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

Method for purifying phosphorylated eledoisin

A technology of muscarin and a purification method, which is applied in the field of chemical purification, can solve the problem of inability to purify a large number of phosphorylated polypeptides and the like, and achieves the effects of good purification, economical efficiency and simple operation.

Active Publication Date: 2014-06-18
中国人民解放军防化学院
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the technologies reported so far are mainly in the field of proteomics to carry out trace relative enrichment of natural phosphorylated polypeptides in biological samples (application numbers 201310253448.X, 200810202189.7, 200810211727.9), in order to improve the detection of phosphorylated polypeptides in mass spectrometry The purification rate still stays at the analytical level, which cannot meet the requirements of a large amount of purification of phosphorylated polypeptides synthesized by chemical methods, especially the purification method of phosphorylated muscotin has not been reported yet.

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 purifying phosphorylated eledoisin
  • Method for purifying phosphorylated eledoisin
  • Method for purifying phosphorylated eledoisin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Purification of a simple sample of phosphorylated muscotin mixed with two other non-phosphopeptides:

[0028]Weigh 100 mg of titanium dioxide into a purified glass tube with a sieve plate, add 10 mL of 70% acetonitrile / 3% TFA solution, shake up and down for 1 minute, filter the solution under reduced pressure at 0.05-0.09 mbar, and wait for the sample; weigh the pure product of phosphorylated polypeptide 1 Pyr-Pro-Ser-Lys-Asp-Ala-Phe-Tyr (PO 3 H)-Gly-Leu-Met-NH 2 and two pure non-phosphorylated polypeptides [Pyr-Pro-Ser-Lys-Asp-Ala-Phe-Phe(Cl)-Gly-Leu-Met-NH 2 and Pyr-Pro-Ser-Lys-Asp-Ala-Phe-Tyr-Gly-Leu-Met-NH 2 ] each 10 mg, after mixing, dissolve in 20 mL of 70% acetonitrile / 3% TFA solution, oscillate up and down for 5 minutes, filter the solution under reduced pressure at 0.05-0.09 mbar; add 60% acetonitrile / 0.1% TFA 20mL of the solution, shake up and down for 1 minute, filter the solution under reduced pressure; add 20mL of the solution of 50% acetonitrile / 0.1% T...

Embodiment 2

[0030] Purification and preparation of phosphorylated muscotin chemically synthesized by monomer method:

[0031] Weigh 1 g of titanium dioxide into a purified glass tube with a sieve plate, add 10 mL of a solution of 70% acetonitrile / 3% TFA, shake up and down for 1 minute, filter the solution under reduced pressure at 0.05-0.09 mbar, and wait for Loading the sample; Weigh the phosphorylated muscarin [Pyr-Pro-Ser(PO 3 H)-Lys-Asp-Ala-Phe-Ile-Gly-Leu-Met-NH 2 ] Crude product, dissolve it in 20mL of a solution of 70% acetonitrile / 3% TFA, shake it up and down for 5 minutes, filter the solution under reduced pressure at 0.05-0.09mbar; add a solution of 60% acetonitrile / 0.1% TFA 20mL, oscillate up and down for 1 minute, filter the solution under reduced pressure at 0.05-0.09mbar; add 20mL of 50% acetonitrile / 0.1% TFA solution, oscillate up and down for 1 minute, filter the solution under reduced pressure; add 20mL of 50% acetonitrile solution, shake up and down for 1 minute, filte...

Embodiment 3

[0033] Purification and preparation of phosphorylated muscotin chemically synthesized by overall method:

[0034] Weigh 1 g of titanium dioxide into a purified glass tube with a sieve plate, add 10 mL of a solution of 70% acetonitrile / 3% TFA, shake up and down for 1 minute, filter the solution under reduced pressure at 0.05-0.09 mbar, and wait for Sample loading; Weigh the phosphorylated muscarin [Pyr-Pro-Ser(PO 3 H)-Lys-Asp-Ala-Phe-Ile-Gly-Leu-Met-NH 2 ] Crude product, dissolve it in 20mL of a solution of 70% acetonitrile / 3% TFA, shake it up and down for 5 minutes, filter the solution under reduced pressure at 0.05-0.09mbar; add a solution of 60% acetonitrile / 0.1% TFA 20mL, shake up and down for 1 minute, filter the solution under reduced pressure at 0.05-0.09mbar; add 20mL of 50% acetonitrile / 0.1%TFA solution, shake up and down for 1 minute, and filter the solution at Filter under reduced pressure; add 20mL of 50% acetonitrile solution, shake up and down for 1 minute, filt...

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 purifying phosphorylated eledoisin, and belongs to the technical field of chemical purification. The phosphorylated eledoisin has very strong antihypertensive activity, and a chemosynthetic crude product of the phosphorylated eledoisin is difficult to purify in quantity, so that the application of the phosphorylated eledoisin is limited. The purification and preparation of the chemosynthetic phosphorylated eledoisin are realized by dint of technologies, namely vacuum filtration and the like, through three key steps of sample loading, cleaning and elution by utilizing a principle that titanium dioxide and the phosphorylated eledoisin generate higher Lewis acid-base effect in an acid environment and the Lewis acid-base effect disappears in an alkaline environment and adopting an independently researched phosphorylated polypeptide purification device (an application number: 201420158316.9). The method disclosed by the invention is easy and convenient to operate, economy and high-efficiency, and provides an effective method for the purification of the phosphorylated eledoisin, thereby promoting the application of the phosphorylated eledoisin in the fields of medicines and the like.

Description

technical field [0001] The invention relates to a method for purifying phosphorylated muscothin, which belongs to the technical field of chemical purification and mainly provides conditions for the effective application of phosphorylated polypeptides in the fields of medicine and the like. Background technique [0002] Eledoisin is a polypeptide containing 11 amino acids extracted from Eledone, and its sequence is Pyr-Pro-Ser-Lys-Asp-Ala-Phe-Ile-Gly-Leu-Met-NH 2 , has strong peripheral vasodilation and increased coronary blood flow, is the most effective antihypertensive substance among known peptides, and its effect is thousands of times stronger than nitroglycerin. Phosphorylated muscotin is obtained by phosphorylating Ser at the 3rd position of its N-terminus, and the sequence is Pyr-Pro-Ser(PO 3 H)-Lys-Asp-Ala-Phe-Ile-Gly-Leu-Met-NH 2 , its activity is more than two times higher than that of muscine (a powerful antihypertensive phosphopeptide and its preparation method...

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): C07K7/06C07K1/22A61P9/12
Inventor 隋少卉董俊军游炎富李建吴为辉李善茂刘基伟张鸣蒋辉刘尚义
Owner 中国人民解放军防化学院
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