Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for preparing N-acetyl bromide-7-aminocephalosporanic acid through enzyme catalysis

A technology for the preparation of aminocephalosporanic acid and catalysis, which is applied in the direction of fermentation, etc., can solve problems such as complicated operations, harsh conditions, and inconformity with sustainable social and environmental development, and achieve simple and easy control of the reaction process, mild reaction conditions, and realization of repeatability The effect of using

Active Publication Date: 2016-08-31
SOUTH CHINA UNIV OF TECH +1
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Or replace bromoacetyl bromide with chloroacetyl chloride, and prepare cefathiamidine through a similar chemical acylation and condensation process (China Pharmaceutical Industry Journal, 2009, 40 (12): 888; CN102285999 B; CN 101486718 B), but chloroacetyl The activity of chlorine in the base is low, resulting in low yield of the second step condensation reaction, so in industry, the first step of cefathiamidine synthesis still uses bromoacetyl bromide as the acyl donor
Another method for synthesizing cefathiamidine is to first synthesize side chain 1,3-diisopropylamidino-2-thio-acetic acid hydrochloride, then after activation by Vilsmeier reagent, finally combine with protected The 7-ACA condensation of base, after deprotection, obtains cefathiamidine (Chinese Journal of Medicinal Chemistry, 2001,11 (5): 293; CN101704827 B); Although the target product yield of this synthetic approach route is relatively high (56% ), but its operation is complicated and harsh, and the silylating reagent used is expensive, and the cost is higher; and the activation process needs to adopt highly toxic active reagents such as triphosgene and triphenylphosphine, but also toluene, xylene, chlorine Highly toxic reagents such as benzene and dichlorobenzene
Therefore, chemical preparation of cefathiamidine not only requires the use of highly active acyl donors (acyl chlorides or acid bromides), volatile organic solvents and bases, but also requires low-temperature reactions. The reaction conditions are harsh, the process is complicated, and the environment is seriously polluted. Current requirements for sustainable social and environmental development
In addition, the yield of N-bromoacetyl-7-ACA, the key intermediate of cefathiamidine, is usually not ideal, the highest is only 73% (China Modern Applied Pharmacy, 2010 27(2):126)

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
  • Method for preparing N-acetyl bromide-7-aminocephalosporanic acid through enzyme catalysis
  • Method for preparing N-acetyl bromide-7-aminocephalosporanic acid through enzyme catalysis
  • Method for preparing N-acetyl bromide-7-aminocephalosporanic acid through enzyme catalysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Add 10mL phosphate buffer (100mM, pH 7.5), 40mM 7-ACA and 120mM methyl bromoacetate into a 50mL Erlenmeyer flask with a cap, mix well, then add 30U penicillin G acylase II (purchased from CSPC), React at 20°C and 200r / min. After 11 h, liquid chromatography analysis showed a 34% yield of N-bromoacetyl-7-ACA.

Embodiment 2

[0031] Add 10mL phosphate buffer (100mM, pH 7.5), 40mM 7-ACA and 120mM methyl bromoacetate into a 50mL Erlenmeyer flask with a cap, mix well, then add 30U penicillin G acylase IV (purchased from CSPC), React at 20°C and 200r / min. After 7 h, liquid chromatography analysis showed that the yield of N-bromoacetyl-7-ACA was 32%.

Embodiment 3

[0033] Add 10mL phosphate buffer (100mM, pH 7.5), 40mM 7-ACA and 120mM methyl bromoacetate into a 50mL Erlenmeyer flask with a cap, mix well, then add 30U penicillin acylase PGA-750 (purchased from Zhejiang Shunfeng Haider Co., Ltd.), react at 20°C and 200r / min. After 3 h, liquid chromatography analysis showed that the yield of N-bromoacetyl-7-ACA was 91%. The liquid chromatograms before and after the reaction are shown in the attached figure 1 and attached figure 2 As shown, the retention times of 7-ACA and N-bromoacetyl-7-ACA were 2.58 and 4.10 min, respectively. Subsequently, the enzyme was filtered out, and hydrochloric acid was added to the filtrate to adjust the pH to 1.0, filtered, washed with water, and dried to obtain N-bromoacetyl-7-ACA with a yield of 88%.

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 invention belongs to the field of enzyme catalysis and biological pharmacy and discloses a method for preparing N-acetyl bromide-7-aminocephalosporanic acid through enzyme catalysis. The method includes the following steps that 7-aminocephalosporanic acid and acyl donors are added into a buffer solution to be mixed evenly, immobilized penicillin amidase is added for reaction, and N-acetyl bromide-7-aminocephalosporanic acid is obtained through separation after reaction. The method has the advantages of being mild in reaction condition, environmentally friendly, simple in process, high in target product yield and the like.

Description

technical field [0001] The invention belongs to the field of enzyme catalysis and biopharmaceuticals, in particular to a method for preparing N-bromoacetyl-7-aminocephalosporanic acid catalyzed by penicillin acylase. Background technique [0002] Cefathiamidine is my country's first self-developed semi-synthetic cephalosporin. Cefathiamidine is a broad-spectrum antibacterial agent, which has a strong killing effect on most Gram-positive bacteria and some Gram-negative bacteria, especially the killing effect on enterococci exceeds that of similar species used in clinical practice. In addition, cefathiamidine also shows certain curative effect on the infection of methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis (Chinese Journal of Antibiotics, 2005,30(2):107). Clinical studies have shown that cefathiamidine has the characteristics of high blood concentration and good clinical curative effect. In recent years, the market of cefa...

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 Applications(China)
IPC IPC(8): C12P35/04
CPCC12P35/04
Inventor 李宁张晓利宗敏华罗春
Owner SOUTH CHINA UNIV OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products