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

Method for synthesizing linaclotide by solid-liquid phase combination

A technology of liralotide and synthetic method, which is applied in the field of peptide synthesis and preparation, can solve the problems of high cost, cumbersome operation, and a lot of waste liquid

Inactive Publication Date: 2018-04-20
SINOPEP ALLSINO BIOPHARMACEUTICAL CO LTD
View PDF15 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This document uses ACM, Trt, PMeOBzl, Mmt and other side chain protecting agents to oxidize disulfide bonds respectively, but still uses the method of disulfide bonds in the liquid phase, and uses a variety of oxidants at the same time, which is not conducive to purification. The waste water problem has not been solved
The document Solid-PhaseSynthesis of the Cys-rich Peptide Linaclotide, Journal of Peptide Science, 2008, 14 (8): 56-56, mentioned the disulfide bond full selective synthesis strategy of 2Trt, 2PMeOBzl, 2StBu, using 6 Trt strategies Oxidize three disulfide bonds at one time, and use four Trt strategies to oxidize two disulfide bonds at one time. There is also a method for final liquid phase synthesis of disulfide bonds. At the same time, a variety of oxidants are used, which is not conducive to purification and cumbersome operation. The waste water problem has not been solved
The existing synthesis method of linaclotide has the disadvantages of cumbersome operation, many waste liquids and many impurities, which is not conducive to environmental protection, requires a large amount of acetonitrile, high cost and is not conducive to large-scale production

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 synthesizing linaclotide by solid-liquid phase combination
  • Method for synthesizing linaclotide by solid-liquid phase combination
  • Method for synthesizing linaclotide by solid-liquid phase combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1, a solid-liquid phase combination synthesis method of liralotide, the method adopts a 6+8 synthesis method, first synthesizes 6 peptide fragments, and then couples the fragments to a solid-phase resin, through Cys Protecting groups at different positions complete the selective synthesis of three disulfide rings on the solid-phase resin, which are then cleaved from the resin and purified directly.

[0069] The specific steps are as follows:

[0070] A. The fragments are condensed into various liralotide intermediates, and the amino acid protecting groups are given as follows:

[0071] (1) Synthesis of R-Cys(R1)-Glu(R2)-Tyr(R4)-Cys(R1)-R3;

[0072] (2) Synthesis of R-Cys(R1)-Cys(R1)-Glu(R2)-Tyr(R4)-Cys(R1)-R3;

[0073] (3) Synthesis of R-Cys(R1)-Cys(R1)-Glu(R2)-Tyr(R4)-Cys(R1)-Cys(R1)-R3;

[0074] (4) Synthesis of R-Cys(R1)-Cys(R1)-R3;

[0075] (5) Synthesis of R-Cys(R1)-Cys(R1)-Glu(R2)-R3;

[0076] (6) Synthesis of R-Cys(R1)-Cys(R1)-Glu(R2)-Tyr(R4)-R3;

[0...

Embodiment 2

[0122] 1. Synthesis of Boc-Cys(Fm)-Cys(Mmt)-Glu(OtBu)-Tyr(Trt)-Cys(Trt)-Cys(Fm)-OH

[0123] (1) Synthesis of Fmoc-Cys(Mmt)-Glu(OtBu)-OH

[0124] Solution 1: Weigh 100 millimolar Fmoc-Cys(Mmt)-OH and 110 millimolar HOSu into a 500ml round bottom flask, add 200ml THF to dissolve, and cool to 0-5°C under magnetic stirring for later use.

[0125] Solution 2: Weigh 110mmol of DCC, put it in a small beaker, dissolve it with 100ml of THF, cool to 0-5°C for later use

[0126] Solution 3: Weigh out 130 mmol of NH 2 -Glu(OtBu)-OH, dissolved in 100ml of 10% sodium carbonate solution, set aside at room temperature

[0127] Slowly add the DCC solution (solution 2) dropwise into the stirred solution 1 at 0-5°C, react for 15 minutes, and react for 2 hours at 25°C, and confirm the end of the reaction by spotting the plate (the specific reaction time is based on the spotting time allow). Remove the white precipitate by filtration, wash the precipitate with 30ml THF and filter, combine the ...

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 relates to a method for synthesizing linaclotide which is a product in the medicine field. The method adopts a solid-liquid phase combination process to synthesize the linaclotide product. A 6+8 synthesizing mode is adopted, firstly six peptide fragments are synthesized, then the fragments are coupled to solid-phase resin, selective synthesis of three dithio-rings is completed on thesolid-phase resin through protecting groups of Cys in different positions, and then the dithio-rings are cut from the resin and are directly purified, so that the synthesizing efficiency and the synthesizing yield are greatly improved, the production of the three wastes (waste water, waste solid and waste gas) is greatly reduced, and furthermore, the production cost is greatly lowered.

Description

technical field [0001] The invention belongs to the field of polypeptide synthesis and preparation, in particular to a synthesis and preparation method of liralotide. technical background: [0002] The structural formula of liralotide is as follows: [0003] [0004] L-cysteinyl-L-cysteinyl-L-glutamyl-L-tyrosyl-L-cysteinyl-L-cysteinyl-Lasparaginyl-L-prolyl-L-alanyl-L-cysteinyl-L-threonyl-glycyl-L-cysteinyl-L- Tyrosine, cyclic (1-6), (2-10), (5-13)-tris (disulfide). [0005] Linaclotide was approved for marketing by the US FDA and EU EMEA in August 2012 and November 2012, respectively. It is a 14-amino acid guanylate cyclase C agonist that, upon binding to intestinal GC-C, leads to increased intracellular and extracellular guanylate (cGMP) concentrations. The increase of intracellular cGMP can stimulate the secretion of intestinal juice, accelerate the migration of gastrointestinal tract, thereby increasing the frequency of defecation, and the increase of extracellular ...

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): C07K7/08C07K1/06C07K1/04C07K1/02
CPCC07K7/08Y02P20/55
Inventor 徐峰柳铎芳谷海涛苏军孙美禄王小涛
Owner SINOPEP ALLSINO BIOPHARMACEUTICAL 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