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Excitotoxicity related injury treatment peptide

An active peptide and nervous system technology, applied in the medical field, can solve problems such as nerve cell damage

Active Publication Date: 2017-11-03
BIOCELLS BEIJING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The neuronal death or injury caused by cerebral ischemia is a damage cascade reaction process. After cerebral ischemia, tissue blood perfusion decreases, excitatory neurotransmitters increase, activate NMDA and AMPA receptors, cause ion channels to open, calcium ions Influx, activation of a large number of enzymes triggers signaling cascades, resulting in multiple pathways of neuronal damage

Method used

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  • Excitotoxicity related injury treatment peptide
  • Excitotoxicity related injury treatment peptide
  • Excitotoxicity related injury treatment peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1: Screening of Active Peptide Molecules

[0093] According to the reported research results, the Tat membrane-penetrating peptide YGRKKRRQRRR (SEQ ID NO: 2) was selected and connected with different numbers of amino acids to form a peptide library. The chimeric peptide molecules in the peptide library interacted with the PDZ1 / 2 domains expressed and purified in vitro, and the peptides were initially screened according to the strength of the interaction.

[0094] The immobilized molecule (ligand) is PDZ1 / 2 protein, molecular weight: ~20kD, concentration: 2mg / ml; molecule (analyte) in mobile phase: polypeptide to be screened, molecular weight: ~2kD, concentration: 10mg / ml. Using Biacore 3000 instrument, CM5 chips were fixed. The electrophoresis buffer was PBS+0.005% Tween 20. Immobilization was performed using an amino-coupling method. The concentration of ligand was 10 μg / ml. Fixation buffer was 10 mM sodium acetate, pH 4.0. Fixed amount: 1400RU, fixed to F...

Embodiment 2

[0108] Example 2: Pull-down experiments detect the interaction between P5 and PDZ1 / 2 domains

[0109] To prove that P5 can interact with the PDZ1 / 2 domain, a Pull-down experiment was performed.

[0110] The column was equilibrated for 5 min with 100 μl of His beads and 1 ml of MCAC-0 buffer. Shake at 4°C. The mixture was centrifuged at 5000g for 1 min at 4°C, and the supernatant was discarded. Add 1 mg of PDZ1 / 2 protein to the mixture and make up to 1 ml with buffer. The mixture was rotated for 1 hour at 4°C. The mixture was centrifuged at 5000g for 1 min at 4°C, and the supernatant was discarded. Wash with 1 ml of MCAC-0 buffer 3 times, 5 minutes each time (washing with shaking at 4°C). Add 1 mg of P5 protein to the mixture and make up to 1 ml with buffer. The mixture was rotated for 2 hours at 4°C. The mixture was centrifuged at 5000g for 1 min at 4°C, and the supernatant was discarded. Wash with 1 ml of lysate for 3 times, 5 minutes each time (washing with shaking a...

Embodiment 3

[0112] Embodiment 3: the therapeutic effect of chimeric peptide on rat MCAO model

[0113] MCAO preparation method and scoring criteria:

[0114] Preparation of focal cerebral ischemia-reperfusion model According to the reversible middle cerebral artery occlusion (MCAO) thread embolization method proposed by Longa and improved according to the anatomical structure diagram of the rat brain, the focal cerebral ischemia-reperfusion model was made, with 10 Intraperitoneal anesthesia with % chloral hydrate 0.3ml / kg, median neck incision, exposure of the common carotid artery (CCA), external carotid artery (ECA) and pterygopalatine artery, 0.5cm of the head end of 0.26mm monofilament nylon fishing line was coated with paraffin, And marked at the 20mm length, all rats were inserted through the right CCA incision, the pterygopalatine artery was clipped temporarily to prevent mis-insertion, the length of the embolized suture was about 18-20mm from the CCA bifurcation, depending on the ...

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PUM

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Abstract

The present invention relates to an excitotoxicity related injury treatment peptide, and provides a peptide containing an amino acid sequence YEKLLDTEI (SEQ ID NO:1) or a functional variant thereof, wherein the peptide is an active peptide for treating central nervous system injury. The invention further provides a chimeric peptide containing the active peptide and an internalized peptide. The present invention further provides a pharmaceutical composition containing the active peptide or the chimeric peptide, and medical applications of the active peptide or the chimeric peptide.

Description

technical field [0001] The present application relates generally to the field of medicine, and in particular, the present application provides peptides, compositions and methods for treating central nervous system injuries. Background of the invention [0002] Stroke is a common acute cerebrovascular disease among middle-aged and elderly people, and it tends to be younger. It is one of the three most harmful diseases (cancer, cardiovascular and cerebrovascular diseases, diabetes) to humans in the world today. Nearly 3 million people die from cerebrovascular diseases in my country every year, which is 4 to 5 times higher than that of European and American countries, 3.5 times that of Japan, and even higher than developing countries such as Thailand and India; the incidence rate is rising at an annual rate of 8.7%, and the recurrence rate More than 30%, the recurrence rate within 5 years is as high as 54%; 75% of stroke survivors are disabled to varying degrees, and 40% are se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K19/00A61K38/08A61K47/42A61P25/00A61P29/00A61P25/28A61P25/22A61P25/08A61P9/10A61P25/02A61P25/04A61P21/00A61P25/16A61P25/14
CPCA61K38/00A61K47/42C07K7/06C07K2319/10
Inventor 芦颖韩化敏童威葛平菊景波
Owner BIOCELLS BEIJING BIOTECH CO LTD
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