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

Method for preparing (1R, 3S)-3-aminocyclopentanol, integrase inhibitor and application

An integrase inhibitor, aminocyclopentanol technology, applied in the field of bioengineering, can solve problems such as complex operation, high production cost, and large environmental pollution, and achieve the effect of simplifying the synthesis steps and mild reaction conditions

Pending Publication Date: 2020-10-02
ENZYMASTER NINGBO BIO ENG CO LTD
View PDF5 Cites 0 Cited by
  • 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 preparing (1R, 3S)-3-aminocyclopentanol, to solve the existing problems in the preparation of (1R, 3S)-3-aminocyclopentanol proposed in the above-mentioned background technology Most of the methods are complicated to operate, the production cost is high, and there is a problem of large environmental pollution

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 (1R, 3S)-3-aminocyclopentanol, integrase inhibitor and application
  • Method for preparing (1R, 3S)-3-aminocyclopentanol, integrase inhibitor and application
  • Method for preparing (1R, 3S)-3-aminocyclopentanol, integrase inhibitor and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] An Escherichia coli expressing transaminase, said transaminase is an enzyme encoded by a transaminase gene, the amino acid sequence of said transaminase is as shown in SEQ ID No.2, and the nucleotide sequence of said transaminase gene is shown in SEQ ID No.1 Show.

[0036] Specifically, the construction and culture method of the Escherichia coli expressing transaminase are as follows:

[0037] 1) The artificially synthesized transaminase gene DNA fragment was double-digested with restriction endonuclease Nco I and restriction endonuclease EcoR I at 37°C for 8 hours, purified by agarose gel electrophoresis, and purified using agarose gel DNA The recovery kit recovers the target fragment;

[0038] 2) Under the action of T4 DNA ligase, the target fragment was ligated with the plasmid pET-28a that was digested with restriction endonuclease Nco I and restriction endonuclease EcoR I at 25° C. overnight, get the connection product;

[0039] It should be noted that when the ...

Embodiment 2

[0042] The recombinant Escherichia coli wet thallus prepared in Example 1 was lysed back with 0.1mol / L aqueous phase buffer (pH=7.0), homogeneously crushed, centrifuged to collect the supernatant of the enzyme solution and freeze-dried to obtain transaminase ( Enzyme powder), wherein, the add-on of described aqueous phase buffer is added according to the ratio of recombinant Escherichia coli wet thallus: aqueous phase buffer=1:5.

Embodiment 3

[0044]A method for preparing (1R, 3S)-3-aminocyclopentanol, specifically comprising the following steps: taking R-3-hydroxycyclopentanone as a substrate, adding transaminase (enzyme powder) prepared in Example 2, and Add amino donor, coenzyme, and aqueous phase buffer to mix to form a reaction system, and then place in a reaction bottle for enzymatic reaction and separation to obtain the (1R, 3S)-3-aminocyclopentanol; specifically, the After adding magnets into the reaction bottle, place it on the reactor preheated to 30°C, adjust the reactor speed to 400rpm for stirring reaction (reaction times are 1h, 2h, 4h, 20h, 24h), and the reaction solution is obtained. Take 50 μL of the reaction solution and add 1 mL of acetonitrile with a volume concentration of 50% to terminate the reaction, inactivate it by shaking, take the supernatant after centrifugation, and detect the conversion rate by HPLC. The specific detection results are shown in Table 1.

[0045] Wherein, the condition o...

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 particularly discloses a method for preparing (1R, 3S)-3-amino cyclopentanol, an integrase inhibitor and application. The method for preparing (1R, 3S)-3-amino cyclopentanol comprises the following steps: by taking R-3-hydroxycyclopentanone as a reactant, adding an amino donor, coenzyme and transaminase, mixing, and performing enzymatic reaction in a water-phase buffer solution, thereby obtaining the (1R, 3S)-3-amino cyclopentanol. Meanwhile, the invention provides an amino acid sequence for catalyzing R-3-hydroxycyclopentanone to generate transaminase of the (1R, 3S)-3-amino cyclopentanol. The invention further provides a nucleotide sequence for encoding the transaminase, a preparation method and a reaction process. The reaction condition for preparing the (1R, 3S)-3-amino cyclopentanol through enzyme catalysis disclosed by the invention is mild, green and environment-friendly, and the problems that the conventional chemical method for preparing the (1R, 3S)-3-amino cyclopentanol is multiple in chemical reaction steps, tedious in operation, high in production cost and severe in environmental pollution are solved.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for preparing (1R,3S)-3-aminocyclopentanol, an integrase inhibitor and an application. Background technique [0002] (1R,3S)-3-Aminocyclopentanol, as an important pharmaceutical intermediate, has a very broad market prospect. The main active ingredient of the new anti-AIDS (Acquired Immune Deficiency Syndrome, AIDS, acquired immunodeficiency syndrome) drug Biktarvy developed by Gilead Sciences is the integrase inhibitor Bictegravir. And Bictegravir is obtained by constructing a bridging ring with (1R, 3S)-3-aminocyclopentanol hydrochloride as a raw material, which is the only chiral source to constitute a chiral molecule. Therefore, (1R, 3S)-3-amino The preparation of cyclopentanol has also become one of the key steps in the synthesis of Bictegravir. [0003] At present, the preparation method of (1R,3S)-3-aminocyclopentanol is usually prepared by a resolution m...

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
IPC IPC(8): C12P13/00A61K31/133A61P31/18
CPCC12P13/001A61K31/133A61P31/18
Inventor 石淑敏章兆琪陈承黄勇开陈海滨
Owner ENZYMASTER NINGBO BIO ENG CO LTD
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

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com