Enzymatic synthesis method of eliglustat intermediate

Abstract

The invention relates to the technical field of biological pharmacy, in particular to an enzyme catalytic synthesis method of an eliglustat intermediate, which comprises the following steps: by taking a compound II as a substrate, carrying out biological catalytic reaction in the presence of carbonyl reductase, coenzyme, an enzyme circulating hydrogen donor, coenzyme circulating enzyme, a cosolvent and a buffer solution to generate the eliglustat intermediate, namely a compound I; and the specific synthesis route is as follows: the synthesis method disclosed by the invention is simple and convenient to operate and environment-friendly, and efficient synthesis of the chiral intermediate is realized.

Description

technical field

[0001] The invention relates to the technical field of biopharmaceuticals, in particular to an enzyme-catalyzed synthesis method of an eliglukast intermediate. Background technique

[0002] Eliglustat was approved by the FDA in 2014. It is a first-line drug for the long-term treatment of type 1 Gaucher disease. The original research was done by the University of Michigan, and it was later licensed to Genzyme, a subsidiary of Sanofi. The trade name is Cerdelga. Gaucher disease occurs in people who do not produce enough of an enzyme called glucocerebrosidase, a deficiency that causes fatty material to collect in the spleen, liver and bone marrow. Key signs include enlarged liver and spleen, low red blood cell count (anemia), low platelet count, and bone problems. Cerdelga is a gelatin capsule, the oral formulation targets glucosylceramide synthase (glucosylceramide synthase, GCS), which can reduce the production of glucosylceramide. Type 1 Gaucher disease is...

Examples

Embodiment 1

[0021] Embodiment 1 Preparation of Eliglukast intermediate

[0022] In a 250ml Erlenmeyer flask, add 60ml of PB buffer (0.2mM, pH7.5), dissolve 800mg of commercially available carbonyl reductase A (commercial enzyme, product number P1B02, source Codexis company), 200mg of isopropanol dehydrogenase successively Enzyme powder (commercial enzyme, product number WT2302, purchasing company: Suqian Alpha Technology Co., Ltd.), 10 mg NADP+. 1 g of the substrate compound II was dissolved in 40 ml of isopropanol (as a coenzyme cycle hydrogen donor and co-solvent), added to the reactor, stirred at 220 rpm, and reacted at 30° C. for 16 h to obtain compound I. The reaction result was detected by HPLC, the conversion rate was 93%, the purity was 98.68%, and the ee value was 99.9%.

[0023]

Embodiment 2

[0024] Embodiment 2 Preparation of Eliglukast intermediate

[0025] In a 250ml Erlenmeyer flask, add 60ml of PB buffer (0.2mM, pH7.5), dissolve 800mg of commercially available carbonyl reductase B (commercial enzyme, product number P1B10, source Codexis company), 200mg of isopropanol dehydrogenase successively Enzyme powder (commercial enzyme, product number WT2302, purchasing company: Suqian Alpha Technology Co., Ltd.), 10 mg NADP+. 1 g of the substrate compound II was dissolved in 40 ml of isopropanol (as a coenzyme cycle hydrogen donor and co-solvent), added to the reactor, stirred at 220 rpm, and reacted at 30° C. for 16 h to obtain compound I. The reaction result was detected by HPLC, the conversion rate was 91%, the purity was 96.77%, and the ee value was 95.5%.

[0026]

Embodiment 3

[0027] Embodiment 3 Preparation of Eliglukast intermediate

[0028] In a 250ml Erlenmeyer flask, add 60ml of PB buffer (0.2mM, pH7.5), dissolve 800mg of commercially available carbonyl reductase C (commercial enzyme, product number KRED130, source Codexis company), 200mg of isopropanol dehydrogenase successively Enzyme powder (commercial enzyme, product number WT2302, purchasing company: Suqian Alpha Technology Co., Ltd.), 10 mg NADP+. 1 g of the substrate compound II was dissolved in 40 ml of isopropanol (as a coenzyme cycle hydrogen donor and co-solvent), added to the reactor, stirred at 220 rpm, and reacted at 30° C. for 16 h to obtain compound I. The reaction result was detected by HPLC, the conversion rate was 89%, the purity was 95.68%, and the ee value was 91.5%.

