Enzyme-catalyzed asymmetric synthesis of key intermediates of dextromethorphan

A kind of technology of dextromethorphan hydrobromide and intermediate, which is applied in the field of preparation of key intermediates of antitussives

Active Publication Date: 2021-11-23
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] We found that in the existing process, there is no enzyme-catalyzed asymmetric reduction to prepare (S)-1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-8 Method for Hydroisoquinoline ((S)-2)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Enzyme-catalyzed asymmetric synthesis of key intermediates of dextromethorphan
  • Enzyme-catalyzed asymmetric synthesis of key intermediates of dextromethorphan
  • Enzyme-catalyzed asymmetric synthesis of key intermediates of dextromethorphan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: the acquisition of highly expressed genetically engineered bacteria

[0024] The whole gene synthesis was completed by Shanghai Xuguan Company.

[0025] According to the gene AtIR of Amycolatopsis thermoflava (WP_027931121.1), the gene StIR of Streptomycesthermoilacinus (WP_023587323.1), the gene PmIR of Prauserella marina (WP_091804541.1), the gene SmIR of Sciscionella marina (WP_020496004.1 of Amyalcolatops gene), WP_020635634.1) and the gene ShIR (SHE96216.1) of Streptoalloteichus hindustanus were respectively codon-optimized in order to enable the gene to be expressed in the E. coli expression host. See the attached table for the sequence. And add corresponding enzyme cutting sites at both ends of the gene, and construct them into corresponding vectors to obtain genetically engineered bacteria IR1, IR2, IR3, IR4, IR5, and IR6.

[0026] Transform the prepared recombinant vector into Escherichia coli BL21, Rosetta or Origami by conventional methods to c...

Embodiment 2

[0027] The cultivation of embodiment 2 genetically engineered bacteria and the preparation of resting cells

[0028]Pick a single colony on the plate and inoculate it into 5ml of fermentation medium containing corresponding antibiotics, cultivate it for about 15 hours as a seed solution, inoculate it into 600ml of fermentation medium according to the inoculation amount of 1%, and cultivate it on a shaker at 37°C and 200rpm to OD 600 = about 0.6-0.8, add IPTG with a final concentration of 0.1 mM to induce for more than 10 h, and collect the bacteria by centrifuging the culture solution at 8000 rpm.

Embodiment 3

[0029] Embodiment 3 Utilizes the asymmetric reduction of resting cells of IR1 to catalyze formula (1) hydrochloride

[0030] Take 2.5g of IR1 resting cells and resuspend in 100mL sodium phosphate buffer (100mM, pH 7.5), add glucose (1g), NADP + (5mg), GDH enzyme powder (50mg), 1-(4-methoxybenzyl)-3,4,5,6,7,8-hexahydroisoquinoline hydrochloride (1) (0.5g) , use 10% sodium carbonate solution to control pH 7.5, react under nitrogen protection at 25°C for 24 hours, HPLC detection shows that the reaction is complete (liquid chromatography column: Daicel CHIRALPAK OJ-H (250mm×4.6mm, 5μm), mobile phase: iso Propanol (0.5% ethanolamine) / n-hexane=10:90, detection wavelength: 230nm, flow rate: 0.5mL / min), 10% sodium carbonate solution to adjust pH to 9.0, ethyl acetate extraction (100mL*3), anhydrous After drying with sodium sulfate, it was spin-dried, separated and purified by column chromatography to obtain (S)-1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquine The morphine ((S)-...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention discloses imine reductase AtIR (WP_027931121.1) derived from Amycolatopsis thermoflava or imine reductase StIR (WP_023587323.1) of Streptomyces thermolilacinus or imine reductase PmIR (WP_091804541.1) of Prauserella marina or Sciscionella marina imine reductase SmIR(WP_020496004.1) or imine reductase AaIR(WP_020635634.1) of Amycolatopsis alba or imine reductase ShIR(SHE96216.1) of Streptoalloteichus hindustanus, and use this imine reductase as a biocatalyst Preparation of the key intermediate (S)-1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-octahydroisoquinoline of the central antitussive drug dextromethorphan . The corresponding imine reductase can catalyze 5-50g / L of the substrate, and the conversion rate is greater than 99%. The method has the remarkable characteristics of high yield, good stereoselectivity, and mild reaction conditions.

Description

technical field [0001] The invention relates to a new method for preparing a key intermediate of antitussive medicine, which belongs to the field of biochemical industry. It specifically relates to the preparation of the key intermediate (S)-1-(4-methoxybenzyl)-1,2,3,4,5,6,7,8-eight of dextromethorphan hydrobromide through enzyme-catalyzed asymmetric reduction A new approach to hydroisoquinolines. Background technique [0002] Dextromethorphan hydrobromide is also called dextromethorphan hydrobromide, the chemical name is (+)-3-methoxy-17-methyl-(9α,13α,14α)-morphinan hydrobromide monohydrate , suitable for colds, acute or chronic bronchitis, bronchial asthma, pharyngitis, tuberculosis and dry cough caused by other upper respiratory tract infections. [0003] [0004] There are many synthetic methods of dextromethorphan hydrobromide, such as Chinese patents CN201310041846, CN201310051880, CN201210405684, CN201310004262, CN201410169133 and literature [Today’s Pharmacy, 2...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/12
CPCC12P17/12
Inventor 姚培圆徐泽菲于珊珊吴洽庆朱敦明马延和
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap