143900-44-1Relevant academic research and scientific papers
Efficient bioreductive production of (S)-N-Boc-3-hydroxypiperidine using ketoreductase ChKRED03
Xu, Guang-Peng,Wang, Hai-Bo,Wu, Zhong-Liu
, p. 881 - 885 (2016)
Ibrutinib is an anticancer drug targeting B-cell malignancies. The key chiral intermediate for ibrutinib synthesis is the alcohol (S)-N-Boc-3-hydroxypiperidine ((S)-NBHP), which can be produced via ketoreductase (KRED)-catalyzed bioreduction. After screening a small inventory of 27 KREDs mined from the genome of Chryseobacterium sp. CA49, ChKRED03 was selected as the best performer, leading to the complete conversion of 100 g substrate/L within 10 h to yield (S)-NBHP with high enantiomeric excess (> 99% ee). The enzyme was NADPH dependent, and the highest enzymatic activity was observed at 30 °C in potassium phosphate buffer (pH 7.0). At a substrate/catalyst ratio of 66.7 (w/w), ChKRED03 catalyzed the complete conversion of 200 g/L substrate within 3 h to yield (S)-NBHP with >99% ee, demonstrating great potential for industrial application.
Characterization of a carbonyl reductase from Rhodococcus erythropolis WZ010 and its variant Y54F for asymmetric synthesis of (S)-N-Boc-3-hydroxypiperidine
Ying, Xiangxian,Zhang, Jie,Wang, Can,Huang, Meijuan,Ji, Yuting,Cheng, Feng,Yu, Meilan,Wang, Zhao,Ying, Meirong
, (2018)
The recombinant carbonyl reductase from Rhodococcus erythropolis WZ010 (ReCR) demonstrated strict (S)-stereoselectivity and catalyzed the irreversible reduction of N-Boc-3-piperidone (NBPO) to (S)-N-Boc-3-hydroxypiperidine [(S)-NBHP], a key chiral intermediate in the synthesis of ibrutinib. The NAD(H)-specific enzyme was active within broad ranges of pH and temperature and had remarkable activity in the presence of higher concentration of organic solvents. The amino acid residue at position 54 was critical for the activity and the substitution of Tyr54 to Phe significantly enhanced the catalytic efficiency of ReCR. The kcat/Km values of ReCR Y54F for NBPO, (R/S)-2-octanol, and 2-propanol were 49.17 s?1 mM?1, 56.56 s?1 mM?1, and 20.69 s?1 mM?1, respectively. In addition, the (S)-NBHP yield was as high as 95.92% when whole cells of E. coli overexpressing ReCR variant Y54F catalyzed the asymmetric reduction of 1.5 M NBPO for 12 h in the aqueous/(R/S)-2-octanol biphasic system, demonstrating the great potential of ReCR variant Y54F for practical applications.
Efficient synthesis of Ibrutinib chiral intermediate in high space-time yield by recombinant E. coli co-expressing alcohol dehydrogenase and glucose dehydrogenase
Chen, Yitong,Ma, Baodi,Cao, Songshuang,Wu, Xiaomei,Xu, Yi
, p. 2325 - 2331 (2019)
The production of (S)-N-boc-3-hydroxy piperidine (NBHP) via asymmetric bioreduction of 1-boc-3-piperidinone with reductase is impeded by the need for expensive coenzymes NAD(P)H. In order to regenerate the coenzyme in situ, the gene of alcohol dehydrogenase from Thermoanaerobacter brockii and glucose dehydrogenase from Bacillus subtilis were ligated into the multiple cloning sites of pRSFDuet-1 plasmid to construct the recombinant Escherichia BL21 (DE3) that co-expressing alcohol dehydrogenase and glucose dehydrogenase. Different culture conditions including the medium composition, inducer and pH etc were systematically investigated to improve the enzyme production. The enzyme activity was increased more than 11-fold under optimal culture condition, from 12.7 to 139.8 U L?1. In the further work, the asymmetric reduction of 1-boc-3-piperidinone by whole cells of recombinant E. coli was systematic optimized to increase the substrate concentration and reaction efficiency. At last, S-NBHP (>99% ee) was prepared at 500 mM substrate concentration without external addition of cofactors. The conversion of S-NBHP reached 96.2% within merely 3 h, corresponding a high space-time yield around 774 g L?1 d?1. All these results demonstrated the potential of recombinant E. coli BL21 (DE3) coupled expressing alcohol dehydrogenase and glucose dehydrogenase for efficient synthesis of S-NBHP.
