the steps of enabling (3S)-3-(tert-Butoxycarbonyl)amino-1-chloro-4-phenyl-2-butanone, carbonyl reductase and excess isopropyl alcohol to be subjected to a biological enzyme catalytic reaction to obtain transformation liquid, and then performing continuous concentrating and crystallizing to obtain target product crystals. The carbonyl reductase has the characteristic of tolerance to high-concentration isopropyl alcohol through directional mutation, so as to realize thepurpose of solubilization of a substrate with the excessive isopropyl alcohol, the addition quantity of the substrate is further increased, the conversion rate is increased, the yield of products isnot lower than 95%, and the chirality purity is not lower than 99.95%. Through continuous concentrating and crystallizing on the transformation liquid, the crystalinity of the obtained products is 95%or above, the crystals are uniform, the purity of impurities is not less than 99.95%, and the content is not lower than 99%.

Description

technical field [0001] The invention relates to the field of biological enzyme catalysis technology, in particular to a preparation method of (2S,3S)-2-hydroxyl-4-phenylbutane derivatives. Background technique [0002] Non-peptide HIV protease inhibitors (PI) are the first choice for the treatment of HIV, such as saquinavir, nelfinavir, fosamprenavir, darunavir, etc. The process of releasing new and mature virus particles from the surface of infected host cells inhibits the protease of the virus. In June 2006, the FDA approved Darunavir in combination with other antiretroviral drugs for the treatment of HIV infection. For adult patients, darunavir was launched in 27 EU member states in March 2007. [0003] The core structures of the above compounds are all (2S,3S)-2-hydroxyl-4-phenylbutane derivative structures. For example, darunavir is mainly prepared through the compound of formula III (chemical name: (2S,3S)-N-tert-butoxycarbonyl-3-amino-1-chloro-2-hydroxy-4-phenylbuta...

AI Technical Summary

Problems solved by technology

[0008] Chinese patent CN102482648B discloses that selection comes from Novosphingobium aromaticivorans Carbonyl reductase gene construction engineering bacteria catalyze (3S)-3-(tert-butoxycarbonyl)amino-1-chloro-4-phenyl-2-butanone, the substrate concentration is 100g / L, the yield is greater than 95%, and the product The diastereomeric excess de is greater than 99.9%, but the patent only provides the technology to prepare the RS configuration, not the SS configuration

For the biological preparation of the SS configuration, Chinese patent CN104745649A discloses the use of (3S)-3-(tert-butoxycarbonyl)amino-1-chloro-4-phenyl-2-butanone as the substrate, and the substrate dosage concentration It is 50g / L, and the fluidity of the conversion system is promoted through the aid of co-solvents and surfactants, and the contact area between the substrate and the enzyme is increased. If no co-solvent is added, the fluidity of the conversion system will become poor, thereby affecting the conversion rate

However, in order to aid dissolution in this patent, additional substances such as toluene and Tween 60 added to the system are not conducive to the final separation and purification of the product, so the product purity is only 99%

Chinese patent CN104745649A also discloses a method for preparing biological enzymes in SS configuration, but the conversion system of this method uses glucose as the hydrogen supply system, the substrate concentration is 100g / L, and the reaction system is almost entirely in the water phase. Due to the substrate and SS configuration The products are hardly soluble in water. With the increase of the substrate concentration and no co-solvent, the fluidity of the system will gradually deteriorate, and the conversion rate will gradually decrease, which will limit the dosage of the substrate concentration, and the reaction will end in the system. Contains by-product gluconate, which brings difficulties to product separation and purification, and is difficult to be used in industrial production

[0009] To sum up, (2S,3S)-2 -Hydroxy-4-phenylbutane derivatives, the substrate concentration generally cannot exceed 100g / L due to solubility problems, or additional co-solvents are required to bring other impurities into the system, which will affect the separation and purification of SS configuration products and purity, the complex production process also increases the production cost

Images