Method for preparing moxifloxacin side chain through biological method

A technology of compound and transaminase, which is applied in the production of bulk chemicals, organic chemistry, fermentation, etc., can solve the problems of long nonane steps, high environmental pressure, and unknown enzyme sources, etc.

Active Publication Date: 2017-02-15
ENZYMEWORKS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The synthetic route of US 5703244 requires 11 steps of reaction, which is difficult to process application
The route of CN201110312411 only needs 5 steps of reaction, but its key step reaction substrate and product need to be separated by column chromatography, the yield is not high (36%-57%), and an equal amount of chiral reaction reagent is required
In summary, the preparation of (S,S)-2,8-diazabicyclo[4.3.0]nonane by chemical methods generally has problems such as long steps, low yield, harsh conditions, and the need for chiral reagents, resulting in the production of High cost and high environmental pressure in the production process
[0014] Another biological asymmetric synthesis route uses transaminase for carbonyl amination (CN201410418987). As shown in the figure, because the ee value and yield of the product are low, and the source of the enzyme used is unknown, it lacks industrial application value

Method used

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  • Method for preparing moxifloxacin side chain through biological method
  • Method for preparing moxifloxacin side chain through biological method
  • Method for preparing moxifloxacin side chain through biological method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Screening of amino protecting groups

[0065] The substrates were 10 mg of 4-(3-chloropropyl)-3-pyrrolidone (4a, 4b, 4c) protected by benzyl group, ethyl formate group and Cbz group, dissolved in 0.2 mL DMSO, and added to 0.8 mL In triethanolamine buffer solution (pH8.0, 0.1M), add transaminase (SEQ ID NO: 18) 10mg, pyridoxal phosphate (PLP) 1mg, isopropylamine 0.01mL, react at 30 degrees for 24 hours, and detect the conversion rate> 99%, adding 1mL ethyl acetate to extract 5 times, combining the organic phases, drying and concentrating to obtain products 2a, 2b, and 2c were 3.44mg, 7.44mg, and 8.37mg respectively, and the molar yields were 40%, 88%, and 95% respectively. %.

[0066]

Embodiment 2

[0067] Embodiment 2: Investigation on the stability of benzyl-protected substrates

[0068] The stability of the benzyl-protected substrate was investigated due to possible problems with the stability of the substrate. Substrate benzyl-protected 4-(3-chloropropyl)-3-pyrrolidone (4a) 10mg, dissolved in 0.2mL DMSO, respectively added 0.8mL triethanolamine buffer solution (pH 8.0, 0.1M), phosphate buffer solution (pH 8.0, 0.1M) and deionized water, add 0.01mL of isopropylamine, shake at 30 degrees for 24 hours, take a sample to detect the change of the peak area of ​​the substrate in HPLC, as shown in the following table:

[0069]

[0070] It shows that the substrate is unstable under the enzyme reaction conditions.

Embodiment 3

[0071] Embodiment 3: reaction temperature selection

[0072] Dissolve 10 mg of substrate 4c in 0.2 mL DMSO, add 0.8 mL of triethanolamine buffer solution (pH 8.0, 0.1 M), add 1 mg of transaminase (SEQ ID NO: 18), 0.1 mg of PLP, 0.01 mL of isopropylamine, different Reaction under the temperature for 24 hours, detection conversion rate, the results are shown in the table below:

[0073]

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PUM

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Abstract

The present invention provides a method for preparing a moxifloxacin side chain through a biological method, particularly to a method for preparing a compound represented by a formula 2. The method comprises: catalyzing a compound represented by a formula 4 with transaminase to form a compound represented by formula 3, and carrying out spontaneous ring closure on the compound represented by the formula 3 to obtain the compound represented by the formula 2, wherein the transaminase is selected from the omega-transaminase in Arthrobacter, Aspergillus terreus, Vibrio fluvialis, Bacillus megaterium, Sphingomonas paucimobilis, Hyphomonasneptunium, and Chromobacterium violaceum, and in the formulas 2, 3 and 4, R is selected from benzyloxycarbonyl, benzyl and ethoxycarbonyl. The formulas 2, 3 and 4 are defined in the specification.

Description

technical field [0001] The invention belongs to the technical fields of biopharmaceuticals and biochemical engineering, and in particular relates to a production method for preparing moxifloxacin side chains by a biological method. Background technique [0002] (S,S)-2,8-Diazabicyclo[4.3.0]nonane(1) and its N-protected derivative(2) are key chiralities for the synthesis of widely used quinolone antibacterial drug moxifloxacin Intermediate, also known as moxifloxacin side chain or moxi small ring: [0003] [0004] Since (S,S)-2,8-diazabicyclo[4.3.0]nonane contains two chiral centers, its synthesis methods usually include resolution method, asymmetric synthesis method and chiral source method. The resolution method is the most important application method. Using pyridine 2,3-dicarboxylate as the starting material, the racemate of benzyl-protected 1 is obtained by high-pressure hydrogenation and other methods, and the product is obtained by resolution using tartaric acid a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/18C07D471/04
CPCY02P20/55
Inventor 陶军华梁晓亮蒋相军乐庸堂
Owner ENZYMEWORKS
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