Engineering an Alcohol Dehydrogenase from Kluyveromyces polyspora for Efficient Synthesis of Ibrutinib Intermediate
Wu, Yanfei,Zhou, Jieyu,Ni, Jie,Zhu, Cheng,Sun, Zewen,Xu, Guochao,Ni, Ye
, (2021/02/26)
(S)-N-Boc-3-hydroxypiperidine [(S)-NBHP] is a key intermediate for the synthesis of mantle cell lymphoma drug, ibrutinib. Here, KpADH, an alcohol dehydrogenase from Kluyveromyces polyspora, exhibits evolutionary potential in the asymmetric reduction of N-Boc-3-piperidone (NBPO) to (S)-NBHP. By screening key residues in substrate binding pocket of KpADH, an excellent variant Y127W was obtained with 6-fold improved activity of 119.3 U mg?1, 1.8-fold enhanced half-life of 147 h and strict S-stereoselectivity (>99% ee). When catalyzed by Y127W, a complete conversion of 600 g L?1 NBPO was achieved at a substrate to catalyst ratio (S/C) of 30 in 10 h. Based on crystal-structure of Y127W, molecular docking and dynamic simulations reveal hydrogen bonds formed between W127 and Boc group of NBPO, as well as improved structural stability mainly contribute to the increased catalytic activity and stereoselectivity of Y127W. This study offers guidance for engineering ADHs for biosynthesis of chiral heterocyclic alcohols, and provides insights into mechanisms in catalytic activity and stereoselectivity toward carbonyl-containing heterocyclic substrates. (Figure presented.).
Preparation method of (S)-1-tert-butyloxycarbonyl-3-hydroxypiperidine
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, (2020/03/25)
The invention discloses a preparation method of (S)-1-tert-butyloxycarbonyl-3-hydroxypiperidine, and relates to the technical field of biological pharmacy. The preparation method comprises the following steps: 1, preparing 3-hydroxypyridine (2) as a raw material; 2, preparing racemic 3-hydroxypiperidine (3) from 3-hydroxypyridine (2); 3, preparing a derivative (2S, 3S)-N-(4-chlorphenyl)-2, 3-dihydroxy succinamic acid (4) from racemic 3-hydroxypiperidine (3) and D-tartaric acid; 4, carrying out resolution reaction on the derivative (2S, 3S)-N-(4-chlorphenyl)-2, 3-dihydroxy succinamic acid (4) prepared from D-tartaric acid to obtain a (S)-hydroxypiperidine salt (5); and 5, subjecting an obtained mixture to extraction, concentration and crystallization, to prepare a (S)-1-tert-butyloxycarbonyl-3-hydroxypiperidine salt (5). The preparation process has the advantages that the synthesis steps are reduced, the yield is increased, an adopted chiral resolving agent can be recycled, and the preparation process is suitable for industrial production.
Preparation method of N-tert-butyloxycarbonyl-3-piperidone and derivative thereof
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Paragraph 0063; 0073-0078, (2020/07/21)
The invention discloses a preparation method of N-tert-butyloxycarbonyl-3-piperidone, and the method comprises the following step: oxidizing N-tert-butyloxycarbonyl-3-hydroxypiperidine by virtue of aNaDCC-TEMPO system so as to generate N-tert-butyloxycarbonyl-3-piperidone. The invention also discloses a method for preparing (S)-N-tert-butyloxycarbonyl-3-hydroxypiperidine through an oxidation reaction and a reduction reaction by virtue of the NaDCC-TEMPO system. The method has the advantages of simple operation steps, high conversion rate and high product purity.
