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Application of nervous excitation damage related polypeptide in preventing, relieving or treating pain

A technique of using and internalizing peptides, which is applied in the medical field and can solve problems such as weakening pain signals

Active Publication Date: 2018-04-17
BIOCELLS BEIJING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the above process, we can see that Ca 2+ Concentration and the interaction of NR2B and PSD-95 are key conditions for amplifying pain signals, so limiting either of these conditions may attenuate pain signals

Method used

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  • Application of nervous excitation damage related polypeptide in preventing, relieving or treating pain
  • Application of nervous excitation damage related polypeptide in preventing, relieving or treating pain
  • Application of nervous excitation damage related polypeptide in preventing, relieving or treating pain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1: Screening of Active Peptide Molecules

[0091] According to the reported research results, the Tat membrane-penetrating peptide YGRKKRRQRRR was selected and linked with different numbers of amino acids to form a peptide library. The 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.

[0092] 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 Flow Cell 2. A flow rate of...

Embodiment 2

[0098] Example 2: Pull-down experiments detect the interaction between P5 and PDZ1 / 2 domains

[0099] To prove that P5 can interact with the PDZ1 / 2 domain, a Pull-down experiment was performed.

[0100] 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

[0102] Example 3: Detection of hot plate analgesic effect of P5 polypeptide

[0103] Experimental procedure

[0104] animal screening

[0105] Select 18 240-260 g male SD rats (purchased from Huafu Kangcheng Co., Ltd.) as experimental mice, and divide them into 3 groups with 6 rats in each group: one group with 0.2 μg / kg dose of P5 polypeptide, one group with normal saline One group of positive drug control group (ibuprofen, 50mg / kg) was applied to the hot plate experiment. Pre-experimental screening: Before hot plate experiment, put candidate rats on a 55°C hot plate. Rats that did not lick their hind feet or lift their feet within 30 seconds were not used in the experiment.

[0106] Animal model establishment

[0107] 4.0 Chromium catgut (purchased from Shandong Boda Medical Supplies Co., Ltd.) needs to be soaked in normal saline (purchased from Shijiazhuang No.4 Medicine Co., Ltd., H13023200) for 30 minutes in advance to soften. The middle part of the thigh of the rat w...

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Abstract

The invention provides an application of a peptide containing an amino acid sequence YEKLLDTEI or functional variants thereof or pharmaceutical composition containing the peptide in preparation of drugs for preventing, relieving or treating pain, particularly nociceptive pain as well as a method for preventing, relieving or treating the pain.

Description

field of invention [0001] The present application generally relates to the medical field, and specifically, the present application provides the application of nerve excitation injury-related polypeptides in preventing, alleviating or treating pain. Background of the invention [0002] Pain is an unpleasant sensation that people often experience throughout their lives. It provides an alarm signal that the body is threatened and is an indispensable special protective function for life. On the other hand, it is the most common symptom of various diseases, and it is also one of the most serious problems plaguing human health today. Pain is a neural activity with a very complex mechanism. Pain research has become one of the important topics of current neuroscience research. [0003] Existing studies have shown that the NR2B subunit of the N-methyl-D-spartate (NMDA) receptor is involved in excitatory synaptic transmission, central sensitization, and chronic Pain and other phy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/08A61K38/10A61P29/00
CPCA61K38/08A61K38/10A61P29/00A61P25/04A61K38/00C07K7/06C07K7/08
Inventor 童威田雨佳芦颖韩化敏
Owner BIOCELLS BEIJING BIOTECH CO LTD
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