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Excitotoxicity related injury treatment method

A nervous system, active peptide 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 method
  • Excitotoxicity related injury treatment method
  • Excitotoxicity related injury treatment method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1: Screening of active peptide molecules

[0092] According to the reported results, the Tat penetrating peptide YGRKKRRQRRR (SEQ IDNO: 2) was selected and connected with different numbers of amino acids to form a peptide library. The chimeric peptide molecules in the peptide library were respectively interacted with the PDZ1 / 2 domains expressed and purified in vitro, and the peptides were initially screened according to the strength of the interaction.

[0093] The immobilized molecule (ligand) is PDZ1 / 2 protein, molecular weight: ~20kD, concentration: 2mg / ml; mobile phase molecule (analyte): polypeptide to be screened, molecular weight: ~2kD, concentration: 10mg / ml. Using Biacore 3000 instrument, CM5 chip was fixed. The running buffer is PBS+0.005% Tween 20. Use amino coupling method for immobilization. The concentration of the ligand is 10 μg / ml. The fixation buffer is 10 mM sodium acetate, pH 4.0. Fixed amount: 1400RU, fixed to flow cell 2. The flow rate us...

Embodiment 2

[0107] Example 2: Pull-down experiment to detect the interaction between P6 and PDZ1 / 2 domain

[0108] To prove that P6 can interact with the PDZ1 / 2 domain, a Pull-down experiment was performed.

[0109] Equilibrate the column 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 minute 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 and combined for 1 hour at 4°C. The mixture was centrifuged at 5000 g for 1 minute at 4°C, and the supernatant was discarded. Wash with 1 ml of MCAC-0 buffer 3 times, 5 minutes each time (wash with shaking at 4°C). Add 1 mg of P6 protein to the mixture and make up to 1 ml with buffer. The mixture was rotated and combined for 2 hours at 4°C. The mixture was centrifuged at 5000 g for 1 minute at 4°C, and the supernatant was discarded. Wash 3 times with 1ml lysate, 5 minutes each time (wash ...

Embodiment 3

[0111] Example 3: Therapeutic effect of the chimeric peptide on the rat MCAO model

[0112] MCAO preparation method and scoring standard:

[0113] 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 of the rat brain, a focal cerebral ischemia-reperfusion model was made. % Chloral hydrate 0.3ml / kg abdominal anesthesia, median neck incision, exposure of common carotid artery (CCA), external carotid artery (ECA) and pterygopalatine artery, 0.26mm monofilament nylon fishing line 0.5cm head end with paraffin coated, All rats were inserted through the right CCA incision, and the pterygopalatine artery was briefly clipped to prevent accidental insertion. The length of the tether line was about 18-20 mm from the CCA bifurcation. It depends on the animal’s weight. Lateral middle cerebral artery, then suture the skin, and fi...

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Abstract

The invention provides a peptide containing an amino acid sequence YEKLTTLDTGGV (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 generally relates to the medical field. Specifically, the present application provides peptides, compositions, and methods for treating central nervous system damage. Background of the invention [0002] Stroke is a common acute cerebrovascular disease in middle-aged and elderly people, and there is a trend of getting younger. It is one of the three most harmful diseases (cancer, cardiovascular and cerebrovascular diseases, diabetes) in the world today. Nearly 3 million people die from cerebrovascular diseases in my country each year, which is 4 to 5 times higher than that of European and American countries, 3.5 times higher than 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 reaches 54%; 75% of stroke survivors lose the ability to work to varying degrees, and 40% are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K19/00A61K38/10A61K47/64A61K47/42A61P25/00A61P25/28A61P25/08A61P25/22A61P9/10A61P25/16A61P25/14
CPCA61K38/00C07K7/08C07K2319/10
Inventor 芦颖韩化敏童威葛平菊景波
Owner BIOCELLS BEIJING BIOTECH CO LTD
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