PROCESSES AND INTERMEDIATES FOR PREPARING A BTK INHIBITOR
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Page/Page column 15; 16, (2018/04/21)
Disclosed is a process for the preparation of certain intermediates, e.g. the following compound: (I) which intermediate and processes are useful in the preparation of a BTK inhibitor, such as ibrutinib.
Methodology Development in Directed Evolution: Exploring Options when Applying Triple-Code Saturation Mutagenesis
Qu, Ge,Lonsdale, Richard,Yao, Peiyuan,Li, Guangyue,Liu, Beibei,Reetz, Manfred T.,Sun, Zhoutong
, p. 239 - 246 (2018/02/09)
Directed evolution of stereo- or regioselective enzymes as catalysts in asymmetric transformations is of particular interest in organic synthesis. Upon evolving these biocatalysts, screening is the bottleneck. To beat the numbers problem most effectively, methods and strategies for building “small but smart” mutant libraries have been developed. Herein, we compared two different strategies regarding the application of triple-code saturation mutagenesis (TCSM) at multiresidue sites of the Thermoanaerobacter brockii alcohol dehydrogenase by using distinct reduced amino-acid alphabets. By using the synthetically difficult-to-reduce prochiral ketone tetrahydrofuran-3-one as a substrate, highly R- and S-selective variants were obtained (92–99 % ee) with minimal screening. The origin of stereoselectivity was provided by molecular dynamics analyses, which is discussed in terms of the Bürgi–Dunitz trajectory.
Preparation of structurally diverse chiral alcohols by engineering ketoreductase CgKR1
Zheng, Gaowei,Liu, Yuan-Yang,Chen, Qi,Huang, Lei,Yu, Hui-Lei,Lou, Wen-Yong,Li, Chun-Xiu,Bai, Yun-Peng,Li, Ai-Tao,Xu, Jian-He
, p. 7174 - 7181 (2017/11/06)
Ketoreductases are tools for the synthesis of chiral alcohols in industry. However, the low activity of natural enzymes often restricts their use in industrial applications. On the basis of computational analysis and previous reports, two residues (F92 and F94) probably affecting the activity of ketoreductase CgKR1 were identified. By tuning these two residues, the CgKR1-F92C/F94W variant was obtained that exhibited higher activity toward all 28 structurally diverse substrates examined than the wild-type enzyme. Among them, 13 substrates have a specific activity over 50 U mg-1 (54-775 U mg-1). Using CgKR1-F92C/F94W as a catalyst, five substrates at high loading (>100 g-1 L-1) were reduced completely in gramscale preparative reactions. This approach provides accesses to pharmaceutically relevant chiral alcohols with high enantioselectivity (up to 99.0% ee) and high space-time yield (up to 583 g-1 L-1 day-1). Molecular dynamics simulations highlighted the crucial role of residues 92 and 94 in activity improvement. Our findings provide useful guidance for engineering other ketoreductases, especially those possessing a similar active pocket to that in CgKR1.
Stereo-complementary bioreduction of saturated N-heterocyclic ketones
Li, Chao,Liu, Yan,Pei, Xiao-Qiong,Wu, Zhong-Liu
, p. 90 - 97 (2017/04/28)
The asymmetric bioreduction of several saturated N-heterocyclic ketones is demonstrated in a stereo-complementary fashion using the ketoreductases READH and ChKRED20 for the production of (S)- and (R)-alcohols, respectively. The reaction accepts substrates with a five-, six- or seven-membered ring, and exhibits excellent stereoselectivity when using 2-propanol as both the ultimate reducing agent and cosolvent, achieve >99% ee in the majority of cases for both enantiomers.
