A kind of efficient preparation method of heterocyclic drug intermediate

Abstract

The invention discloses a high-efficiency preparation method of a heterocyclic drug intermediate. The present invention couples alcohol dehydrogenase mutants and glucose dehydrogenases to catalyze heterocyclic substrates to generate heterocyclic drug intermediates. The alcohol dehydrogenase mutants have an amino acid sequence as shown in SEQ ID NO.2 Tyrosine at position 127 of the parent alcohol dehydrogenase shown is mutated to tryptophan. The mutant Y127W of the present invention can alleviate the product inhibition effect in a single aqueous phase system without adding any co-solvent, so that the conversion rate can reach more than 99% within 12 hours. Coupling the mutant Y127W with glucose dehydrogenase (BmGDH) achieved a 50 mL scale in a single aqueous phase system without adding any exogenous coenzymes and organic co-solvents, and the substrate concentration was as high as 600 g L ‑1 Gram-level preparation of , the catalyst loading was 3.3%. The e.e. value of the final product (S)-NBHP is as high as 99.4%, and the space-time yield is about 1400g L ‑1 d ‑1 , the product purity is 99.58%.

Description

technical field [0001] The invention relates to a high-efficiency preparation method of a heterocyclic drug intermediate, belonging to the technical field of biocatalysis. Background technique [0002] (S)-N-B OC -3-Hydroxypiperidine [(S)-NBHP] is a key chiral intermediate in the synthesis of the drug Ibrutinib (IBRUTINIB) for the treatment of lymphoma. The method of biosynthesis of (S)-NBHP is more green and environment-friendly, thus attracting more and more attention. However, previous studies have shown that the biosynthesis of (S)-NBHP requires the addition of organic co-solvents and expensive coenzymes, and the high concentration of substrates will lead to substrate or product inhibition, which will increase the cost of (S)-NBHP synthesis. [0003] (1) 2009A CHERETZ et al., used the method of biocatalytic synthesis for the first time, using the reductase in carrot cubes for catalysis. The catalyst was cheap and environmentally friendly, and provided a new idea for t...

AI Technical Summary

Problems solved by technology

[0013] (11) Patent CN201310173088.2 discloses a method of using recombinant ketoreductase (KRED) enzyme powder to asymmetrically reduce N-B OC -3-piperidone, but did not disclose the gene sequence or amino acid sequence of ketone reductase (KRED)

Patent CN201310054684.9 discloses an asymmetric synthesis of (S)-1-tert-butoxycarbonyl-3-hydroxypiperidine using alcohol dehydrogenase PAR, but the organic reagent isopropanol is used for the coenzyme cycle, and the organic reagent There is a greater degree of damage to the enzyme activity, and it has obvious inhibitory effect

Patent CN201610132936.9 discloses a method using carbonyl reductase R E The CR enzyme asymmetrically reduces ketoreductase (KRED), but this enzyme requires N I -NTA purification, and it uses 2-octanol-water two-phase reaction, which is not conducive to large-scale production or relatively high production cost

Pichia pastoris P reported in patent CN108220358A ICHIA SP.SIT2014 can be used as a biocatalyst for preparing (S)-NBHP, but too much catalyst addition increases the production cost

CN10822061A reports that ketoreductase MT-KRED is used for preparing (S)-NBHP, but needs to add expensive coenzyme in reaction process

[0014] Although the ketoreductase reported above can be used to prepare (S)-NBHP, the reaction process requires the addition of expensive coenzyme, a large amount of enzyme and organic solvents, which is not conducive to the scale-up application in the actual industry.

Images