Patent ID: 9127075
Filing Date: 2015-09-08
Classification: A61K,A61P,C07K

Abstract:
1. A process for purifying an analgesic peptide, wherein said analgesic peptide is SEQ ID NO: 2, comprises the following steps: (1) dissolving a scorpion in distilled water or acid solution or alkaline solution, and removing the insoluble impurities by a centrifuge, and taking the supernatant as a leaching liquid for further purification, wherein the pH is between 2 and 12, the temperature is between 0 and 45° C.; (2) using a hydrophobic chromatography column to separate an analgesic active component I from the leaching liquid, wherein the pH is between 5 and 9, the concentration of neutral salt in elute is between 0.5 to 2 M, the neutral salt is selected for the group consisting of (NH (3) removing salt and non-essential proteins from said analgesic active component I by ultrafiltration, and obtaining an analgesic active component II with concentration by an ion exchange chromatographic column, wherein a ultrafiltration membrane is selected from the group consisting of the membranes permeating 10-50 kDa of peptides and the membranes retaining 1-5 kDa of peptides; (4) obtaining an analgesic active component III from the analgesic active component II by a hydrophobic chromatographic column; (5) obtaining an analgesic active component IV from the analgesic active component III by an ion exchange chromatographic column, wherein the pH is between 2 and 12, the concentration of salt in elute is between 0.1 to 0.5 M, the salt is selected for the group consisting of (NH (6) obtaining the scorpion analgesic peptide VGG from the analgesic active component IV with a gel filtration chromatography, wherein the gel filtration chromatography is selected from the group consisting of Sephacryl S-100 HR or Sephacryl S-200 HR or Sephadex G-50 or Sephadex G-75 or Sephadex G-100 or Sephadex G-150 or Superose 12 prep grade or Superose 6 prep grade or Superdex 30 prep grade or Superdex 75 prep grade or Superose 12 HR or Superose 6 HR or Superdex Peptide HR or Superdex75 HR and Superdex Peptide PE, the ion concentration of eluent is 0.15-1M; (7) further purifying the analgesic active peptide VGG by a reversed phase chromatographic column, wherein the organic solvent of the eluent is selected from the group consisting of acetonitrile, methanol and tetrahydrofuran, the concentration of organic solvent in the eluent is 20%-90%.