Carbonyl reductase and application thereof in preparation of Chiral N-protecting-hydroxy nitrogen heterocycle

Abstract

The invention relates to a carbonyl reductase and application of the carbonyl reductase in the catalysis of asymmetric reduction of N-tert butoxycarbonyl-carbonyl nitrogen heterocyclic compound to generate N-tert butoxycarbonyl-hydroxy nitrogen heterocyclic compound having high optical purity and corresponding (S)-configuration, wherein the carbonyl reductase is a carbonyl reductase mutant which has improved reduction activity of N-tert butoxycarbonyl-carbonyl piperidine constituted by a novel amino acid sequence formed by substituting a 90th, 92nd, 94th, 99th, 138th or 174th amino acid residue of the amino acid sequence shown in SEQ ID No. 2 with amino acid. Compared with the prior art, the engineering carbonyl reductase provided by the invention has advantages of high activity and broad substrate spectrum, and has excellent industrial application prospects in the preparation of (S)-N-Boc-hydroxy nitrogen heterocycle.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an engineered carbonyl reductase and its application in the preparation of chiral N-protected hydroxyl nitrogen heterocyclic rings and derivatives thereof. Background technique [0002] Many natural and non-natural drugs contain hydroxy nitrogen heterocyclic structure, optically pure hydroxypiperidine and hydroxypyrrolidine and their derivatives are important precursors for the synthesis of these drugs, and have broad application prospects. Among them, (S)-N-Boc-3-hydroxypiperidine (wherein Boc is the abbreviation of tert-butoxycarbonyl) is an important precursor for the synthesis of new anticancer drug BTK inhibitor Ibrutinib (Ibrutinib), using (S) -N-Boc-3-Hydroxypiperidine Synthetic technology of Ibrutinib is as follows: [0003] [0004] At present, there are three main preparation methods of chiral N-Boc-3-hydroxypiperidine, which are chemical resolution method, ...

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Problems solved by technology

[0005] In general, the activity of the reported N-Boc-3-carbonylpiperidine reductase catalysts is not high, and in the reported patent documents, the catalysts used are all free enzyme liquid or enzyme powder catalysts, and the catalyst preparation process has many steps , the cost is high, and it is difficult to apply to industrial production on a large scale; due to the use of enzyme preparations to catalyze the reaction, an additional expensive coenzyme must be added during the reaction process, resulting in a further increase in production costs

In addition, the substrate spectrum of the reported N-Boc-3-carbonylpiperidine reductase catalyst is relatively narrow, although the alcohol dehydrogenase PAR disclosed in the patent 201310054684.9 can also catalyze the reduction of some other carbonyl substrates, but no catalytic The reports of N-Boc-carbonylpiperidine and its structural analog N-Boc-carbonylpyrrole substituted by carbonyl groups at different positions limit the scope of application of this type of carbonyl reductase

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