Enzymatic preparation method of ketoreductase and S-1-BOC-3 hydroxypiperidine

Abstract

The invention discloses an enzymatic preparation method of S-1-BOC-3 hydroxypiperidine. The enzymatic preparation method comprises the following steps: by using 1-BOC-3 piperidone as a substrate, performing reacting in the presence of a coenzyme, a coenzyme circulation system and ketoreductase to obtain a reaction solution containing the S-1-BOC-3 hydroxypiperidine; The nucleotide sequence of a ketoreductase encoding gene is shown as SEQ ID NO: 1 in a sequence table, and the amino acid sequence of the ketoreductase is shown as SEQ ID NO: 2 in the sequence table. Compared with other preparationmethods of the S-1-BOC-3 hydroxypiperidine, the route for catalytic synthesis of the S-1-BOC-3 hydroxypiperidine by the ketoreductase provided by the invention is simple, tedious steps of a chemicalsynthesis method are avoided, and the problems of large coenzyme dosage, low unit enzyme activity and low tolerance concentration of a substrate 1-BOC-3 piperidone in the existing enzyme catalysis technology are solved. The substrate concentration can be increased to 400 g / L, the coenzyme dosage is 0.1 g / L, the 24-hour reaction conversion rate is as high as 99.9%, the optical purity e.e% is 99.9%,the production cost of the S-1-BOC3- hydroxypiperidine is reduced, and the method is economical and environmentally-friendly and has good industrial application prospects.

Description

technical field [0001] The invention belongs to the technical field of biocatalysis, in particular to an enzyme-catalyzed preparation method of ketone reductase and S-1-BOC-3 hydroxypiperidine. Background technique [0002] S-1-BOC-3 hydroxypiperidine is a very important intermediate for the synthetic drug ibrutinib. Ibrutinib (ibrutinib) is a new targeted anti-cancer drug jointly developed by Johnson and Pharmacyclics. It is used in the treatment kit Cellular lymphoma, its drug effect is very strong from the current situation. At present, the market demand for ibrutinib is growing strongly, from the sales of US$500 million just launched on the market to the current sales of more than US$5 billion, and the market prospect is very broad. [0003] At present, the synthesis of S-1-BOC-3 hydroxypiperidine mainly adopts chemical resolution or enzymatic synthesis: [0004] The notification number is CN105801518B "A Synthetic Method for (S)-1-Boc-3-Hydroxypiperidine". The invento...

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Problems solved by technology

[0004] The notification number is CN105801518B "A Synthetic Method for (S)-1-Boc-3-Hydroxypiperidine". The inventor uses R-glyceraldehyde acetone as the starting material, and through witting reaction, palladium carbon reduces the double bond. S-1-BOC-3 hydroxypiperidine is synthesized by catalytic hydrogenation and other steps. This synthetic method utilizes the chemical resolution method. Although compared with the conventional resolution method, the starting chiral material is changed and the cost is reduced, but This method has many steps and uses more reagents, which is a very uneconomical and environmentally friendly method.

[0005] In recent years, companies at home and abroad have developed a technology for the stereoselective reduction of 1-BOC-3-piperidone to (S)-1-BOC-3-hydroxypiperidine with ketoreductase, and the announcement number is CN105200089B "(S) -1-tert-butoxycarbonyl-3-hydroxypiperidine preparation method and its device” patent, using ketone reductase to catalyze 1-BOC-3 piperidone to generate S-1-BOC-3 hydroxypiperidine, and using isopropyl Alcohols are used as hydrogen donors. Compared with traditional chemical synthesis methods, this method is more environmentally friendly and has advantages in the synthesis of chiral products. However, the concentration of 1-BOC-3 piperidone in a single reaction in this scheme is 125g / L, the coenzyme concentration is high, and it is difficult to realize industrial application in the face of high raw material prices of 1-BOC-3 piperidone and coenzyme

[0006] The notification number is CN106520856B "(S)-N-tert-butoxycarbonyl-3-hydroxypiperidine enzymatic preparation method" patent, using ketoreductase catalytic technology to catalyze N-tert-butoxycarbonyl-3-piperidone Carry out reduction reaction to obtain (S)-N-tert-butoxycarbonyl-3-hydroxypiperidine, but the substrate concentration in the reaction process can reach up to 250g / L, although it can be applied industrially, but because the single reaction substrate concentration is low increase production cost

[0007] Therefore, although there are many studies on the catalytic synthesis of S-1-BOC-3 hydroxypiperidine using ketoreductase, most of them show that the concentration of the single-reaction catalytic substrate 1-BOC-3 piperidone is low, and the enzyme and The high amount of coenzyme increases the cost, and at the same time brings a lot of inconvenience to the post-treatment purification steps, making it difficult to realize industrial production

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