[0029]

technical field

[0001] The invention relates to the technical field of biopharmaceuticals, in particular to an enzyme-catalyzed synthesis method of an eliglukast intermediate. Background technique

[0002] Eliglustat was approved by the FDA in 2014. It is a first-line drug for the long-term treatment of type 1 Gaucher disease. The original research was done by the University of Michigan, and it was later licensed to Genzyme, a subsidiary of Sanofi. The trade name is Cerdelga. Gaucher disease occurs in people who do not produce enough of an enzyme called glucocerebrosidase, a deficiency that causes fatty material to collect in the spleen, liver and bone marrow. Key signs include enlarged liver and spleen, low red blood cell count (anemia), low platelet count, and bone problems. Cerdelga is a gelatin capsule, the oral formulation targets glucosylceramide synthase (glucosylceramide synthase, GCS), which can reduce the production of glucosylceramide. Type 1 Gaucher disease is...

Examples

Embodiment 1

[0021] Embodiment 1 Preparation of Eliglukast intermediate

[0022] In a 250ml Erlenmeyer flask, add 60ml of PB buffer (0.2mM, pH7.5), dissolve 800mg of commercially available carbonyl reductase A (commercial enzyme, product number P1B02, source Codexis company), 200mg of isopropanol dehydrogenase successively Enzyme powder (commercial enzyme, product number WT2302, purchasing company: Suqian Alpha Technology Co., Ltd.), 10 mg NADP+. 1 g of the substrate compound II was dissolved in 40 ml of isopropanol (as a coenzyme cycle hydrogen donor and co-solvent), added to the reactor, stirred at 220 rpm, and reacted at 30° C. for 16 h to obtain compound I. The reaction result was detected by HPLC, the conversion rate was 93%, the purity was 98.68%, and the ee value was 99.9%.

[0023]

Embodiment 2

[0024] Embodiment 2 Preparation of Eliglukast intermediate

[0025] In a 250ml Erlenmeyer flask, add 60ml of PB buffer (0.2mM, pH7.5), dissolve 800mg of commercially available carbonyl reductase B (commercial enzyme, product number P1B10, source Codexis company), 200mg of isopropanol dehydrogenase successively Enzyme powder (commercial enzyme, product number WT2302, purchasing company: Suqian Alpha Technology Co., Ltd.), 10 mg NADP+. 1 g of the substrate compound II was dissolved in 40 ml of isopropanol (as a coenzyme cycle hydrogen donor and co-solvent), added to the reactor, stirred at 220 rpm, and reacted at 30° C. for 16 h to obtain compound I. The reaction result was detected by HPLC, the conversion rate was 91%, the purity was 96.77%, and the ee value was 95.5%.

[0026]

Embodiment 3

[0027] Embodiment 3 Preparation of Eliglukast intermediate

[0028] In a 250ml Erlenmeyer flask, add 60ml of PB buffer (0.2mM, pH7.5), dissolve 800mg of commercially available carbonyl reductase C (commercial enzyme, product number KRED130, source Codexis company), 200mg of isopropanol dehydrogenase successively Enzyme powder (commercial enzyme, product number WT2302, purchasing company: Suqian Alpha Technology Co., Ltd.), 10 mg NADP+. 1 g of the substrate compound II was dissolved in 40 ml of isopropanol (as a coenzyme cycle hydrogen donor and co-solvent), added to the reactor, stirred at 220 rpm, and reacted at 30° C. for 16 h to obtain compound I. The reaction result was detected by HPLC, the conversion rate was 89%, the purity was 95.68%, and the ee value was 91.5%.

[0029]

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