Method for synthesizing R-3-(2-chloro-1-hydroxyethyl) phenol, phenylephrine and eye drops

A hydroxyethyl, R-3- technology, applied in chemical instruments and methods, botanical equipment and methods, preparation of organic compounds, etc., can solve problems such as complex steps

Pending Publication Date: 2020-07-07
ENZYMASTER NINGBO BIO ENG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the embodiments of the present invention is to provide a method for synthesizing R-3-(2-chloro-1-hydroxyethyl)phenol,

Method used

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  • Method for synthesizing R-3-(2-chloro-1-hydroxyethyl) phenol, phenylephrine and eye drops

Examples

Experimental program
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Effect test

Embodiment 1

[0071] An Escherichia coli expressing ketoreductase, said ketoreductase is a reductase encoded by a ketoreductase gene, the amino acid sequence of said ketoreductase is shown in SEQ ID No.2, said ketoreductase gene The nucleotide sequence is shown in SEQ ID No.1.

[0072] Specifically, the construction and culture method of the Escherichia coli expressing ketoreductase are as follows:

[0073] 1) Digest the artificially synthesized DNA fragment of the ketoreductase gene with restriction endonuclease NdeI and restriction endonuclease EcoRI at 37°C for 8 h, purify by agarose gel electrophoresis, and use agarose gel DNA The recovery kit recovers the target fragment;

[0074] 2) Under the action of T4 DNA ligase, the target fragment is ligated with the plasmid pET24a that has also been digested with restriction endonuclease NdeI and restriction endonuclease EcoRI at 25°C, and the ligated product is transformed overnight Obtain recombinant Escherichia coli;

[0075] 3) Inoculate...

Embodiment 2

[0077] The thalline prepared in Example 1 was lysed back with 0.1 mol / L PBS buffer solution (pH=7.0), homogeneously crushed, centrifuged to collect the supernatant of the enzyme solution and freeze-dried to obtain ketoreductase (enzyme powder) , wherein, the added amount of the PBS buffer is added according to the weight ratio of bacteria: PBS buffer = 1:5.

Embodiment 3

[0079] The ketoreductase prepared in Example 2 is used as a catalyst to catalyze the synthesis of R-3-(2-chloro-1-hydroxyethyl)phenol, specifically, the catalytic synthesis of R-3-(2-chloro-1- The method of hydroxyethyl) phenol comprises the following steps: with 2-chloro-1-(3-hydroxyphenyl) ethanone as substrate, with the ketoreductase prepared in embodiment 2 as catalyst, adding hydrogen donor, buffer Liquid and coenzyme NAD form a reaction system to carry out catalytic reaction in a reaction bottle. Specifically, add magnets to the reaction bottle and place it on a reactor preheated to 30°C, and adjust the reactor speed to 400rpm for stirring reaction. 24 After 1 hour, 5 mL of acetonitrile was added to terminate the reaction to obtain a reaction solution.

[0080] In the embodiment of the present invention, the phenylephrine intermediate is R-3-(2-chloro-1-hydroxyethyl)phenol.

[0081] In the embodiment of the present invention, the total volume of the catalytic system is ...

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Abstract

The invention relates to the field of bioengineering and particularly discloses a method for synthesizing R-3-(2-chloro-1-hydroxyethyl) phenol, phenylephrine and eye drops. The method for synthesizingthe R-3-(2-chloro-1-hydroxyethyl) phenol comprises the steps of taking 2-chloro-1-(3-hydroxyphenyl) ethanone as a substrate and ketoreductase as a catalyst, adding a hydrogen donor, a buffer solutionand a coenzyme to form a reaction system for catalytic reaction and obtaining a phenylephrine intermediate. The ketoreductase provided by the embodiment of the invention has high activity and stereoselectivity. The synthesis method provided by the embodiment of the invention is enzymatic synthesis of the phenylephrine intermediate by adopting the ketoreductase, just one-step reaction is needed and the amount of a used reagent is small, so that the synthesis step is greatly simplified, the problem of complicated steps in an existing method for synthesizing the R-3-(2-chloro-1-hydroxyethyl) phenol is solved, and the method provided by the invention is mild in reaction condition, high in optical purity, green and environment-friendly.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for synthesizing R-3-(2-chloro-1-hydroxyethyl)phenol, phenylephrine and eye drops. Background technique [0002] Phenylephrine is a potent vasoconstrictor widely used in hypotension first aid or as a pupil dilator in eye examinations, because the agonistic effect of its R-isomer is much higher than that of the S-isomer , so chiral selectivity needs to be considered when preparing phenylephrine, wherein R-3-(2-chloro-1-hydroxyethyl)phenol is a common intermediate of phenylephrine, which belongs to the R-type isomer. [0003] At present, the synthetic method of R-3-(2-chloro-1-hydroxyethyl)phenol is usually synthesized by resolution method, that is, the synthesis of phenylephrine intermediate by splitting and flipping two steps, which cannot be directly Solving the chirality problem in the synthesis process of phenylephrine involves many reagents, complicated operation, and...

Claims

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Application Information

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IPC IPC(8): C12P7/22C12N9/04C12N15/53C07C39/24A61K31/137C07C215/60C07C213/02
CPCC12P7/22C12N9/0006C07C39/24A61K31/137C07C215/60C07C213/02Y02P20/52
Inventor 胡虎马克·博科拉格雷戈日·库比克陈海滨金炉萍
Owner ENZYMASTER NINGBO BIO ENG CO LTD
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